Milk Synthesis

Suporn Katawatin
Mammary Gland: the special gland

„ Offer no advantage to dam

„ Huge physiological demand from dam

„ Get high priority on nutrients, on expense of

dam health
Demand metabolic change

„ Increase metabolic rate

„ Increase blood flow

„ Increase nutrient supply

„ Inability to meet demand result in metabolic

disorder: milk fever (hypocalcemia), ketosis
(hypoglycemia)
Three major functions in milk synthesis

„ First, breaks down substrates to provide energy:

mitochondria
„ Second, synthesize components of milk: lipids,
casein, lactose
„ requires substrates, enzymes, and environment
„ Third, regulate non-mammary synthesized
constituents: water, vitamins, and minerals
blood precursors for milk

„ Glucose
„ Acetate
„ β-hydroxybutyrate
„ triglyceride fatty acids
„ Amino acids

„ Two major sources of energy are glucose and

acetate
Energy: one of three pathways

„ Embden-Meyerhoff glycolytic pathway

„ Citric acid cycle: final common pathway of

metabolism

„ Pentose phosphate pathway, or shunt

 Energy Production in Mammary Gland

Glucose is utilized by mammary cells

„ Glycolysis (Embden-Meyerhof glycolytic pathway) allows
cytoplasmic anaerobic conversion of glucose to pyruvic acid and
ATP, then coupled to citric acid cycle, and yield 38 ATP
„ Utilization of pentose phosphate pathway or shunt to yield
NADPH+ H+ to yield 35 ATP

In cow: of blood glucose taken up by mammary cell

„ Sixty to 70% utilized for lactose synthesis in alveolar cell
„ Twenty to 30% oxidized via the pentose phosphate shunt for
fatty acid synthesis
„ Less than 10% utilized via glycolysis and Krebs Cycle
Milk Synthesis Process
Milk synthesis processes

„ Precursors leave blood and enter extracellular fluid,

through basolateral membrane and to appropriate

synthetic pathway

„ Immunoglobulins transported intact through cell

Milk synthesis processes

5 routes to enter alveolar cells :

„ For uptake amino acids
„ For uptake sugars and salts
„ For uptake milk fat precursors
„ For uptake preformed proteins
(immunoglobulins)

„ paracellular pathway
 Fat droplet
Alveolar lumen casein

Tight junction

pathway

Salts glucose FA, glycerol

Precursors from blood aa immunoglobulin
leukocyte acetate OH-
butyrate
Lactose
Products form in Salts glycerol
aa lipids immunoglobulin
lumen leukocytes FA
 Fat
Lumen
droplet

Protein

Fat
Tight
junction

Basal membrane
Amino acids to proteins

„ Aa absorbed via basal membrane by specific aa

transport systems
„ Inside cell, aa covalently bound together to form
proteins at polysomes on RER
„ transferred from RER to Golgi apparatus to post-
translational processing and packaging as secretory
vesicle
„ Proteins synthesized include casein, β-lactoglobulin,
α-lactalbumin and membrane bound proteins or
enzymes
Amino acids to proteins

„ Secretory vesicles of milk proteins (also

lactose) move to apical membrane

„ secretory vesicle fuses with inner

surface of apical membrane, opening

and vesicle contents discharged into

alveolar lumen
Glucose to lactose

„ Glucose enters cell via basolateral membrane by specific

transport mechanism
„ Some converted to galactose, both glucose and
galactose enter Golgi and forming lactose
„ lactose drawing water into cell, Golgi, and becoming
part of milk
„ Thus, Golgi apparatus involved in processing of milk
proteins, synthesis of lactose, and osmotic draw for
water
„ Lactose secreted via secretory vesicles with milk
proteins
Milk fat precursors to milk fat

„ Precursors taken up via basolateral membrane

„ Acetate and β-hydroxybutyrate, precursors of FA

synthesis in ruminants mammary cells

„ Preformed fatty acids, glycerol, and

monoacylglycerides absorbed into synthesis of

triglycerides of milk
Milk fat precursors to milk fat

„ Milk fat triglycerides synthesized on SER and form

small droplets

„ Small droplets fuse and moves toward apical

membrane and pinches off to alveolar lumen

„ Thus, inside cell nonmembrane-bound lipid

droplet but in alveolus lumen, milk fat globule

surrounded by membrane
Transport of Milk Components Not
Synthesized in the Epithelial Cells

Immunoglobulins

„ Ig bind to specific receptors on basolateral

„ "into" cell in endocytic vesicles

„ transported to fuses with inner apical membrane

and releases into alveolus lumen

„ no serum albumin receptor, however, serum

albumin may internalized along with Ig
Paracellular Pathway

„ way through Tight junctions between

epithelial cells

„ little or no "flow" of anything via Tight

junctions let except water and some ions

„ Tight junctions become ‘leaky’ during

mastitis or involution, or when oxytocin
causing milk ejection
Paracellular Pathway: leukocytes

„ leukocytes pass between epithelial cells and

"break open" tight junctions
„ allows other extracellular components like
salts to diffuse into the lumen and milk
components to diffuse out of the lumen into
the extracellular fluid
„ More in Mastitis Module
Paracellular Pathway

„ allow lactose and K+ move from lumen into

extracellular space, and Na+ and Cl- move opposite

„ Results in changing electrical conductivity of milk

(as used in detecting mastitis), increase lactose

and other milk-specific components in blood

„ Lactose found in urine of a cow during peripartum

period

„ Milk proteins found in cow's blood during

lactation and early involution

Synthetic Activity in Mammary Cells

„ Cells within an alveolus synchronized in

synthetic activity
„ some alveoli cells full of lipid droplets
and secretory vesicles, while other
none of those structures
Synthetic Activity in Mammary Cells

Secretory activity occur in two phases:

„ Formation of intracellular secretory structures like lipid

droplets and secretory vesicles

„ tall and columnar cells

„ Release products into lumen

„ Cuboidal cells , lumen fills with milk

„ Intracellular synthesis may decrease during this time

Synthetic activity in mammary cell

incubate mammary tissue with labeled aa,

„ Tracer moves through cell as a pulse

„ Stays in cytoplasm ~3-15 min. [newly
synthesized proteins]
„ Appears in Golgi within 15-30 min. [newly
synthesized proteins being processed]
„ increased in lumen 30-60 min. later [secreted
proteins]
Synthetic activity in mammary cell

inject radiolabeled precursors of milk components into

animal

„ found in milk at varying times

„ Those enter by equilibrating across the apical or Golgi

membranes (Na+, K+, Cl-) take ~1 hr to reach max.

specific activity in the milk

„ Those enter by synthesis in the Golgi (lactose, casein,

Ca, citrate, phosphate) take 2-3 hr.

„ Those enter as part of milk fat synthesis take 5-7 hr.

Further readings

„ DASC Ch.VII:

„ Milk Biosynthesis: MilkBiosynthesis.doc

„ Biochemistry of the Mammary Gland:
MamBiochem.doc