description): The investigators plan to produce canine clones for genes that are known to be expressed in testis development and define the temporal and spatial expression patterns of these genes in normal canine embryonic gonads.
The specific aims are: 1) To clone the entire coding sequence of canine SOX-9. The investigators are currently screening a canine genomic DNA library with a human SOX-9 that hybridizes to canine DNA. Homologous regions will be subcloned and characterized by sequence analysis. 2) To clone canine genomic DNA sequences of MIS (Amh), Wt-l, and SF-1. They will use standard library screening methods and polymerase chain reaction (PCR) to identify clones in a genomic DNA library. Regions of homology will be subcloned and their identity confirmed by sequence analysis. 3) To characterize the temporal and spatial expression patterns of Sry, Sox 9, MIS, Wt-1 and SF-1 in normal canine embryonic gonads, using reverse transcriptase PCR and in situ hybridization with canine probes. At the end of this study, they will be poised to examine the temporal and spatial expression patterns of these genes in affected embryos, which will be critical to understanding the genetic mechanisms responsible for abnormal testis development.
| Meyers-Wallen, Vicki N (2006) Genetics, genomics, and molecular biology of sex determination in small animals. Theriogenology 66:1655-8 |
| Meyers-Wallen, Vicki N (2005) Sf1 and Mis expression: molecular milestones in the canine sex determination pathway. Mol Reprod Dev 70:383-9 |
| Meyers-Wallen, V N (2003) Sry and Sox9 expression during canine gonadal sex determination assayed by quantitative reverse transcription-polymerase chain reaction. Mol Reprod Dev 65:373-81 |
| Meyers-Wallen, V N; Schlafer, D; Barr, I et al. (1999) Sry-negative XX sex reversal in purebred dogs. Mol Reprod Dev 53:266-73 |
