Female Anatomy

Female Reproductive Tract Anatomy

Functions of the female reproductive tract

  • reproductive organs controlled hormonally

    • endocrine function: ovary, uterus, fetus, placenta, pituitary gland, hypothalamus, pineal gland

    • exocrine function: female gamete production

  • houses the oocyte (egg)

    • the female gamete

  • transports sperm and oocyte/embryo via stimulation of:

    • myometrial contractions

    • oviductal contractions

  • facilitate the events of fertilization

    • assists with the passage of sperm from the site of deposition to the oviduct

  • provide environment for embryo and fetus

    • oviduct

      • very early in pregnancy

    • uterus

      • the remainder of pregnancy

  • give birth to fetus

    • dilation (effacement) of the cervix

    • strong uterine contractions

  • “recovery” to become pregnant again

    • uterine involution

    • return to a state of positive energy balance

  • provide nutrients to young

    • lactation

      • stimulate mammary gland development

Tubular Layers of the Tract

  • Layers of tract tubes (superficial to deep)

    • serosa

      • simple squamous connective tissue

      • cover the surface

    • muscularis

      • double layer of smooth muscle

        • outer longitudinal layer

        • inner circular layer

      • gives the ability to contract

        • contractions are necessary for:

          • transport of secretory products, gametes, early embryos, expulsion of fetus and fetal membranes during parturition

    • submucosa

      • varying thickness

      • houses blood vessels, nerves, and lymphatics

      • supporting tissue for mucosa

    • mucosa

      • varying type of mucosal epithelium

        • dependent on location

      • lines the lumen

Supporting Structures

Broad Ligament

  • supports and suspends the ovaries, oviduct, uterus, cervix and anterior vagina

  • developed by partial fusion of peritoneum to form a double layered connective tissue sheet

  • houses vascular supply, lymphatic drainage, and nerves

  • components

    • mesovarium

      • anterior/cranial portion

      • attaches to and supports the ovary

      • houses the blood and lymphatic vessels and nerves that supply the ovary

      • forms the hillus of the ovary

      • in the bitch:

        • Ovarian Bursa

          • an extension of the mesovarium that almost surrounds the ovary in dogs

          • fat-covered peritoneal lining

    • mesosalpinx

      • thin, serous part

      • surrounds and supports the oviduct (salpinx)

      • serves as a bursa-like pouch that surrounds the ovary

      • helps to orient the infundibulum so that ova released at ovulation have a high probability of being directed to the oviduct

    • mesometrium

      • largest and most visible part

      • supports the uterine horns (cornua) and uterine body

      • dorsal portion is continuous with the dorsal peritoneum

        • thus the uterus “hangs” from the dorsal body wall

Utero-Ovarian Ligament

  • attaches ovary to uterus

  • other name:

    • proper ligament of the ovary

Blood Supply

General

  • presence of a functional CL will increase blood flow

  • example: ewe

    • blood flow increases from 1mL/minute to 3-7mL/min as CL develops

  • ovarian pedicle

    • ligament containing blood vessels that nourish the ovary

Ovarian Artery (Utero-Ovarian Artery)

  • supplies: ovaries, oviducts, potion of uterine horn

  • return route: utero-ovarian vein)

Middle Uterine Artery

  • supplies: remainder of uterine horns and uterine body

  • in the cow:

    • ~5 months gestation

    • enlarges during middle and late pregnancy

    • can be palpated as an aid in pregnancy detection

      • a palpable vibration - fremitus “swoosh”

    • cannot feel fremitus in mares, doesn’t seem to develop

Hypogastric Artery

  • supplies: cervix, vagina, vulva

External Organs/Structures

Perineum

  • region surrounding the anus and vulva and covers the pelvic outlet

  • dorsal to caudal

    • Anus → cutaneous bridge → dorsal commissure → labia → ventral commissure

Vulva

  • consists of two labia (major and minor) and meet in the medial portion of tract to form the dorsal and ventral commissures

    • the labia form a closure to keep foreign material out of vagina

      • labia majora (external)

      • labia minora (internal)

      • labia constituents

        • skin (integument)

          • sebaceous and sweat glands, and hair follicles

        • mainly adipose tissue

          • imbedded in small bundles of smooth muscles known as constrictor vulvae

            • these insure the labia stay side by side

    • clitoris

      • female homologue of the penis

      • erectile tissue covered with stratified squamous epithelium

        • erection caused by onset of estrus with high estrogen levels

      • highly innervated (sensory)

      • not known to have a functional significance in domestic animals

      • cow: buried in mucosa

      • mare: well developed

      • sow: long, sinus

  • functions:

    • external opening to the female reproductive tract

      • first effective barrier to combat external environmental infections

      • protects the internal reproductive organs from ascending infections

    • passage of urine

    • opening for mating

    • portion of birth canal

  • in a freemartin vulva is not open, but rather fused

  • Caslick’s procedure

    • surgical closing of the upper part of the vulva of a mare

      • performed on mare’s with abnormal or disfigured vulvar anatomy

    • required reading:

Caudal Portion of the Internal Tract

Vestibule

  • otherwise known as the caudal/posterior vagina, its serves as the “opening” to tract

  • functions:

    • common area for both urinary and reproductive tract

    • stimulates male for copulation

    • passage for fetus during parturition

  • develops from the ventral part of the cloaca

  • no secretory glands, but epithelial does produce some secretions

  • both a part of the urinary and genital systems

  • extends from the external urethral orifice to the labia

  • contains:

    • gartner’s tubules (ducts)

      • open directly into the vestibule and are blind sacs

      • no apparent function

      • known as an embryonic remnant of the male reproductive system of the embryo (Wolffian duct)

        • Wolffian duct becomes vas deferens in the male and degenerates in female

    • major vestibular glands

      • also known as Bartholin’s glands

      • located in the caudal portion of vestibule in the submucosa

      • actively secrete mucus-like material and pheromones during estrus

        • pheromones: airborne chemical messages

      • cow:

        • left and right glands

        • single excretory duct

      • mare and sow:

        • multiple minor glands

        • large number of small excretory ducts

    • suburethral diverticulum

      • not present in mare & bitch

        • present in domestic ruminants, pigs and camelids

      • small, blind pouch in the floor of vestibule that lies immediately ventral to the urethral opening

        • blind pocket ventral to orifice

      • sits below shelf that separates it between the urethra

      • function unknown

        • cautions when AI’ing, don’t want to stick rod in pouch

        • can be used as a landmark for the insertion of a urinary catheter

    • urethral tubercle

      • bitch ONLY

      • bulb or ridge-like structure protruding caudally from vagina into the vestibule directly above urethral opening

      • varies in size

      • covers and contains urethral opening

    • transverse vestibulovaginal fold

      • mare ONLY (“open” tract)

      • also known as the transurethral fold or intravaginal transverse fold

      • overlies the external urethral orifice and separates the vagina from the vestibule

      • fold of tissue in the vagina that blocks and prevents the backflow of urine into the RT

      • serves as a secondary barrier for the uterus against external contaminants

        • seals vagina

        • resistance against speculum indicates good tone and function

  • vulvo-vaginal sphincter muscle

    • between vagina and vestibule, behind urethra

    • contracts (closes off) when urinating so urine does not breach reproductive tract

    • cow, ewe, goat, pig (mare)

      • mare has transurethral fold to help direct urine outward

  • hymen

    • membrane at the junction of vagina and vestibule

Vagina

  • functions:

    • copulatory organ, site of semen deposition in some species (fornix vagina), site for expulsion of urine during micturition, passive birth canal

  • poorly organized and ill-defined muscular layer

  • well developed and highly adaptive mucosal epithelium

  • no glands

    • secretions come from passage of plasma components as well as from cervix

    • pH is acidic (5.7)

      • bacteriostatic

    • stimulates glans penis in some species

      • temperature and pressure

      • artificial vagina (AV)- correct water temperature and water pressure

  • caudal vagina (vestibule)

    • originates from an invagination of the urogenital sinus

    • stratified squamous epithelium

    • secretory and thickness of epithelium changes with endocrine status

      • estrogen dominance (estrus) - thickens dramatically

        • mechanically protects vagina during copulation

        • thickened mucosa prevent microorganisms from entering the vasculature in the submucosa

  • cranial vagina

    • originates from paramesonephric ducts and fuses with caudal vagina

    • high degree of secretory activity from columnar epithelium and ciliated columnar epithelium, (luminal epithelium)

    • fornix vagina

      • NOT in the sow

      • protrusion of the cervix into the cranial vagina

      • forms a crypt, or pocket

      • spermatozoa are deposited in the fornix vagina by the bull during live-cover

      • composed of columnar epithelial cells

        • as an extension of the cervix, it secretes high amounts of mucus during estrus

Cervix

  • relatively thick-walled, non-compliant

  • made of muscle and connective tissue

    • surface covered with columnar and some ciliated columnar epithelium

  • layers:

    • serosa, outer muscular layer, deep stromal layer, epithelial and superficial stromal layer, cervical lumen with corresponding structure

  • functions:

    • barrier to and transports sperm

      • mucus and anatomy of cervix act as a sperm filter

      • prevents large numbers of sperm from reaching oviduct

      • first sperm barrier in cow, ewe, doe, bitch and queen

      • not involved as a barrier in the sow or mare

    • produces long strands of mucus

      • mucus production for lubrication, prevention of MO’s from entering uterus and transport of sperm

    • reservoir for sperm until capacitation

      • sperm enter the crypts and folds of cervix

        • crypts and folds are covered with columnar epithelium, some ciliated

        • protects sperm for a long period of time

        • provides a favorable environment for sperm as they move into the uterus

          • conditions in cervical mucus favor sperm motility

      • majority of sperm (~90%) is lost out of the vagina

        • retrograde transport

    • blocks bacterial invasion during pregnancy

      • mucus is highly cross-linked

      • cervix constricts tightly

      • leukocytes are present to kill bacteria (MO’s)

    • cervical seal of pregnancy

      • cervical mucosa produces a mucus secretion which forms a mucus plug that helps close the cervical canal

    • birth canal

      • cervical plug liquifies

      • cervix dilates to permit fetal expulsion at parturition

    • very dynamic structure

      • remodels itself at the end of gestation to allow parturition

      • stimulates the development of maternal behavior

    • isolates uterus from external environment during pregnancy

      • by forming a barrier made of highly viscous mucus

        • mucus flows from cervix towards exterior

        • lubricates vagina during copulation

        • mucus flushes out foreign material from copulation

          • including sperm

          • minimizes microorganism contamination in uterus

        • biochemical and physical properties change depending on stages of estrous cycle

    • transportation vesicle of spermatozoa

    • responsible for isolation of the conceptus within uterus from external environment

      • progesterone causes mucus to become viscous

        • forms the cervical seal of pregnancy

          • viscous mucus “glues” folds of cervix together

            • thus, foreign material cannot enter uterus during pregnancy

            • disruption of this seal will typically cause abortion

              • because microorganisms are able to access the interior uterus

                • causing infection and subsequent embryonic death

  • Cervical Mucus

    • under estrogen control = thin and watery

    • conditions in cervical mucus favor sperm motility

      • sperm enter parallel strands of mucus in the vagina and are transported to the cervical crypts and folds

    • muscular action of vagina and cervix may aid in sperm movement

    • two types:

      • sialomucin

        • low viscosity, long strands

        • “privileged pathway”

          • direct pathway for sperm

          • most don’t make it here

      • sulfomucin

        • high viscosity, ferning pattern

        • forms a trap/selective

    • during estrus:

      • clear, watery long parallel strands (estrogen)

    • during diestrus:

      • little mucus, jelly-like mucus cross-linked by disulfide bonds (progesterone)

  • cervical ripening

    • complex process of modification of cervical tissue that enables dilation of the cervix at parturition

    • cervical smooth muscle tissue

      • may play a role

      • collagen fibrils are denatured and digested as part of an inflammatory cascade

    • characterized by:

      • further decrease in collagen concentration

      • increase in collagen denaturation and water imbibition

        • results in cervical tissue with an extremely soft, watery and well-distensible (swelling) character

    • cervical tissue is able to dilate

      • give way to pressure exerted by the fetal membranes or the fetus when the uterus contracts

  • dynamic characteristics

    • cervix is very hard and rigid during estrus

    • composed of thick connective tissue

    • crypts and folds increase surface area for sperm reservoir

    • cervix tightly closed except during estrus (standing heat)

  • anatomy varies between species

    • cervical canal (lumen) fold(s) or ring(s) protruding into the cervical canal

      • single- bitch, queen

        • no elaborate folds or rings

        • relatively smooth

        • portion of cervix protrudes into cranial vagina

          • bitch ONLY

        • quite small, thick-walled compared to uterus and vagina

          • queen ONLY

      • multiple- cow, ewe, sow, mare

        • several rings form interlocking finger-like projections

          • cow and ewe ONLY

          • called cervical rings/annular rings

            • protrude into the lumen

          • primary function of cervix is to produce mucus during estrus

          • cow has 3, 4-5 rings

          • in beef cows ~17% of all cases of dystocia are of cervical origin

          • ewe cervical os classifications:

            • duckbill, slit, rose, papilla, flap

        • Interdigitating pads

          • sow ONLY

          • also called interdigitating prominences

          • smaller quantity of mucus produced

          • interdigitations require special penile adaptations in boar

            • boar has corkscrew or spiral twist in glans penis so it becomes “locked” in cervix

            • thus, semen deposition occurs in cervix of sows

            • large volume of semen → quickly enters uterus

        • longitudinal folds

          • conspicuous, loose folds of mucosa protruding into the vagina

            • mare ONLY

            • no rings

              • but many longitudinal cervical folds

                • continuous with endometrial folds of uterus

            • smaller quantity of mucus produced

            • mare’s cervix is soft and pliable during estrus and flattens on floor of pelvis

              • during pregnancy is tight and closed

              • there are no obstacles in the mare after the fornix vagina

        • cervical folds

          • doe only

          • cervical os classifications:

            • star, duckbill, crescent, spiral, cluster, bump

  • semen deposition of stallion and boar

    • ejaculate through the cervix

    • stallion:

      • glans penis bells out and expands cervix

      • force of ejaculation sends semen directly into the uterine body

    • boar:

      • corkscrew penis

      • glans penis locks into interdigitating pads of the cervix

      • volume of semen (300-400mL) moves into the uterus

      • UTJ is more functional in sow to control sperm numbers entering the oviduct

Cranial Portion of Internal Tract

Uterus

  • organ of pregnancy

  • connects cervix to oviducts

  • consists of two uterine horns (cornua)

  • primary functions:

    • sperm transport and motility

      • transports sperm when female is in estrus

        • estrogen stimulates myometrial contractions so that sperm move to site of fertilization

      • viable sperm (motility) are important so that they are not absorbed

    • absorption and phagocytosis

      • occurs by uterine epithelium and leukocytes (immune cells) which fight infection

    • luteolysis and control of cyclicity

      • involved with regression of the CL

        • if not pregnant

          • uterine endometrium releases PGF2a to cause CL to regress

        • if pregnant

          • embryo provides chemical signal which allows CL to be maintained

            • "maternal recognition of pregnancy”

    • environment for preattachment and sustainment of embryo

      • uterine secretions stimulated by estrogen and progesterone

      • proper timing of embryo and uterus is important for embryo transfer

    • maternal contribution to the placenta and development of fetus

      • quiescent myometrium (due to progesterone)

      • immunological protection from rejection by maternal immune system

    • muscle contractions

      • expulsion of the fetus and fetal placenta

        • strong rhythmic myometrial contractions (progesterone is low)

      • move towards oviduct during estrus (standing heat)

      • following ovulation:

        • towards cervix until progesterone increases from CL

    • recovery from pregnancy

      • uterine involution

      • myometrial contractions and enzymatic activity shrink uterus back to normal size

  • components of the uterus:

    • uterine horns

      • two cornua (horns)

      • length:

        • Pig- 4-5ft.

        • Cow- 7-8in.

    • uterine body

      • fusion between horns (common opening to both)

      • allows transuterine migration

        • in ewe, mare, pig, dog, cat

      • site of semen deposition sow and mare

        • also during artificial insemination…

  • tissue composition of the uterus

    • serosal layer

      • perimetrium

        • part of the peritoneum

        • continuous with serosal layer covering the mesosalpinx

        • quite thin, almost transparent

        • above muscle layer

        • outer serous layer continuous with peritoneum

        • blocks adhesions

    • muscularis layer

      • myometrium

        • beneath serosal layer

        • two layers

          • outer longitudinal layer of smooth muscle

            • has creases or small ridges that run cranial → caudal

          • inner circular layer of smooth muscle

            • beneath longitudinal layer

            • cells wrap around uterine horn in circular fashion

        • several physiologic responsibilities

          • provide motility for the uterus

            • form of contraction

            • peristalic contractions “worm-like”

          • uterine tone

            • has high degree of tone (partial state of contraction)

              • NOT in the mare

              • influenced by high estrogen

            • can be palpated

              • estrogen dominance → turgidity/hardness

              • progesterone dominance → soft/flaccid

                • EXCEPT in early stage of mare pregnancy

                  • appropriate because during this time, embryo will enter uterus for attachment

                  • if it was highly motile or hard in tone, it would minimize successful attachment of conceptus

            • related to transport mechanisms for sperm and mucus-like material produced by uterus

          • parturition

            • during parturition, muscle is a major driving force for expulsion of fetus and fetal membranes

    • mucosa + submucosa layer

      • endometrium

        • cells of this inner uterine layer produce prostaglandin F2a

          • causes luteolysis

            • corpus luteum regression if animal is not pregnant

        • provides point of placental attachment and glands provide secretions for embryo development

        • layer 1- submucosal layer

          • predominantly connective and supporting tissue

          • origin of uterine glands

            • functionality changes during estrous cycle

            • “waxing and wanting”

              • secretory activity of glands changes as a function of the current stage of the estrous cycle

        • layer 2- mucosal epithelium

          • responsible for secreting materials into lumen of uterus

            • enhance embryo development and sperm viability

          • uterine glands

            • develop from mucosal epithelium

            • produce materials important to the survival and function of the preimplantation embryo

            • penetrate into submucosa and begin to coil under influence of estrogen

              • reach full secretory capacity under progesterone dominance

            • bitch ONLY

              • uterine glands serve as a temporary storage site of sperm following insemination

              • not known if these sperm are later released to accomplished fertilization

        • endometrial species differences

          • human

            • endometrium of uterus is sloughed to the exterior

          • domestic mammals

            • endometrial glands are not sloughed

        • species differences of placenta formation

          • ruminants

            • caruncles

              • small protrusions from surface of endometrium

              • highly vascularized, nonglandular

              • develop maternal side of placenta if embryo attaches

          • sow and mare

            • many endometrial folds

              • folds provide uterine surface for development of placenta

          • queen

            • placental “scars”

              • caused by marginal hematomas (hematophagous zones)

                • bands of maternal hemorrhage at the margins of the zonary placenta

                • product of hemoglobin breakdown

                  • from hemosiderin macrophages

                    • iron-storage complex

              • pigmented regions of the endometrium

              • represent sites of previous placental attachment

              • appear as bands around luminal surface

                • indicative of zonary placentation

              • not truly scars

                • rather, zones of uterine repair

                • less noticeable several months postpartum

  • three distinct anatomical uterus types

    • duplex uterus

      • two cervical canals that separate each uterine horn into distinct compartments

      • two types

        • advanced duplex (marsupial (opossum, kangaroo)

          • single vaginal canal opening to the exterior

          • in the interior, bifurcates into two vaginas and two cervices

          • two cervices, two (three) vaginas, two uterine horns

        • duplex (rabbits, mouse)

          • two uterine horns, two distinct cervical canals connected to a single vaginal canal

          • able to deposit semen of two studs into separate horns

            • produce two sets of offspring with different genetic types

          • one vagina, two cervices, two uterine horns

    • bicornate uterus

      • two uterine horns, small uterine body

      • uterus opens into vagina through a single cervical canal

      • internal and external uterine bifurcation of horns can be distinguished

      • length of uterine horns dependent on degree of fusion between the paramesonephric ducts in developing female fetus

        • high degree of fusion

          • mare

          • short uterine horns, relatively large uterine body

        • moderate degree of fusion

          • cow, ewe, goat

          • uterine horns of intermediate (smaller) length

          • one vagina, one cervix, smaller uterine horns

        • little fusion

          • sow, bitch, queen

          • long uterine horns

      • mare and cow

        • poorly to moderately developed uterine horns

      • bitch, queen, sow

        • highly developed uterine horns

      • sow

        • two long uterine horns (litter bearing), common uterine body, one cervix, one vagina

    • simplex uterus

      • single uterine body

      • primates (humans)

      • small rudiments of a uterine horn structure, no uterine horns

      • almost complete fusion of paramesonephric ducts

        • resulting in single-chambered uterus without horns

      • no uterine horn, all uterine body

  • uterine environment

    • estrogen

      • increase vascularity

      • thickening of the uterus endometrium

      • stimulates growth of endometrial glands

    • progesterone

      • cause endometrial glands to coil and branch

        • to secrete histotroph “uterine milk”

          • nutritive material derived from maternal tissue other than the blood, utilized by the early embryo

Oviduct (Salpinx)

  • connects uterus to ovary

  • main functions:

    • final maturation of gametes

    • fertilization of oocyte

    • transport and facilitate early embryo and egg development

      • after fertilization, zygote remains at AIJ for 2-5 days (cow), until lumen of isthmus dilates to allow passage into uterine horn

      • muscular contractions move embryo into uterus very rapidly

      • embryos held in oviduct allow time for uterus to prepare for implantation

    • reduce sperm numbers to prevent polyspermy

    • ligaments contract

      • mesovarium supports ovary

      • mesosalpinx supports infundibulum and oviduct

    • transport oocyte to site of fertilization

      • ampullary cilia and infundibulum fimbria

    • provide proper environment for oocyte, sperm and fertilization

      • secretions from ampullary epithelial cells help prepare sperm for fertilization (capacitation)

      • secretions stimulated by estrogen are unique to estrus phase of cycle

  • two portions (in Stein’s world)

    • isthmus

      • smaller diameter, very muscular

      • few folds and few ciliated epithelium in lumen

      • serves as a sperm reservoir until it is released to AIJ

      • connected directly to uterus

        • uterotubal junction (UTJ)

          • point of uterine junction

          • functions to block polyspermy

          • in the cow

            • regulate movement of embryo into uterus

            • high estradiol → UTJ forms kink

              • blocking movement of embryos

            • decreasing estradiol → UTJ unkinking, straightening

              • isthmus lumen no longer blocked

              • thus, embryos can enter uterine lumen

          • in the sow

            • constriction of UTJ serves as a major barrier

              • to sperm transport

              • prevents excessive number of sperm from reaching ampulla

              • important in the prevention of polyspermy

          • other species

            • oviduct attaches to uterus without a kink-like constriction

    • ampulla

      • occupies 1/2 or more of oviductal length

      • site of fertilization

      • merges with isthmus of oviduct

      • large diameter, not very muscular

      • internal portions

        • many fern-like mucosal folds with ciliated epithelium

      • ampullary-isthmic junction

        • point of ampulla and isthmus junction

        • generally ill-defined

        • site of fertilization

        • in the mare

          • A-I junction serves as a control point

            • allows only fertilized oocytes to pass into isthmus, thus uterus

        • Fate of Unfertilized Oocytes:

          • sow, cow, ewe, doe

            • UFO will pass from oviduct into the uterus

          • mare

            • UFO will not pass into the uterus

            • embryos will bypass unfertilized oocytes and pass into the uterus

            • UFO’s will be resorbed

    • infundibulum (not technically part of the oviduct in Stein’s World)

      • terminal end of the oviduct

      • delicate, membrane-like component in close apposition to ovary

      • funnel-shaped opening

        • forms a pocket that “captures” the newly ovulated oocyte

        • directs ovulated oocyte into the oviduct

        • cumulus oophorus cells surrounding ovum aids in pick-up by the fimbria lining the infundibulum

      • surface covered with many velvety, finger-like projections (cilia)

        • fimbrae

          • greatly increase surface area of infundibulum

          • beat towards the opening of ampulla

          • cause it to glide or slip over entire surface of ovary near time of ovulation

            • this maximizes chance that oocyte will be “captured” after ovulation and transported through an opening

              • ostium

                • opening from infundibulum and ampulla

  • layer functions:

    • muscularis layer

      • transport newly ovulated oocytes and sperm to site of fertilization (ampulla-isthmic junction)

        • spermatozoa and ova move in opposite directions to meet at AI Junction

        • Isthmus

          • contractions toward the ovary

            • some sperm will continue out through oviduct

        • Ampulla

          • contractions and beating of cilia towards the uterus

    • mucosal layer

      • secrete substances

        • provide optimum environment for the free-floating, unfertilized oocyte

        • sustains spermatozoal function until oocyte arrives after ovulation

        • epithelium produce substances to support fertilizing capability of sperm

      • after fertilization, houses newly formed zygote

        • for a few days, before entering uterus

        • composition of fluid secretions from oviductal epithelium vital to providing suitable environment for development of early embryo

      • cilia on fimbria come in contact with COC and occyte

Ovary

  • egg-shaped, dense, turgid structure

  • easily palpated via rectum

    • cow, mare, camel

  • exocrine functions

    • female gamete production (ova)

    • house oocyte

    • diploid (2N)

  • endocrine functions

    • hormone production

      • estrogen

        • produced by follicle

        • secondary sex characteristics

      • progesterone

        • produced by corpus luteum

        • prepares uterus for pregnancy

        • steroid hormone

        • required for maintenance of pregnancy

  • size and shape between species:

    • cow

      • 30 x 20 x 40 mm

      • oval shaped

    • queen

      • 15 mm

      • round shaped

    • sow

      • 35 mm

      • lobulated

    • mare

      • 50 x 40 x 25 mm

      • kidney shaped

  • structures

    • germinal epithelium

      • named from the original belief of this is where germ cells were produced

      • single layer of cuboidal cells, serosal layer

      • continuous with peritoneal lining

      • prevents adhesions, does not produce germ cells, ruptures at ovulation

      • responsible for over 85% of ovarian events in humans

    • tunica albuginea

      • layer beneath the germinal epithelium

      • made of dense white connective tissue

      • provides structure to the ovary

    • ovarian cortex

      • houses the population of oocytes (germ cells)

      • cells surrounding oocytes will develop and produce follicles that will mature and eventually ovulate

        • oocytes within follicle

          • follicles produce estrogen

        • forms corpus hemorrhagicum, which luteinizes to form →

        • corpus luteum produces progesterone

          • degenerating CL = corpus albicans

        • ovulation occurs at random locations on the exterior of the cortex

          • everything BUT the mare

      • houses the:

        • functional corpus luteum

          • relatively large, visible structure

          • produces progesterone

          • can be palpated in the cow

        • degenerating corpora albicans

          • visible in most species

          • white, scar-like structure

          • represent corpus lutea in various stages of degeneration from previous estrous cycles

          • white from increasing ratio of connective tissue to secretory tissue

            • as corpus luteum degenerates undergoes a gradual transition from orange/yellow to white scar-like structure

    • ovarian medulla

      • houses the vasculature, nerves and lymphatics

      • supports cortex

      • made up of dense connective tissue

    • the mare’s ovarian cortex and medulla

      • cortex and medulla are reversed

        • cortex inside, medulla outside

      • function does not differ

      • ovulation only occurs at one location in the ovary

        • ovulation fossa

        • can’t superovulate because there is no space

      • follicles can be palpated per rectum in mare, but not corpus lutea

        • corpus lutea do not protrude from ovarian surface but penetrate into ovarian tissue

        • CL is internal because the cortex is internal

Oocyte and Follicular Development

Follicle Development

General:

  • located in the cortex

  • various types represent different stages of follicular development and maturity

  • four main stages, undergo:

    • foliculogenesis

      • process of immature follicles develop into more advanced follicles, to eventually be candidates for ovulation

        • atretic follicle

          • degeneration of a follicle

          • granulosa cells are fragmenting

          • cloudy appearance

Stages

  • primordial follicles

    • microscopic

    • most immature, smallest of all

    • oocyte surrounded by a single layer of squamous cells

    • stored, for recruitment

    • immature and smallest follicle in the cortex

  • primary follicle

    • “resting stage”

    • developed as a slightly more advanced primordial follicle

    • oocyte surrounded by simple cuboidal epithelium or follicular cells

    • stage of the majority of follicles

    • do not divide into other primary follicles

      • either develop into secondary follicles or degenerate

  • secondary follicle

    • developed as a more advanced primary follicle

    • oocyte surrounded by more than one layer of follicular (granulosa) cells

    • no antrum

      • follicular fluid-filled cavity

    • oocyte surrounded by the zona pellucida

      • relatively thick, translucent layer

  • antral follicle

    • blister-like structure, varying in size

      • size depends on stage of development or regression, and species

    • contains an antrum

      • differentiation of several distinct cell layers in the follicle wall

    • produces estrogen

    • also referred to as a tertiary follicle

    • graafian follicle

      • a tertiary follicle that is the dominant preovulatory follicle

      • unknown as to what makes a dominant preovulatory follicle

      • in cattle, sheep and horses

        • develop in sequential waves during both the follicular phase and luteal phase of the cycle

      • in primates, pigs and rodents

        • only develop during the follicular phase of the cycle

    • some are able to be observed with naked eye

      • on surface of ovary

    • three distinct layers

      • theca externa

        • composed of loose connective tissue

        • completely surrounds and supports follicle

      • theca interna

        • directly below theca externa

        • cells are responsible for production of androgens under the influence of lutenizing hormone

      • granulosal cell layer

        • also called membrana granulosa

        • beneath theca interna

          • separated by a thin basement membrane

        • produce variety of materials and have follicle stimulating hormone receptors

          • estrogen, inhibin, follicular fluids

          • FSH stimulates growth of follicle

        • govern the maturation of oocyte

Follicular Components

Stroma

  • includes more general components:

    • immune cells, blood and lymphatic vessels and nerves

Thecal Cell Layer

  • Theca Externa

    • line the outside layer of the follicle wall

    • provides structural integrity and support

    • composed of connective tissue and blood vessels

  • Theca Interna

    • outside the basement membrane of an antral follicle

      • middle layer of cells on the follicle wall

    • LH receptors here

    • produce testosterone under the influence of LH

      • converted to estrogen by granulosa cells

      • (two-cell theory)

Basement Membrane

  • separates the theca interna from the granulosa cell layer

Granulosa Cell Layer

  • line the inside of the follicle wall, surround the oocyte

  • support and control oocyte development

    • immature: contains FSH receptors

    • mature: contain FSH and LH receptors

      • LH and FSH are released from anterior pituitary

      • (tonic and surge)

  • two-cell theory: produce estrogen from testosterone

  • released with the oocyte during ovulation

  • important for maturation and transport in the oviduct

Hillock

  • stalk of granulosa cells supporting the ovum

Cumulus Oophorus (COC cells)

  • _______ cells surrounding the _

  • layer of cells outside of the Zona Pellucida

  • released with oocyte in/after ovulation

  • important for __ in the oviduct

  • a determining factor in oocyte selection for IVF

  • inhibit oocyte maturation

    • for fertilization to occur, the communication between COC cells and oocyte must be broken

      • “cumulus expansion” in oviduct

Corona Radiata

  • a single layer of granulosa cells

  • in direct contact with the zona pellucida of the oocyte

    • attaches cumulus cells to the zona pellucida

Zona Pellucida

  • a thick, translucent mucoprotein surrounding the oocyte and early embryo

Antrum

  • a cavity that contains follicular fluid

    • serum-like (liqour folliculi)

Two Cell Theory

  • “2-Cell, 2-Gonadotropin Model” for Estradiol Synthesis

  • Cholesterol(C27) → Progesterone(C21) → Testosterone(C19) → Estrogen(C18)

    • aromatase: enzyme converting testosterone to estrogen

Steps

  • Theca Interna Cells

    • produce testosterone

    • contain receptors for LH from the blood

      • the binding of LH initiates a cascade of intracellular events

        • net effect is conversion of cholesterol to testosterone

          • LH → cAMP → protein kinase (progesterone) takes cholesterol → testosterone

          • androstenedione is a weak androgen and is technically testosterone in this case

            • causes hair growth in women: axillary and pubic

        • testosterone then diffuses out of theca cells to granulosa cells

  • Granulosa Cells

    • produce estrogen (estradiol)

    • contain receptors for FSH from the blood

      • binding of FSH causes the conversion of testosterone to estradiol

        • FSH → cAMP, → protein kinase (aromatase) takes testosterone → estradiol

        • estradiol to blood capillaries

Estradiol

  • predominant estrogen produced b dominant follicles during the follicular phase of the estrus cycle

  • estradiol targets after diffusion into blood capillaries

    • brain

      • increase lordosis, phonation, physical activity

    • reproductive tract

      • increase blood flow, edema of tissues, secretions (mucus), leukocytes, smooth muscle motility, growth of uterine glands

    • increases blood flow (hyperemia) to all the organs

      • facilitates secretory activity

      • allows for delivery of leukocytes into submucosal region of RT (leukocytosis)

        • so invading foreign materials(including sperm) are engulfed after copulation

      • swelling of vulva (from elevated blood flow)

  • BRCA1 and BRCA2 are genes encoding DNA repair proteins

    • their mutants predispose a person to breast cancer

      • especially in the presence of estradiol

        • which is anti-apoptosis

Ovulation

  • definition:

    • release of an egg(s) from an ovary

      • from follicle rupturing

Steps Prior to Ovulation:

  • surge of LH

    • resume meiosis

  • collagenase causes the basement membrane to deteriorate

    • collagenase: causes the breakdown of collagen (major component of CT)

    • gap junctions between the granulosa cells and oocyte deteriorate

      • cells begin to intermingle

      • before meiotic resumption

    • they physical separation between Theca (interna) and Granulosa cells become incomplete

  • ovulation of antral follicles caused by surge of LH

    • small blood vessels rupture

    • causing local hemorrhage

    • can be seen by naked eye

During Ovulation:

  • loss of follicular fluid from antrum

    • causes walls of follicle to collapse into many folds

      • implosion

        • collapsed follicle contains blood, lymph, granulosa and thecal cells

    • granulosa and theca layers pushed to the apex of the follicle

      • theca interna and granulosa cells mix together

After Ovulation:

  • formation of a corpus hemorrhagicum

    • blood clot forms (blood filled “follicle” cavity)

    • walls

    • theca interna and granulosa cells mix together

    • basement membrane constitutes the connective tissue

  • luteal phase begins during formation of the corpus hemmorrhagicum

Types of Ovulators:

  • Induced

    • species - rabbit, cat, alpaca, camel, lion

    • cat:

      • tomcat has penile spines on glans penis to stimulate ovulation in the female

    • alpaca:

      • the seminal fluid (ovulation-inducing factor) stimulates ovulation in the female

    • lioness:

      • mates up to 100 times a day

      • average interval between mating: 17 minutes

      • each mating lasts for: ~21 seconds

  • Spontaneous

    • species- cow, sow, ewe, mare and woman

    • ovulation controlled by events of the hormone cycle

      • not affected by intromission

Luteal Phase

  • Definition: phase of the estrous cycle characterized by progesterone dominance and the presence of a functional corpus.

    • begins after ovulation and ends after lysis of the corpus luteum

  • luteal tissue is composed of luteal cells that originate from granulosa and thecal cells

  • consists of :

    • luteinization

    • synthesis and secretion of large quantities of progesterone

    • luteolysis

The Four L Words:

  • Luteinization

    • process where granulosa and theca cells are transformed into luteal cells

      • governed by luteinizing hormone (LH)

  • Luteotropic

    • a material having a stimulating action on the development or assisting in the maintenance of the CL

      • LH is a luteotropic material

  • Luteolysis

    • process where luteal tissue undergoes regression and cell death

      • (CL dies)

  • Luteolytic

    • a material that promotes luteolysis

      • ex. PGF2a (prostaglandin)

        • PGF2a plasma clearance in cattle was fivefold greater than in mares

          • mares = 1/5 dose of PGF2a as cows

Corpus Hemmorrhagicum

  • definition: a small, blood clot that results from rupture of blood vessels during ovulation

  • “bloody body”

  • appears immediately after ovulation

  • is a local hemorrhage resulting from the rupture of many small blood vessels

    • vascular breakage from ovulation

  • appears as a blood clot on the surface of the ovary

    • small, pimple-like structure

  • the basement membrane forms the connective tissue substructure of the CH to then form into the CL

    • in cows, process from which a CH turns to a CL takes 5 days

Corpus Luteum

  • definition: an orange, yellow or white colored transient endocrine structure formed after ovulation from granulosa and thecal cells of the ovarian follicle. responsible for producing progesterone an oxytocin. originates from an ovulatory follicle

  • forms from the corpus hemmorrhagicum and is developed via luteinization

    • during formation, the CL increases in size and loses its hemorrhagic appearance

    • increases in mass until the middle of the cycle

      • when size is maximal and coincides with the maximum secretion of progesterone during diestrus

    • the basement membrane forms the connective tissue substructure/network

    • in some instances, there is a remnant of the follicular antrum that forms a small cavity in the center of the CL

    • cow and mare: yellow color

    • sow and ewe: white color

  • CL Vigor

    • depends on:

      • number of luteal cells

      • the degree to which the CL becomes vascularized

  • cells of theca interna and granulosa cells differentiate into luteal cells

    • large luteal cells (LLC)

      • formerly granulosa cells

      • contain FSH and LH receptors

        • LH only at maturity

      • produce progesterone

      • contain a PGF2a receptor

        • necessary for actual prostaglandin

      • produces relaxin

      • produces oxytocin

        • signals for PGF2a to be released from the uterus

        • also produced by hypothalamus

      • during luteinization:

        • undergo hypertrophy

      • total number of granulosa cells determine the CL’s steroidogenic potential

    • small luteal cells (SLC)

      • formerly theca interna cells

      • contain LH receptor

      • produce progesterone

      • during luteinization:

        • undergo hyperplasia

      • contain fibroblasts around steroidogenic cells

Progesterone:

  • caps the surge → ovulation will not occur → maintains homeostasis

  • responsible for:

    • inhibits estrus and parturition

    • blocks myometrial contractions

    • stimulating endometrial secretion of nutrients for embryo

    • indirectly stimulating the production of a luteolytic agent

      • if no embryo present → prostaglandinF2a to regress the CL

    • decreases basal GnRH amplitude and frequency

      • blocking the preovulatory surge of luteinizing hormone

      • GnRH- Gonadotropin Releasing Hormone

        • releases LH and FSH

  • Progesterone is an inhibitor because it:

    • reduces GnRH pulse frequency

    • prevents behavioral estrus

    • stops the preovulatory LH surge

    • reduces myometrial tone

      • except in the mare

Corpus Albicans

  • regressed or degenerated corpus luteum

  • causes a drop in blood progesterone

  • composed of connective tissue (scar tissue)

    • no function

    • white, scar-like structure

  • continues to get smaller under no longer visible

  • luteolysis results in:

    • cessation of progesterone secretion

    • structural regression to form a corpus albicans

    • removal of negative feedback by progesterone upon GnRH secretion

      • resulting in a new follicular phase

PGF2a Control of Luteolysis

  • Uterus releases PGF2a

    • controls life of the CL

    • PGF2a is rapidly metabolized in some species

      • vasoconstrictor

    • cow (~90%) and pig (~40%) destroyed in one passage through pulmonary system

    • delivered to ovary to regress CL

  • Evidence of counter-current circulation pattern

    • exchange between vein and artery

    • PGF2a synthesis by uterine endometrium is released into Uterine Vein

    • PGF2a diffuses from the utero-ovarian vein into the ovarian artery

      • moves into artery → through counter-current exchange

      • PGF2a pickup up by ovarian artery and delivered back to ovary

        • causes lysis of CL

  • During pregnancy

    • embryo produces chemical signals to either…

      • inhibit PGF2a production

      • release to prevent CL death

  • sow, cow & ewe

    • artery is very close to vein

    • very tortuous

    • increasing contact between vein and artery

    • progesterone from CL stimulates production of uterine PGF2a after day 15 in the cow

  • mare

    • ovarian artery is straight and caudal to the vein

    • limited contact area between vein and artery

  • ewe example:

    • in the intact uterus

      • CL lifespan same as in normal cycle (5-17d)

    • total uterectomy

      • CL lifespan similar to normal gestation length (148d)

        • no PGF2a produced

    • partial uterectomy (contralateral to CL)

      • CL lifespan similar to a normal cycle (15-17d)

    • partial uterectomy (ipsilateral to CL)

      • CL has a longer lifespan than normal (35d)

Oocyte Development/Functionality

Background

  • females are born with a defined number of oocytes

  • no regeneration

  • germinal vesicle

    • the nuclear membrane around the chromosomes

  • ooplasma: oocyte cytoplasm

  • vitelline membrane: cell membrane around oocyte

  • last mitotic division completed close to time of birth

  • germ cells become located in gonad as the embryo/fetus develops

    • differentiate in fetal ovary

    • proliferate to form female germ cell population

    • primordial = first created

  • stages:

    • primary oocyte → secondary oocyte → ootid (haploid) → zygote

Cell Division (Meiosis I & 2)

  • phases of cell division

    • prophase, metaphase, anaphase, telophase

  • meiosis I

    • begins during fetal development

    • stops in prophase I

      • DNA enters resting stage (dictyotene)

      • can last for several years

    • oocyte is in meiosis I until ovulation

      • ovulation is a result of completion of meiosis I

    • cow ovulates secondary oocytes

    • mare and bitch ovulates primary oocyte

  • meiosis II

    • begin following ovulation

    • secondary oocyte + 1 polar body

      • polar body removes excess DNA

      • expulsion of first 1/2 of meosis I

    • not complete until egg is fertilized

      • secondary oocyte will stop in metaphase II until fertilization occurs

      • when fertilization occurs, meiosis II will resume and be completed

    • completion of meiosis II produces an → ootid + 2nd polar body

      • ootid contains 1 male and 1 female pronuclei

    • when male and female pronuclei break down nuclear membranes and DNA comes together (fuses) a zygote is formed

Oocyte Development

  • primary oocyte (2N)

    • single-cell

    • the largest cell of the body

      • ~180mm

    • suspended in dictyotene stage during Prophase I of Meiosis I since birth

    • mare ovulates primary oocytes

  • Secondary Oocyte (1N)

    • 1 polar body present

    • completion of Meiosis I

      • MI completed at time of ovulation

    • 2nd stage of oocyte (meiotic) arrest

      • MII suspended in Metaphase II

      • completion just prior to/at time of fertilization

  • Ootid

    • oocyte after meiotic divisions in which polar bodies are present

      • 2 polar bodies are present

      • 2nd polar body is released to remove “excess DNA”

    • mature ovum after penetration by sperm but before the formation of a zygote

      • i.e. before the fusion of the male an female pronuclei

  • Zygote

    • diploid cell resulting from the fusion of the male and female pronuclei

    • one cell organism

Timeline of Oogenesis

  • Cattle:

    • birth: 50,000 to 100,000 oocytes/ovary

    • puberty: 20,000 to 30,000 oocytes/ovary

    • 2 yr. old: 5,000 oocytes/ovary

  • Human:

    • midway through gestation: 6 to 7 million oocytes/ovary

    • birth: 1 to 2 million oocytes/ovary

    • puberty: 30,000 oocytes/ovary

    • 37 yr. old: 25,000 oocytes/ovary

      • atresia rate increases

Follicular/Ovarian Cyst

  • a follicle greater than 17mm to 25mm in diameter

    • usually accompanied by one or more other large follicular structures

    • no CL is present

    • persist at least 10 days in the absence of a CL

      • very thin walled

  • a follicle becomes cystic when:

    • it fails to ovulate

      • and persists on the ovary

  • common signs an animal may be cystic

    • anestrus → lack of standing heat

    • abnormal estrous behavior patterns

      • including persistent estrus or shortened estrus intervals

  • dairy cattle and cystic follicles

    • incidence rate: 2.7-30%

      • peak incidences between 14-40 days postpartum

    • 70% occur between 16-50 days postpartum

      • highest occurrence between 30-40 days postpartum

    • before day 16 and after day 50 postpartum = lowest incidence of ovarian cysts

  • causes:

    • stress

      • usually in high producing dairy cows

    • seasonal changes

      • altered hormone patterns

      • higher incidence during winter

    • genetics

      • can select against

    • other

      • retained placenta, milk fever, uterine infection, twins, etc.

  • what happens:

    • hypothalamus, pituitary and or an ovarian malfunction

      • insufficient LH released to cause ovulation

        • low levels of GnRH released

      • ovary does not respond well to LH

  • effects/treatment:

    • grow & regress (repeatedly)

      • causes nymphomania and short cycles

    • what is needed: increased LH levels to make cysts ovulate

      • administer GnRH to increase LH level

      • follow with PGF2a ~9 dyas later to regress luteal cyst

  • luteal cyst:

    • thick walled luteal tissue

      • CL with a fluid-filled cavity

        • not a problem with cyclicity…

    • LH induces luteinization of follicle wall, high levels of progesterone are produced and anestrus occurs

    • give PFG2a to make cysts regress

  • multiple transitional follicles

    • cystic ovaries in mares do not occur as often as dairy cows

    • the presence (or absence) of cystic ovaries in the mare, as described in dairy cattle has been a subject of debate for years