Lactation, reproduction, and oviposition 🍼

Egg laying (oviposition)

What are the hormones for bone remodeling and egg shell calcification?
● Before laying eggs:
○ liver secretes (stimulated by estrogen) a Ca2+ binding protein into the blood →
increases plasma [Ca2+] from 2.5mmol/L to 6
● During egg shell formation:
○ in domestic hens, 2-4g (10% of total calcium content of their bones → need
Ca2+ in diet) of Ca2+ transferred to oviduct (so, decrease in Ca2+
concentration in hen ECF); levels in the hen normalize when egg shell
formation is complete
○ increase in PTH & increase in calcitriol production in kidneys → increase Ca2+
absorption from intestine
● During egg white formation:
○ phosphate binds to proteins and lipids
● Hormone estrogen and testosterone
○ estrogen and testosterone → additional bone tissues deposited in bone
cavities 2 weeks before egg laying
■ vitamin D also involved
○ estrogen → stimulates synthesis of Ca2+ binding protein in the liver

Describe the reproductive system in female birds.

🥚
● In most species, only the left ovary and oviduct develop to maturity
● Passageway of the egg
○ ovarium sinister
○ oviductus sinister
■ infundibulum: 15 minutes; distal glands that produce coating that
condenses to form chalaza (also where fertilization occurs); also
smooth muscle cells
■ magnum: 3 hours; glands that secrete components of albumen (egg
white)
● glands are stimulated by estrogen and progesterone (like how
these hormones stimulate endometrial glands in mammals)
■ isthmus: 1.5 hours; glands that produce sulfur-containing keratin
proteins that form the double-layered shell membrane
■ uterus: 20 hours; glands that add water to albumen and produce
cuticle (non-calcified layer made of polysaccharides, lipids,
antimicrobial proteins; protects against drying); calcium carbonate
(CaCO3) and calcium salts that produce calcareous shell; pigment
from heme for camouflage
● results of shell synthesis:
○ decrease in Ca2+ in ECF
 ○ increase in PTH synthesis
■ 1) minor effect on Ca2+ reabsorption in the distal
tubules and collecting ducts
■ 2) Inhibits HPO4 2- (phosphate) reabsorption →
decrease in plasma [HPO4 2-] → increases ionized
calcium bc prevents HPO4 2- from forming
water-insoluble salts with calcium
■ 3) Stimulates conversion of inactive vitamin D3
to active vitamin D3 (calcitriol) → calcitriol
increases the absorption of Ca2+ from the
intestines (See below)
■ 4) Stimulates osteoclasts to break down bone →
releases Ca2+ (resorption) into plasma
○ increase in calcitriol synthesis
■ activated form of vitamin D3 from the kidneys
that travels through the blood
■ + Ca2+ absorption from intestines → increased
plasma [Ca2+]
■ vagina: 5-10 minutes; stores sperm for 2-3 weeks in fossulae
spermaticae (utero-vaginal junction; goes into infundibulum by
antiperistaltic movement for fertilization); location of copulation;
transports egg to cloaca
○ urodeum of cloaca
■ via ostium cloacale oviducti sinister
● No corpus luteum
● But ovary still produces other steroids
○ estrogen, testosterone, progesterone
● Follicles develop in a series & the larger yolk contains the yolk
○ white yolk has a smaller no. of lipid droplets but larger sized droplets
○ yellow yolk has a larger no. of lipid droplets but smaller sized droplets

Describe the oviposition process.

● Ovulation & egg laying hormones:
○ Ovulation also controlled by hypothalamic-pituitary-ovarian axis (like in
mammals)
■ LH surge (only in dark/low light envmt) triggered by progesterone
increase (4-6 hours before ovulation), not estrogen as in mammals!
■ In commercial egg production, 8-9 hours of darkness (LH release) and
15-16 hours of light
○ Also regulated by prostaglandins and arginine vasopressin
● Only one egg produced per day
● 9-11 days for a follicle in the ovary to develop until ready for ovulation
● Oviduct has a folded wall that expands to transport the egg
● Laying occurs by:
○ relaxation of abdominal muscles and sphincters between uterus and vagina
 ○ contraction of uterine smooth muscles
○ vagina protrudes through the cloaca; egg does not come in contact w/ the
cloaca (to prevent contamination of egg shell w/ feces)

Describe the composition of egg white and egg yolk.

● Components of egg white
○ ovalbumin is the main protein in egg white; provides nutrition and binds
digestive enzymes; 55% of total protein in egg white
○ ovotransferrin is an iron-binding protein
○ ovomucoid
○ ovomucin
○ globulins
○ lysozyme
○ magnesium, vitamin B2, B3, potassium, sodium, zinc, etc
● Components of egg yolk
○ contains water, proteins, lipids, minerals, vitamins, inorganic ions and
enzymes; few carbs
○ lipoproteins (transport form of fats bc they are water insoluble) of the yolk are
formed in the liver under influence of estrogen – transferred to the ovary,
deposited in the developing follicles
■ fosvitin (binds iron)
■ livetins (alpha, beta, and gamma)
● foaming, emulsifying, works like globulin?
■ vitellogenin

Lactation
What is milk?
● The secretory product of the mammary gland (modified apocrine, tubulo-alveolar
sweat gland)
● An aqueous solution of colloidal suspended proteins (particularly casein and whey),
lipids, lactose, minerals, and vitamins (e.g. vitamin A)
○ As lactation progresses, more fat & less lactose

Describe the biosynthesis of lactose.

● Glucose + galactose + lactose synthase (enzyme) = lactose
○ glucose comes directly from diet in monogastrics; from propionic acid →
gluconeogenesis in ruminants
○ when prolactin (PRL) binds to the membrane receptors of mammary epithelial
cells → synthesis of α-lactalbumin
■ α-lactalbumin binds to the regulatory unit of lactose synthase (allowing
it to function)
● Takes place in the Golgi apparatus
● Released by exocytosis in secretory vesicles (which also contain synthesized proteins)
How do ruminants conserve glucose for lactation (not wasting it in other tissues)?
● Other tissues preferentially use other metabolites:
○ non-esterified FAs
○ ketone bodies

Describe the synthesis of milk proteins.

● Synthesized in the RER using AAs from the blood (AAs come from protein in feed,
microbial protein in ruminants, or body protein catabolism)
○ synthesized proteins are transported from RER to Golgi apparatus (where they
are packaged into secretory vesicles; these vesicles also contain the
synthesized lactose)
○ secretory vesicles are released into alveolar lumen by exocytosis
● Excellent AA profile, highly digestible

Describe the synthesis and secretion of lipids for milk.

● 40-50% of FAs in milk lipids are synthesized in the mammary epithelial cells (de novo
FA synthesis)
○ i.e. using acetate (ruminants) or glucose (monogastrics)
○ the remaining 50% comes from “preformed FAs” = FAs from triglycerides in
circulating chylomicrons or VLDL
■ these triglycerides in chylomicrons/VLDL are degraded into FAs by
lipoprotein lipase in the capillary wall
● De novo FA synthesis occurs in the cytosol (like with all other cells)
● Triglyceride synthesis in the SER → accumulate as fat droplets in the cytosol
○ transported to apical cell membrane, which envelops the fat droplet →
released into the lumen
○ the apical membrane gives
■ the droplet a polar surface that allows it to remain stable in the
aqueous milk
■ milk the white color
■ cholesterol and phospholipids for the neonate
In which species (monogastrics or ruminants) does the fat in feed reflect the fat in the
milk?
● In monogastrics – bc there is no biohydrogenation of USFA in the DS
○ so the FA composition of of milk fat is relatively = FA composition of feed
Vs. in ruminants, most USFA are biohydrogenated in the rumen → static acid

Do ruminants or monogastrics use glucose for FA synthesis in milk?

● Ruminants = no; use acetate
● Monogastrics = yes; metabolism of glucose → acetyl coA → used as carbon skeleton
for FA synthesis

What is colostrum?
● The first secretion of the mammary gland
● High in proteins (e.g. Igs and components of innate immunity like complement),
solids, fat, minerals, vitamins
○ lower in lactose (compared to milk)
● Important for transfer of passive immunity through maternal Igs
 ○ Must be ingested w/in 1-2 days (highest Igs during first 6 hours after birth)
○ Maternal Igs persist in newborn for 4-6 weeks after birth

Why is colostrum essential in some species and relatively less important in others?
● Due to differences in placental types
○ Syn/epitheliochorial (cattle, sheep, pigs, horse)
■ Cannot transfer Ig to fetus
■ Which Ig is dominant in their colostrum?
● IgG
○ Endotheliochorial (Dogs, cats)
■ Moderate Ig transfer
■ Which Ig is dominant in their colostrum?
● IgG
○ Hemochorial (humans, rodents)
■ Capable of Ig transfer
■ Which Ig is dominant in their colostrum?
● mostly IgA
Describe the anatomy of the mammary complex (gland).
 ● alveolus: a single layer of secretory epithelial cells bound together by tight junctions
in a cylindrical fashion
● Each alveolus is surrounded by myoepithelial cells which contract like smooth
muscle in response to oxytocin (“milk letdown”) → milk released from alveoli to duct
system
● The duct system is lined by two layers of cuboidal epithelium
○ becomes stratified squamous starting at Fürstenburg’s rosette
○ Fürstenburg’s rosette: a mucosal fold/venous plexus that divides pars
glandularis and pars papillaris
■ contains lymph protection against bacteria

🐄🐐 🐎🐄🐑🐐
Describe species differences in the anatomy of the mammary gland.
● : have specialized areas for holding milk (cisterns) into which all ducts empty
● udder: in , in which pairs of mammary glands are closely apposed to each
other
● Suspensory apparatus allows 🐄 to hold large amts of milk; formed by superficial and
deep layers of the external fascia of the trunk; suspends glands from the ventral
aspect of the trunk
○ median suspensory ligament: composed of elastic CT
○ lateral (non-elastic) suspensory ligament: composed of dense CT; enters the
glands laterally to become part of the interstitial CT framework of the udder
● Location of mammary glands & number

🐘
Species Location

axillary
🤰 thoracic

🐈 thoracoabdominal

🐕🐖 thoracoinguinal

🐎🐄🐑🐐 inguinal

Species No. of No. of Picture

mammary mammary
complexes units
(glands) (ducts)
per

🐎
complex

2 2

🐄 4 1
 🐑🐐 2 1

🐈 8 5-10

🐕 10 5-10

🐖 14 2-3

Describe the blood supply to the 🐄udder.

● Arterial supply:
○ 1) Left ventricle → 2) abdominal aorta → 3) external iliac artery → 4) external
pudendal artery → 5) caudal and cranial mammary arteries
 ● Venous supply:
○ 1) caudal and cranial mammary veins → 2) external pudendal vein → 3)
external iliac vein → 4) caudal v. cava → right atrium
○ *when lying down (recumbency), use the superficial cranial epigastric vein or
subcutaneous abdominal vein → internal thoracic vein

What is the difference between mammogenesis, lactogenesis, galactopoiesis, and

involution?
● mammogenesis: growth and development of the mammary gland
○ begins during fetal dvlpmt and continues (differentiating & regressing;
cyclical) throughout reproductive life

🐄🐑🐐🐁🐀🤰
○ GH, adrenocorticoids, IGFs, progesterone, estrogen, prolactin (late pregnancy),
lactogen (acts like prolactin; in )
● lactogenesis: initiation of lactation; when alveolar cells differentiate into milk
producing/secreting cells
○ progesterone, cortisol (induces growth of RER and Golgi apparatus), prolactin,
estrogen
● galactopoiesis: maintenance and enhancement of established lactation
○ mostly the same hormones as lactogenesis
● involution: regression of the alveolar cells following cessation of milking
 ○ FIL

Describe the phases of mammogenesis.

● Impt bc of correlation btwn mammary growth and milk yield
● 1st phase (fetal dvlpmt)
○ Important hormones: GH, adrenocorticoids, IGFs
○ Thickening of ectoderm, condenses into cell masses that define where future
mammary glands wilk form
○ Excretory duct system develops during middle third of fetal dvlpmt
● 2nd phase (birth to puberty)
○ No secretory or glandular parts, but well-developed teats
○ Isometric growth (= same as body rate)
■ except in cow, which has allometric growth (= faster than body rate)
starting at 3-4 months old until a few months after puberty
■ overfeeding during allometric growth → excess fat deposition →
inhibits parenchyma growth in mammary → inhibits future milk
production
● 3rd phase (puberty to conception)
○ Important hormones: estrogen, progesterone, GH, glucocorticoids, prolactin
○ Allometric growth for the first few estrous cycles, then isometric until
conception
○ Parenchyma growth mostly results from
■ Elongation of ductal system
■ Branching of ductal system
■ *due to estrogen
 ○ Dvlpmt of milk ducts
■ due to GH and glucocorticoids
○ Dvlpmt of alveoli
■ due to GH, glucocorticoids (cortisol), progesterone, prolactin
● 4th phase (conception to parturition)

🐄🐑🐐🐁🐀🤰
○ Important hormones: progesterone, estrogen, prolactin (late pregnancy),
lactogen (acts like prolactin; in )
○ 45-95% of total mammary growth during this period
○ Exponential growth (most growth occurs during last trimester)
○ Early pregnancy
■ Elongation and brunch of ductal system
○ Mid-late pregnancy
■ Alveolar dvlpmt

What are the phases of lactogenesis?

● 1) partial enzymatic and cytological differentiation of alveolar cells w/ limited
secretion
● 2) copious secretion of all milk components shortly before parturition & extends
several days postpartum

What does galactopoiesis entail? Maintaining… (3)

● alveolar cell number
● synthetic activity in each cell
● efficacy of milk ejection reflex

How do mammary epithelial cells involute?

● apoptosis
● autophagy
● phagocytosis by neutrophilic granulocytes & macrophages
● destruction of tight junctions between mammary epithelial cells
● water & lactose in the alveoli diffuse back into the blood

What are the two main processes of lactation?

● Milk secretion + milk removal

What is the role of prolactin in lactation?

● Produced by lactotrophs in the anterior pituitary
● Acts on alveolar cells → stimulates them to make more milk components (milk
secretion)

What is the role of prolactin-inhibiting hormone (PIH; dopamine) in lactation?

● Produced by arcuate nucleus in the hypothalamus
● Travels down the hypophyseal portal system to the anterior pituitary
 ○ Binds to lactotrophs → inhibits prolactin production

What is the role of thyrotropin-releasing hormone (TRH) in lactation?

● Produced by paraventricular nucleus in the hypothalamus
● Travels down the hypophyseal portal system to the anterior pituitary
○ Binds to lactotrophs → stimulates prolactin production

What is the role of oxytocin in lactation?

● Functions
○ Causes myoepithelial cells surrounding the alveoli to contract → “milk
letdown”
○ Reduces resistance in the small excretory ducts and teat canal
○ Causes smooth muscle cells in sphincter at the tips of the teats to reax
○ Stimulates lactotrophs and the paraventricular nucleus ( → TRH)
● Steps
○ 1) Stimulation of oxytocin secretion
■ Tactile stimulation of teat sensory receptors
● e.g. suckling, udder preparation during machine milking
■ Conditioned responses from higher brain centers to external stimuli
● e.g. cry of offspring, approaching the milking parlor, etc
○ 2) Oxytocin produced by the paraventricular and supraoptic nuclei of the
hypothalamus, released into the bloodstream by the posterior pituitary
○ 3) Oxytocin takes effect within 45-60 seconds from the stimulation

What is the role of estrogen in lactation?

● Produced by the placenta and ovaries
● Stimulates prolactin production by acting on lactotrophs directly
● Inhibits arcuate nucleus (so it inhibits PIH production) → indirectly stimulates
prolactin production

Does estrogen always stimulate lactation?

● No; very high levels of estrogen and progesterone inhibit prolactin → prevents
wasting milk before offspring is born
○ How do they inhibit? by blocking the transcription of genes for prolactin
receptors
● At the end of gestation/after parturition, estrogen and progesterone levels drop →
prolactin can act on alveolar cells
Histologically, how would you describe the mammary gland during lactation vs. non
lactation?
● Non lactation
○ large adipocytes
○ large amt of CT
○ narrow alveolar lumen
● Lactation
○ dedifferentiated adipocytes
○ minimum amount of CT
○ wide alveolar lumen

What happens when milking stops?

● Cessation of milking → increased pressure in udder → increased Feedback Inhibition
of Lactation (FIL) hormone → involution (loss of secretory activity)

What is the Feedback Inhibition of Lactation (FIL)?

● A whey protein secreted by mammary epithelial cells working in a negative feedback
mechanism
● Suppresses enzymes in alveolar cells → slows milk component secretion
 ● Increase [FIL] over time →
○ Breakdown of intracellular casein
○ Reduced no. of PRL receptors on cells
○ Inhibition of cell differentiation

Questions on the final exam (2022):

● Mammary gland (anatomy, structure, dvlpmt and growth during puberty and
pregnancy, hormonal control, milk letdown, colostrum, milk differences, etc)
● Colostrum
● MIlk letdown (milk ejection reflex)