La fecundación

One can speak of a successful fertilization when the genetic

material of the male spermatozoon has united with that of the female
oocyte thereby creating a new, unique individual.
Los gametos: el espermatozoide

centriole

Maduración, activación y capacitación

Capacitación = el espermatozoide adquiere
competencia para la fecundación Involucra la
pérdida de colesterol de la membrana, una
hiperpolarización y la activación de cAMP/PKA
 Los gametos: El oocito
• El oocito ha madurado a lo largo del ciclo y se ha parado en la metafase de la 2ª división
meiótica
• El oocito maduro está rodeado por la zona pelúcida y las células foliculares (corona radiata)

Zona pelúcida 1er cuerpo polar

The oocyte in the cloud of

cumulus cells following
ovulation Células foliculares
The fluid that lies in between is sticky and stringy (effect of the
G 7.7
hyaluronic acid) with a high concentration of progesterone (to
attract the spermatozoa).
 Las fases de la fecundación

• Reconocimiento y
binding del
espermatozoide
• Reacción
acrosómica
• Penetración en la
zona pelúcida
• Fusión de los
gametos
• Reacción cortical
• Fusión de los pro-
núcleos
• Reaccioes zigóticas
W 12.22
 Reconocimiento, binding y reacción acrosómica

• La actividad
hialuronoidasa del
espermatozoide permite
atravesar la barrera celular GalT: ZP3
• La segunda capa es la
zona pelúcida formada por
glicoproteínas.
• El reconocimiento está
mediado por la interacción
de proteínas del
espermatozoide (ZP-
binding proteins) con las
proteínas ZP de la zona Fertilin:
pelúcida, de manera
intergin
especie-específica
• La reacción acrosómica
libera enzimas que
destruyen localmente la
ZP.
G 7.8
 La reacción acrosómica

ZPbs (sperm)
Β-1,4-galactosyltranf-I
GalT

Reconocimeinto y binding ZPbp:ZP3

ZP3

Reacción acrosómica:exocitosis acrosin

Lisis y penetración de la ZP Inner membrane of

sperm free for
recognition

Corona radiata Zona pellúcida Membrane of the ovule

Fusión de los gametos

• La fusión de las
membranas de los
gamétos se debe a la
interacción entre :
fertilin : integrins
(ADAM proteins and integrin-
asociated CD9)

centriolo

S 10.2
G 7.20
 Reacciones rápidas: Onda(s) de calcio
• La fusión de los gametos determina una onda de calcio intracelular
• La onda de calcio produce la liberación por exocitosis de enzimas
protelíticos, la reacción granular cortical en toda la membrana del oocito.
 Reacciones rápidas: La reacción granular cortical
reestructura la ZP y previene la poliespermia
• La onda de calcio
produce la liberación
por exocitosis de
enzimas protelíticos,
la reacción granular
cortical en toda la
membrana del oocito
(N-acetyl-glucosaminidases
fecundado. + proteases)
 Iniciación de las respuestas zigóticas

La activación de la vía IP3/DAG de la

membrana y la onda de Ca2+
determina las respuestas zigóticas

Respuestas tempranas (segundos-

minutos):
Reacción granular cortical y
prevención de la polispermia

Respuestas tardías (horas):

Fin de la meiosis del oocito,
expulsión del 2º cuerpo polar
Migración y fusión de los
pronúcleos
Inicio de la división celular
Aumento de pH, activación
metabólica y respiratoria, activación
y síntesis de proteínas y mRNAs
¡ojo!
En algunas especies la actividad
transcripcional no empieza hasta que
hayan ocurrido varias divisiones: La Pronúcleos antes de la fusión, y los
mid-blastula transition núcleos tras la primera división. El
pronúcleo masculino ha sido marcado
con BrdU
 Inicio de la segmentación

G 11.21

Early blastomers Somatic cells

MPF

G 8.2
 Algunas preguntas para pensar

• When the spermatozoon leaves the testicle it is not yet capable of

fertilizing an oocyte. What changes must it undergo to make it
capable of uniting with the oocyte (capacitation)?

• How does the oocyte get from the ovary into the fallopian tube?
• What makes it possible for the spermatozoon to penetrate into the
oocyte?

• How is it made sure that a spermatozoon from another species is

unable to deposit its genetic material?

• When the fertilization is completed, the genes of the two parents

have united. How does this union of the two genomes take place?
 Referencias
LANGMAN´S, Lippincot Wiliams & Wilkins 9-10th editions, 2004,
2006

LARSEN´S HUMAN EMBRYOLOGY, Churchill Livinstone,

Elsevier, 2008

EMBRYOLOGY.CH
http://www.embryology.ch/genericpages/moduleembryoen.html

ALBERTS ET AL. MOLECULAR BIOLOGY OF THE CELL.

GARLAND SCIENCE, CAP. 20

SCOTT F.GILBERT, DEVELOPMENTAL BIOLOGY. Sinauer, 6th

ed. 2000. CAP. 11

PATHOPHYSIOLOGY OF THE REPRODUCTIVE SYSTEM:

http://www.vivo.colostate.edu/hbooks/pathphys/reprod/index.html
 Más información:
gametos
Maturation steps of the spermatozoa in the
epididymis:
Through the deposition of new proteins in the nucleus,
the DNA becomes more condensed. The sperm
head becomes smaller, thereby, and more compact.
This is an important step for the later correct
decondensation of the paternal DNA in the maternal
oocyte. The already meager cytoplasma is further
reduced, making the sperm cells more slender.
The ability for motility is achieved but at the same
time inhibited by the milieu. The structure of the
plasma membrane is altered. This has effects on the
motility, the capacitation ability and the ability for the
acrosome reaction.
The ejaculation represents the second step in the
maturation of the sperm cells. In this process, the
motile capabilities of the spermatozoa are abruptly
activated. The precise mechanisms of this activation
are still unclear. The result of the interaction of the
mechanical stimulation by the ejaculation and the
chemical effects of the glandular secretions on the
sperm cells correspond with a physiological activation
of the sperm cells so that they are ready to go through
the upcoming capacitation and the acrosome reaction -
both are preconditions for an approach and binding of
the spermatozoa to the pellucid zone of the oocyte.
Oogenesis begins in roughly the 7th week (stage 20). The
secondary germinal cords that have grown into the ovarian
cortex decompose into individual groups of cells. A lively
proliferation result, whereby the oogonia remain connected with
each other via cellular bridges, permitting a synchronization of
the mitosis and the subsequent meiosis steps (prophase).
As soon as these oogonia have commenced with meiosis, they
are named primary oocytes (12th week). All oocytes are arrested
in the first meiosis at the end of the prophase. The primary
oocytes loose themselves from their cellular binding; they
become surrounded by flat, somatic cells (follicle or granulosa
cells) and are now called primordial follicles. In the 20th week
nearly 7 million germ cells are formed and the whole cortex
consists of these primordial follicles. After birth, only around 2
million are present and after puberty only around 250,000 remain
in each ovary
Roughly a week before the midpoint of the menstrual cycle the
dominant follicle develops in one of the two ovaries. This grows
faster than the other follicles and prepares itself for ovulation. It
reaches a diameter of up to 25 mm and is also known then as the
graafian follicle. Approximately one-and-a-half days before the
midpoint of the cycle, the concentration of the luteinizing hormone
(LH) rises steeply: triggering ovulation and creating an optimal
milieu for a successful fertilization, With the end of the first
meiosis the name of the oocyte changes from primary oocyte to
secondary oocyte.
The follicle that is about to rupture the granulosa cells also
secrete an extra-cellular matrix, mainly hyaluronic acid, into the
follicle fluid. Before ovulation the follicle fluid volume increases
markedly. The cumulus cell bonds loosen further. In this way,
together with the enclosed oocyte, they free themselves from
where they were attached to the follicular wall and now swim in
the follicle fluid. The wreath of granulosa cells that enclose the
oocyte is called the corona radiata.
 Más información: La fecundación tiene lugar en el tracto
genital femenino
Mechanism of oocyteFimbriae
acquisition:
apposed
• Maduración y Initial positions
ruptura del folículo

The oocyte now "waits" in the

fallopian tube on fertilization by the
sperm. The matrix of hyaluronic acid
holds it "captive" there, so to speak.
After a number of hours the matrix
liquefies more and more and the
oocyte is gradually transported
towards the uterus by the ciliary
beats of the tube's epithelium cells.
Since after ovulation the oocyte can
only be fertilized within a few hours,
the fertilization must almost
inevitably take place in the ampullary
part of the fallopian tube.
 Más información: El recorrido del espermatozoide
Female genital tract - path the sperm cells travel
The path the sperm cells travel is marked in yellow.
The triangles indicate those places along the path
where it has been shown that the sperm cells can wait
for longer periods of time. They are the crypts in the
cervix, the region of the tube isthmus and the
ampullary part of the fallopian tube.
In order, though, that a sufficient number of sperm cells appear
in the ampulla at the right time, a large number of sperm cells
must be present in the ejaculate. Of the roughly 200 million
ejaculated sperm cells only a few hundred are able to traverse
the long way through the cervix, the uterus, and past the fallopian
tube isthmus to the tube's ampullary region to there meet oocyte.
Along the way whole groups of sperm cells can halt at certain
places and enter a phase of reduced activity. That is why a portion
of the sperm cells can retain their fertilizing capability for up to 4
days.
With the restructuring of the cervical barrier the mucus becomes
thinner and more fluid. Therein meandering passages coated
with specific chemotactic molecules form that the sperm cells
will prefer in order to pass through the cervix.
The passage through the cervical canal is an important step for the
selection of the sperm cells. The cervical mucus barrier
functions as a filter in which atypical sperm cells remain hanging.
They are hindered in ascending by means of a hydrodynamic
effect. Through this simple mechanism it is assured that only
normally formed and highly mobile sperm cells are able to
overcome the cervical mucus barrier.
Capacitation is a functional maturation of the spermatozoon.
The changes take place via the sperm cell membrane in which it
may be that receptors are made available through the removal of a
glycoprotein layer. The area of the acrosomal cap is also so
altered thereby that the acrosome reaction becomes possible.
Through the membrane alterations, the motile properties of the
spermatozoon also change. Discharging whipping movements
of the tail together with larger sideways swinging movements of
the head take place. This type of motility is designated as
hyperactivity. One can therefore say that the visible consequences
of capacitation consist in hyperactivity of the spermatozoon.
 Más información: Las proteínas de la Zona Pelúcida ZP 1-3

• Las primeras proteínas reconocidas por

el espermatozoide son las ZP3

• Tras esta unión sucede un cambio

conformacional y un binding secundario
con las ZP2

• El acrosoma libera acrosina (proteasa) y

acetylglucosaminidasas que degradan
las glicoproteínas de la ZP exponiendo la
membrana del oocito a la interacción con
la membrana del espermatozoide

G 7.17
 Más información: la migración y fusión de los
núcleos
Figure 1. Distribution of paternal chromatin in
early mouse embryos. BrdU-treated male mice
were mated with untreated females and the
resulting embryos stained with FITC-conjugated
anti-BrdU antibody (green). Nuclei and
chromosomes were counterstained with DAPI
(blue). a, Highly condensed sperm nucleus and
fertilized egg. b, Male and female pronuclei at 10 h
after fertilization. The somewhat larger male
pronucleus shows a nearly uniform BrdU staining,
indicating that the entiresp erm DNA is substituted
with BrdU. c, After nuclear envelope breakdown
the two chromosome sets form a single diploid
nucleus. d, First metaphase at 20 h after
fertilization. e, Twocell embryo during G1 phase at
22 h. The second polar body remains completely
BrdU negative. f, Two-cell embryo during G2
phase at 32 h. The male chromatin occupies
approximately half of the nuclear volume. Bars, 10
mm.
From Mayer et al. (2008), J.Cell Biol.