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Search for specific expressed genes in chicken sperm storage tubules by differential display polymerase chain reaction

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Abstract:

The objective of this study was to find the specific expressed genes in chicken sperm storage tubules. One hundred and fifty dual-purpose hens at 30 weeks of age were inseminated with 1 × 108 spermatozoa on two consecutive days. Eggs were collected and set daily during the 24 days following the latter of two inseminations. Five hens with fertility below 60% were inseminated with fresh semen. Fifteen hens with fertility above 80% were divided into three groups. Fresh semen, frozen dead semen and 0.9% NaCl, respectively, were injected into the vagina of five hens in each group. RNA from the utero-vaginal junction (UVJ) of the hens was reverse transcribed and differentially displayed. Four bands of interest which were expressed in UVJ from hens inseminated with fresh semen or frozen semen but not from hens inseminated with saline were selected, re-amplified, cloned and sequenced. BLAST search showed that band A is similar to part of chromosome 3 (the homology is 98%) and the nearest gene to the fragment is 1,722 base pairs up-stream whose product is mitogen-activated kinase. Band B is similar to phosphatidylinositol glycan anchor biosynthesis gene (PIGK) and the homology is 98%. PIGK is a subunit of glycosylphosphatidylinositol transamidase (GPI8). GPI-linked proteins participate in the release of membrane proteins. Band F is similar to a hypothetical protein gene and the homology is 97%. This hypothetical protein belongs to chicken neurexophilin. Band G is similar to fat3 and the homology is 97%. Neurexophilin and fat3 are proteins that function in the nervous system. Taken together these data indicate that hens may regulate sperm storage in UVJ through the nervous system and may release sperm from the UVJ through GPI8.

Keywords: CHICKEN; DDRT-PCR; SSTS; UVJ

Document Type: Research Article

DOI: http://dx.doi.org/10.3184/175815509X12474776260634

Publication date: August 1, 2009

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