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In Situ Hybridization of High Density Lipoprotein (Scavenger, Type 1) Receptor Messenger Ribonucleic Acid (mRNA) during Folliculogenesis and Luteinization: Evidence for mRNA Expression and Induction by Human Chorionic Gonadotropin Specifically in Cell Types that Use Cholesterol for Steroidogenesis¹

Departments of Obstetrics and Gynecology and Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan 48109

Address all correspondence and requests for reprints to: Dr. K. M. J. Menon, University of Michigan, 6428 Medical Science Building I, 1301 East Catherine Street, Ann Arbor, Michigan 48109-0617.

The present studies were undertaken to examine the expression of the high density lipoprotein (HDL) receptor, SR-B1 messenger RNA (mRNA) in ovarian cell types during folliculogenesis and luteinization using in situ hybridization histochemistry and to examine its hormonal regulation using Northern blots. For the in situ study for HDL receptor mRNA localization, 21-day-old rats were treated with 50 IU PMSG, and ovaries were collected 0, 24, and 56 h postinjection. At 56 h, animals were treated with a single dose of hCG, and ovaries were subsequently collected at 6-, 12-, 24-, and 72-h and 5-day intervals. In addition, on day 4 of pseudopregnancy, a second dose of 50 IU hCG or saline was administered, and ovaries were collected at 12, 24, and 48 h to determine the induction of the expression of HDL receptor mRNA. The results of in situ hybridization histochemistry showed that in the immature ovary, HDL receptor mRNA is associated with theca interna and interstitial cells (stroma). The mRNA expression in these cell types increased with PMSG treatment, but no signal was detected in the granulosa cells. Northern blot analysis also showed a marked increase in mRNA content in thecal and interstitial cells during follicular development. During luteinization, the intensity of the signal began to appear in the luteinized granulosa cells. With the completion of luteinization, the signal in the corpus luteum tissue became more intense. Further treatment with hCG increased the HDL receptor mRNA content compared with that in the saline-treated control. These results demonstrate that the cholesterol-using cell types of the ovary, namely the interstitial cells, thecal cells, and fully luteinized granulosa cells are endowed with the HDL receptor mRNA, which provides credence to the functional significance of the role of HDL receptor SR-B1 in cholesterol transport and ovarian steroidogenesis.

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