Ovary-selective genes I: the generation and characterization of an ovary-selective complementary deoxyribonucleic acid library.
The importance of several ovary-selective/specific genes, i.e. genes preferentially or exclusively expressed in the ovary, has been established. Indeed, null mutant female mice for the c-mos, growth and differentiation factor-9, alpha-inhibin, and zona pellucida-3 genes proved sterile. A loss of function mutation of the human FSH receptor gene established its critical role in ovarian function. These data support the hypothesis that genes expressed selectively or specifically in the ovary are probably essential for the normal functioning of this organ system. We have used the differential screening technique suppression subtractive hybridization to systematically isolate and clone genes that are expressed in an ovary-selective/specific manner. The resultant target complementary DNA (cDNA) library has been exhaustively screened to a point at which additional sequencing was increasingly unlikely (< or = 4%) to yield additional previously unencountered cDNAs. In toto, 844 clones were sequenced and analyzed for homology to known genes using the Basic Local Alignment Tool (BLAST). Of those, 342 were determined to be independent (nonredundant). One hundred and fifty-nine independent clones proved identical to previously characterized genes, whereas an additional 100 independent clones proved significantly homologous (but not identical) to previously characterized genes. Yet 83 other independent clones did not display significant homology to previously characterized genes now listed in the publicly accessible nonredundant databases. As such, these latter genes were deemed novel. Of these 83 novel genes, a total of 36 displayed ovary-specific/selective expression, as determined by probing mouse multitissue Northern blots with 32P-labeled/PCR-amplified cDNA inserts. Under these circumstances, the false positive rate was minimal, as only one novel clone was expressed at a higher level in nonovarian tissues relative to ovary. Of the 36 ovary-specific/ selective novel genes, 22 proved subject to hormonal regulation during a simulated estrous cycle. In this communication we focus on 2 such novel ovary-specific/hormonally-dependent genes, the full-length sequences of which were isolated using rapid amplification of 3'-cDNA ends technology. Taken together, the present study accomplished systematic identification of those genes that are restricted in their expression to the ovary. These ovary-selective genes may have significant implications for the understanding of ovarian function in molecular terms and for the development of innovative strategies for the promotion of fertility or its control.
Hennebold
J D
JD
Department of Obstetrics and Gynecology, University of Utah Health Sciences Center, Salt Lake City 84132, USA.
Tanaka
M
M
Saito
J
J
Hanson
B R
BR
Adashi
E Y
EY
D015067:17-Hydroxysteroid Dehydrogenases / Q000235:genetics; D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001141:Aromatase / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D055430:Bone Morphogenetic Protein 15; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004527:Egg Proteins / Q000235:genetics; D005136:Eye Proteins; D005260:Female; D015723:Gene Library; D055429:Growth Differentiation Factor 9; D006133:Growth Substances / Q000235:genetics; D006801:Humans; D007265:Inhibins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000235:genetics; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D010053:Ovary / Q000737:chemistry*; D010455:Peptides / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics; D011506:Proteins / Q000737:chemistry / Q000235:genetics; D017231:Proto-Oncogene Proteins c-mos / Q000235:genetics; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; D000098682:Steroidogenic Acute Regulatory Protein
C525901:BMP15 protein, human; C525902:Bmp15 protein, mouse; D055430:Bone Morphogenetic Protein 15; D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D004527:Egg Proteins; D005136:Eye Proteins; C525897:GDF9 protein, human; C525898:Gdf9 protein, mouse; D055429:Growth Differentiation Factor 9; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; C411842:M7-A10 protein, mouse; C485853:MGARP protein, human; C509731:MGARP protein, mouse; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D010455:Peptides; D010750:Phosphoproteins; D011506:Proteins; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; C094377:inhibin-alpha subunit; D000098682:Steroidogenic Acute Regulatory Protein; D007265:Inhibins; D015067:17-Hydroxysteroid Dehydrogenases; D015096:3-Hydroxysteroid Dehydrogenases; D001141:Aromatase; D017231:Proto-Oncogene Proteins c-mos