This article was downloaded by: [Florida Atlantic University]

On: 17 November 2014, At: 20:04

Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer
House, 37-41 Mortimer Street, London W1T 3JH, UK

African Journal of Herpetology

Publication details, including instructions for authors and subscription information:
http://www.tandfonline.com/loi/ther20

Folliculogenesis and ovarian histology of the

oviparous gecko, Hemidactylus mabouia (Sauria:
Gekkonidae)
a b
Ganas K. Moodley & Johannes H. van Wyk
a
School of Biological & Conservation Sciences , University of KwaZulu‐Natal ,
(Howard Campus), Durban, 4000, South Africa
b
Department of Botany & Zoology , University of Stellenbosch , Stellenbosch, 7600,
South Africa E-mail:
Published online: 31 Aug 2010.

To cite this article: Ganas K. Moodley & Johannes H. van Wyk (2007) Folliculogenesis and ovarian histology of the
oviparous gecko, Hemidactylus mabouia (Sauria: Gekkonidae), African Journal of Herpetology, 56:2, 115-135, DOI:
10.1080/21564574.2007.9635558

To link to this article: http://dx.doi.org/10.1080/21564574.2007.9635558

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”)
contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors
make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any
purpose of the Content. Any opinions and views expressed in this publication are the opinions and views
of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources of information. Taylor
and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses,
damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection
with, in relation to or arising out of the use of the Content.

This article may be used for research, teaching, and private study purposes. Any substantial or systematic
reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any
form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://
www.tandfonline.com/page/terms-and-conditions
 African Journal of Herpetology, 2007 56(2): 115-135. ©Herpetological Association of Africa

Original article

Folliculogenesis and ovarian histology of the oviparous

gecko, Hemidactylus mabouia (Sauria: Gekkonidae)

GANAS K. MOODLEY1 AND JOHANNES H. VAN WYK2

1
School of Biological & Conservation Sciences, University of KwaZulu-Natal (Howard Campus),
Durban, 4000, South Africa
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

2
Department of Botany & Zoology, University of Stellenbosch, Stellenbosch, 7600, South Africa
jhvw@sun.ac.za

Abstract.—Histological changes during the ovarian cycle of the oviparous gekkonid lizard,
Hemidactylus mabouia are described. The ovaries each contained a single germinal epithelium (bed)
with numerous oogonia and primordial follicles. Similar to other squamates, the ovarian stroma was
characterized by follicles in different stages of development, and may contain corpora lutea, and corpo-
ra atretica. The seasonal study revealed that vitellogenic ovarian hypertrophy commenced in spring
(September) and ovulation of the first clutch (two eggs) occurred in summer (November). A second
vitellogenic cycle followed, the second clutch being ovulated in late-summer (January). Ovarian quies-
cence, marked by predominance of pre-vitellogenic (hydration stage) follicles, was evident in females
collected from February to July. Although oocyte growth was accompanied by changes in the granulosa
layer, zona radiata and thecal layers, the zona pellucida remained unchanged throughout. The small cells
of the granulosa layer differentiated into three distinct cell types - small, intermediate and the conspicu-
ous large pyriform cells. Similar to most other squamates, the intermediate and pyriform cells at the
onset of vitellogenesis regressed to a single cuboidal epithelium. Following ovulation, the granulosa
layer hypertrophied forming luteal tissue. The theca layer differentiated into two layers and septal inva-
sion of the corpus luteum took place. At the time of oviposition, corpora lutea regressed to form ovari-
an scars (corpora albicantia). Follicular artresia occurred in previtellogenic follicles (hydration stage)
and seldom in vitellogenic follicles. The highest incidence of atresia occurred in the post-breeding peri-
od (February). The gross morphology, oogenesis and folliculogenesis of the ovaries of H. mabouia cor-
respond to the general squamate pattern described for oviparous reptiles.

Key words.—Gekkonidae, Hemidactylus mabouia, female reproduction, oogenesis.

he cellular dynamics of oogenesis and fol-
T liculogenesis remains central in studies
describing the female ovarian cycle in reptiles.
Macroscopic changes associated with ovarian
cycles are commonly reported in studies of
temporal changes in the reproductive cycles of
both oviparous and viviparous species (Norris
1996). Detailed description of cytological
changes during the ovarian cycle, however, has
received much less attention (Van Wyk 1984;
Guraya 1989; Uribe et al. 1996; Uribe &
Guillette 2000; Gomez & Ramirez-Pinilla
2004; Hernandez-Franyutti et al. 2005). This
has often lead to oversimplification in the liter-
ature when relating macroscopic changes to
functional aspects associated with the ovarian
cycle, particularly in the case of oviparous rep-
tilian species. More detailed histological stud-
ies are needed to substantiate/validate such
relationships.
Folliculogenesis in both oviparous and vivipa-
rous reptiles is characterized by the surround-
ing of the naked oocyte by ovarian stromal
cells, followed by dramatic changes in these
layers as well as the ooplasm during yolk depo-

115
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
sition (process of vitellogenesis) (Guraya
1989). Although the sequence of folliculogene-
sis and subsequent preovulatory follicular
growth follow the same general pattern in
lizards (Hernandez-Franyutti et al. 2005), the
detailed histology of the follicular epithelium
not only varies among different groups of rep-
tiles, but, greatly varies during follicular
growth within the temporal limits of the ovari-
an cycle (Guraya 1989; Uribe & Guillette
2000). It is generally acknowledged that more
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
detailed histological studies would contribute
towards understanding the functional signifi-
cance of the diversity of cell types known from
the granulosa and thecal layers, as well as the
role of the corpus luteum and atretic follocles.
Such studies should include a greater array of
species from different reptilian clades and
reproductive modes (oviparous vs viviparous).
Oogenesis in squamates occurs throughout life
in discrete regions known as germinal beds -
the number of germinal beds varies among
species (Jones 1978; Klosterman 1983; Van
Wyk 1984; Klosterman 1987; Guraya 1989).
The process of folliculogenesis and subsequent
migration into the stroma of the ovarian medul-
la, is essentially similar among reptilian species
studied so far (Guraya 1989; Hernandez-
Franyutti et al. 2005). The surrounding layer of
pre-follicular cells in oocytes, newly emerged
from the germinal bed, differentiate into at least
three distinct cell types (Guraya 1989). Of
these, the large flask shaped, pyriform cells,
characterized by the narrow protoplasmic
extension that transverse the homogeneous,
acellular, zona pellucida membrane formed
between the oocyte are well-described features
in the granulosa layer in squamates (Van Wyk
1984; Guraya 1989; Uribe et al. 1996; Gomez
& Ramirez-Pinilla 2004; Hernandez-Franyutti
et al. 2005). As follicular development pro-
gresses, the zona pellucida (located between
the outer granulosa and inner oocyte) is subdi-
vided into an outer homogeneous layer and an
inner striated layer lying against the oolemma
116
(Van Wyk 1984; Andreuccetti & Carrera 1987;
Guraya 1989). Surrounding the ovarian follicle,
including the granulosa layer, are the thecal
layers, the theca interna and theca externa,
composed of connective tissue, blood vessels,
fibroblasts and secretory cells (Van Wyk 1984;
Guraya 1989).
Postovulatory follicles are transformed into
corpora lutea in most reptiles, which are histo-
logically very similar to corpora lutea in mam-
mals (Fox & Guillette 1987; Gomez &
Ramirez-Pinilla 2004). Although the role of the
corpus luteum during gestation has been
acknowledged in several viviparous reptile
species, the functional significance of these
short-lived, temporary endocrine organs in
oviparous species remains enigmatic.
The regression (artresia) of ovarian follicle dur-
ing the ovarian cycle has been described in all
vertebrate classes (Byskov 1978; Van Wyk
1984; Guraya 1989). Oocytes are known to
become atretic at any stage of development,
previtellogenic or vitellogenic ovarian follicles
(Van Wyk 1984; Hernandez-Franyutti et al.
2005), although more commonly observed in
previtellogenic follicles (Gomez & Ramirez-
Pinilla 2004). Artretic follicles may a play
important part in limiting the number of eggs
produced during a particular reproductive cycle
(Hernandez-Franyutti et al. 2005).
Studies regarding the reproductive biology of
southern African lizards have recently
increased (Van Wyk 1991; Flemming 1993;
Flemming 1994; Van Wyk 1994; Van Wyk &
Mouton 1995; Van Wyk 1995; Van Wyk &
Mouton 1998; Flemming & Branch 2001;
Flemming & Mouton 2002; Du Toit et al. 2003;
Nkosi et al. 2004), but detailed descriptions of
folliculogenesis occurring during the ovarian
cycle has only been reported for a single
species, the rock lizard Agama atra (Van Wyk
1984). Most of the mentioned studies are on
cordylid or agamid lizards - only one detailed
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
report exists on the reproductive cycle of a
South African gekkonid (Flemming & Bates
1995).
The present study describes the histological
changes during the ovarian cycle (including
folliculogenesis, artretic follicle and corpus
luteum development) of the oviparous tropical
house gecko, H. mabouia and consider mor-
phological similarities to ovarian morphology
of other oviparous species. This species, known
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
from the tropical regions of Africa,
Madagascar, other islands in the Indian Ocean,
and east coast of South and central America,
has colonised many areas of southern Africa
(Bourquin 1987; Branch 1998; Carranza &
Arnold 2006). It is a nocturnal, scansorial,
gecko found living on trees, buildings, and rock
crevices. H. mabouia females produces at least
two clutches per breeding season and has a
fixed clutch size of two eggs (Branch 1998).
MATERIALS AND METHODS
Female Hemidactylus mabouia (N = 115) were
collected monthly in Durban (290 58’S/300
57’E), South Africa, and surrounding areas
over a period of one year from October 1987 to
September 1988. The geckos were killed using
Ether, weighed on an electronic balance and
snout-vent lengths measured to the nearest
0.01cm with the aid of vernier callipers. To
standardize sampling procedure, only the right
hand side ovaries were removed and weighed.
The ovaries were first fixed in Smith’s fixative
(Humason 1967) for 24 hrs and then transferred
through two changes of 10% buffered formalin.
After embedding in paraffin wax, the tissues
were sectioned at 7-10 μm on a Jung rotary
microtome. The sections were stained with
Carazzi’s haematoxylin and alcoholic eosin
(H&E) (Humason 1967). For histological
descriptions, we used size categories (1.5 mm
intervals) dividing folliculogenesis into seven
stages. We recognized four corpus luteum
117
development stages and two types of follicular
atresia, previtellogenic atresia and vitellogenic
atresia (after van Wyk 1984). Measurements of
the ovarian follicles were carried out with the
aid of either a micro-projector (for large folli-
cles) or an ocular micrometer (for smaller folli-
cles) to the nearest 0.01mm. For each ovary
examined, four measurements of the largest
follicle in four different planes were made and
the mean obtained. Other data, for example, the
presence or absence of corpora lutea, atretic
follicles, the size and state of the largest folli-
cle, for each specimen were routinely recorded.
Since the females collected during the breeding
season were not all synchronized in their repro-
ductive condition, they were grouped accord-
ing to ovarian activity rather than in month of
collection (see Table 1).
Cellular dimensions were compiled from the
mean of ten measurements of the structure
under consideration and values were presented
as the mean ± one standard deviation. Similar
data for each month and or reproductive stage
was averaged and expressed as the mean ± 1
SD. Whenever the means of similar variables
between different sized follicles were com-
pared, the Students t-test for unpaired samples
was used (NWA Statpak 4.1; Northwest
Analytical Inc. Portland, Oregon). Seasonal
variation in follicle diameters during the annu-
al cycle was analyzed using one-way ANOVA
procedure (Super ANOVA VI. I, Abacus
Concepts, Inc., California) and Duncan’s mul-
tiple range test employed to indicate significant
differences between monthly means.
RESULTS
Changes in follicle size during the ovarian
cycle
Seasonal variation in mean diameter of the
largest follicle in each ovary during the ovarian
cycle of Hemidactylus mabouia is presented in
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007

Table 1. Ovarian Stages used to group female Hemidactylus mabouia (modified from van Wyk 1983).

i. Winter condition: Hydration stage ovarian follicles (diameter smaller than 1.5 mm); no oviducal eggs,
corpora lutea or fresh remnants of corpora lutea.

ii. Pre- or early vitellogenic - P1: Onset of vitellogenesis (follicular diameter larger than 1.5 mm but less
than 3.0 mm); no oviducal eggs, corpora lutea or fresh remnants of corpora lutea.

iii. Preovulatory I (Pre I) - P2: Late vitellogenic follicles (diameter larger than 3 mm); no oviducal eggs,
corpora lutea or fresh remnants of corpora lutea.
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

iv. Postovulatory I (Post I) - P3: Hydration stage follicles (diameter smaller than 1.5 mm); presence of
oviducal eggs and functional corpora lutea (good blood supply); no remnants of corpora lutea.

v. Late postovulatory (Post II) - P4: Early vitellogenic follicles (diameter larger than 1.5 mm but small-
er than 3 mm); presence of oviducal eggs and corpora lutea; no remnants of corpora lutea.

vi. Preovulatory II (Pre II) - P5: Late vitellogenic follicles (diameter larger than 3 mm); no oviducal eggs
or functional corpora lutea (poor blood supply); fresh remnants of corpora lutea (pigmented).

vii. Postovulatory III (Post III) - P6: Hydration stage follicles (diameter smaller than 1.5 mm); presence
of oviducal eggs, functional corpora lutea and fresh remnants of corpora lutea.

viii. Postoviposition: Hydration stage follicles (diameter smaller than 1.5 mm); no oviducal eggs or func-
tional corpora lutea; presence of corpora lutea remnants.

Figure 1. Onset of ovarian growth commenced
in September since 10% of the sample collect-
ed possessed follicles larger than 3 mm in
diameter. A rapid increase in mean follicular
size occurred in October (P1 vs P2; P < 0.05,
50% of sample included follicles > 3mm).
These oocytes exhibited a yellowish appear-
ance due to the deposition of yolk. Following
ovulation (P3), the ovary was characterized by
small ovarian follicles (with diameters less
than 1.5 mm) and functional corpora lutea. The
presence of late-vitellogenic follicles (diameter
> 3mm) together with the absence of oviducal
eggs as well as functional corpora lutea indi-
cated a second vitellogenic peak (stage P5) (P4
vs P5; P < 0.05). Subsequently, stage P6 was
characterised by small hydration stage follicles
(diameter < 1.5 mm), and the presence of ovid-
ucal eggs, functional corpora lutea or fresh
remnants of corpora lutea. During February, no
females contained vitellogenic follicles, and
ovarian quiescence followed until the onset of
yolk deposition in September.
Ovarian elements and germinal bed
The ovary is covered externally by a thin peri-
toneal epithelium with deeply basophilic, flat-
tened, elongate nuclei. The ovarian elements
comprise the following: follicles in different
stages of development, corpora lutea, atretic
follicles and a single discretely located germi-
nal epithelium in the ovarian stroma. The ovar-
ian stroma is small, consisting of fibroblasts,
collagen fibres and several small blood vessels
indicating a high vascularity. The germinal

118
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

Figure 1. Seasonal variation in the mean diameter of the largest follicle in each ovary during the ovarian
cycle of female Hemidactylus mabouia. Nine or ten females were included in each monthly sample. Vertical
lines indicate 1 standard deviation. Monthly mean values with the same alphabetic letter are not significantly
different (P < 0.05). For explanation of reproductive stages see Table 1.

epithelium is composed of an elongate region
of cells on the dorsal surface of the ovary. One
side of the germinal epithelium is continuous
with the peritoneal epithelium of the ovary
whilst the other side is curled (Fig. 2). The ger-
minal epithelium consists of epithelial somatic
cells, oogonia (proliferating by mitotic divi-
sions) and oocytes in different stages of devel-
opment and (meiosis) (Fig. 3). Adjacent to the
germinal epithelium is the germinal stroma
which is clearly distinguishable from the ovar-
ian stroma by its histological appearance. The
nuclei of cells in the germinal stroma appear
elongate whilst the nuclei of ovarian stromal
cells are more rounded (Fig. 4). The migrating
primary oocytes have large spherical, nuclei
and the cytoplasm is much more homogeneous
than that of the oogonia (Fig. 4). As the prima-
ry oocyte moves toward the ovarian stroma, it
is surrounded by a single layer of germinal
stromal cells (Fig. 5). Prior to entering the
ovarian stroma, however, the cells surrounding
the oocyte become multi-layered (Fig. 6). At
this stage the primary oocyte projects into the
ovarian stroma (Fig. 7).
Histological changes during oogenesis
Stage 1: Previtellogenesis: 0.1-0.3 mm oocyte
diameter (Figs. 8 & 9).—Follicles of this size
emerge from the germinal stroma and migrate
into the ovarian stroma. Chromatin material is
discernible in the eccentrically located nucleus

119
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

Figure 2. Germinal epithelium of the ovary of Hemidactylus mabouia collected in November. Note the curl on one side
of the ovary and the loosely organized ovarian stroma. (ge = germinal epithelium, gs = germinal stroma, o = oogonia, os
= ovarian stroma, po = primary oocyte).
Figure 3. Germinal epithelium of the ovary of Hemidactylus mabouia collected in October. Note the meiotic activity in
oogonia (arrows). (ge = germinal epithelium).
Figure 4. Migrating primary oocytes in the ovary of Hemidactylus mabouia collected in October. Note the vesicular,
spherical nucleus of the primary oocyte and the difference in histological appearance between the germinal and ovarian
stroma. (gs = germinal stroma, os = ovarian stroma, po = primary oocyte).
Figure 5. Germinal epithelium with a primary oocyte being surrounded by germinal stromal cells. (ge = germinal epithe-
lium, gsc = germinal stromal cells, o = oogonia, po = primary oocyte).
Figure 6. Micrograph of primary oocyte beginning to enter the ovarian stroma. Note the multi-layered appearance of the
surrounding stromal cells and meiotic activity in the oogonia. (gs = germinal stroma, o = oogonia, os = ovarian stroma).
Figure 7. Primary oocyte projecting into ovarian stroma. (ge = germinal stroma, o = oogonia, os = ovarian stroma).

120
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
of the oocyte (Fig. 8). Small stromal cells sur-
round the primary oocyte at this stage. The
basement membrane is located on the outer
border of the stromal cells, which are multi-
layered, and together with the basement mem-
brane now comprise the zona granulosa or
granulosa layer (Figs. 8 & 9). Soon after the
emergence of the primary oocyte into the ovar-
ian stroma, two types of granulosa cells are evi-
dent: small cells and large intermediate cells
(Figs. 8 & 9). The theca is poorly constituted
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
and comprises a single layer of flattened
fibroblasts with elongate nuclei (Fig. 9). The
zona pellucida, although poorly developed, is
also evident (Fig. 9).
Stage 2: Previtellogenesis: 0.3-0.5 mm oocyte
diameter (Fig. 10).—Small cytoplasmic vac-
uoles begin to show on the ooplasmic periphery
in the oocyte (Fig. 10). The nucleus is granu-
lated with a single nucleolus. The granulosa
differentiates further forming three distinct cell
types: small cells, intermediate cells and large
pyriform cells (Fig. 10). During this phase the
granulosa is thick and measures 66.0 ± 8.4 μm.
The intermediate cells are located more
towards the inner border of the granulosa (Fig.
10). The largest cell type of the granulosa layer
is the flask-shaped pyriform cells. Chromatin
material is evident in the nucleoplasm of both
intermediate and pyriform cells. The small
granulosa cells located near the zona pellucida
are termed apical cells while those close to the
theca are called the basal cells (Fig. 10). Small
granulosa cells are also found amongst the
pyriform and intermediate cells of the granu-
losa. The zona pellucida is visible as a thin
homogeneous layer adjacent to the zona granu-
losa (Fig. 10). The theca (10.0 ± 0.1 μm in
thickness) forms a capsule around the develop-
ing follicle and is composed of collagenous
fibres and fibroblasts.
Stage 3: Previtellogenesis: 0.5-1.0 mm oocyte
diameter (Fig. 11).—The ooplasm of the
oocyte is finely vacuolated while the nucleus is
121
granulated. Acidophilic granules are evident
towards the ooplasmic periphery (Fig. 11). A
reduction in granulosa thickness is observed
(43.0 ± 4.8 μm P < 0.001; Stage 2 vs Stage 3).
As in the previous stage, the granulosa is com-
posed of small cells, intermediate cells and
pyriform cells. Cytoplasmic extensions of the
pyriform cells communicate with the ooplasm
(Fig. 11). Besides their shape, the intermediate
cells can also be distinguished from the pyri-
form cells by their darkly staining cytoplasm
and smaller nuclei (Fig. 11). Small granulosa
cells located adjacent to the zona pellucida are
drastically reduced in number while numerous
intermediate granulosa cells are now apparent
in this region (Fig. 11). The zona pellucida (1.0
± 0.1 μm), located between the zona granulosa
and zona radiata, appears as a thin colloidal
layer. Immediately adjacent to the zona pelluci-
da the zona radiata (2.0 ± 0.1 μm) appears as a
radiately striated zone. During this stage only a
few small cells are found amongst the pyriform
cells and intermediate cells of the granulosa.
The theca appears very much the same as in
Stage 2 without any change in thecal thickness
(10.0 ± 0.1 μm).
Stage 4: Previtellogenesis: 1.0-1.5 mm oocyte
diameter (Fig. 12).—The ooplasm appears
more vacuolated than in stage 3 with the
peripheral areas of the oocyte being less vacuo-
lated than the central region. As in Stage 3, aci-
dophilic inclusions are found in a narrow zone
near the periphery of the oocyte. The granulosa
layer (32 ± 4.2 μm) undergoes further reduc-
tion in thickness when compared to Stage 3 (P
< 0.001 Stage 3 vs Stage 4). Concomitant with
the reduction in the thickness of the granulosa
there is a reduction in the height of the pyri-
form cells (Fig. 12). Some of the pyriform cells
exhibit very granular eosinophilic cytoplasm.
Intercellular spaces are more frequently
observed in the zona granulosa during this
stage indicating a reduction in the size of the
cells (Fig. 12). In contrast to the zona pellucida
which shows no change in dimensions (1.0 ±
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

Figure 8. Ovarian follicle (0.1 - 0.3 mm) in ovarian stroma. Note the eccentrically placed nucleus in the ooplasm and the
already formed zona granulosa around the oocyte. (n = nucleus of oocyte, op = ooplasm of oocyte, os = ovarian stroma,
p = peritoneum, zg = zona granulosa).
Figure 9. Higher magnification of the ovarian follicle showing the differentiation of the granulosa into intermediate and
small cells. (ic = intermediate cells, n = nucleus of oocyte, op = ooplasm of oocyte, sc = small cells, t = theca, zp = zona
pellucida).
Figure 10. Ovarian follicle (0.3 - 0.5 mm) showing differentiation of the granulosa into small, intermediate and pyriform
cells. Note the vacuoles in the ooplasm and the distinct thecal layer. (ac = apical cells, bc = basal cells, ic = intermediate
cells, pc = pyriform cells, t = theca, v = vacuole, zp = zona pellucida).
Figure 11. Ovarian follicle (0.5 - 1.0 mm) showing the appearance of the zona radiata. Note the cytoplasmic extensions
of the pyriform cells which communicate with the ooplasm (small arrows) and the acidophilic inclusions (thick arrows)
in the peripheral ooplasmic region. (ic = intermediate cells, pc = pyriform cells, zp = zona pellucida, zr = zona radiata).

0.1 μm) both the zona radiata (2.4 ± 0.5 μm P < 0.05 Stage 3 vs Stage 4) and the theca (17.0 ±
4.8 μm P < 0.001 Stage 3 vs Stage 4) exhibit an increase in thickness. Occasionally small capil-
laries and erythrocytes are observed in the thecal layer.

Stage 5: Vitellogenesis: 1.5-2.0 mm oocyte diameter (Fig. 13).—The ooplasm is more heavily vac-
uolated due to an increase in both the number and size of the vacuoles (Fig. 13). The vacuoles
found on the periphery are much smaller than those found towards the middle of the ooplasm.
However, the peripheral ooplasmic zone immediately adjacent to the zona radiata is homogeneous
and devoid of vacuoles (Fig. 13). Inside this zone, additional acidophilic inclusions are present in
a narrow band around the oocyte. Strands of dense ooplasm ramify throughout the ooplasmic
mass, between the vacuoles. The granulosa now appears as a single, monomorphic cellular layer

122
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
(Fig. 13) due to a drastic reduction in thickness
(3.9 ± 0.9 μm P < 0.001 Stage 4 vs Stage 5).
In this stage the pyriform nature of the cells is
not evident any longer and the thickness of the
zona pellucida remains unchanged (1.0 ± 0.1
μm). The zona radiata, however, is twice as
thick when compared to the previous stage (4.7
± 0.5 μm P < 0.001 Stage 4 vs Stage 5) but
appears less striated (Fig. 13). During this stage
the theca attains its maximum thickness (20.4 ±
1.5 μm P < 0.05 Stage 4 vs Stage 5) but no
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
distinct morphological differences are
observed.
Stage 6: Vitellogenesis: 2.0-2.5 mm oocyte
diameter (Figs. 14 & 15).—The ooplasm
remains vacuolated and a reduction in the size
of the vacuoles towards the periphery is still
observed (Figs. 14 & 15). The acidophilic sub-
stances, forming a narrow band on the ooplas-
mic periphery, are still present. Yolk spheres
are now evident in the growing oocyte with a
gradual increase in size of the spheres from the
periphery towards the centre (Figs. 14 & 15).
The granulosa (3.9 ± 0.7 μm) remains
monomorphic with flattened, deeply staining
basophilic nuclei. There is no difference in the
thickness of the granulosa (3.9 ± 0.7 μm) and
the zona pellucida (1.0 ± 0.1 μm) when com-
pared to the preceding stage. The zona radiata
on the other hand, although still possessing
large spaces between striations, is significantly
reduced (3.5 ± 0.5 μm P < 0.001 Stage 5 vs
Stage 6). During this stage the theca undergoes
a drastic reduction in thickness (8.2 ± 1.2 μm P
< 0.001 Stage 5 vs Stage 6). Blood vessels are
observed in the epithelium covering the ovary.
Stage 7: Vitellogenesis: larger than 3 mm
oocyte diameter (Fig. 16).—The oocyte is
completely filled with large yolk spheres which
decrease in size towards the periphery (Fig.
16). The granulosa, composed of a single layer
of flattened cells with elongate basophilic
nuclei, undergoes further reduction in thickness
(3.1 ± 0.7 μm P < 0.05 Stage 6 vs Stage 7). In
123
this stage the zona pellucida is barely visible
between the zona granulosa and zona radiata
(Fig. 16). The zona radiata maintains its thick-
ness (3.4 ± 1.1 μm P > 0.05 Stage 6 vs Stage 7)
with no appreciable change in structure. The
theca increases slightly in thickness (10.7 ± 3.3
μm P < 0.05 Stage 6 vs Stage 7) and becomes
more vascular with an abundance of thecal ery-
throcytes. Furthermore, large blood vessels and
sinuses are observed in the epithelium covering
the ovary. This stage depicts the mature oocyte.
Within the ovary, the oocyte increases in size
until it is ovulated. The largest preovulatory
follicle in H. mabouia measures 6.29 mm in
diameter.
Postovulatory follicles (Corpora lutea).—After
ovulation, the somewhat collapsed postovula-
tory follicles undergo a series of changes. In
this gecko, four stages of the corpus luteum
were observed.
Corpus luteum - Stage 1 (Fig. 17).—The early
corpus luteum is oval. Shortly after ovulation,
the central cavity of the follicle becomes filled
with cells (Fig. 17). The thecal layer is much
thicker than in the preovulatory follicle and is
composed of fibroblasts, collagen fibres and
numerous blood capillaries. The central mass
of cells, termed the lutein cells, possess round
to oval deeply basophilic nuclei.
Differentiation of the theca into an outer and
inner thecal layer is not evident (Fig. 17). The
border of the theca, close to the lutein mass, is
marked by the presence of fibroblasts which in
places form groups of cells that are concentri-
cally arranged (Fig. 17). In some corpora lutea
of this stage, collagenous fibres and fibroblasts,
from the thecal layer, begin to invade the cen-
tral lutein mass.
Corpus luteum - Stage 2 (Fig. 18).—The cor-
pus luteum is oval to elongate in outline with
the thecal layer having undergone a reduction
in thickness and in fibroblast numbers (Fig.
18). The theca is highly vascular. The lutein
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

Figure 12. Micrograph of ovarian follicle (1.0 - 1.5 mm). The pyriform cell height is reduced. Note acidophilic inclusions
(thick arrows) in the peripheral ooplasm. (ic = intermediate cells, pc = pyriform cells, t = theca, zp = zona pellucida, zr =
zona radiata).
Figure 13. Theca, zona granulosa, zona radiata and zona pellucida of ovarian follicle (1.5 - 2.0 mm). The zona granulosa
is reduced to a monomorphic, single cellular layer. Note the vacuoles and acidophilic inclusions (thick arrows) in the
ooplasm. (t = theca, v = vacuole, zg = zona granulosa, zp = zona pellucida, zr = zona radiata).
Figure 14. Micrograph of ovarian follicle (2.0 - 2.5 mm). Note the increase in size of the yolk spheres from the periphery
to the centre. (t = theca, y = yolk spheres).
Figure 15. Higher magnification of ovarian follicle showing the reduced thecal layer. (t = theca, y = yolk spheres, zg =
zona granulosa, zp = zona pellucida, zr = zona radiata).
Figure 16. Ovarian follicle (6.29 mm) showing the large yolk spheres and the barely visible zona pellucida. (t = theca,
y = yolk spheres, zg = zona granulosa, zp = zona pellucida, zr = zona radiata).
Figure 17. Postovulatory follicle (Stage 1). Note the hypertrophied theca, lutein mass within the central cavity of the fol-
licle, blood capillaries (large arrows) in the outer thecal region and cell "whorls" (small arrows) in the inner thecal region.
(lm = lutein mass, t = theca).

124
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
cells, which are randomly distributed within
the central mass, begin to hypertrophy and their
vesicular nuclei are basophilic. Copious
amounts of a yellow-brown pigment substance
fill the non-vacuolated lutein mass. Numerous
connective tissue strands from the thecal layer
penetrate deeper into this mass (Fig. 18).
Corpus luteum - Stage 3 (Fig. 19).—The cor-
pus luteum is smaller and more rounded than in
the previous stage and the thecal layer is very
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
much reduced forming a thin covering around
the corpus luteum. In this stage the presence of
pigment granules gives the corpus luteum a
yellow-brown appearance. The corpus luteum
undergoes morphological changes associated
with cellular atrophy. Large vacuoles make
their appearance in the luteal cell mass (Fig.
19). The cells appear more variable in size and
shape with some cells displaying pyknotic
nuclei. Septa of connective tissue from the the-
cal layer form a network within the luteal mass
(Fig. 19).
Corpus luteum - Stage 4 (Fig. 20).—There is a
drastic reduction in the size of the corpus
luteum and an increase in the number and size
of vacuoles within the luteal mass when com-
pared to Stage 3 (Fig. 20). The nuclei of the
luteal cells are pyknotic and pigment granules
still persist. Connective tissue forms a thin cov-
ering around the corpus luteum (Fig. 20). This
stage represents remnants of corpora lutea
which are present in ovaries well after oviposi-
tion has occurred. Corpora lutea remnants also
persist in ovaries that produce a second clutch
of eggs.
Follicular atresia.—Follicular atresia is char-
acterized by changes in both the granulosa and
the ooplasm and seems to occur more frequent-
ly in quiescent ovaries. Of the specimens cap-
tured during February, 44.4% exhibited atresia;
this figure being the highest recorded for the
year (Fig. 21). With the exception of one spec-
imen collected in November, atretic follicles
125
were absent in specimens collected between
October and January. No atretic follicles were
observed in specimens collected in May,
August, October, December and January.
Besides February not more than 11.1% of any
sample possessed atretic follicles. Atresia may
occur either in the presence or absence of a
vitellogenic follicle in the ovary. Although two
types of atresia are generally recognized
(Guraya 1989) viz.: (i) hydration stage atresia
occurring in small non-vitellogenic follicles
and (ii) vitellogenic stage atresia occurring in
large vitellogenic follicles, only hydration
stage atresia was observed in H. mabouia. The
irregularly distributed ooplasm, which is aggre-
gated into masses in certain areas, has an
eosinophilic, granular appearance (Fig. 22).
Within the atretic follicle, numerous amoeboid
macrophages are present (Fig. 22). These cells
are presumably responsible for phagocytoses as
evidenced by the accumulation of ooplasmic
granules within the cytoplasm. The zona pellu-
cida and zona radiata are absent. The zona
granulosa is polymorphic and consists of large,
irregularly shaped cells with distinct round to
oval nuclei (Fig. 22). The granulosa is not of
uniform thickness. This is probably due to the
phagocytic activity of the macrophages. The
hypertrophied thecal layer is distinct from the
granulosa layer. As atresia progresses, the gran-
ulosa hypertrophies and begins to invade the
oocyte (Fig. 23). Bits of ooplasm are observed
within the lightly staining cytoplasm of the
polymorphic granulosa. Some of the nuclei of
the granulosa cells exhibit mitotic figures.
Ooplasmic aggregations display eroded edges,
possibly indicating amoeboid macrophage
phagocytic activity. The theca undergoes
hypertrophy and thin strands of thecal connec-
tive tissue penetrate the granulosa (Fig. 23). As
atresia reaches an advanced stage, the number
and thickness of strands of thecal connective
tissue and fibroblasts invading the oocyte
increases. The granulosa cells are eventually
distributed throughout the oocyte.
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
Figure 18. Postovulatory follicle (Stage 2). Note the non-
vacuolated lutein mass, the reduced theca and the septa of
thecal connective tissue (small arrows) invading the lutein
mass. (lm = lutein mass, t = theca).
Figure 19. Postovulatory follicle (Stage 3) showing vac-
uoles and connective tissue network in the lutein mass.
Note the pyknotic nuclei of the luteal cells. (lm = lutein
mass, v = vacuole).
Figure 20. Postovulatory follicle (Stage 4). Note the
increase in size and the number of the vacuoles in the lutein
mass, the pyknotic nuclei of the luteal cells and the thin
connective tissue (small arrows) covering the corpus
luteum. (v = vacuole).
126
DISCUSSION
The gross morphology, folliculogenesis and
oocyte growth of the ovaries of H. mabouia
correspond to the general reptilian pattern
described to date (Van Wyk 1984; Guraya
1989; Uribe et al. 1996; Gomez & Ramirez-
Pinilla 2004; Hernandez-Franyutti et al. 2005).
Histological changes in the follicular wall sug-
gested a synchronized recruitment and devel-
opment of ovarian follicles during ovarian
cycles. Similar to other seasonal breeding
lizards, vitellogenic ovarian hypertrophy
marked the onset of the ovarian cycle in H.
mabouia. Although the production of a second
clutch in late summer extended the breeding
season, a clear seasonal cycle in ovarian mor-
phology is exhibited in H. mabouia.
Similar to spring-summer breeding lizards liv-
ing in the southern hemisphere (Van Wyk 1983;
Van Wyk 1984; Flemming & Bates 1995; Van
Wyk & Ruddock 2000; Nkosi et al. 2004),
vitellogenic activity in H. mabouia commenced
in spring (September) through late-spring
(October) with ovulation of the first clutch in
early-summer (November)(Moodley 1991). A
second vitellogenesis cycle, not uncommon in
oviparous lizards occurring in warm temperate
climates (van Wyk 1984; Flemming & Bates
1995; Nkosi et al. 2004), occurred in mid-sum-
mer (December) with the second clutch being
ovulated in late-summer (January)(Moodley
1991). Ovarian quiescence, i.e. previtellogenic
stage ovaries, containing small hydration stage
follicles, characterise true seasonal breeders
(Van Wyk 1984; Flemming & Bates 1995). In
H. mabouia such ovarian quiescence occurred
during the autumn- winter period (February to
August)(Moodley 1991). The basic reproduc-
tive cycle corresponded to that of the female
reproductive cycle of Pachydactylus bibronii
(Flemming & Bates 1995), the only detailed
study regarding seasonal reproduction activity
published for a South African gekkonid, as well
as the reproductive cycle reported for the
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014

Figure 21. The presence of atretic follicles in the ovaries of Hemidactylus mabouia expressed as percentages
of the monthly sample.

Indian wall lizard, Hemidactylus flaviviridis
(Guraya & Varma 1976). Flemming & Bates
(1995) reported that a few early vitellogenic P.
bibronii females were collected during winter
(June/July) and also suggested the production
of a second clutch in late-summer. Guraya &
Varma (1976) reported strong seasonal repro-
ductive activity in H. flaviviridis with two eggs
being produced in the spring-summer period.
Similar to P. bibronii (Flemming & Bates
1995) and H. flaviviridis (Guraya & Varma
1976) H. mabouia ovulated two eggs per
clutch, one from each ovary.
Folliculogenesis.— The germinal epithelium in
the H. mabouia ovary is composed of a single,
discrete, elongate region of cells (Boyd 1940;
Van Wyk 1984; Guraya 1989) located on the
dorsal surface of the ovary. The germinal bed is
the location containing oogonia that proliferate
by mitotic divisions to increase the number of
germ cells. Jones et al. (1982) and Gomez &
Ramirez-Pinilla (2004) suggested that the num-
ber of germinal beds per ovary correlated with
clutch size (instantaneous fecundity). Several
species are known to exhibit two general beds
producing groups of new oocytes (Guraya
1989; Uribe et al. 1996; Gomez & Ramirez-
Pinilla 2004; Hernandez-Franyutti et al. 2005).
The single germinal bed reported in H.
mabouia agrees with the report of (Guraya &
Varma 1976) for H. flaviviridis. It seems that a
single germinal bed does not exclude the possi-
bility of producing a second clutch in H.

127
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
Figure 22. Early hydration stage follicular atresia.
Note the hypertrophied theca and granulosa layers
and the absence of the zona radiata and zona pellu-
cida. (am = amoeboid macrophage, op = ooplasm of
oocyte, t = theca, zg = zona granulosa).
Figure 23. Advanced hydration stage follicular atre-
sia showing the granulosa cells invading the oocyte,
the hypertrophied theca and the thecal connective
tissue (arrows) penetrating the granulosa. (op =
ooplasm of oocyte, t = theca, zg = zona granulosa).
mabouia. This was also the case in Agama atra
(Van Wyk 1984).
The germinal stromal cells in H. mabouia sur-
round the primary oocyte as it moves towards
the ovarian stromal patch. This pattern of fol-
liculogenesis also occurs in several other squa-
mates (Van Wyk 1984; Guraya 1989; Uribe et
al. 1996; Gomez & Ramirez-Pinilla 2004;
Hernandez-Franyutti et al. 2005) The naked
primary oocyte is selectively encircled by a sin-
gle layer of granulosa cells which increase in
number, become multi-layered to eventually
differentiate into the three different cell types,
128
viz: small cells, intermediate cells and pyriform
cells (Van Wyk 1984; Guraya 1989;
Andreuccetti 1992; Motta et al. 1995). Rahil &
Narbaitz (1973) have reported the presence of
two types of granulosa cells in the turtle
Pseudemys scripta elegans. However, as this
difference is based on the densities of the cyto-
plasmic matrix of the cells, the follicular
epithelium is composed essentially of only one
cell type. Thus, while a non-differentiated fol-
licular epithelium appears to be characteristic
of turtles (Gupta & Duda 1985) and alligators
(Lance 1989; Uribe & Guillette 2000), squa-
mates, on the other hand, exhibit a well-differ-
entiated follicular epithelium.
There seems to be some controversy in the
literature regarding the derivation of the
follicular granulosa cells in reptiles. Some
workers including Boyd (1940), Bragdon
(1952) and Betz (1963) have suggested that the
intermediate and large pyriform cells arise
separately by differentiation from the small
cells while others (Neaves 1971; Van Wyk
1984; Saha et al. 1984; Andreuccetti 1992;
Motta et al. 1996; Loredana et al. 2004) have
suggested that the pyriform cells are derived
from the intermediate cells, which in turn are
derived from the small or basal cells. The flask-
shaped pyriform granulosa cells in H. mabouia
possess narrow protoplasmic prolongations
which traverse the zona pellucida. The
cytoplasm of the oocyte and the pyriform cells
are confluent. At the point of communication, a
notch appears in the zona pellucida. Similar
observations have been made in many reptiles
(Neaves 1971; Lance & Lofts 1978; Van Wyk
1984; Saha et al. 1984; Guraya 1989; Uribe et
al. 1996). Andreuccetti (1992), with the aid of
electron microscopy, has shown that
intercellular bridges are first established
between individual small cells of the early,
monolayered granulosa and the oocyte. True
intercellular bridges between pyriform cells
and the ooplasm are characteristic of squamates
only (Neaves 1971; Motta et al. 1995; Motta et
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
al. 1996). The intercellular canals in the
follicles of H. mabouia were observed more
frequently in large hydration stage oocytes. In
the cobra, Naja naja (Lance & Lofts 1978) and
the lizards Sceloporus jarrovi (Goldberg 1970),
Anolis carolinensis (Neaves 1971) and Agama
atra (Van Wyk 1984) an increase in the
pyriform cell-oocyte bridges occur
immediately prior to yolk deposition. Other
workers, however, have failed to detect any
seasonal changes in the appearance of the
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
pyriform cells (Boyd 1940; Betz 1963; Saha et
al. 1984).
The precise function of the reptilian granulosa
remains uncertain. This is especially true for
the pyriform cells. Some workers have pro-
posed that the pyriform cells are involved in
yolk production (Betz 1963; Goldberg 1970;
Saha et al. 1984). Lance & Lofts (1978) have
suggested that the pyriform cells are not
involved in yolk production since, in Naja, they
disappear just before the onset of yolk deposi-
tion. However, there is abundant evidence in
reptiles, as well as in other oviparous verte-
brates, that yolk is formed from a precursor,
vitellogenin, produced in the liver and taken up
from the blood by the developing oocyte (Gapp
et al. 1979) via extra follicular spaces (Neaves
1972). The vitellogenin thus deposited in the
follicle is converted into the yolk proteins
phosvitin and lipovitellin (Gapp et al. 1979).
Electron microscopic observations of pyriform
cell bridges in Anolis have shown nothing
resembling yolk in either the pyriform cell
body or in the various parts of the bridge
(Neaves 1971; Neaves 1972) thus contradicting
the yolk transport hypothesis. In addition,
Neaves (1971) has shown that the bridges dis-
appear prior to the onset of yolk deposition.
Although this discredits the theory that the
bridges are involved in yolk transport, the
study by Andreuccetti (1992) present evidence
that pyriform cells may indeed have similar
vitellogenic functions as the oocytes. Whether
the uptake of exogenous yolk precursors by
129
pyriform cells occurs in all squamates is still
largely open to question.
Potential extracellular routes across the
follicular epithelium exist in most vertebrates
which have been studied (Neaves 1971).
Cytoplasmic communication between pyriform
cells and oocytes during previtellogenic
development in Anolis suggests that support of
the young oocyte may depend on exchange
with the follicular epithelium (Neaves 1971;
Motta et al. 1995; Motta et al. 1996). During
vitellogenesis, however, the incorporation of
extrafollicular materials by the oocytes
suggests that yolk accumulation in rapidly
growing oocytes may be relatively independent
of the follicle cells. Moto et al., (1995; 1996)
and Loredana et al., (2004) confirmed
intracellular exchange between pyriform cells
(in particular the cytoplasmic content) and the
oocyte suggesting a nurse function during
follicular growth and development. Nuclear
material seems to be recycled by the small
cells, the only cells remaining in the granulosa
layer during vitellogenesis (Motta et al. 1996).
The regression of the intermediate and large
pyriform cells at the end of previtellogenesis
has been suggested to be associated with
programmed cell death (apoptosis)(Motta et al.
1996). All in all, the available evidence seems
to suggest that both extracellular and pyriform-
oocyte intercellular bridges are important
pathways for the transport of substances which
are responsible for the maintenance of the
growing oocyte, preceding the previtellogenic
stages (Motta et al. 1996; Loredana et al. 2004;
Hernandez-Franyutti et al. 2005).
The granulosa thickness varies according to the
stage of follicular development in H. mabouia.
From the result obtained, the granulosa rapidly
increases in thickness and reaches its maximum
width (66 μm) early in the development of the
follicle (0.3-0.5 mm oocyte diameter). In
Agama atra, the granulosa is thickest (52.1
μm) in follicles ranging in size from 0.5-1.0
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
mm (Van Wyk 1984) while Wilhoft (1963)
reports that the granulosa measures 35 μm
before yolk deposition in Leiolopisma. In two
Mabuya species, the granulosa measured 42.2
μm (Hernandez-Franyutti et al. 2005) and 30
μm (Gomez & Ramirez-Pinilla 2004). In the
Mexican lizard, Ctenosaurus pectinata, the
granulosa reached an average thickness of 65.5
μm in prevetellogenic follicles (Uribe et al.
1996).
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
In H. mabouia, the granulosa becomes drasti-
cally reduced prior to the onset of vitellogene-
sis and, in oocytes measuring 1.5-2.0 mm in
diameter, the granulosa is reduced to a
monomorphic, single cellular layer. Although
the degeneration of the granulosa layer (disap-
pearance of intermediate and pyriform cells)
has been made in most studies concerning
detailed ovarian histology of snakes and lizards
(Hernandez-Franyutti et al. 2005), the actual
degenerative process, per se, was not observed
in the granulosa of H. mabouia. This seems to
indicate that the degenerative process occurs at
a fairly rapid rate in this gecko. The marked
reduction in the granulosa thickness from 32
μm to 3.9 μm (Stage 4 vs Stage 5; P < 0.001),
the disappearance of the pyriform and interme-
diate cells during this stage and the rapid
appearance of a single-layered, monomorphic
granulosa support this contention.
During the vitellogenic stage, the zona pelluci-
da reaches maximum thickness in
Hoplodactylus (Boyd 1940), Anolis (Neaves
1971), Naja (Lance & Lofts 1978) Agama (Van
Wyk 1984), Ctenosaurus (Uribe et al. 1996),
Mabuya (Gomez & Ramirez-Pinilla 2004;
Hernandez-Franyutti et al. 2005). In H.
mabouia, however, the zona pellucida main-
tains a relative constant thickness throughout
follicular development. The reason for this is
uncertain and requires further investigation.
The zona radiata of H. mabouia reaches maxi-
mum thickness during the vitellogenic and pre-
ovulatory stages and is consistent with other
130
studies (Gomez & Ramirez-Pinilla 2004). The
increase in both zones are generally attributed
to the dramatic increase in microvilli on the
surface of the oocyte and the cytoplasmic
extensions of the surrounding follicular cells
that interdigitate (Hernandez-Franyutti et al.
2005). This increase in absorptive surface and
dramatic increase in the number of coated pits
and vesicles along the inner surface of the
oocyte microvilli (Klosterman 1987; Guraya
1989; Andreuccetti 1992), seems to be related
to the transfer of yolk precursors and therefore
best developed in oviparous species compared
to microlecital viviparous species, for example,
Mabuya mabouia (Gomez & Ramirez-Pinilla
2004). Hernandez-Franyutti et al. (2005) point-
ed out that these Mabuya species mimic
oogenic patterns seen in mammals (lacking a
polymorphic granulosa) but in contrast to
megalecital species only show limited yolk pre-
cursor exchange but still exchange of cytoplas-
mic components of the pyriform cells.
Unlike some squamates which possess follicles
with differentiated thecae (Boyd 1940; Betz
1963; Guraya 1989), the thecal layer of H.
mabouia, like those described in other squa-
mates (Lance & Lofts 1978; Van Wyk 1984) is
undifferentiated. A distinctive increase in vas-
cularization of the theca is exhibited by large
ovarian follicles of H. mabouia. This is in
accordance with observations made on Agama
atra (Van Wyk 1984) and Anolis carolinensis
(Gerrard et al. 1973). Although thecal mast
cells in maturing follicles have been reported
(Jones et al. 1975; Van Wyk 1984), these cells
were not observed in H. mabouia. This could
be due to the different staining procedures
employed.
Postovulatory follicles.—All reptiles, whether
oviparous or viviparous, develop true corpora
lutea whose life span is correlated closely with
retention of eggs or young in the female genital
tract (Van Wyk 1984; Fox & Guillette 1987;
Guraya 1989; Guarino et al. 1998; Gomez &
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
Ramirez-Pinilla 2004; Hernandez-Franyutti et
al. 2005). The formation of corpora lutea in H.
mabouia is essentially similar to those
described for other reptilian species. After ovu-
lation, the follicular epithelium of the post-ovu-
latory follicle in H. mabouia undergoes rapid
hypertrophy (luteinization) to form the granu-
losa lutein cells which fill the follicular cavity.
The lutein cells originate from the granulosa
probably by differentiation since, firstly, there
is a lack of mitotic figures in the granulosa as it
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
undergoes luteinization and secondly, the luteal
cell size is greater than the small flattened cells
which make up the single-layered, monomor-
phic granulosa of the mature follicle.
The thecal layer of the corpus luteum of H.
mabouia is thicker than in the preovulatory fol-
licle. The theca does not undergo luteinization
to form paralutein cells, thus contradicting the
findings of Altland (1951) and Bragdon (1952).
Unlike the condition reported in Agama atra
(Van Wyk 1984), where thecal differentiation
was observed in the corpus luteum but not in
the preovulatory follicle, both the corpus
luteum and the preovulatory follicle exhibit
undifferentiated thecae in this gecko. While
distinct differentiation of the theca into an
externa and interna has been reported in sever-
al species (Guraya 1989; Gomez & Ramirez-
Pinilla 2004), thecal differentiation was not
observed in Hoplodactylus maculatus (Boyd
1940) and in Xenochrophis piscator (Saha et al.
1984). The thecal layer of the corpus luteum of
most reptiles possess blood vessels (Fox 1977;
Guraya 1989). Although the theca of the corpus
luteum of H. mabouia is vacularized, invasion
of blood vessels into the central luteal mass
does not occur. Septa of connective tissue,
however, invade the luteal cells. The absence of
blood vessels in the luteal mass therefore leads
to the assumption that secretion material pro-
duced by this glandular organ must diffuse into
the thecal layer in order to gain access to the
vascular system.
131
With regard to the degeneration of the corpus
luteum in H. mabouia, the first sign is an
increase of connective tissue inside the luteal
cell mass. This is followed by the luteal cell
nuclei becoming pyknotic, an increase in vac-
uoles within the luteal cell mass, the accumula-
tion of pigment and the general reduction in
size of the gland. In its essential features, cor-
pus luteum degeneration in the gecko corrobo-
rates reports on other reptiles (Altland 1951;
Betz 1963; Cyrus et al. 1978; Van Wyk 1984;
Guraya 1989; Gomez & Ramirez-Pinilla 2004).
Reptilian corpora lutea persist for several days
to several months depending on the species and
the mode of reproduction (Xavier 1987). The
life-span of the corpora lutea has been correlat-
ed to some extent with the period during which
eggs are retained in the uterus (Xavier 1987;
Villagran Santa Cruz & Mendez de la Cruz
1999; Gomez & Ramirez-Pinilla 2004). In sev-
eral viviparous squamate species studies have
shown that maximum activity corresponds to
progesterone secretion, usually associated with
maintenance of gestation or for a short period
following ovulation in oviparous species
(Xavier 1987; Gomez & Ramirez-Pinilla
2004). In the present study, septal invasion of
the corpus luteum takes place after ovulation
but its complete degeneration occurs immedi-
ately after oviposition. Corpora lutea remnants,
however, persist in the ovarian stroma long
after oviposition and can be used to identify
second clutches in a particular season (Van
Wyk 1984).
Follicular atresia.—Follicular atresia or degra-
dation of ovarian follicles has been described in
all vertebrate classes (Guraya 1989). Van Wyk
(1984) distinguished two types of follicular
atresia in the lizard Agama atra, including pre-
vitellogenic hydration stage atresia and vitel-
logenic stage artresia. In this study, only hydra-
tion stage atretic follicles were observed.
Similarly, Varma (1970) did not observe atretic
vitellogenic follicles in the post-ovulatory
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
ovaries of Calotes versicolor. Although atretic
follicles have been observed in many reptiles
(Van Wyk 1984; Guraya 1989; Gomez &
Ramirez-Pinilla 2004), conflicting reports on
the frequency of atresia have emerged. Boyd
(1940) reports that atresia is a rare phenomenon
in the gecko, Hoplodactylus maculatus.
Recently, polyovular follicles reported in the
ovaries of Mabuya (Gomez & Ramirez-Pinilla
2004; Hernandez-Franyutti et al. 2005) have
been recognized as atretic follicles in early fol-
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
liculogenesis similar to the phenomenon found
in Alligator mississippiensis (Guillette et al.
1994) and mammals (Iguchi et al. 1990) in
response to environmental estrogens.
Follicular cells play an important role in the
digestion and removal of ooplasmic contents
(Betz 1963; Guraya 1989). The invasion of the
oocyte by granulosa cells during advanced atre-
sia and the presence of ooplasmic granules
within the cytoplasm of the granulosa cells in
H. mabouia tend to support this finding. This
phenomenon has been reported by van Wyk
(1984) and Uribe et al. (1995). In addition,
macrophages, probably derived from the follic-
ular epithelial cells of atretic follicles, also play
a role in phagocytosis (Betz 1963; Guraya
1965). However, examination of corpora atret-
ica in H. mabouia could neither support nor
strengthen this contention. The possibility that
phagocytes may be transported by the blood
from an extra- follicular source, like they are
transported in mammals to sites of infection
and necrosis, remains probable. The highly
vascular corpora atretica reported in Anolis
carolinensis (Crews & Licht 1974) and in
Agama atra (Van Wyk 1984) tends to support
this view. According to some previous workers
(Altland 1951; Betz 1963) the theca interna
cells also appear to play a minor role in yolk
phagocytosis. Betz (1963) further pointed out
that the theca interna plays an important role in
the atresia of large follicles, where the granu-
losa is composed of a single layer of small cells
at the onset of atresia. This view, also, could
132
not be substantiated in the present investigation
since, in atretic follicles of the gecko, the theca
remains undifferentiated and the theca interna
cells, as such, are not distinguishable.
Jones & Swain (2000) suggested that clutch
size in squamates is regulated either through
recruitment (Sinervo & Licht 1991) or atresia.
The increased incidence of atresia in postovu-
latory ovaries has been reported previously
(Goldberg 1970; Varma 1970; Crews & Licht
1974; Guraya & Varma 1976; Callard et al.
1978; Van Wyk 1984). In accordance with the
above observations, the present study records
the highest incidence of atresia in the post-
breeding period (February). In Agama agama,
however, atretic follicles are present in ovaries
of females in advanced stages of vitellogenesis,
all sexually regressed adults and in some gravid
animals (Eyeson 1971). Gomez & Ramirez-
Pinella (2004) reported high incidence of artre-
sia in growing follicles in Mabuya mabouia,
rather than in preovulatory (vitellogenic) folli-
cles. Therefore, in keeping with the suggestion
of Jones & Swain (2000), they suggest that
clutch size is determined early during folliculo-
genesis. Although Crews & Licht (1974) have
proposed ways in which corpora atretica may
affect ovarian sensitivity, the functional signif-
icance of atretic follicles remains enigmatic
and further research, both in vitro and in vivo
on different reptilian types may in the future
reveal the secrets of a very interesting ovarian
component.
In this study we observed several aspects of
gekonid folliculogenesis and ovarian histology
observed during the natural ovarian cycle cor-
responded to the general ovarian morphology
pattern reported for other oviparous reptilian
species (see Guraya 1989).
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
ACKNOWLEDGEMENTS
We would like to thank the following: Dr H.
Baijnath & Mr K.S. Ganga for assistance with
the collection of specimens, Mr A. Rajh for
help with the photography, Mr S. Naidoo for
assistance with the statistical computations and
Dr. R. Biseswar for his helpful comments on
the draft manuscript. Mr K.S. Ganga also
assisted with the microtomy and microtech-
nique. Financial assistance was provided to
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
GKM. by the University of Durban Westville
(lately amalgamated with Natal University).
LITERATURE CITED
ALTLAND, P.D. 1951. Observations on the structure of
the reproductive organs of the box turtle. Journal of
Morphology 89: 599 - 621.
ANDREUCCETTI, P. 1992. An ultrastructural study of dif-
ferentiation of pyriform cells and their contribution
to oocyte growth in representative Squamata.
Journal of Morphology 212: 1 - 11.
ANDREUCCETTI, P. & CARRERA, M. 1987. The differenti-
ation of the zona pellucida (vitelline envelope) in
the lizard Tarentola mauritanica. Development
Growth and Differentiation 29: 113 - 122.
BETZ, T.W. 1963. The ovarian histology of the dia-
mond-backed water snake, Natrix rhombifera dur-
ing the reproductive cycle. Journal of Morphology
113: 245 - 260.
BOURQUIN, O. 1987. The recent geographical range
extension of Hemidactyulus mabouia mabouia.
Lammergeyer 38: 12 - 14.
BOYD, M.M.M. 1940. The structure of the ovary and the
formation of the corpus luteum in Hoplodactylus
maculatus. Quarterly Journal of Microscopical
Science 82: 337 - 376.
BRAGDON, D. 1952. Corpus luteum formation and fol-
licular atresia in the common garter snake,
Thamnophis sirtalis. Journal of Morphology 91:
413 - 445.
BRANCH, W.R. 1998. Field Guide to Snakes and Other
Reptiles of Southern Africa. Struik Publishers.
Cape Town.
BYSKOV, A.G. 1978. Follicular atresia. Pp. 533 - 561. In
R. Jones (Eds.), The Vertebrate Ovary:
Comparative Biology and Evolution. Plenum, New
York.
133
CALLARD, I.P., LANCE, V.A., SALHANICK, A.R., &
BARAD, D. 1978. The annual ovarian cycle of
Chrysemys picta: Correlated changes in plasma
steroids and parameters of vitellogenesis. General
and Comparative Endocrinology 35: 245 - 257.
CARRANZA, S. & ARNOLD, E.N. 2006. Systematics, bio-
geography, and evolution of Hemidactylus geckos
(Reptilia: Gekkonidae) elucidated using mitochon-
drial DNA sequences. Molecular Phylogenetics and
Evolution 38: 531 - 545.
CREWS, D.P. & LICHT, P. 1974. Inhibition by corpora
atretica of ovarian sensitivity to environmental and
hormonal stimulation in the lizard, Anolis caroli-
nensis. Endocrinology 95: 102 - 106.
CYRUS, R.V., MAHMOUD, I.Y., & KLICKA, J. 1978. Fine
structure of the corpus luteum of the snapping turtle
Chelydra serpentina. Copeia 1978: 622 - 627.
DU TOIT, A., MOUTON, P. LE F.N., & FLEMMING, A. .
2003. Aseasonal reproductin and high fecundity in
the Cape grass lizard, Cordylus anguinus,in a fire-
prone habitat. Amphibia-Reptilia 24: 471 - 482.
EYESON, K.N. 1971. Pituitary control of ovarian activi-
ty in the lizard, Agama agama. Journal of Zoology,
London 165: 367 - 372.
FLEMMING, A.F. 1993. The female reproductive cycle of
the viviparous lizard Pseudocordylus m. melanotus
(Sauria: Cordylidae). Journal of Herpetology. 27:
103 - 107.
FLEMMING, A.F. 1994. The male and female reproduc-
tive cycles of the viviparous lizard Mabuya capen-
sis (Sauria: Scincidae). Journal of Herpetology 28:
334 - 341.
FLEMMING, A.F. & BATES, M.F. 1995. Male and female
reproductive cycles of Bibron’s gecko
Pachydactylus bibronii (Sauria: Gekkonidae) in the
Free State province of South Africa. Journal of
African Zoology 109: 409 - 422.
FLEMMING, A.F. & BRANCH, W.R. 2001. Extraordinary
case of matrotrophy in the African skink Eumecia
anchietae. Journal of Morphology 247: 264 - 287.
FLEMMING, A.F. & MOUTON, P. LE F.N. 2002.
Reproduction in group-living lizard, Cordylus cat-
aphractus (Cordylidae), from South Africa. Journal
of Herpetology 36: 691 - 696.
FOX, H. 1977. The urogenital system of reptiles. In C.
Gans & T. S. Parsons (Eds.), Biology of the
Reptilia: Morphology.E. Volume: 6. Academic
Press, London.
FOX, S.L. & GUILLETTE, L.J. 1987. Luteal morphology,
atresia, and plasma progesterone concentrations
during the reproductive cycle of two oviparous
lizards, Crotaphytus collaris and Eumeces obsole-
tus. American Journal of Anatamy 179: 324 - 332.
 AFRICAN JOURNAL OF HERPETOLOGY 56(2) 2007
GAPP, D.A., HO, S.M., & CALLARD, I.P. 1979. Plasma
levels of vitellogenin in Chrysemys picta during the
annual gonadal cycle. Endocrinology 104: 784 -
790.
GERRARD, A.M., JONES, R.E., & ROTH, J.J. 1973. Thecal
vascularity in ovarian follicles of different size and
rank in the lizard Anolis carolinensis. Journal of
Morphology 141: 227 - 234.
GOLDBERG, S.R. 1970. Seasonal ovarian histology of the
ovoviviparous Iguanid lizard Sceloporus jarrovi
Cope. Journal of Morphology 132: 265 - 276.
GOMEZ, D. & RAMIREZ-PINILLA, M.P. 2004. Ovarian
histology of the placentotrophic Mabuya mabouya
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
(Squamata, Scincidae). Journal of Morphology
256: 90 - 105.
GUARINO, F.M., PAULESU, L., CARDONE, A., BELLINI, L.,
GHIARA, G., & ANGELINI, F. 1998. Endocrine activ-
ity in the corpus luteum and placenta during preg-
nancy in Chalcides chalcides (Reptilia, Squamata).
General and Comparative Endocrinology 111: 261
- 270.
GUILLETTE, L.J., GROSS, T.S., MASSON, G.R., MATTER, J
M., PERCIVAL, H.F., & WOODWARD, A.R. 1994.
Developmental abnormalities of the gonads and
abnormal sex hormone concentrations in juvenile
alligators from contaminated and control lakes in
Florida. Environmental Health Perspectives 102:
680 - 688.
GUPTA, V.K. & DUDA, P.L. 1985. Histogenesis of the
follicular wall in the Indian softshell turtle Lissemys
punctata punctata. Folia Morphologica 33: 22 - 26.
GURAYA, S.S. 1965. A histochemical study of follicular
atresia in the snake ovary. Journal of Morphology
117: 151 - 170.
GURAYA, S.S. 1989. Ovarian Follicles in Reptiles and
Birds. In D. S. Farner (Eds.), Zoophysiology.
Volume: 24. Springer-Verlag, Berlin.
GURAYA, S.S. & VARMA, S.K. 1976. Morphology of
ovarian changes during the reproductive cycle of
the house lizard, Hemidactylus flaviviridis. Acta
Morphologica Neerlando-Scandinavica 14: 165 -
192.
HERNANDEZ-FRANYUTTI, A., URIBE, M., & GUILLETTE,
L. 2005. Oogenesis in the viviparous matrophic
lizard Mabuya brachypoda. Journal of Morphology
265: 152 - 164.
HUMASON, G.L. 1967. Animal Tissue Techniques. W. H.
Freeman and Co. San Francisco.
IGUCHI, T., FUKASAWA, Y., UESUGI, Y., & TAKASUGI, N.
1990. Polyovular follicles in mouse ovaries
exposed neonatally to diethylstilbestrol in vivo and
in vitro. Biololgy of Reproduction 43: 478 - 484.
JONES, R.E. 1978. Control of follicular selection. Pp.
763 - 787. In R. E. Jones (Eds.), The Vertebrate
134
Ovary: Comparative biology and evolution.
Plenum Press, New York.
JONES, R.E. & SWAIN, R. 2000. Effects of exogenous
FSH on follicular recruitment in a viviparous lizard
Niveoscincus lugubris. Comparative Biochemistry
and Physiology A 127: 487 - 493.
JONES, R.E., TOKARZ, R.R., ROTH, J.J., PLATT, J.E., &
COLLINS, A.C. 1975. Mast cell histamine and ovar-
ian follicular growth in the lizard Anolis carolinen-
sis. Journal of experimental Zoology. 193: 343 -
352.
JONES, R., SWAIN, T., GUILLETTE, L.J., & FITZGERALD,
K.T. 1982. The comparative anatomy of lizard
ovaries, with emphasis on the number of germinal
beds. Journal of Herpetology 16: 248 - 252.
KLOSTERMAN, L.L. 1983. The ultrastructure of germinal
beds in the ovary of Gerrhonotus coeruleus
(Reptilia: Anguidae). Journal of Morphology 178:
247 - 265.
KLOSTERMAN, L.L. 1987. Ultrastructural and quantita-
tive dynamics of the granulosa of ovarian follicles
of the lizard Gerrhonotus coeruleus (Family
Anguidae). Journal of Morphology 192: 125 - 144.
LANCE, V.A. 1989. Reproductive cycle of the American
alligator. Amer. Zool. 29: 999 - 1018.
LANCE, V.A. & LOFTS, B. 1978. Studies on the annual
reproductive cycle of the female cobra, Naja naja
IV. Ovarian histology. Journal of Morphology 157:
161 - 180.
LOREDANA, R., VACCARO, M.C., PRISCO, M.,
CAROTENUTO, R., LIGUORO, A., & ANDREUCCETTI,
P. 2004. a and b Spectrin distribution during the
differentiation of pyriform cells in follicles of lizard
Podarcis sicula. Molecular Reproduction and
Development 67: 101 - 107.
MOODLEY, G.K. 1991. The reproductive biology of the
house lizard, Hemidactylus mabouia (Sauria:
Gekkonidae). Unpubl. Masters Thesis, University
of Durban-Westville. Durban, South Africa
MOTTA, C., CASTRIOTA-SCANDERBEG, M., FILOSA, S., &
ANDREUCCETTI, P. 1995. Role of pyriform cells dur-
ing the growth of oocytes in the lizard Podarcis sic-
ula. Journal of experimental Zoology 273: 247 -
256.
MOTTA, C., FILOSA, S., & ANDREUCCETTI, P. 1996.
Regression of the epithelium in the late previtel-
logenic follicles of Podarcis sicula: a case of apop-
tosis. Journal of experimental Zoology 266: 233 -
241.
NEAVES, W. 1971. Intercellular bridges between follicle
cells and oocyte in the lizard, Anolis carolinensis.
Anatomical Record 170: 285 - 302.
 MOODLEY & VAN WYK — Folliculogenesis in Hemidactylus mabouia
NEAVES, W. 1972. The passage of extracellular tracers
through the follicular epithelium of lizard ovaries.
Journal of experimental Zoology 179: 339 - 364.
NKOSI, W.T., HEIDEMAN, N.J.L., & VAN WYK, J.H.
2004. Reproduction and sexual size dimorphism in
the Lacertid Lizard Pedioplanis burchelli (Sauria:
Lacertidae) in South Africa. Journal of Herpetology
38: 473 - 480.
NORRIS, D.O. 1996. Vertebrate Endocrinology.
Academic Press. New York.
RAHIL, K.S. & NARBAITZ, R. 1973. Ultrastructural stud-
ies in the relationship between follicular cells and
growing oocytes in the turtle Pseudemys scripta.
Downloaded by [Florida Atlantic University] at 20:04 17 November 2014
Journal of Anatomy 115: 175 - 186.
SAHA, B., MANNA, D., & SIRCAR, A. 1984. Ovarian his-
tology of the water snake Xenochrophis piscator.
Annals of Zoology 21: 47 - 58.
SINERVO, B. & LICHT, P. 1991. Proximate constraints on
the evolution of the egg size, number and total
clutch mass in lizards. Science 252: 1300 - 1302.
URIBE, M.C.A. & GUILLETTE, L. J. 2000. Oogenesis and
ovarian histology of the American alligator
Alligator mississippiensis. Journal of Morphology
245: 225 - 240.
URIBE, M.C.A., OMANA, M.E.M., QUINTERO, J.E.G., &
GUILLETTE, L.J. 1995. Seasonal variation in ovarian
histology of the viviparous lizard Sceloporus
torquatus torquatus. Journal of Morphology 226:
103 - 119.
URIBE, M.C.A., PORTALES, G.B., & GUILLETTE, L.J.
1996. Ovarian folliculogenesis in the oviparous
Mexican lizard Ctenosaura pectinata. Journal of
Morphology 230: 99 - 112.
VAN WYK, J.H. 1983. Seasonal breeding in the female
rock lizard, Agama atra (Sauria: Agamidae) in the
south western Cape Province with special reference
to possible enviromental controlling factors.
Navorsinge van die Nasionale Museum,
Bloemfontein 4: 193 - 208.
VAN WYK, J.H. 1984. Ovarian morphological changes
during the annual breeding cycle of the rock lizard
Agama atra (Sauria: Agamidae). Navorsinge van
die Nasionale Museum Bloemfontein,
Bloemfontein 4: 237 - 275.
Received: 9 October 2006;
135
VAN WYK, J.H. 1991. Biennial reproduction in the
female viviparous lizard, Cordylus giganteus
(Sauria: Cordylidae). Amphibia-Reptilia 12: 329 -
342.
VAN WYK, J.H. 1994. Physiological changes during the
female reproductive cycle of the viviparous lizard,
Cordylus giganteus (Sauria: Cordylidae).
Herpetologica 50: 480 - 493.
VAN WYK, J.H. 1995. The male reproductive cycle of
the lizard, Cordylus giganteus, (Sauria:
Cordylidae). Journal of Herpetology 29: 522 - 535.
VAN WYK, J.H. & MOUTON, P. LE F.N. 1995. The
reproductive cycles of the oviparous lizards
Platysaurus capensis and P. minor: evidence
suporting a viviparity-oviparity reversal in the
Cordylidae. Amphibia-Reptilia 69: 1 - 15.
VAN WYK, J.H. & MOUTON, P. LE F.N. 1998. The
reproductive cycles of the montane viviparous
lizard Pseudocordylus capensis (Sauria:
Cordylidae). S. Afr. J. Zool. 33: 156 - 165.
VAN WYK, J.H. & RUDDOCK, L. 2000. The male repro-
ductive cycle of the lizard Agama atra (Sauria:
Agamidae) in central South Africa. Amphibia-
Reptilia 22: 119 - 123.
VARMA, S.K. 1970. Morphology of ovarian changes in
the garden lizard, Calotes versicolor. Journal of
Morphology 131: 195 - 210.
VILLAGRAN SANTA CRUZ, M. & MENDEZ DE LA CRUZ,
F.R. 1999. Corpus luteum through the gestation of
Sceloporus palaciosi (Sauria: Phrynosomatidae).
Copeia 1999: 214 - 218.
WILHOFT, D.C. 1963. Gonadal histology and seasonal
changes in the tropical Australian lizard,
Leiolepisma rhomboidalis. Journal of Morphology
113: 185 - 204.
XAVIER, F. 1987. Functional morphology and regulation
of the corpus luteum. Pp. In D. O. Norris and R. E.
Jones (Eds.), Hormones and Reproduction in fish-
es, amphibians and reptiles. Plenum Press, New
York.
Final acceptance: 16 November 2007.