Evidence for ovarian tumor necrosis factor

Thesis/Dissertation · Sun Jan 01 04:00:00 EST 1989

OSTI ID:6159111

Roby, K F

Ovarian folliculogenesis and luteal formation occur concomitantly with the development of new blood vessels that function in nutritional support of the developing follicles. As follicles undergo atresia and the corpus luteum regresses, blood vessels supplying these tissues degenerate. The first study determined if the ovary contained factors that might regulate ovarian angiogenesis. The bovine ovary was subjected to ammonium sulfate (AS) precipitation and the precipitates (ppt.) were assayed in vitro for effects on endothelial cell (CPAE) and fibroblast (3T3 and L929) incorporation of {sup 3}H-thymidine. Heparin sepharose (HS) chromatography of the 80% AS ppt. revealed the inhibitory activity on CPAE and L929 cells did not bind to HS but was found in the HS column breakthrough (80% BT). Sizing chromatography of the 80% BT indicated thymidine incorporation inhibitory activity exhibited a molecular weight of 30,000-50,000 Daltons. TNF was immunohistochemically localized in the human, bovine and rat ovary. Frozen sections were incubated with polyclonal antibody to human recombinant TNF. Antigen-antibody binding was visualized using a Biotin-StreptAvidin peroxidase technique. Immunoreactive TNF (I-TNF) was localized in corpora lutea and the more antral layers of granulosa cells in antral follicles. Incubation of sections with anti-TNF in the presence of excess TNF resulted in lose of immunostaining. Cell blotting and ELISA further indicated I-TNF was present in granulosa cells. In order to determine whether TNF had an effect on follicular steroidogenesis, preovulatory follicles from cyclic proestrus rats were incubated in vitro for up to 24 hours with various doses of human recombinant TNF. Stepwise increases in progesterone (P) accumulation in the incubation media were observed with 30-300 pM TNF.

• Research Organization: Kansas Univ., Lawrence, KS (USA)
• OSTI ID: 6159111
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
AZINES
BIOASSAY
BIOLOGICAL FUNCTIONS
BODY
CATTLE
CELL DIFFERENTIATION
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DOMESTIC ANIMALS
ENDOTHELIUM
ENZYME IMMUNOASSAY
FEMALE GENITALS
FIBROBLASTS
FUNCTIONS
GONADS
GROWTH FACTORS
HETEROCYCLIC COMPOUNDS
HORMONES
HYDROGEN COMPOUNDS
IMMUNOASSAY
ISOTOPE APPLICATIONS
KETONES
MAMMALS
MAN
MITOGENS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OVARIES
PREGNANES
PRIMATES
PROGESTERONE
PROTEINS
PYRIMIDINES
RATS
RIBOSIDES
RODENTS
RUMINANTS
SEPARATION PROCESSES
SOMATIC CELLS
STEROID HORMONES
STEROIDS
THYMIDINE
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES