Endocrinology, Vol 117, 1490-1499, Copyright © 1985 by Endocrine Society

ARTICLES

A monoclonal antibody to a mammary cell line recognizes two distinct subtypes of ovarian granulosa cells

GF Erickson, C Hofeditz, M Unger, WR Allen and R Dulbecco

When screening supernatant fluids from hybridoma clones, Dulbecco and co-workers found that a mouse monoclonal antibody generated against a mammary tumor cell line showed rather striking high binding to rat oocytes and granulosa cells. In this study we have specifically investigated the reactivity of the monoclonal antibody (designated anti- OA-1) with granulosa cells during the differentiation process. This was accomplished using the two-step indirect immunocytochemical technique. When a primordial follicle is recruited to initiate growth, intense immunoreactivity appears in the surface membrane of the granulosa cells. As a follicle proceeds through the preantral stages, the plasma membrane of the granulosa cells is strongly positive for anti-OA-1 reactivity, and the granulosa appear as a homogeneous population. However, once Graafian follicle development is initiated, a major shift in anti-OA-1 immunoreactivity occurs among the granulosa cells. As the antrum expands, 75% of the granulosa in the mural region (those nearest the basal lamina) elongate and become negative for anti-OA-1. This is in contrast to the periantral and cumulus granulosa, which remain rounded and show strong anti-OA-1 reactivity up to the preovulatory stage. The disappearance of anti-OA-1 reactivity in the subpopulation of mural cells is specifically initiated by FSH and occurs very rapidly after a 12-h lag phase. After the loss of anti-OA-1 reactivity, the elongated mural granulosa cells express their terminal differentiated state by acquiring LH/hCG receptor, 3 beta-hydroxysteroid dehydrogenase activity and cytoplasmic lipid inclusions. By contrast, the periantral and cumulus granulosa, which remain positive for anti-OA-1, do not express these differentiated functions; however, they do differentiate ultrastructurally, indicating that they respond to the FSH signal. These results strongly suggest that a monoclonal antibody recognizes a major surface differentiation antigen in the granulosa cell. This antigen is under hormonal control and is inversely linked to expression of the terminal differentiation program in the granulosa cells. We anticipate that the monoclonal antibody will be a valuable probe to aide in the analysis of structure/function relationships in subpopulations of granulosa cells.

This article has been cited by other articles:

				F. J Diaz, K. Sugiura, and J. J Eppig Regulation of Pcsk6 Expression During the Preantral to Antral Follicle Transition in Mice: Opposing Roles of FSH and Oocytes Biol Reprod, January 1, 2008; 78(1): 176 - 183. [Abstract] [Full Text] [PDF]

				M. Matsui, B. Sonntag, S. S. Hwang, T. Byerly, A. Hourvitz, E. Y. Adashi, S. Shimasaki, and G. F. Erickson Pregnancy-Associated Plasma Protein-A Production in Rat Granulosa Cells: Stimulation by Follicle-Stimulating Hormone and Inhibition by the Oocyte-Derived Bone Morphogenetic Protein-15 Endocrinology, August 1, 2004; 145(8): 3686 - 3695. [Abstract] [Full Text] [PDF]

				E. Rajpert-De Meyts, N. Jorgensen, N. Gram, J. Muller, R. L. Cate, and N. E. Skakkebak Expression of Anti-Mullerian Hormone during Normal and Pathological Gonadal Development: Association with Differentiation of Sertoli and Granulosa Cells J. Clin. Endocrinol. Metab., October 1, 1999; 84(10): 3836 - 3844. [Abstract] [Full Text] [PDF]

				K. E. Latham, F. D. M. Bautista, Y. Hirao, M. J. O'Brien, and J. J. Eppig Comparison of Protein Synthesis Patterns in Mouse Cumulus Cells and Mural Granulosa Cells: Effects of Follicle-Stimulating Hormone and Insulin on Granulosa Cell Differentiation In Vitro Biol Reprod, August 1, 1999; 61(2): 482 - 492. [Abstract] [Full Text]