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Restricted Expression of WT1 Messenger Ribonucleic Acid in Immature Ovarian Follicles: Uniformity in Mammalian and Avian Species and Maintenance during Reproductive Senescence¹

^a Division of Reproductive Biology, Department of Gyn/Ob, Stanford University Medical School, Stanford, California 94305-5317 ^b Department of Ob/Gyn, University of California School of Medicine, Los Angeles, California 90095 ^c Department of Animal Sciences, University of Illinois, Urbana, Illinois 61801 ^d Institut National de la Sante et de la Recherche Medicale, U-355 Clamart, France ^e Department of Obstetrics, Gynecology and Cell Biology, Duke University Medical Center, Durham, North Carolina 27710

WT1 is a zinc finger protein with transcriptional repressor activity on several growth factor and growth factor receptor genes. In the ovary, a potential role for WT1 in the suppression of the development of immature follicles has been demonstrated. Here, gel retardation assays further showed that recombinant WT1 protein interacted with consensus DNA sequences in the inhibin- gene promoter. We investigated the pattern of WT1 expression in a wide variety of species and also over the reproductive life span in rats. In chicken ovaries, Northern blot analysis revealed the presence of WT1 transcript in small healthy white follicles (1–5 mm in diameter) and its absence in small yellow (6–12 mm in diameter) or larger follicles (F₁–F₅). In pig and monkey ovaries, WT1 expression was limited to granulosa cells of preantral follicles, as shown by in situ hybridization analysis. In rats, Northern blot analyses demonstrated the presence of WT1 transcript in the ovaries of young (3-mo-old) and middle-aged (9-mo-old) rats on the proestrous day, with a decrease in old (12-mo-old) rats in persistent estrus. In situ hybridization analysis further suggested that the decrease in WT1 expression in aging ovaries was associated with fewer immature follicles. Thus, WT1 expression is restricted to immature follicles in diverse avian and mammalian species and over the reproductive life span in rats. These data demonstrated that WT1 is a marker for immature follicles and suggested a potential role of this transcriptional repressor in the slow growth of early follicles.

¹ This work was supported by NIH grant HD31398 (A.J.W.H.) and HD11827 (D.W.S.). E.A.M. is an American Society for Reproductive Medicine-NICHHD Fellow of the Reproductive Scientist Development Program.

² Correspondence. FAX: 650 725 7102; aaron.hsueh{at}stanford.edu

³ Current address: Hormone Research Center, Dept. of Biology, College of Natural Sciences, Chonnam National University, Kwangju, 500–757, Republic of Korea.

⁴ Current address: Department of Biology&Microbiology, California State University, Los Angeles, CA 90032.

⁵ Current address: INSERM U-407, Faculte de Medecine, Lyon-Sud, BP 12, 69 921 Oullins Cedex, France.

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