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|標題:||Transcripts of enriched germ cells responding to heat shock as potential markers for porcine semen||作者:||Gau, B.H.
|關鍵字:||microarray;heat shock;single nucleotide polymorphism (SNP);hsp105;boar semen;single nucleotide polymorphisms;regulated expression;a spermatogonia;gene-expression;thap domain;rat testis;protein;sperm;boars;temperature||Project:||Theriogenology||期刊/報告no：:||Theriogenology, Volume 69, Issue 6, Page(s) 758-766.||摘要:||
A cDNA microarray-assisted experiment was conducted to survey genes that respond early to heat shock in enriched immature porcine germ cells; the 5'-UTR flanking the highest upregulated gene, heat shock 105/110 kDa protein I (Hsph1 or Hsp105), in response to heat shock was also investigated. We established a porcine testis cDNA microarray with 9944 transcripts from two libraries constructed from the testes of mature boars, with or without heat shock. After a mild heat shock treatment (39 degrees C for I h and recovered at 34 degrees C for 2 h), 380 transcripts demonstrated significant gene expression in enriched immature germ cells; 326 were upregulated and 54 were downregulated. Ten transcripts of interest exhibiting significance analysis of microarrays (SAM) scores higher than the median were subjected to quantitative real-time PCR; three (Hsp105, Hspa4l and Thap4) were upregulated > 1.5-fold. The sequence of the 5'-UTR of Hsp105, the highest upregulated transcript, was cloned and analyzed. A single nucleotide polymorphism (SNP) was found at position -762 (C or T) upstream of the translational start site (ATG codon). Only two genotypes (CC or TC) were found in the mature boars that were studied (n = 3 1). A heterozygous genotype (TC) at this SNP site revealed an elevated percentage of morphologically normal sperm during hot and cold seasons; this SNP may be a useful marker for semen quality in boars. Furthermore, the cell-model established from enriched primitive germ cells has potential for the study of reproduction in mature animals. (c) 2008 Elsevier Inc. All rights reserved.
|Appears in Collections:||微生物暨公共衛生學研究所|
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