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Metallothionein-1 Messenger RNA Transcription in Steroid-Secreting Cells of the Rat Ovary During the Periovulatory Period¹

Department of Biology,³ Trinity University, San Antonio, Texas 78212 Department of Obstetrics and Gynecology,⁴ Kumamoto University Medical School, Kumamoto 860-8556, Japan Department of Molecular and Cellular Biology,⁵ Baylor College of Medicine, Houston, Texas 77030

An increase in metallothionein 1 (MT-1) mRNA was detected in the ovaries of immature Wistar rats that were primed with s.c. injection of 10 IU eCG followed 48 h later by 10 IU hCG s.c. to initiate the ovulatory process. Ovarian RNA was extracted at 0, 2, 4, 8, 12, 24, 72, 144, and 288 h after the primed animals were injected with hCG. These extracts were used for reverse transcription polymerase chain reaction (RT-PCR) differential display and Northern analyses that yielded complementary gene fragments for MT-1. Expression of MT-1 mRNA increased significantly by 24 h after hCG treatment and reached a peak at 144 h after hCG. In contrast, a disintegrin and metalloproteinase with thrombospondin motifs and a tissue inhibitor of metalloproteinase 1, which were also detected by the RT-PCR differential display procedure, reached a peak at 12 h after hCG and returned to control levels in the ovaries by 72 h after hCG. In situ hybridization indicated that most of the MT-1 mRNA was expressed in the vicinity of the theca interna of preovulatory follicles and in the lutein granulosa of postovulatory follicles. Thus, MT-1 mRNA expression is primarily in the vicinity of steroid-secreting areas of the ovary. The substantial increase in MT-1 mRNA expression might be important in protecting the ovarian tissues from oxidative stress generated by ovarian inflammatory events during the ovulatory process and luteinization.

¹ This work was supported by NSF grant 9870793 (to L.L.E.), a grant to support T.U. as a Research Fellow of The Lalor Foundation, Providence, Rhode Island (to L.L.E.), and NIH grants HD-16229, HD-16272, and SCCPRR-HD07495 (to J.S.R.).

² Correspondence. FAX: 210 999 7229; lespey{at}trinity.edu