Submitted to: BARC Poster Day
Publication Type: Abstract Only
Publication Acceptance Date: March 29, 2002
Publication Date: April 15, 2002
Citation: Long, E.L., Sonstegard, T.S., Long, J.A., Zuelke, K.A. 2002. Transcript profiling in turkey sperm storage tubules using sage. BARC Poster Day. Technical Abstract: The sperm storage tubules (SST) are unique epithelial structures within the avian oviduct that provide for prolonged storage of fertile sperm following natural mating or artificial insemination (AI). In turkeys, spermatozoa can remain fertile after being stored in the SST for upwards of 70 days. We hypothesized that specific and characteristic gene expression events enable and regulate prolonged sperm storage within the turkey SST. In order to elucidate the molecular basis for prolonged sperm storage, we applied Serial Analysis of Gene Expression (SAGE) to identify and characterize the relative levels of gene expression in the SST of laying turkey hens following a sham (no sperm) AI. Total RNA was prepared from turkey SST isolated at 48h following a sham AI with PBS. A SAGE tag library was then constructed from the SST RNA as per the protocol provided with the I-SAGE Kit (InVitrogen). The DNA sequences obtained from 18,903 tags represented 9526 putative unique transcripts from within the mRNA population that was sampled; 2484 tags were represented at least twice within the sample. Combined, approximately 10% of the total SAGE tags in this library represented 27 different putative mRNA transcripts. One tag (5'-ACACTGAACT-3') appeared with the highest frequency of 1.48 % (280 out of 18,903 tags). These early SAGE tag data begin to reveal the relative frequencies of individual mRNA transcripts within turkey SST in the absence of sperm and provide a useful starting point for identifying and characterizing genes that may be involved in SST function. Due to the lack of available 3' cDNA sequence data from turkey oviduct, the identity of the specific genes to which these tags corresponded remains undetermined. Ongoing sequencing of this library along with another comparable SAGE library from post-AI SST will provide a comparison between SST gene expression in the presence and absence of sperm.