Developmental changes of heat-shock proteins in porcine testis by a proteomic analysis.
Heat-shock proteins (HSPs) are important in spermatogenesis. This study investigated developmental changes in the expression of major HSPs in porcine testis. The testis from five immature (mean age 2.9+/-0.1 months) and five mature boars (35.7+/-14.0 months) were examined. Two-dimensional polyacrylamide gel electrophoresis was conducted and proteins were identified by Western blotting and/or matrix-assisted laser desorption/ionization mass spectrometry. Moreover, the 90, 70, and 60 kDa HSPs, 70 kDa heat-shock cognate protein (HSC 70), tubulin, and actin were quantified on two-dimensional gels. Protein spots were quantified by densitometry, combined with a computer-assisted image analysis system. Immunohistochemistry was performed to analyze the expression pattern of major HSPs and beta-tubulin in testis. One isoform of HSP 90 (HSP 90 alpha), two isoforms of HSC 70 (HSC 70a and HSC 70c), one isoform of HSP70 (HSP 70e), and tubulin increased after sexual maturation (P<0.05). A testis-specific HSP70 (P70t) was markedly increased in the testes of sexually mature boars. Meanwhile, levels of actin and some isoforms of HSPs including 60 kDa HSP remained similar in both groups. These observations were further confirmed by immunohistochemistry; therefore, the upregulation of protein expression in the adult testis could be attributed to a higher level of protein expression and the number of cells that were HSPs-positive already resided in the immature testis. The differential expression of major HSPs suggested that they may be important in porcine spermatogenesis.
Huang
San-Yuan
SY
Division of Biotechnology, Animal Technology Institute Taiwan, PO Box 23, Chunan 350Miaoli, Taiwan, ROC.
Tam
Ming-F
MF
Hsu
Yu-Tsin
YT
Lin
Jyh-Hung
JH
Chen
Hsin-Hsin
HH
Chuang
Chin-Kai
CK
Chen
Ming-Yu
MY
King
Yueh-Tsu
YT
Lee
Wen-Chuan
WC
D000199:Actins / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D018834:Chaperonin 60 / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D050883:HSC70 Heat-Shock Proteins / Q000032:analysis; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis; D018841:HSP90 Heat-Shock Proteins / Q000032:analysis; D006360:Heat-Shock Proteins / Q000032:analysis*; D007150:Immunohistochemistry; D008297:Male; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013091:Spermatogenesis; D013552:Swine / Q000254:growth & development*; D013737:Testis / Q000737:chemistry* / Q000254:growth & development*; D014404:Tubulin / Q000032:analysis
D000199:Actins; D018834:Chaperonin 60; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D014404:Tubulin