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The Journal of Clinical Endocrinology & Metabolism Vol. 84, No. 8 2744-2750
Copyright © 1999 by The Endocrine Society


Original Studies

Human Growth Differentiation Factor 9 (GDF-9) and Its Novel Homolog GDF-9B Are Expressed in Oocytes during Early Folliculogenesis1

Johanna Aaltonen2, Mika P. Laitinen2, Kaisa Vuojolainen, Risto Jaatinen, Nina Horelli-Kuitunen, Laura Seppä, Henna Louhio, Timo Tuuri, Jari Sjöberg, Ralf Bützow, Outi Hovatta, Leslie Dale and Olli Ritvos3

Department of Bacteriology and Immunology, Haartman Institute (J.A., M.P.L., K.V., R.J., L.S., O.R.), Haartmaninkatu 3, University of Helsinki, FIN-00014 Helsinki; the Laboratory of Molecular Genetics, Helsinki University Central Hospital (N.H.-K.), Haartmaninkatu 4, FIN-00290 Helsinki; the Infertility Clinic, Family Federation of Finland (H.L., T.T., O.H.), Kalevankatu 16, FIN-00100 Helsinki, Finland; and the Department of Obstetrics and Gynecology, University of Helsinki (J.S., R.B.), Haartmaninkatu 2, FIN-00290, Helsinki, Finland; and the Department of Anatomy and Developmental Biology, University College London (L.D.), London, United Kingdom WC1E 6BT

Address all correspondence and requests for reprints to: Dr. J. Aaltonen, Department of Bacteriology and Immunology, Haartman Institute, P.O. Box 21, Haartmaninkatu 3, University of Helsinki, FIN-00014 Helsinki, Finland. E-mail: johanna.aaltonen{at}helsinki.fi

Growth differentiation factor 9 (GDF-9) is a transforming growth factor-ß family member that is required for normal folliculogenesis in female mice, but its role as a regulator of human fertility is still unclear. We determined here by in situ hybridization and immunohistochemical analyses the localization of the GDF-9 messenger ribonucleic acid (mRNA) and protein during human folliculogenesis. The GDF-9 transcripts were not detected in primordial follicles, but they are abundantly expressed in primary follicles in frozen sections of ovarian cortical tissue material obtained at laparoscopic surgery. We raised antipeptide antibodies against GDF-9 and showed by immunohistochemical studies on paraffin sections of whole human ovaries that the GDF-9 protein is most abundantly expressed in primary follicles. We recently demonstrated that a novel GDF-9-related factor, GDF-9B, is coexpressed with GDF-9 during murine folliculogenesis. We now isolated human GDF-9B complementary DNA and genomic clones and report the unusually restricted expression pattern of human GDF-9B. The human GDF-9B transcript can be detected only in the gonads by RT-PCR analysis, and in situ hybridization studies indicate that it is not expressed in small primary follicles but, rather, in the oocytes of late primary follicles. Functional studies using the Xenopus laevis embryo model indicate that unlike the transforming growth factor-ß family members activin and bone morphogenetic protein-4, neither GDF-9 nor GDF-9B affects mesoderm induction, suggesting that they may use signaling pathways distinct from those well defined for activin and bone morphogenetic protein-4.

We conclude that 1) both GDF-9 mRNA and protein are abundantly expressed in oocytes of primary follicles in human ovary, suggesting that the GDF-9 transcript is translated at this early stage of folliculogenesis; 2) human GDF-9B is specifically expressed in gonads at low levels; and 3) the expression of GDF-9 mRNA begins slightly earlier than that of GDF-9B in the human oocytes during follicular development. Our results are consistent with the suggestion that GDF-9 and GDF-9B may regulate human folliculogenesis in a manner specific to the ovary.




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