10

Order Dactylopteriformes
Order Gadiformes. Cod, whiting
Order Gasterosteitormes. Sticklebacks
Order Lampridiformes
Order Lophiiformes. Anglerfish
Order Pegasilormes
Order Perciformes. Perch, bass, angelfish, gouramis,
sunfish, wrasses
Order Pleuronectiformes. Flounder, sole
Order Salmoniformes. Pike, salmon, trout
Order Scorpaeniformes. Greenlings, lingeods, sculpins
Order Synbranchiformes. Rice eels
Order Tetraodontiformes. Boxtish, putfers, triggertish
Order Zeiformes. John Dories
Subclass Sarcopterygil. Lobe-finned fishes
Order Coelacanthini. The Costacanth
Order Dipnol. Lungfishes
Class Amphibia (2,400+ species)
Subclass Lissamphibia
Order Anura (Salientia). Frogs, toads
Order Gymnophiona (Apoda). Caecilians
Order Urodela (Caudata). Salamanders
Class Reptilia (6,000+ species)
Subclass Anapsida
Order Chelonia. Turtles
Subclass Lepidosauria. Primitive diapsids
Order Sphenodonta. Tuatara
Order Squamata
Suborder Amphisbaenia. Amphisbaenians
Suborder Sauria. Lizards
Suborder Serpentes. Snakes
Subclass Archosauria. Advanced diapsids
Order Crocodylia. Crocodiles, alligators
Class Aves (8,700+ species)
Subclass Neornithes
Superorder Paleognathas
Order Apterygiformes. Kiwis
Order Causariformes. Emus
Order Rheitormes. Rheas
Order Struthioniformes. Ostriches
Order Tinamiformes. Tinamous
Superorder Neognathae
Order Apoditormes. Hummingbirds, swifts
Order Anseriformes. Ducks, geese, swans
Order Caprimulgiformes, Goatsuckers, nighthawks
Order Charadrilformes, Gulls, terns, sandpipers
Order Ciconiiformes. Flamingos, storks
Order Coliiformes. Mousebirds
Order Columbiformes. Doves, pigeons
Order Coraciiformes, Hombilis, kingfishers
Order Cuculiformes. Cuckoos
Order Falconiformes. Eagles, hawks, vultures
Order Galliformes. Fowl
Order Gaviltormes. Loons
Order Gruiformes. Coots, moorhens, rails

11

Order Passeriformes. Songbirds

Order Peticaniformes. Pelicans, cormorants, gannets
Order Piciformes. Sapsuckers, woodpeckers
Order Podicipediformes. Grebes
Order Procellariltormes. Albatroses, petrels, shearwaters
Order Psittaciformes. Cockatoos, parrots
Order Spheniscitormes. Penguins
Order Strigiformes. Owls
Order Frogoniformes. Trogons
Class Mammalia (4,500+ species)
Subciass Prototheria (8 species)
Order Monotremata. Duckbill platypus, echidnas
Subciass Theria

Infraclas$ Metatheria (Marsupialia)(242 species)

Order Caenolestidea. Rat opossums
Order Diprotodontia. Kangaroo, koata, wallabies, wombat
Order Peramelina. Bandicoots
Order Polyprotodontia. Opossums, Tasmanian devil,

numbat

Infraclass Eutheria (4,300+ species)

Order Artiodactyla. Camels, catlle, deer, pigs, sheep
Order Carnivora. Bears, dogs, cats, seals
Order Cetacea. Whales, dolphins
Order Chiroptera, Bats
Order Dermoptera. Flying lemur
Order Edentata. Anteaters, armadillos, sioths
Order Hyracoidea. Conies
Order insectivora. Shrews, moles
Order Lagomorpha. Rabbits, pikas
Order Perissodactyla. Horses, rhinoceroses, tapirs
Order Pholidota. Pangolins
Order Primates. Apes, humans, lemurs, monkeys
Order Proboscidea. Elephants
Order Rodentia. Rodents
Order Scandentia, Tree shrews
Order Sirenia. Dugongs, manatees
Order Tubulidentata. Aardvark
EHAPTER 2
SEX AND SEXUAL DIFFERENTIATI

ON

Qutline
Introduction and Overview
Acquiring the Sexual Phenotype
Primary Sex Determination
Formation of the Indifferent Gonad
Gonadal Differentiation
Secondary Sex Determination
Primary Sex Characteristics
Secondary Sex Characteristics
Genetic Basis of Sex Determination
Hormonal Influences on Sex Determination
Environmental Influences
Social Factors
Temperature-Dependent Sex Determination
Endocrine Disrupters
The Sex Ratio
Sex Allocation
Gonochorism
Hermaphroditism

... for strange effects and extraordinary combinations we must go

to life itself, which is always far more daring than any effort of the

imagination.”
«Sir Arthur Conan Doyle

introduction and Overview

* The sexual phenotype, or gender, of vertebrates is determined

through either one of two main mechanisms. In most forms, it is determined
by genetic factors such as the presence of certain genes that reside on
particular combinations of specialized chromosomes. In others, maleness or
femaleness is not as clearly rooted in genotype and is acquired
independently of chromosomal composition. The sexual phenotype of such
forms is' under epigenetic control and is the result of environmental
influences that act on the genes and regulatory mechanisms involved in
sexual differentiation. Many species have evolved ways of taking
advantage of this epigenetic nature of sexual differentiation. in a few
species, individuals are even capable of switching from one gender to
another according to the environmental conditions that they encounter.

This chapter explores the ways in which fundamental

characteristics of sexual phenotype are initially acquired during embryonic
development and how they are generally organized in the various
vertebrate groups. In addition, I will consider the influences that direct the
14

development of sexual phenotype and the mechanisms by which the

environment can alter sexual phenotype independently of genotype.
Finally, 1 will focus on the ways in which these developmental and
environmental inftuences result in the distribution of gender between
individuals within populations.

Acquiring the Sexual Phenotype

The sexual phenotype of an individual is commonly associated

with a wide variety of structural and functional characteristics. In our
common experience, we identify gender by considering one or more
characteristics that we have learned to associate with a particular sexual
phenotype. Coloration, body size/mass, behavior, scent, and appearance of
external genitalia as well as other outward modifications of body form are
characteristics that we commonly take into account in assessing the gender
of an individual, Still, many of these characteristics may be shared by
both genders or may even be absent altogether within some taxonomic
groupings. For example, the members of some vertebrate taxa simply do not
possess external genitalia, making it impossible to assess gender on that
basis alone. In others, external genitalia are present yet virtually
indistinguishable on the basis of gender. When it comes to studying
sexuality in vertebrates, then, we must establish a more broadly applicable
method of assessing gender.

When performing a comparative analysis of vertebrate sexuality,

there is one reliable indicator of sexual phenotype in an individual — the
type of gametes that it is capable of producing. The male gender is
identified on the basis of its capacity to produce sperm, or spermatozoa, by
the process of spermatogenesis. The female gender produces ova through
cogenesis. Both types of gametes are produced by specialized structures
called gonads. in males, functional gonads are referred to as the testes
while in females they ate called the ovaries.

Primary Sex Determination

Primary sex determination is the process by which gonads are

formed and then differentiate into either testes or ovaries. For this reason,
primary sex determination is considered to be the ultimate basis for
acquiring sexual phenotype in most vertebrates. ln most taxa, the type of
gonad to be formed is determined early in embryonic development. In some
amphibians and reptiles, however, gonads do not differentiate during
embryonic development and as a consequence, gender is not established until
well into the first year of independent life. In the case of some fishes,
gonadal differentiation and primary sex determination is delayed for
several years.1

las in the American eel, Anguilla rostrata.

15

indifferent gonad
Mesenchyme

Coeélornic epitheliura Dorsal mesentery

A Coélomic space

Genital ridge

Primary sex cords

Coelomic epithelium.

Primary sex cords i

Tunica albuginea —

Primary sex cords

(degenerating)

Testicular cords AL Tunica aibuginea

tdegenerating)
Secondary sex cords

Coelomic epithelium

Tunica albugines

Rete testis Remnant of primary

. sex cords
Seminiferous tubules

. . Ovarlar: follicles
Tunica albuginea Coelomie epithelium

Testis Ovary

Figure 2.1 Early gonadal development and differentiation in mammals. Gonadal

differentiation begins when the coelomic epithelium of the gonadal ridge thickens
and
forms primary sex cords which extend into the underlying mesenchyme. Primary sex
corás are then separated from the surface epithelium by connective tissue elements
of
the tunica albuginea. En males, primary sex cords differentiate into seminiferous
tubules, In females, primary sex cords degenerate as secondary sex cords form at
the
surface epithelium. Secondary sex cords give rise to ovarian follicles.

Formation of the lndifferent Gonad

" Most of what we know of gonadal differentiation is based on studies

of reptiles, birds, and mammals (see Figure 2.1). Gonadal differentiation in

these amniotes begins with a paired lateral thickening of the splanchnic

mesoderm along the posterior wall of the coelom and mesial to the
opisthonephros or mesonephros.? This mesodermal thickening is referred to
as the genital ridge or gonadal blastema3 Scon after this ridge is

2 Sas Chapter 4 for a description of these structures.

3 This structure has also been referred to as the germinal epitheHuam because it
was mistakenly
thought that the germ cells originated within the epithelium.
16

established, the basal lamina of its overlying coelomic epithelium begins

to drop down into the subjacent mesenchymal tissues at numerous locations
along its length. As the lamina descends, it carries with it cells derived
from the proliferating coelomic epithelium. This leads to the formation of
numerous epithelially-derived tissue cords which extend deep within the
underlying mesenchyme and remain surrounded by a basal lamina. These
cords are referred to as the primitive sex cords and are configuous with the
coelomic epithelium of the genital ridge.

While the primitive sex cords are forming, the presumptive gonad
of both males and females is considered sexually undifferentiated and
referred to as an indifferent gonad. This structure consists of two basic
layers. The outermost layer is the cortex and is comprised of a thickened
epithelial layer facing the coelom. The inner layer is the medulla and is
cormprised of the primitive sex cords and their surrounding mesenchymal
tissues. Subsequent structural and functional differentiation of the gonad
involves a further modification of the basic organization established in
this indifferent gonad,

Gonadal Differentiation

The following description of gonadal differentiation is based on the

mamumalian condition.

During testicular differentiation, the primitive sex cords

established in the indifferent gonad undergo further growth and are
separated from the germinal ridge by an invading layer of mesenchymal
cells which differentiate into the connective tissues of the tuamica
albuginea. Establishment of this tunica severs continuity between the
coelomic epithelium and its derivative sex cords. Shortly after their
formation in the indifferent gonad, the primitive sex cords are invaded by
migrating primordial germ cells (PGCs), or gonocytes that originated
elsewhere in the body. Primordial germ cells (PGCs) are relatively large
celis that, depending on the taxon, migrate from different points of origin
into the tissues of the secondary sex cords. Ín fishes, PGCs originate in the
gut endoderm while in amphibians they originate from the vegetal
endoderm or marginal mesoderm. Among the nonmammalian amniotes, they
derive from either the anterior, medial or posterior? extraembryonic
tissues (Hubert, 1976) while in mammals they may even arise from the
hindgut endoderm.

4 As in birds and snakes.

5 As in some lizards.
6 As in most lizards including the tuatara, Sphenodor, and turtles.

17

A Epididymal duct

Deterert duct 3 + 1
(vas deferens) A (+ mesonephric duce)
Epididymal head

Efferent duet

Epididymaf body
Tunica albuginea

Rete testis

Tubulus rectus

Coelomic epithetum
Comnestive tissue septum

Seminiterous tubules
within fobule

Epididymal tail

Figure 2.2 Organization of the mammalian (human) testis.

The presence of the PGCs is essential to the completion of

gametogenesis since they are components of the germ cell line that
ultimately give rise to the gametes. Ín addition, mesenchymal cells
derived from tissues of the degenerating mesonephric tubules migrate into
the areas between the growing and elongating sex cords where they
differentiate into testosterone producing interstitial cells, or Leydig cells,
and elements of the testicular interstitial connective tissue, As the gonad
continues to differentiate, the sex cords eventually give rise to the greatly
elongated testicular cords which in turn differentiate into the seminiferous
tubules comprised of epithelial cells and cells derived from PGCs (Figure
2.2).

In females, the primary sex cords and medullary mesenchyme of the

indifferent gonad undergo progressive degeneration to form tissues of the
ovarian hilam. However, the overlying coelomic epithelium continues to
proliferate, giving rise to numerous of secondary sex cords, or cortical cords,
that remain close to the outer surface of the ovary. These structures then
subdivide into small clusters of cells known as ovarian follicles or primary
follicles that eventually are surrounded by connective tissue of the ovarian
stroma (Figure 2.3).

Once within the secondary sex cords, PGCs divide and give rise to a
population of interconnected gonial celis. Eventually, each of these is
surrouded by epithelial cells of the newly formed follicle and their
intercellular connections are severed. These epithelial cells are also
referred to as granulosa cells. Mesenchymai cells derived from the
18

Late secondary
folficie Theca cells

Early secondary
tollicie
Primary follicie
Cagonia
Early tertiary
Secondary Tofiole
sex cords "heca cols
Í - ce
Ovarian artery hs
Ovarian vein Cocyte within

cunsulus cophorus
Antrum

Corpus abicans Granutusa cells

of mature tertlary

follicie
Ruptured tollicie

Mature corpus
luteum

Oboyte within:
adherent cumulus
Hilum oophorus
Ovarian stroma
Lymphatic space

Early corpus luteur

Figure 2.3 Organization of the mamumalian (human) ovary. Successive stages of

follicular development are represented in a clockwise manner. H should be noted
that
the stages represented do not occur concurrently.

medullary mesenchyme differentiate into theca cells which adhere to the

outward facing basal lamina of granulosa cells. Newly formed ovarian
follicles therefore consist of a single gonial cell surrounded by a layer of
folicular epithelium and an overlying layer of mesenchymally-derived
theca cells. Though there is some taxon-specific variation in the patterns
described above, it can be said that with the exception of teleosts, it is the
medullary tissue and primary sex cords that are differentiated into the
functional tissues of the male gonad while in females it is primarily the
cortical tissue.

Comparative Aspects

Since so much of what we know about gonadal development in

vertebrates is based on mammalian studies and so little is known of this
process in the nonmamunalian vertebrates, we are forced to make deductions
about gonadal development in nonmammalian vertebrates based not on
direct observations of that development but rather on observations of their
adult gonadal configuration. Still, despite this obvious disadvantage, it is
possible to gain some general insights into the mechanisms of their
development by considering the position and relative functional
differentiation of gonads im members of the major vertebrate groups.
Observations of adult gonadal configuration can also help us to appreciate

the many ways in which these structures were subject to modification during
their evolutionary history.

19

In fishes, the general configuration of gonads is highly variable. In

cyclostomes, paired gonads begin their development on either side of the
body and then fuse to form a single median structure. ln the sea lamprey,
Petromyzon marinus, this single fused gonad is either testicular or ovarian
in structure. However, in the hermaphroditic hagfish such as the Atlantic
hagfish, Myxina glutinosa, the anterior region of the single gonad
differentiates into an ovary while the posterior region becomes testicular in
organization. Because individual hagfish alternate between the production
of sperm and ova, onty one of these areas is functional at any given time.
Paired dorsal testes characterize all male elasmobranchs. Among female
elasmobranchs, there are, however, some differences in gonadal configu-
ration. In zygoparous” elasmobranchs, ovaries are paired while in embryo-
parous species? only the right ovary becomes functional. This situation is
reversed in the exclusively embryoparous rays in which it is the left gonad
that becomes functional, In teleosts, testes are paired and dorsally situated
while ovaries may be either paired or fused to varying degrees along the
midline. In the indifferent teleost gonad, both male and female germ cells
are grouped into areas containing germ cells of similar sex or, even more
often, they are scattered throughout its tissues. During testicular
differentiation, the number of female germ cells is reduced. Differentiation
of the ovary is marked by a similar reduction in male germ cells, Most of
the tissues of the functional gonad of both sexes are derived from the cortex
of the indifferent gonad. This common origin of gonadal tissues may be
related to the apparent facility by which hermaphroditism has
independently evolved among many teleosts (Dodd et al., 1960).

Basic overall configuration of the gonad takes several forms among

the tetrapods. In amphibians, testes tend to be paired lobular structures
while ovaries are paired and hollow, In reptiles, testes and ovaries are
both, paired and solid. In male birds, the left testis generally develops more
extensively than the right testis, while in most species of female birds,
only the right ovary develops. Experimental studies have provided some
insights into the proximate origins of gonadal asymmetry among the birds.
Unequal development of the gonads is related to the degree to which the
PGCs migrate into the presumptive gonad. In most reptiles and mammals,
PGCs migrate equally into the two gonads. In birds however, the PGCs are
unequally distributed between the right and left genital ridges beginning at
about 96 hours of development. PGCs preferentially migrate to the left
genital ridge in a ratio of about 5:1. The resulting inequality of PGC
distribution appears to affect the development of the germinal ridges and
prevents functional differentiation of the right gonad in most species. The
same pattern of differentiation may also occur during gonadal development

7 Those with no significant period of embryo retention.

8 Those with significant periods of embryo retention,
20

in other lineages in which one gonad develops to a greater extent than the
other?

The gonads of amphibians, reptiles, birds, and manwmnais differ in

anatomical position, organization, and in the manner of their functional
differentiation. Although they share the basic form of the vertebrate
gonad, the processes that underlie gonadal development must have been
subject to very different selective pressures during the course of each group's
unique evolutionary history. Each group's unique history led to a obvious
differences in its mature gonad, and yet despite these differences, there are
marked similarities among the groups at the level of tissue interactions and
modes of differentiation. Any specific differences between gonadal
development in different taxa, then, may be thought of as variations on the
basic, ancestral theme of vertebrate gonadal formation. For a more detailed
account of the organization of adult vertebrate gonads, see Chapter 4.

Secondary Sex Determination

Gonadal differentiation during primary sex determination is only

the first step toward establishing sexual phenotype. Adult sexual
phenotype also involves features associated with the transport and storage
of gametes within the body as well as adaptations which heip to ensure
that the gametes of opposite sexes are brought together. Sexual phenotype
can involve as well certain adaptations that facilitate the care of
developing ofíspring.

Secondary sex determination involves the development of such

extragonadal characteristics. Differentiation of such structures usually
takes place under the influence of hormones secreted by the gonad. Thus,
the adult sexual phenotype of an individual can be thought of as the
product of gonadal differentiation followed by the action of gonadally-
produced hormones ún extragonadal tissues. The process of secondary sex
determination occurs in two phases. The first phase of secondary sex
determination occurs during embryonic or larval development and gives rise
to overt extragonadal features of sexual phenotype. These primary sex
characteristics include basic features of the urogenital system such as sex-
specific urogenital ducts and external genitalia. The second phase occurs
during the transition from juvenile to adult and leads to the development of
sex-specific characteristics known as secondary sex characteristics.

Primary Sex Characteristics

Primary sex characteristics are initially established through the
differential development of paired Wolffian ducts (=mesonephric ducts)
and Miillerian ducts (=paramesonephric ducts) present in the embryos of
both sexes (Figure 2.4). Differences in how these mesodermally derived
ducts develop lead to fundamental differences between the primary sex

9 As in the ovaries of embryoparous sharks and rays.

21

characteristics of males and females. The following description of primary

sex characteristics and how they arise in vertebrate males and females
adopts the mammalian system as its model.

In male mamumals, primary sex characteristics are established

through atrophy of the Múllerian ducts and continued development of the
Wolffian ducts. This is accomplished through the action of two

Sexually indifferent

Mesonephric tubute Gonad

Mesonephric duet Paramesonephric duct
(primary utinary duct) (Múllerian duet)

Urogenital sinus

Female

Oviduot

Duectus deterens Uterus and vagina

Urogenital sinus Urogenital sinus

Primordium of prostate

Figure 2.4 Differentiation of the urogenital ducts in manuals.

22

AUTOSOMES co o 10983 Promo ing

factors

INDIFFERENT |
GONAD ccanaco!

TESTIS OVARY

ovary-praducing E——— 7
actora

Y-CHROMOSOME ———> tistis-determining

factors

INTERSTTIAL — SUSTENTACULAR THECA AND GRANULOSA

CELLS CELLS SELLS
, j j
ANTI-MULLERIAN
DUCTHORMONE
WOLFFIAN
DUOT
W
TESTOSTERONE MULLERIAN BREAST
buer TISSUE
EPIDIDYMIS, VAS
DEFERENS. mOGEN
SEMINAL VESIOLES ES
UTERUS, OVIQUOY, ARMAR
OERVIX, UPPER PONER

5 -OIHYOROTESTOSTERONE VAGINA

GENITAL PENIS
TUBEROLÉ

Figure 2.5 Control of urogenital development in mammals,

masculinizing hormones, testosterone and anti-Múllerian duct hormone

(AMH). Testosterone is produced by interstitial cells of the testes and
promotes Wolffian duct development. Anti-Múllerian duct hormone which
dis also known as Miillerian inhibiting substance (MIS) is produced by
-sustentacular cells of the testis and is indirectly regulated by a gene
productl0 responsible for initiating the cascade of interactions that result in
a wide range of male phenotypic characteristics (Zwingman et al., 1993)11
In males, AMB causes atrophy of the Milllerian ducts thereby preventing
development of characteristically female structures such as the oviducts
(Fallopian tubes), uterus, and upper vagina (Figure 2.5).

la contrast to the amount of information we have about the

hormonal regulation of male secondary sex determination during embryonic
development, our understanding of female secondary sex determination is
limited. Differentiation of Múllerian ducts into the oviducts, uterus, and
upper vagina is promoted by the influence of estrogen secreted by the cells
associated with the ovarian follicle (both the theca and granulosa cells)
(Figure 2.5).

10 The SRY gene product.

11 See the ensuing section on the genetic basis of sex determination.

23
Undifterentiated
Genital tubecie
Genital told:
Genital groove
Genital swelling
Aral git
Gians
Glans
Shatt
Jrogenital groove Shatt
Genital fold Urogeaital groove
Labio-scrotal Genital told
Swelling Labio-Scrotal
Perineal raphé: swelling

Anal tubercle Perineal rapné

nm Anal tunerale

Urethral meatus Shatt of clitoris

Glans penis o
Glans clitoris
Prepuce (retracted)

s .
Shaft of penis Uretbral meatus.

Peno-scrotal raphé Labia minora

Labia majora

Perineal raphé Perineal rapné

Anus AUS ce,

Male Female

Figure 2.6 Differentiation of the genital tubercle in humans.

Primary sex characteristics not only involve differential

development of Múllerian and Wolffian ducts but they also involve
differences in the development and final configuration of external
genitalia. During early development in many mammals, both the male and
female external genitalia originate from three common primordia in the
sexually indifferent embryo. These are the genital tubercle, genital folds,
and the genital swellings (Figure 2.6). Depending on the gender of the
individual, the genital tubercle differentiates to form most of the male
penis or the female clitoris. Genital folds give rise to the penile urethra
and surrounding tissues in males and the labia minorae and tissues of the
vestibule in females. Genital swellings form the male scrotal tissues, while
in females they give rise to the tissues of the labia majorae. Primordia
eventually differentiate into female structures, unless the hormone 5 Q%-
dihydrotestosterone is present. A product of testosterone conversion by 5 0%-
ketosteroid reductase, this androgenic, or male-producing hormone, is
24

responsible for differentiation of the mamumalían urogenital sinus and its

surrounding genital primordia into male structures. A more comprehensive
treatment of vertebrate urogenital systems is presented in Chapter 4.

Secondary Sex Characteristics

Features of adult sexual phenotype that show a high degree of

structural modification during the transition from juvenile to adult are
known as secondary sex characteristics. These include all post-embryonic or
post-larval changes in sexual phenotype and are primarily directed by the
hormonal influences of the gonads (see Figure 2.5). The resulting differences
between external appearance of adult males and females are the basis of a
species” sexual dimorphism and may either be permanent or temporary.
Permanent features, such as plumage color in some male birds or facial hair
growth in male humans, persist throughout adult life while temporary ones
appear only during the reproductive season in seasonally breeding forms.

Because genetically-based variations in degree of development and

differentiation of secondary sex characteristics may offer individuals a
selective advantage, natural selection operates to maintain such variations
within a population. The mechanism by which sexual characteristics are
responsible for an individuals differential fitness is known as sexual
selection. In some cases, sexual selection results in quite extraordinary
adaptations — adaptations that demonstrate a statistically significant
relationship with mating success (Andersson, 1994). However, the same
traits that increase mating success may also reduce individual

- survivability (Lande, 1980). Therefore, these traits evolve as stable

features of reproduction within a given population only if the advantages

accrued by their development outweigh the disadvantages.

Vertebrate secondary sex characteristics often include modifi-

cations of general body size and proportions (i.e., changes in allometry),
post-larval or post-juvenile changes in the functional differentiation of
specific body parts, changes in coloration, and changes in behavior
patterns, in female lampreys, secondary sexual characteristics involve a
seasonal swelling of the perianal and pericloacal tissues as well as
enlargement of the urogenital papilla and ventral fin. In chondrichthyan
fishes, sex-specific differences in body size and proportions are common
types of secondary sexual characteristics. Osteichthyan fishes exhibit
numerous examples of sex-specific modifications of body size, morphology
and coloration. Body shape and coloration are often used in mating and
aggressive displays, while differences in body size and proportion are
moderately pronounced in salmonids (e.g. the pink salmon, Oncorhynchus

12 Sex-specific modifications in osteichthyan: fishes include dorsal and caudal

fins of male
siamese fighting fish, the gemmeous dragonet, Callionymus lyra, and the modified
caudal fin of
the poeciliid Xiphopkorus helleri.

25

gorbuscha) and can be quite striking in some deep-sea ceratioid anglerfishes

(e.g. Edriolychnus schmidti).

In some instances, secondary sex characteristics include structures

associated with internal fertilization, embryo retention, or the care of
young. In fishes, they may even involve tissues of the gonad. Modification
of the ovary as an organ of post-copulatory sperm storage and site of
embryo-retention occurs in some teleosts.23 In teleost species with internal
fertilization, the males develop secondary sexual characteristics that
include modified anal or pelvic fins used as copulatory organs. in other
species, modified male pelvic and pectoral fins are used in brood
maintenance24 In an interesting instance of male secondary sexual
characteristics, the ventral surface of the male seahorse, Hippocampus,

becomes a highly specialized brood pouch, or marsupium, used to protect

and nourish developing young.

Perhaps the most remarkable example of sexual dimorphism,

however, occurs in deep-sea ceratioid anglerfish (eg., Linophryne,
Neoceratias and Edriolychnus)(see Figure 2.7b). Males possess a host of
traits that help make their search for females more efficient. The male
deep-sea ceratioid anglerfish is distinguished from the female by his
larger eyes, smaller body size, and more well-developed tail (Berthelsen,
1951). In some ceratioid anglerfishes, males also possess an extremely well
developed nasal organ they use to locate females in the vast and sparsely

Figure 2.7 Secondary sex characteristics. A, Salmon, Oncorhynchus (after Walker

and Lie) B, Ceratioid anglerfish, Linophryne (after Regan and Trewavas). C, Newt,
Triturus vittatus (after Raxworthy).

13 Modification of the ovary occurs in some members of the Poeciliidae, for

example.

14 Modified pelvic and pectoral fins appear, for example, in the ovuliparous South
American
lungfish, Lepidosiren paradoxus.
26

populated darkness of the deep-sea (see Jumper and Baird, 1991).15 Once
the diminutive adult male anglerfish locates a female, he uses bis jaws to
clamp himself onto her body to which he remains permanently attached.
For the rest of his life, he lives by drawing sustenance from the females
tissues at the site of attachment. In some species (e.g. Edriolychnus
schmidti), the permanent association of female and male involves an
interaction between the female and male vascular systems, in which case,
all the male's organ systems degenerate with the important exception of
his reproductive organs. Thus, the male reproductive organs continue to
supply sperm when the female releases her ova. In effect, the male's
identity as an individual is lost and the female of the species becomes a
hermaphrodite. For a detailed discussion of hermaphroditism, see below.

Many amphibians also exhibit marked secondary sex

characteristics. Sex-specific differences in body size and proportions,
increased bulk of brachial musculature and the development of spines or
abrasive nuptial pads are common in many anurans (e.g., Leptodactylus
ocellatus). The males of some anuran species even develop an extra digit
called the prepollux that helps maintain a grasp on the female during the
mating association of amplexus. Other examples of amphibian sexual
dimorphism include the relatively large tympanic membranes and
pronounced vocal sacs of male anurans (e. g., Rana catesbiana) and the male
coloration patterns of several species of urodeles (e.g., the brightly colored
tail crest in Triturus)(see Figure 2.7c). In some taxa, sexual dimorphism also
involves adaptations associated with internal fertilization, embryo
retention, or the brooding of young.16 Secondary sex characteristics among
amphibians may also involve differences in male and female calling and
locomotory behaviors.

In the reptiles, secondary sex characteristics are generally less

overt than in other vertebrate groups. The most obvious non-behavioral
examples of secondary sex characteristics include the concave ventral
plastron of turtles which allows the male to fit over the female during
copulation. Sex-specific differences in body size and body part proportions
are most pronounced in crocodilians. In the snakes, scale configuration is the
most obvious outward manifestation of gender? although in some species
of snakes, the production and detection of scent likely plays the most
important role in gender identification. In lizards, throat appendages,
horns (e.g., Chamaeleon bifurcus), or other such modifications of the body
surface and overall configuration serve as sex-specific visual signals for use
mating. Behavioral examples of secondary sex characteristics are mumerous

15 Relative to body size, the males of some species of ceratioid' anglerfish have
the largest nasal
organ of any vertebrate (see Berthelsen, 1951).

16 To be discussed in subsequent chapters.

17 Butis likely not subject to sexual selection.
27

among the reptilian taxa and involve stereotypic movements of the head
and body during mating. In some species, they may even involve
vocalization.18

The appearance and maintenance of secondary sex characteristics in

birds is controlled either directly by the pituitary or by gonadal steroid
secretion (Witschi, 1962). While secondary sex characteristics are evident
in the elaborate behavioral patterns associated with mating, nest building,
and parental care, birds are perhaps best known for their sex-specific
patterns of plumage coloration and song. The brilliantly colored tail
feathers of the male peacock, Pavo cristatus, the ornamental tail feathers
of the male hummingbird, Spathura underwoodi, and the fantastic
phimage of the male bird of paradise are only a few of the more extreme
traits that sexual selection is capable of producing. Species of fowl are
known for other secondary sex characteristics including combs and throat
appendage, while sex-specific patterns of vocalization are apparent in
species of songbirds. The male songbird possesses a specialized sound
generating tissue at the region of the juncture between the trachea and
bronchi, Known as the syrinx, it is responsible for helping generate the
complex song of such birds as the male sedge warbler (Catchpole, 1980).
Still, this is not to say that all species of birds exhibit dramatic sexual
dimorphism of this kind. In fact, the females and males of certain species
are virtually indistinguishable.1*

Secondary sex characteristics are well developed in many

mammnals. In males, most of these characteristics develop under the
influence of testosterone and include sex-specific differences in body size
and proportion,2% differences in muscle mass? presence of either horns,
antlers, or enlarged dentition,22 as well as patterns of hair distribution and
color. In female mamuals, estrogen along with other hormones has an
influence on the sex-specific differentiation of a number of characteristics
including the mammary glands.23

It is interesting to note that, within a given species, overt secondary

sex characteristics are often far more pronounced in males than they are in
females. One reason for this may be that females are subject to far less
intrasexual competition owing to the fact that ova are usually more limited
a resource than sperm, This is directly related to sex-based differences in

18 As when male crocodyles roar during mating.

19; : :
2 Dimorphisms are not apparent in the gannet, Sula, diver, Gavia, grebe, Podiceps,
and puffin
Fratercula arctica. "

20 As in the large size of male elephant seals, Mirounga anguistirostris.

21 Asin some primates including humans.

22 Asi
in the tusks of bull elephants, narwals, and walruses or the prominent canines of
some
primates.

23 See Chapter 11 for discussion of the regulation of mammary development.

28

gamete size and the differential costs associated with their production,
Due to these differences, the pressures of sexual selection tend to be stronger
on the male sexual phenotype of most vertebrates.2%

Although overt examples of secondary sex characteristics abound in

vertebrates, some may be less obvious and therefore more difficult to
classify. For instance, differences in male and female behaviors may be
based in the different structures of the male and female nervous system, but
the role that the central nervous system plays in vertebrate behavior is so
complex that it defies easy explanation. The sex-specific behaviors of
birds, for example, are known to be established and maintained by gonadal
hormones. Male song production in birds is made possible by steroids like
testosterone which influence the size of groups of cell bodies or nuclei,
within the central nervous system (CNS) that are associated with male
song. These cell bodies are larger in male zebra finches, Poephia guttata,
and canaries, Serinus canaria, than they are in the females of the species,
implying that song is the behavioral manifestation of secondary sex
determination of the central nervous system. For this reason, female birds
rarely sing while male songbirds are capable of producing elaborate
vocalizations. As a consequence, the relationship between testosterone,
brain morphology, and compiex behavior patterns appears correlated with
cyclic changes in serum testosterone levels, and so the relationship is
evidently reversible (Nottebohm, 1981),

It is slightly more difficult to identify the morphological bases of

sex-specific behaviors in mammals since, despite the amount of attention
they have received recently, only a few sex-specific behaviors have been
clearly identified. Still, testosterone does appear to have a significant
impact on mammalian CNS structure and function. For example, spinal
nuclei involved in the coordination of penile thrusting are larger in male
rats than the corresponding structures in females of the species (Tobin and
Joubert, 1992). In species where males must search for females during
mating, there are sex-based differences in spatial learning ability that are
correlated with differences in brain structure. Since hippocampal structures
are involved in spatial learning, these structures are larger in the male
than they are in the female of the species, for example (Gaulin and
FitzGerald, 1989; Jacobs et al., 1990). These differences in the structure and
function of CNS tissues are often attributed to critical periods of CNS
sensitivity during development when the levels of testosterone circulating
in males differ from the levels circulating in females.

The functionality of the mammalian female brain is often regarded

as a default condition arising from low serum testosterone levels during
embryonic and post-natal development. However, such an assessment may

4 The evolutionary consequences of limited availability of ova in relation to

spermatozoa will be
more carefully discussed in relation to the evolution of mating systems in Chapter
13,
29

not be entirely accurate, since neuronal sensitivity to estrogen occurs in CNS

structures of adult female rats and may also be operating during
embryogenesis. Structures affecied by estrogen include the amygdala,
hypothalamus and pituitary which are implicated in the coordination of
typically female behaviors%* (Rubin and Barfield, 1980). Functional
differentiation of the female brain may also be due to the action of
estradiol formed through the intracellular aromatization of testosterone
(MacLusky and Naftolin, 1981).

The developing female brain's sensitivity to the. action of

testosterone poses interesting problems for some embryoparous species. In
some mammals, it appears that serum testosterone levels in female embryos
can be affected by uterine position with respect to cohort male embryos.
Though normally present at low levels in the circulatory system of
developing females, testosterone titers may be elevated through its
diffusion from a male cohort to the maternal uterine circulation. In this
way, female embryos situated near one or more male embryos are often
exposed to elevated levels of testosterone. When this occws, these
individuals develop into adults exhibiting more masculine morphology and
greater incidences of male” behaviors than do female embryos situated
between two female cohorts (Goy and McEwen, 1980).

In some mammals, the necessity of development within what is

essentially a female uterine environment may pose potential problems for
male embryos as well Elevated levels of the hormones estrogen and
placentally-derived progesterone have the capability of feminizing male
embryos. To protect against female steroids that might cross the placenta,
male embryos produce specialized serum proteins which bind and
inactivate them - serum proteins that include albumin, a -acid glyco-
protein, o-fetoprotein, corticosteroid binding globulin, progesterone-binding
globulin, sex hormone-binding globulin, and several other forms of steroid
binding protein. Though their operation is not as well understood, similar

proteins likely play an equivalent role in controlling hormone titers during

the development of females.

Genetic Basis of Sex Determination

“In a good many vertebrates, the sex or gender of an individual is

directly determined by the activity of certain genes that reside on specific
chromosomes. This mode of sex determination is known as genetic sex
determination. In vertebrates with genetic sex determination, chromosomal
complement is usually predictive of sexual phenotype. We will now
consider the chromosomes that are responsible for sexual phenotype and
discuss a number of the genes contained within these chromosomes.

25 Behaviors such as lordosis, or arching of the back, to accommodate penetration

of the male
phallus during copulation in rodents.
30

Most vertebrates possess one of two combinations of a specialized

set of heteromorphic chromosomes called homologous heterosomes or sex
chromosomes. Unlike the homomorphic autosomes which comprise the
remainder of the chromosomal complement, homologous heterosomes are
structurally distinct from one another. It is through different combinations
of the homologous heterosomes, established at the time of fertilization,
that the sex of an individual with genetic sex determination is established.

Thus, in forms with genetic sex determination the sex of an

individual is deterministic and usually due to the presence of the sex-
determining chromosome. Á heterogametic individual has a 50% chance of
producing a haploid gamete containing either one of the two types of sex
chromosomes that it possesses. Generally speaking, in forms with genetic
sex determination, it is the heterogametic parent's gamete that determines
the sex of the offspring. If the two heterosomes of an individual are
different from one another, that individual is said to be heterogametic. But
if they are similar, the individual is said to be homogametic. In most
mammals, the heterogametic condition (usually notated as XY) results in
the differentiation of a males In most reptiles and birds, females are
heterogametic, and therefore to avoid confusion a different system of
notation (ZW) is applied. Thus, the notation of XX/XY or XX/XO is used to
represent the sex chromosomes for forms in which females are
homogametic, while ZZ/ZW or ZZ/ZO have been used to represent forms
in which the males are homogametic. The letters used in the XY system are
loosely based on their similarity to the form of the chromosomes they
represent. Unfortunately, the letters used in the ZW system have been
arbitrarily assigned. When the notation "O" is used, it indicates that one
of the heterosomes is absent. Though sex chromosomes are heteromorphic in
most vertebrates, it should be noted that in at least one species of crocodile
(Singh and Ray-Chaudhuri, 1973) and three species of turtles they are
outwardly homomorphic.

Translocation of genes between the sex chromosomes, or from sex

chromosomes to autosomes, can lead to some anomalous chromosomal
combinations in certain individuals. XX males or other unusual
combinations arise when fragments of the Y chromosome are translocated to
either the X chromosome or to the autosomes (Simpson, 1982; Burgoyne,
1984). However, such anomalous arrangements seidom persist within
populations.

Nondisjunction of sex chromosomes during meiosis can lead to the

formation of aneuploid offspring lacking a normal chromosomal
complement. Aneuploids may either lack a particular chromosome, or
possess an extra copy of that chromosome. For example, human males who
suffer from Klinefelter's syndrome, possess an extra X chromosome derived
from a gamete that contained an extra copy of the X heterosome. Humans
with this syndrome have a genotype of XXY and are phenotypically male.
31

However, presence of the extra X chromosome usually results in mental

retardation and an inability to undergo spermatogenesis.

Other aberrant heterosomal combinations in humans include XYY,

AXXY, XXXXY, XO, XXX, XXXX, and XXXXX and are also the product of
nondisjunction, XO females occur when an heterosome-deficient ovum is
fertilized by a sperm carrying an X chromosome. These individuals suffer
from Turners syndrome and lack the ability to differentiate functional
gonads, XXX females are known as super females are phenotypically
normal but usually sterile. "These examples illustrate that either a Y or at
least two X sex chromosomes are required for functional differentiation of
the gonad and that regardless of the degree of aneuploidy in mammals, the
male phenotype is always expressed if a Y sex chromosome is present.
Supermamerary sex chromosomes are normal features of the nucleus in
marsupials (e.g. XYY males and XXXX females)(Hayman and Rofe, 1977)
as well as in some birds, amphibians, and fishes with a ZW chromosomal
system.

Recent studies on humans and mice indicate that the expression of a

gene on the Y sex chromosome is ultimately responsible for genetic sex
determination in mammals. This gene, called the SRY (sex-determining
region of the Y) gene in humans” is located within a 35 kb region on the
short arm of the Y sex chromosome. Expression of SRY/Sry by primary sex
cords of the indifferent gonad for a brief period of time results in the
production of a testis-determining factor (TDF) (McLaren et al, 1984).
Action of TDF initiates a developmental cascade resulting in functional
differentiation of the male phenotype. Thus, SRY/Sry gene expression qn
the Y chromosome in mammalis is the first step in a developmental cascade
associated with male differentiation that ultimately involves the
expression of autosomal genes (McLaren et al., 1988). Presence of the Y-
linked SRY/Sry gene may explain why certain aneuploid individuals
exhibit a male phenotype. Genotypes such as XXY, XXXY and XXXXY
result in male phenotype because of the developmental cascades initiated
by the Y-linked SRY /S+y gene. Absence of the SRY /Sry gene in individuals
with XO, XXX, XXXX, and XXXXX genotypes would therefore lead to
differentiation of the female phenotype as in normally XX individuals.

- Mapping of the human Y-chromosome has recently led to the

discovery of other genes that may also be involved in sex determination.
One of these, the WTI gene, encodes a zinc-finger protein similar to
members of the early growth response family of polypeptides (Wolf, 1995).
WTI takes part in the early commitment of gonadal tissue and
establishment of the bipotential gonad. It also is involved with the
development of nephric structures (Wolf, 1995). In adults, WTI expression is

26 kagwn as Sry in mice.

32

limited to somatic cells of the male seminiferous epithelium and the

sexually homologous epithelial cells of the ovarian follicle in females.
Another recently identified gene is the steroidogenic factor 1 (SF-1) gene
that is expressed during early stages of gonadal development. SF-1 is
expressed in male and female urogenital ridges during early development.
Once the gonad has developed into an ovary or testis, SF-1 expression
ceases in females but continues in the interstitial and sustentacular cells of
the testis (Smith, 1994). SF-1 appears to be involved in both the regulation
of steroid hydroxylases and in transduction of the SRY signal to the MIS
promoter in somatic cells of the seminiferous epithelium (Wolf, 1995),

It has also recently been proposed that in females, testis

differentiation is inhibited by an autosomal gene, Z, whose product
stimulates ovarian differentiation and inhibits testicular formation. In
genetic males, it appears that the Y-linked SRY/Sry gene is inhibitory to
this autosomal gene or its product (McElreavey et al, 1993). Thus, there
seem to be sex-specific genes that are responsible for eliciting patterns of
functional differentiation in females as well as in males. The existence of a
Z gene would suggest that, in developmental and genetic terms, the female
phenotype may be no more passively attained or incompletely formed than
that of the male (see Eicher and Washbuzn, 1986).27

Since the male phenotype appears initiated by the action of a

single gene, it is interesting to ask why sex determination in mammals
involves heteromorphic chromosomes. The genes controlling sexual
differentiation would be expected to reside just as easily on homomorphic
as on heteromorphic chromosomes. Certainly, Mendelian patterns of
inheritance would be able to maintain a 1:1 sex ratio if sex chromosomes,
like the autosomes, were homomorphic. On the surface it would appear
that genetic sex determination could have involved some heterozygous or
homozygous allele combination resulting in a 1:1 phenotypic ratio as a basis
for sex determination. This, however, is not the case.

Departure from such a system by heteromorphic sex chromosomes

may be related to the action of SRY/Sry which initiates male phenotype in
the hemizygous state. Fleteromorphism of the mammalian sex
chromosomes is thought to have originated through a transfer of a portion
of a biramous precursor Y chromosome to the X chromosome. Such a transfer
would result in a smaller uniramous Y chromosome and a larger X
chromosome containing additional material derived from the Y precursor.
Once such a chromosomal rearrangement took place, there may have been
no selective advantage for reversion since the SRY/Sry gene would still be
capable of functioning in the hemizygous state. Without any advantage to

27 According to these findings, it appears that the long held Aristotelian view
that females are
the “default” condition of sexual differentiation, and that the male sexual pattern
is
superimposed onto that of the female, may be in jeopardy.

33
reversión, it would be expected that such rearrangements of the sex
chromosomes could be maintained within the lineage. The fact that
heteromorphism of the sex chromosomes has independently evolved in
several teleost lineages raises the possibility that a structurally distinct Y
sex chromosome may, in fact, offer some form of selective advantage.

In nonmammalians, the heterogametic condition can give rise to

males or females depending on the specific lineage in question. In
cyclostomes and elasmobranchs, the genetic basis of sex determination is at
present unclear. Ín teleosts, the mechanism is either XX/XY or ZZ/ZW.28
Heterogamety also occurs among the amphibians. Most reptiles exhibit a
ZZ/ZMN system. in birds, a ZZ/ZO system is present with a ZO genotype
being normal for females. As we have already seen, mammals have what is
generally referred to as a XX/XY system with the male as the
heterogametic sex. All of these differences between the chromosomal
systems of reptiles, birds, and mammals suggest that the genes responsible
for sex determination did not associate with particular chromosomes until
late in vertebrate history.

Within several vertebrate lineages, there are some very interesting

departures from the normal distributions and actions of sex chromosomes.
These departures all seem to have been caused by nondisjunction of the sex
chromosomes during meiosis. Although nondisjunction tends to result in
impaired or nonviable ofíspring, in some mamunals it has given rise to some
extraordinary sex chromosome combinations retained as regular features of
the species” reproductive biology.

Chromosomal nondisjunction has led to the persistence of normally

occurring supernumerary sex chromosomes within two species of spiny
anteaters, the shortnosed spiny anteater, Tachyglossus aculeatus, and the
longmnosed spiny anteater, Zaglossus bruijmii. In both cases, the male
genotype is XXY and the female genotype is XXXX. In the European mole
rat, Ellobíus lutescens, and the Japanese spiny rat, Tokudaia osimensis, as
well as several other rodents, both males and females possess a structurally
similar X chromosome within the context of an XO genotype (Ohno, 1977;
Fredga, 1983). This condition undoubtedly involves two forms of the X
chromosome within populations of these species — one in which the Sry gene
is present and one in which dt is not. Ín the North American creeping vole,
Microtus oregoni, males ate heterogametic (XY) while females possess
either an XO or XX genotype. This condition comes about through routine
chromosomal nondisjunction during spermatogenesis resulting in viable OY
and non-viable XXY sperm. In these voles, homogametic females are
produced through nondisjunction of the X chromosome during oogenesis and

28 Both of these mechanisms are described in Xiphophorus helleri (=

maculatusKKallman, 1965).
34

subsequent fertilization of the ovum by an O containing sperm (Ohno et al.,

1966).

In the Eurasian wood lermming, Myopus schisticolor (Fredga et al.,

1976), and similarly in Dichrostonyx torquatus (Beneson, 1983), there is an
X-linked gene of high frequency that is capable of inhibiting testicular
development. Since males are the heterogametic sex (XY), presence of this
Xlinked gene in males leads to suppression of the male phenotype and
subsequent formation of a fertile XY female. Such females are capable of
producing XX and XY daughters that carry the X-linked gene. However,
fernales carrying this gene are incapable of producing male offspring.
Because males are produced only when an X chromosome lacks this gene,
the number of males within a population is far lower than the number of
females.2

Hormoñal Influences on Sexual Differentiation

hh a proximate sense, sexual phenotype may be thought of as the

result of hormonal signals that are produced and received according to a
precise schedule. In vertebrates with genetic sex determination, the genes
initiate the cascade of hormonal interactions leading to the acquisition of
sexual phenotype. However, any artificial or environmentally-induced
alteration of one or more of the hormonal signals involved in such a cascade
has the potential of altering the sexual phenotype expected from genotype.
Such epigenetic inftuences on sexual phenotype have the potential to take
place either during embryonic development or during adult life. Many
nonmammalian vertebrates have an enormous susceptibility to such
influences — so large in fact that they can be induced quite easily to reverse

gender under the influence of experimental hormonal exposure during

development.

Environmental Influences

By eliciting subtle changes in the production, reception, and timing

of the hormonal signais involved in sex differentiation, environmental
influences can act as epigenetic factors affecting the acquisition of sexual
phenotype (Pieau, 1982) Chromosomal complement is not necessarily
entirely determinative of sexual phenotype, since the phenotype may be
altered by environmentally-mediated changes in endocrine activity, for
example. Sexual phenotype can be affected not only by environmental

influences from endocrine disrupters to temperature changes, but by social

considerations as well.
29 ¡ ¡ ¡
However, it does not appear that this peculiar genetic system of sex determination
is an
adaptation to allow a female-biased sex ratio in response to population structure
(Bulmer, 1988).

35

Social Factors

In some fishes, social environment is known to trigger a change in

sexual phenotype. Though usually considered a form of sex reversal, sexual
differentiation in some male coral reef fishes occurs through a socially
induced alteration of the female phenotype if males are absent from the
immediate environment. Such a socially induced change in sexual
phenotype has likely evolved as a mechanism for maintaining specific sex
ratios in a population (Shapiro, 1984),

Sex reversals among some fishes may be induced by changes in

social interactions (Shapiro, 1984), population density (Chan and Yeung,
1983) or other environmental cues. For example, in the protogynous
Australian cleaner-fish, Labroides dimidiatus, it ás typical for a single
large male to control a group of up to six females.% The presence of a large
male within the immediate environment suppresses sex reversal in the
females. However, if the male dies or is removed from the area, the largest
female within the group undergoes sex reversal and replaces him, Thus, for
many vertebrates, chromosomal complement or even the presence of specific
gene combinations cannot be correlated with a specific sexual phenotype
since epigenetic factors related to a variety of environmental influences
often play a significant role in regulating the expression and maintenance of
sexual phenotype.

Temperature-Dependent Sex Determination

Specific temperature regimes during embryonic development can

influence the sexual phenotype of several vertebrates. The effects of
temperature on the hormonal milieu of the developing embryo effectively
establishes two distinct mechanisms of sex determination among
vertebrates. One involves the original mechanisms associated with
genotypic sex determination, and the other involves epigenetic control in
the form of temperature-dependent sex determination. The superimposition
of temperature dependent sex determination onto pre-existing mechanisms
of genotypic sex determination allows environmental conditions to directly
impact the ratio between males and females within populations of many

species. :

Certain vertebrates undergo labile sex determination where

temperature acts on sexual phenotype by directly altering hormonal
influences during early development. Known as temperature-dependent sex
determination (TSD or TDS), this process involves temperature-dependent
changes in hormona! interactions, and brings about sexual differentiation
quite independently of genotype. In the self-fertilizing ovuliparous
30 Such a group of females controlled by one male is known as a harem.
36

cyprinodont fish Rivulus marmoratus, the proportion of males hatching

from a clutch of “eggs” can be experimentally increased by decreasing the
temperature at which they develop (Harrington, 1961, 1971; Conover and
Kynard, 1981). TSD also occurs in other fish (e.g., the sole, turbot, and
Atlantic silverside, Menidia menidid), some frogs (Rana catesbeiana, Rana
jeponica, Rane sylvatica, Rana temporaria, Bufo vulgaris, and Bufo
vulgaris formosa), and urodeles (Hynobius retardatus and Pleurodeles
waltlii). In the amphibians, temperature increases during development
often result in male sexual differentiation, while decreased temperatures
promote female sexual differentiation.

Ín some turtlesói and all crocodilians, sexual phenotype is also

subject to the influences of temperature. In turtles, lowered developmental
temperatures result in a male phenotype while in crocodilians lower
temperatures produce females. ín the snapping turtle, Chelydra serpentina,
a top to bottom temperature gradient within the nest (Wilhoft et al., 1983)
is known to influence the production and action of estrogens which induces
female development (Rhen and Lang, 1994). Male sexual phenotype is
acquired in the absence of estrogenic influence. Among many crocodilians,
both male and female sexual phenotypes arise when developing young are
maintained at an intermediate temperature regime.

Among the alligators, pre-hatching developmental temperature

regimes are highly dependent on nest location. In such forms, it would
appear that nest site choice could be considered a mechanism of sex
determination and might constitute an important mechanism for
maintaining or skewing the ratio of males and females within a population
(Ferguson and Joanen, 1982). The dramatic 10:1 ratio of adult females to
adult males seen in populations of some crocodiles (Woodward and Murray,
1993) is likely due in part to maternal nest site selection and its consequent
effects cn embryonic development. However, in green sea turtles it is not
nest site as much as the position of the embryo within a clutch of 'eggs' that
determines the sex of the ofíspring. Because of the metabolic heat
generated by the developing clutch, “eggs' situated at the center of a clutch
are warmer than those at the periphery and therefore differentiate as
females while those at the periphery differentiate as males.

There is evidence that the ratio of male to female ofíspring among

other birds and reptiles may also be skewed by temperature.32
Temperature-induced labile sex determination may take place in a limited
number of birds, % while in some geckos, temperature-induced changes in
hormone interaction may even be responsible for regulating gender
31 Chelonia mydas (Morreale et al, 1982) and Lepidochelys olivacea (Ruiz et al,
1981).

32 e.g. The zygoparous lizard, Agata agama (Charnier, 1966), and the 2ygoparous
leopard gecko,
Eublepharis macularius (Bull, 1980).

Be, 8. the black-capped chickadee, Parus atricapillus (see Cooper, 1992).

37

switching during adulthood, Labile sex determination does not appear to

influence sexual phenotype in snakes or marmals.

Long-term changes in environmental temperature have the

potential to adversely affect the ratio of male to female offspring within
those populations that have evolved this kind of temperature sensitivity.
For example, prolonged cold temperatures could have the effect of leading
to an overproduction of one sexual phenotype and an ecologically
significant underproduction of the other. Over time, this could severely
disrupt the reproductive ecology of forms undergoing temperature-
dependent sex determination and it has even been implicated as a

contributing factor in the extinction of dinosaurs (see Ferguson and Joanen,

1982).

Endocrine Disrupters

A wide variety of naturally and artificially occurring compounds

are also capabie of affecting sexual phenotype in a wide range of
vertebrates by mimicking the presence of sex hormones during development.
These compounds are referred to as endocrine disrupters because they are
capable of interrupting the endocrine mechanisms underlying normal
reproductive function. Many such endocrine disrupters act as estrogen
agonists% and can profoundly affect the endocrine interactions involved in
sexual differentiation and reproductive function.

The effects of such environmental contaminants on wildlife

populations were first described in alligator populations from
contaminated and uncontaminated lakes in central Florida. Sexual
differentiation of pre-hatching alligators from contaminated waters is
permanently modified by exposure to dicofol and DDT (see Guillette et al.,
1994). Because of their direct action on hormone receptors, estrogen agonists
are capable of eliciting abnormal gonadal morphology and patterns of
steroidogenesis. lt appears that contamination by such compounds has the
effect of significantly altering embryonic sexual development by modifying
the ability of the developing gonad to produce sex hormones. The
possibility that environmental estrogen mimics have a significant impact
on the acquisition of sexual phenotype and fertility in other species
including humans) has led to much research attention in recent years.

The Sex Ratio

The sex ratio of a population is an expression of the relative

proportion of male and female phenotypes within that population. There
are several different types of sex ratios as well as means of expressing
them. The primary sex ratio is only expressed for forms with genetic sex
34 Also referred to as estrogen mimics or estrogenic xenabiotics.
38

determination and is the ratio of male and female phenotypes produced at

the time of fertilization.

The secondary sex ratio is the ratio following completion of the

period of parental investment in the offspring. This ratio can reflect the
effects of temperature-dependent sex determination during development,
differential pre-partum mortality in embryos of live-bearing forms, or
differential mortality that occurs during periods of post-hatching or post-
partum parental care. For example, in some rodenis (Florida packrat,
Neotoma floridana), post-partum sex ratios are adjusted by the culling of
female juveniles by the maternal organism. This is a form of sex-selective
infanticide and in rodents is often accomplished through food deprivation
(see McClure, 1981).

The tertiary sex ratio, or operational sex ratio, is the most

commonly considered sex ratio and represents the relative numbers of adult
males and females within a given population (Greenwood and Adams, 1987;
Sutherland, 1987). Differences in tertiary sex ratio may be due to
differential rates of predation on sexual phenotypes as well as differential
rates of mortality arising from sex-specific effects of other environmental
factors. In the case of Richardsonm's ground squirrel, Spermophilus
richardsonii, for example, there appear to be differences in the rates at
which the males and the females succumb to predators. The sex-based
differences in predation rates may be the result of the different sex-specifie
activities undertaken by male and female ground squirrels. When sex-
specific activities help to preserve one sex while exposing the other sex to
predation the stage is set for the emergence of some pretty extreme
differences between the secondary and tertiary sex ratios in the affected
population.

Sex ratios are often expressed according to one of several

conventions. Most commonly, sex ratio is expressed as the number of males
divided by the number of females in a given population. However, this
proportion is often reported in a variety of ways. For example, consider a
hypothetical population of six individuals consisting of one adult male and
five adult females. This population is most commonly described as having a
tertiary sex ratio of 1:5, However, the same population might also be
described as 1 in 6 male, or 16.67% male and 83.33% female.

Sex Allocation

The partitioning of male and female reproductive function among

different individuals within a population is a reflection of that

population's sex allocation.3 Different patterns of sex allocation reflect

35 Since the coverage of sex allocation theory is beyond the scope of this book,
the reader is
referred to Charnov (1982) for an excellent and comprehensive review of the
subject.

39

the specific environmental pressures that have operated during the

evolutionary history of particular lineages (Policansky, 1987).

Gonochorism

The most common pattern of sex allocation among the vertebrates

involves the allocation of male and female roles to different individuals
within a population. Vertebrates that allocate sex in this way are
considered gonochoristic or bisexual. Sexual phenotype is determined early
in the life of these vertebrates and, once established, persists throughout
adult Hfe,%

Hermaphroditism

Certain populations contain individuals that are capable of

functioning as both a male and a female, expressing both sexual phenotypes
during the course of their life. Populations that have evolved this
alternative form of sex allocation are known as hermaphroditic or cosexual.

Hermaphrodites are either capable of alternating between the

production of male and female gametes over the course of their life, as in
sequential or serial hermaphroditism, or of producing functional male and
female gametes at the same time, as in simultaneous or synchronous
hermaphroditism. Adult sexual phenotype in sequential hermaphrodites
is labile and not irreversibly established during embryonic development. In
fact, such forms commonly undergo sex reversal. En some species, sex
reversal can occur more than once during an individual's life history. When
sex reversal occurs more than once in a lifetime (male to female to male, for
instance), this double sex reversal is known as diandry or biandry.36

Sequential hermaphroditism is most common among the fishes. It

occurs in hagfish as well as in several families of ovuliparous teleosts,
where it can take place well into adulthood (Warner, 1975; Fischer,
1988).22 ln the black hamlet, Hypoplectrus nigricans, monogamously mated
pairs are capable of a coordinated alternation of sexual phenotype with
each member of the pair switching between the production of ova and sperm
in a process known as egg trading (Fischer, 1980).40 As a consequence, each
individual contributes to the production of the more energetically
expensive ova during the course of their relationship. Sequential
hermaphrodites that initially differentiate into females undergo

36 Though sex reversals have been experimentally induced in birds, the

differentiation of sexual
phenotypes in gonochoristic species is for the most part irreversible under natural
conditions.
97 Also known as gender change or sex inversion.
38 Double sex reversal occuzs in the piscine families Labridae and Scaridae
(Reinboth, 1967).

39 Ovuliparous teleosts capable of sequential hermaphroditism include the sea

basses,
Epinephelinae and Anthiinae.

20 Members of this species do not self-fertilize nor provide any form of parental
care.
40

protogynous hermaphroditism, or protogyny% This type of

hermaphroditism is favored when male size impacts fitness and tends to
occur in species where smaller males are less competitive than larger
males. in such cases, fitness is more readily achieved early in life as a
small female than it would be as a smaller male. In the protogynously
hermaphroditic East Asian swamp eel, Monopterus albus, the young fish
begins life as á female and starts to produce ova by its second year. The
following year, testicular tissue begins differentiating and, following a
brief intersexual period, the eel switches over to the male sexual
phenotype (Liem, 1963). By contrast, sequential hermaphrodites that
initially differentiate into males undergo protandrous hermaphroditism,
or protandry, which occurs among the members of eight families of teleosts
including porgies and sea breams (Shen et al, 1979) Protandrous
hermaphroditism is considered adaptive when large males are not
successful at monopolizing females and when larger females produce more
ova than do smaller females (Demski, 1987).

There are far fewer instances of synchronous hermaphroditism than

of protandrous hermaphroditism: among the vertebrates. Most synchronous
hermaphrodites possess an ovotestis containing oogenic as well as
spermatogenic tissues (Figure 2.8), The synchronous hermaphrodite,
therefore, is simultaneously male and female, unlike the protandrous
hermaphrodite which must switch from male to female or from female to
male, Synchronous hermaphroditism is believed to occur in the abyssal
aulepiform fishes and the benthic fishes of the Alepisauridae,
Chlorophalmidae, Evermannelidae, Ipnopidae, Serranidae, and
Paraleninidae.2 It has also been described in the shallow-water fishes of
the Cyprinodontidae (Peters, 1975).

Most synchronous hermaphrodites are not self-fertilizing,

however. The internally selffertilizing cyprinodont Rivulus
marmoratus(Harrington, 1971) is the only known self-fertilizing, or selfing,
synchronous Hhermaphrodite. In populations where selfing might
occasionally occur, it would tend to produce offspring that are highly prone
to the deleterious effects of recessive alleles — a fact that may be
responsible for the limited distribution of selfing among synchronous
hermaphrodites. Because selfing could be considered the ultimate form of
inbreeding, its persistence in Rívulus marmoratus has yet to be adequately
explained.

Another form. of synchronous hermaphroditism is known as

acquired hermaphroditism and occurs in the deep-sea ceratioid anglerfish,
4l For example, the black hamlet, Hypoplectrus nigricans, Australian cleaner-fish,
and East Asian
swamp eel, Monopterus albus.

22 The Serranidae is the only shallow water family in which simultaneous

hermaphrodism is
cominon.

41

Edriolychnus schmidti. As noted in an earlier discussion and in a slightly

different context, the male deep-sea created anglerfish attaches himself to
the female body and fuses with it. The male shares the female's blood
supply and all his organ systems degenerate — with the important exception
of the male reproductive system. In effect, this remarkable fusion creates a
single individual possessing both male and female genital structures (see
Demski, 1987).

Hermaphroditism may be an important mechanism of sex

allocation in environments where individuals are dispersed over large
areas (Reinboth, 1970). A synchronous hermaphrodite has twice the chance
of meeting a potential mate than does a gonochorist. This is because any
encounter between two adult synchronous hermaphrodites in a widely
seattered low-density population has the potential of yielding sexually
recombined offspring (Fischer, 1980, 1988). This might serve to explain why
synchronous hermaphroditism occurs in so many deep water and pelagic
species of fish, Still, individuals of some hermaphroditic species do not
eccur in low density populations. lt may be that these taxa initially
evolved under low density conditions and that a hermaphroditic mating
system simply remains a vestige of this evolutionary history.

Ovarian lametña

Cacyte

Entovarian duct

Yesticular tobe

Figure 2.8 Diagrammatic representation of ovotestis organization as it occurs in

one
of its lobes in sea basses of the family Serranidae (after D'Ancona).
CHAPTER 3
MODES OF REPRODUCTION

Outline
The Vertebrate Lite Cycle
Sexual Reproduction
Metosis
Genetic Recombination
Mating and Fertilization
Costs of Sexual Reproducilon
Benefits of Sexual Reproduction
Unisexual Reproduction
Parthenogenesis
Gynogenesis
Hybridogenesis
Cytological Mechanisms of Unisexuality
Polyembryony

“Sexual reproduction ís . . . the masterpiece of nature”

-Erasmus Darwin

introduction and Overview

By far, sexual reproduction is the more common pattern among

hiving vertebrate forms and its widespread occurrence suggests that it is the
plesiomorphic, or primitive, reproductive mode among the vertebrates. In
ali sexually reproducing forms, parental genomes that lie within an
individual recombine to produce specialized sex cells containing one half
the chromosomal complement of other cells in the body. As part of
successful reproduction, these haploid cells fuse to form a single diploid
zygote. Thus, sexual reproduction involves combination of genetic
information between two genetically recombined and distinct individuals.

Since it is likely that gender reflects differences in the

characteristics of sperm and ova, it is likely as well that fundamental
differerices exist in the relative contributions of each gender to the next
generation. For example, a single sperm is much smaller than an ovum and
primarily contributes only its genome to the zygote. By contrast, ova are
usually larger than sperm by one or more orders of magnitude. They provide
both nuclear and mitochondrial genes as well as much of the cellular
environmentl necessary to embryonic development and, in some cases, into
post-embryonic life. Thus, for most vertebrates, the energy needed to
produce a single gamete is far greater for a female than it is for a male.
1 Cytoplasm, organelles, nutrient reserves, riborrucleic acids, etc.