Serial analysis of gene expression in turkey sperm storage tubules in the presence and absence of resident sperm.
Turkey sperm lose viability within 8-18 h when stored as liquid semen using current methods and extenders. In contrast, turkey hens maintain viable, fertile sperm in their sperm storage tubules (SST) for 45 or more days following a single insemination. Our long-term objectives are to identify and characterize differentially expressed genes that may underlie this prolonged sperm storage and then use this information to develop improved methods for storing liquid turkey semen. We employed serial analysis of gene expression (SAGE) to compare gene expression patterns in turkey SST recovered from hens after artificial insemination (AI) with extended semen (sperm AI) or extender alone (control AI). We constructed two separate SAGE libraries with SST RNA obtained from sperm and control AI hens. We used these libraries to generate 95,325 ten-base pair SAGE tags. These 95,325 tags represented 27,430 unique genes. The sperm and control AI libraries contained 47,663 and 47,662 tags representing 18,030 and 19,101 putative unique transcripts, respectively. Approximately 1% of these putative unique genes were differentially expressed (P<0.05) between treatments. Tentative annotations were ascribed to the SAGE tag nucleotide sequences by comparing them against publicly available SAGE tag and cDNA sequence databases. Based on its SAGE tag nucleotide sequence, we cloned a partial turkey avidin cDNA and confirmed its up-regulation in the sperm AI SST. The bioinformatics and experimental procedures employed to clone turkey avidin and confirm its differential expression represent a useful paradigm for analyzing SAGE tag data from relatively uncharacterized model systems.
Long
Ezhou L
EL
Biotechnology and Germplasm Laboratory, Animal and Natural Resources Institute, USDA-ARS, Beltsville, Maryland 20705, USA.
Sonstegard
Tad S
TS
Long
Julie A
JA
Van Tassell
Curtis P
CP
Zuelke
Kurt A
KA
D000818:Animals; D001360:Avidin / Q000096:biosynthesis / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005784:Gene Amplification; D015870:Gene Expression / Q000235:genetics*; D015723:Gene Library; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012661:Semen / Q000502:physiology; D012671:Seminiferous Tubules / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology*; D014422:Turkeys / Q000502:physiology*
D018076:DNA, Complementary; D012333:RNA, Messenger; D001360:Avidin