Although the one-cell zygote has a large complement of mRNAs synthesized during oogenesis, the extent to which these maternal genes modulate early development is unknown. We have focused our investigations on understanding the molecular details of oocyte-specific genes and have chosen as our model system the genes that code for three sulfated glycoproteins of the mouse zona pellucida (ZP-1, ZP-2, and ZP-3). We have isolated cDNAs coding for ZP-3, the mouse sperm receptor, from an ovarian cDNA library cloned in the Lambdagt-11 expression vector. The identity of these clones was confirmed by a comparison of their nucleic acid sequence with the amino acid sequence of the ZP-3 protein. We have shown that the ZP-3 message is expressed as a 1.6 kb poly-adenlyated mRNA which is found uniquely in ovarian tissue. Furthermore, from Northern analysis and in situ hybridizations, it appears that ZP-3 is expressed only in oocytes and not in surrounding granulosa cells. Based on our in situ data, the gene does not appear to be transcribed in resting oocytes (17 Mum) but rapidly becomes a very abundant message in growing oocytes (50 Mum before being turned off in fully grown oocytes (70 Mum). Thus, the transcription of the gene appears to closely parallel our previous obsrevations on zona protein biosynthesis during oogenesis. Somatic cell hybrids were used to localize the ZP-3 locus to mouse chromosome 6 where it appears to exist as a single copy gene. Although the structure of the gene is the same in germ-line and somatic tissues, the ZP-3 gene is hypomethylated in ovarian tissue (where it is expressed) compared to somatic tissue.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
U.S. National Inst Diabetes/Digst/Kidney
Department
Type
DUNS #
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Country
United States
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Hoodbhoy, Tanya; Aviles, Manuel; Baibakov, Boris et al. (2006) ZP2 and ZP3 traffic independently within oocytes prior to assembly into the extracellular zona pellucida. Mol Cell Biol 26:7991-8
Hoodbhoy, Tanya; Joshi, Saurabh; Boja, Emily S et al. (2005) Human sperm do not bind to rat zonae pellucidae despite the presence of four homologous glycoproteins. J Biol Chem 280:12721-31
Hoodbhoy, Tanya; Dean, Jurrien (2004) Insights into the molecular basis of sperm-egg recognition in mammals. Reproduction 127:417-22
Dean, Jurrien (2004) Reassessing the molecular biology of sperm-egg recognition with mouse genetics. Bioessays 26:29-38
Zhao, Ming; Boja, Emily S; Hoodbhoy, Tanya et al. (2004) Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. Biochemistry 43:12090-104
Rankin, Tracy L; Coleman, Jenell S; Epifano, Olga et al. (2003) Fertility and taxon-specific sperm binding persist after replacement of mouse sperm receptors with human homologs. Dev Cell 5:33-43
Zhao, Ming; Gold, Lyn; Dorward, Heidi et al. (2003) Mutation of a conserved hydrophobic patch prevents incorporation of ZP3 into the zona pellucida surrounding mouse eggs. Mol Cell Biol 23:8982-91
Boja, Emily S; Hoodbhoy, Tanya; Fales, Henry M et al. (2003) Structural characterization of native mouse zona pellucida proteins using mass spectrometry. J Biol Chem 278:34189-202
Takasaki, N; Rankin, T; Dean, J (2001) Normal gonadal development in mice lacking GPBOX, a homeobox protein expressed in germ cells at the onset of sexual dimorphism. Mol Cell Biol 21:8197-202
Rankin, T L; O'Brien, M; Lee, E et al. (2001) Defective zonae pellucidae in Zp2-null mice disrupt folliculogenesis, fertility and development. Development 128:1119-26

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