Transcripts of enriched germ cells responding to heat shock as potential markers for porcine semen quality.
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 1 (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 1h and recovered at 34 degrees C for 2h), 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=31). 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.
Gau
B-H
BH
Department of Chemical Engineering, National Tsing Hua University, 101, Sectio 2, Kuang-Fu Road, Hsinchu 300, Taiwan, ROC.
Chu
I-M
IM
Huang
M-C
MC
Yang
K-T
KT
Chiou
S-H
SH
Fan
Y-H
YH
Chen
M-Y
MY
Lin
J-H
JH
Chuang
C-K
CK
Huang
S-Y
SY
Lee
W-C
WC
D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D012333:RNA, Messenger / Q000032:analysis*; D012661:Semen / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry*; D013552:Swine; D013737:Testis / Q000166:cytology
D020121:5' Untranslated Regions; D015415:Biomarkers; D006360:Heat-Shock Proteins; D012333:RNA, Messenger