Localization of growth differentiation factor-9 (GDF-9) mRNA and protein in rat ovaries and cDNA cloning of rat GDF-9 and its novel homolog GDF-9B.
Although targeted gene disruption of GDF-9, an oocyte derived growth factor, leads to an arrest of folliculogenesis and causes infertility in female mice, little is known on the expression of GDF-9 protein in the ovary. We show that GDF-9 protein is expressed in rat oocytes during folliculogenesis from the early primary follicle stage onwards but the most intensive immunostaining was seen in primary and preantral follicles. Northern blot analyses of the ontogeny of GDF-9 gene expression in postnatal rat ovaries showed that the GDF-9 transcript levels are clearly increased on the second postnatal day concomitant with the appearance of primary follicles. Interestingly, Northern blot and in situ hybridization analyses indicate a similar expression pattern for GDF-9B, the rat ortholog of a mouse GDF-9 like factor for which we recently reported the partial amino acid sequence. The polypeptide sequences deduced from isolated ovarian cDNAs indicate that the rat GDF-9 prepropeptide is 440 amino acids (aa) in length and the putative mature peptide is 135 aa whereas rat GDF-9B is 391 aa long and the mature region is 125 aa. We conclude that (1) the GDF-9 protein is highly expressed in the oocytes of primary follicles of rat ovaries suggesting that it plays a role mainly in early folliculogenesis and that (2) the full-length polypeptide sequence of GDF-9B suggests that this novel TGF-beta family member is likely to be a secreted growth factor that may regulate folliculogenesis at similar developmental stages as GDF-9.
Jaatinen
R
R
Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Finland.
Laitinen
M P
MP
Vuojolainen
K
K
Aaltonen
J
J
Louhio
H
H
Heikinheimo
K
K
Lehtonen
E
E
Ritvos
O
O
D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055430:Bone Morphogenetic Protein 15; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D018507:Gene Expression Regulation, Developmental; D055429:Growth Differentiation Factor 9; D006133:Growth Substances / Q000737:chemistry / Q000235:genetics*; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D006080:Ovarian Follicle / Q000378:metabolism; D010053:Ovary / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D051381:Rats; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000235:genetics
C525902:Bmp15 protein, mouse; C525904:Bmp15 protein, rat; D055430:Bone Morphogenetic Protein 15; D018076:DNA, Complementary; C525898:Gdf9 protein, mouse; C525899:Gdf9 protein, rat; D055429:Growth Differentiation Factor 9; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta