Proteomic and genomic analyses of female subfertility in chickens and turkeys

Previous research has identified a novel 22 kD protein associated with low sperm binding and consequent low female fertility in an experimental lines of chickens selected for growth rate. The objective of the following work was to assess the generalizability of the apparent 22kD protein in birds and to identify the biochemical mechanism responsible.; Two commercial populations of chickens were screened for the presence of the 22kD protein. An immunoreactive 22kD protein was found in both of the commercial lines. Commercial Line 1 (CL-1) exhibited a 25% frequency of hens expressing the variant protein, resulting in a 25% decrement in fertility of those hens. Commercial Line 2 (CL-2) had a 9% incidence of an immunoreactive 22kD protein, with 16% lower fertility than normal hens. These results suggest that occurrence of the 22kD protein happened early in the evolution of Gallus Domesticus and that the occurrence of the protein and its association with fertility is not unique to small selected populations of birds.; Eighty-six percent of the reported 22kD protein sequence is homologous to the chicken ZP3/ZPC protein. The purpose of the following experiment was to assess the possibility of the occurrence of the 22kD protein in species other than chicken by comparing ZP3/ZPC homology across species, with reference to potentially conserved 22kD domains. Novel tissue expression of ZP3/ZPC was shown in the immature white follicles of the adult ovary, infundibulum of both turkeys and chickens. Using antibodies developed to specific amino acid sequences from chicken ZP3/ZPC (chZP3), western blot analysis of the perivitelline membranes (PVM) of laid turkey eggs revealed an immunoreactive band with the molecular mass of approximately 45 kD—which is larger than the 42kD chZP3 protein. Coomassie stained turkey PVM revealed a candidate 25kD turkey PVM protein, western blot analysis with the anti-chZP3/ZPC-peptide antibody showed no immunoreactive band, indicating a lack of homology to the 22kD protein.; rtPCR analysis and sequencing of the PCR product showed that full sequence wild type ZP3/ZPC mRNA is expressed in the granulosa cells of 22kD carrier hens, suggesting that the 22kD protein was not the result of a mutation of wild type ZP3/ZPC. A 600 by rtPCR product was sequence from wild type ZP1/ZPB obtained from the liver of 22kD (+) hens. This partial ZP1 sequence coded for 25 of the 27 ZP1/ZPB-like sequence reported in the 22kD protein and also coded for an additional 167 ZP1/ZPB AA. (Abstract shortened by UMI.)...