Modulation of cultured porcine granulosa cell responsiveness to follicle stimulating hormone and epidermal growth factor

Thesis/Dissertation · Wed Jan 01 04:00:00 EST 1986

OSTI ID:6974528

Buck, P A

Ovarian follicular development is dependent upon the coordinated growth and differentiation of the granulosa cells which line the follicle. Follicle stimulating hormone (FSH) induces granulosa cell differentiation both in vivo and in vitro. Epidermal growth factor (EGF) stimulates granulosa cell proliferation in vitro. The interaction of these two effectors upon selected parameters of growth and differentiation was examined in monolayer cultures of porcine granulose cells. Analysis of the EGF receptor by /sup 125/I-EGF binding revealed that the receptor was of high affinity with an apparent dissociation constant of 4-6 x 10/sup -10/ M. The average number of receptors per cell varied with the state of differentiation both in vivo and in vitro; highly differentiated cells bound two-fold less /sup 125/I-EGF and this effect was at least partially induced by FSH in vitro. EGF receptor function was examined by assessing EGF effects on cell number and /sup 3/H-thymidine incorporation. EGF stimulated thymidine incorporation in both serum-free and serum-supplemented culture, but only in serum-supplemented conditions was cell number increased. EGF receptor function was inversely related to the state of differentiation and was attenuated by FSH. The FSH receptor was examined by /sup 125/I-FSH binding. EGF increased FSH receptor number, and lowered the affinity of the receptor. The function of these receptors was assessed by /sup 125/I-hCG binding and progesterone radioimmunoassay. If EGF was present continuously in the cultures. FSH receptor function was attenuated regardless of FSH receptor number. A preliminary effort to examine the mechanism of this interaction was performed by analyzing hormonally controlled protein synthesis with /sup 35/S-methionine labeling, SDS polyacrylamide gel electrophoresis and fluorography. FSH promoted the expression of a 27,000 dalton protein. This effect was attenuated by EGF.

• Research Organization: Duke Univ., Durham, NC (USA)
• OSTI ID: 6974528
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
AZINES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
BODY
CARBOXYLIC ACIDS
CELL CULTURES
CELL DIFFERENTIATION
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
EPIDERMIS
EPITHELIUM
EVEN-ODD NUCLEI
FEMALE GENITALS
FSH
GONADOTROPINS
GONADS
GROWTH
HETEROCYCLIC COMPOUNDS
HORMONES
IMMUNOASSAY
IMMUNOLOGY
IN VITRO
IN VIVO
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KETONES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MEMBRANE PROTEINS
METHIONINE
MOLECULAR WEIGHT
NUCLEI
NUCLEOSIDES
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
OVARIES
PEPTIDE HORMONES
PITUITARY HORMONES
PREGNANES
PROGESTERONE
PROTEINS
PYRIMIDINES
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RESPONSE MODIFYING FACTORS
RIBOSIDES
SKIN
STEROID HORMONES
STEROIDS
SULFUR 35
SULFUR ISOTOPES
SWINE
SYNTHESIS
THYMIDINE
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES