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Transforming Growth Factor ß Receptor Expression in Hyperstimulated Human Granulosa Cells and Cleavage Potential of the Zygotes¹

^a Departments of Obstetrics and Gynecology and ^b Physiology and Biophysics, Division of Reproductive Endocrinology and Infertility, Olson Center for Women's Health, University of Nebraska Medical Center, Omaha, Nebraska 68198–4515

A possible relationship between transforming growth factor ß receptor type I (TßRI) and type II (TßRII) protein expression in human granulosa cells and the quality of preimplantation embryo development in vitro was studied using immunoblot analysis of TßRI and TßRII in hyperstimulated granulosa cells and morphological assessment of the cleavage potential of the zygotes in vitro. Washed granulosa cells were collected from 35-yr-old women with either tubal defects or mild endometriosis who were undergoing controlled ovarian hyperstimulation prior to oocyte retrieval for in vitro fertilization. TßRI and TßRII were immunoprecipitated from 100 000 g soluble and crude membrane fractions using receptor-specific antibodies and analyzed by Western immunoblotting, and the relative expression was quantitated from the luminographs. The gross morphology (embryo grade) of the preimplantation embryos developed in vitro was determined using a stereomicroscope. Both TßRI and TßRII are expressed in the soluble and membrane fractions of granulosa cells. Most notably, the zygote always developed into a grade 1 quality preimplantation embryo when the oocyte originated from a follicle that expressed a low amount of TßR protein in the granulosa cell membrane. Reduced expression of TßR in the granulosa cell membrane may form a mechanism critically regulating TGFß action on granulosa cells, and the latter in turn precisely control oocyte development, hence, the subsequent cleavage potential.

¹ This study was supported by grants from the National Institute of Child Health and Human Development, Bethesda, MD, USA (HD 28165), and Olson Foundation for Women's Health, Omaha, NE, USA, to S.K.R. A part of the study was presented in the Annual Meeting of American Society of Reproductive Medicine, 1996.

² Correspondence: Shyamal K. Roy, Departments of OB/GYN and Physiology/Biophysics, University of Nebraska Medical Center, BH 5005, 600 S. 42nd Street, Omaha, NE 68198–4515. FAX: 402 559 6164.

³ Current address: Amy M. Carlson, Department of Psychiatry, University of Colorado Health Sciences Center, Denver, CO 80220.

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