Molecular cloning of porcine growth differentiation factor 9 (GDF-9) cDNA and its role in early folliculogenesis: direct ovarian injection of GDF-9 gene fragments promotes early folliculogenesis.
Growth differentiation factor-9 (GDF-9) is a growth factor secreted by oocytes in growing ovarian follicles. To investigate the ovarian function of GDF-9 in pigs, we first cloned porcine GDF-9 complementary DNA (cDNA), and then injected its gene fragments into the ovary in gilts. Porcine GDF-9 has open reading frame (ORF) homologies of 81.4%, 84.6%, 84.2%, 72.7% and 72.6% with its human, bovine, ovine, rat and mouse counterparts respectively. Regarding the deduced amino-acid sequence of the mature protein, the corresponding homologies reach 92.1%, 97.8%, 97.0%, 89.6% and 88.1% respectively. To investigate the role of GDF-9 in early folliculogenesis, the ovaries of 2-month-old prepubertal gilts were injected with GDF-9 gene fragments. The injection of porcine GDF-9 gene fragments resulted in an increase in the number of primary, secondary and tertiary follicles, concomitant with a decrease in the number of primordial follicles. These results indicated that exogenous GDF-9 can promote early folliculogenesis in the porcine ovary, and that a technique for direct ovarian injection of GFD-9 gene fragments may contribute to a novel therapy for prevention and treatment of infertility associated with ovarian dysfunction.
Shimizu
Takashi
T
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan. shimizut@idac.tohoku.ac.jp
Miyahayashi
Yasunori
Y
Yokoo
Masaki
M
Hoshino
Yumi
Y
Sasada
Hiroshi
H
Sato
Eimei
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055430:Bone Morphogenetic Protein 15; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000008:administration & dosage* / Q000235:genetics; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D055429:Growth Differentiation Factor 9; D007267:Injections; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000737:chemistry / Q000502:physiology*; D010053:Ovary; D016649:Primary Ovarian Insufficiency / Q000628:therapy; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012756:Sheep; D013552:Swine
C525901:BMP15 protein, human; C525902:Bmp15 protein, mouse; C525904:Bmp15 protein, rat; D055430:Bone Morphogenetic Protein 15; D018076:DNA, Complementary; C525897:GDF9 protein, human; C525898:Gdf9 protein, mouse; C525899:Gdf9 protein, rat; D055429:Growth Differentiation Factor 9; D036341:Intercellular Signaling Peptides and Proteins