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Publication : Inhibition of zona pellucida gene expression by antisense oligonucleotides injected into mouse oocytes.

First Author  Tong ZB Year  1995
Journal  J Biol Chem Volume  270
Issue  2 Pages  849-53
PubMed ID  7822321 Mgi Jnum  J:23006
Mgi Id  MGI:70639 Doi  10.1074/jbc.270.2.849
Citation  Tong ZB, et al. (1995) Inhibition of zona pellucida gene expression by antisense oligonucleotides injected into mouse oocytes. J Biol Chem 270(2):849-53
abstractText  During murine oogenesis, the zona pellucida proteins (ZP1, ZP2, and ZP3) are synthesized and secreted to form an extracellular matrix that surrounds the oocyte and mediates specific biological functions essential to mammalian fertilization and early development. To investigate the relationship among the zona proteins during zona matrix assembly, we have undertaken to inhibit de novo biosynthesis of specific zona proteins with antisense oligonucleotides complementary to the 5'-ends of ZP2 (nucleotide position 19-42) and ZP3 (nucleotide 21-44) mRNAs. When injected into the cytoplasm of growing mouse oocytes, the antisense oligonucleotides targeted specific zona mRNAs for degradation, as confirmed by a RNase protection assay. Individual zona pellucida protein synthesis was followed by immunoprecipitation with ZP2- and ZP3-specific monoclonal antibodies. New zona protein synthesis from the targeted mRNA was abolished, but nontargeted zona protein continued to be synthesized. Interestingly, abolishment of either ZP2 or ZP3 protein synthesis prevented the incorporation of the other protein into the extracellular zona matrix. These results suggest that ZP2 and ZP3 proteins are independent of each other in their biosynthesis but are dependent upon each other for their incorporation into the zona pellucida matrix. This study provides an experimental system in which destruction of a targeted mRNA generates a transient loss-of-expression phenotype during mouse oocyte growth.
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