1374021

pubmed:Citation

5

To further the identification and characterization of insulin-like growth factor binding proteins at the level of the immature rat ovary, we have set out to study the ovarian expression, cellular localization, and hormonal regulation of the insulin-like growth factor binding protein (IGFBP)-3. To this end, use was made of a solution hybridization/RNAse protection assay wherein ovarian total RNA from immature (21-23 days old) female rats was hybridized with a 343 bases-long [32P]-labeled rat IGFBP-3 riboprobe. As in liver, a single protected fragment (315 bases-long) corresponding to IGFBP-3 transcripts was identified in whole ovarian material. Cellular localization studies revealed the IGFBP-3 gene to be exclusively expressed in the theca-interstitial rather than the granulosa cell compartment. To confirm presence and cellular distribution of the IGFBP-3 protein, media conditioned by cultured granulosa cells, theca-interstitial cells, and whole ovarian dispersates were subjected to Western Ligand Blotting. Importantly, media conditioned by cultured theca-interstitial (but not granulosa) cells displayed an IGFBP the size of rat IGFBP-3 (46kDa) as determined by comigration with a rat serum standard. A similarly-sized band was apparent in media conditioned by cultured whole ovarian dispersates reflecting in all likelihood the contribution of the theca-interstitial cell component. Significantly, deglycosylation of media conditioned by cultured theca-interstitial cells revealed the glycosylated nature of the 46kDa IGFBP species as judged by the apparent reduction in its molecular size to 35kDa. Similar alterations were noted in corresponding rat serum samples. Hypophysectomy of immature rats resulted in a modest but statistically insignificant decrease in the relative (densitometrically-quantified) abundance of ovarian IGFBP-3 transcripts, an effect further augmented by the systemic provision of either FSH or diethylstilbestrol (DES). In contrast, systemic treatment of hypophysectomized rats with GH produced a marked (3.2-fold) increase (P less than 0.05) in the steady state levels of ovarian (as well as hepatic) IGFBP-3 transcripts. However, the concurrent provision of either FSH or DES resulted in substantial (P less than 0.05) attenuation (78 and 57% inhibition, respectively) of the upregulatory GH effect. These findings document the highly compartmentalized expression of the IGFBP-3 gene at the level of the immature rat ovary, implicate the theca-interstitial cell as the sole source of its generation, reveal its pituitary dependence, and disclose its diametrically-opposed (indeed antagonistic) regulation by FSH (or estrogens) and GH.(ABSTRACT TRUNCATED AT 400 WORDS)

eng

AIM

MEDLINE

0013-7227

Print

NLM

3092-4

pubmed-meshheading:1374021-Animals, pubmed-meshheading:1374021-Blotting, Western, pubmed-meshheading:1374021-Carrier Proteins, pubmed-meshheading:1374021-Cells, Cultured, pubmed-meshheading:1374021-Diethylstilbestrol, pubmed-meshheading:1374021-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:1374021-Female, pubmed-meshheading:1374021-Follicle Stimulating Hormone, pubmed-meshheading:1374021-Gene Expression, pubmed-meshheading:1374021-Gonadotropins, pubmed-meshheading:1374021-Granulosa Cells, pubmed-meshheading:1374021-Growth Hormone, pubmed-meshheading:1374021-Hypophysectomy, pubmed-meshheading:1374021-Insulin-Like Growth Factor Binding Proteins, pubmed-meshheading:1374021-Ovary, pubmed-meshheading:1374021-Rats, pubmed-meshheading:1374021-Rats, Inbred Strains, pubmed-meshheading:1374021-Sexual Maturation, pubmed-meshheading:1374021-Somatomedins, pubmed-meshheading:1374021-Theca Cells, pubmed-meshheading:1374021-Transcription, Genetic

Rat ovarian insulin-like growth factor binding protein-3: a growth hormone-dependent theca-interstitial cell-derived antigonadotropin.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't