Role of UCH-L1/ubiquitin in acute testicular ischemia-reperfusion injury.
The most significant complication of testicular torsion is loss of the testis, which may lead to impaired fertility. Molecular mechanisms how spermatogenesis impairs owing to testicular torsion remain unknown. This investigation, by using mouse model of testicular torsion, was undertaken to gain insight into the cellular and molecular mechanism underlying torsion-induced germ cell loss. Male mice were subjected to 2h ischemia-inducing torsion, and testes were examined at 24, 48, and 72h after the repair of torsion (reperfusion). Ischemia-reperfusion (IR) of the testes resulted in germ cell, mostly in spermatogonia, apoptosis, which was revealed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) technique. At 24h after torsion repair germ cell apoptosis reached peak, then decreased until 72h repair. Western blots showed that apoptotic proteins (p53, Caspase-3 and -9) gradually were upregulated at 48h reperfusion, however, anti-apoptotic proteins (Bcl-2 and BDNF) were downregulated in the relevant IR treatment. IR injury induced CHOP protein appearance with maximum expression at 24h of reperfusion. Furthermore, the germ cell apoptosis triggered downregulation of ubiquitin carboxyl-terminal hydrolase-L1 (UCH-L1) at both mRNA and protein levels. To test further whether ubiquitination was involved in IR stress, both mono- and poly-ubiquitin levels in IR stress condition were examined, which showed that both mono- and poly-ubiquitin expression significantly impaired. These results provide evidences of UCH-L1/ubiquitination signaling to the testis IR injury in vivo.
Sun
Jinhua
J
Department of Genetics and Center for Developmental Biology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China.
Ying
Ming
M
Li
Hui
H
Shang
Xuan
X
He
Yan
Y
Chen
Ke
K
Cheng
Hanhua
H
Zhou
Roujia
R
D000818:Animals; D008297:Male; D051379:Mice; D015427:Reperfusion Injury / Q000378:metabolism*; D013086:Spermatic Cord Torsion / Q000378:metabolism*; D013733:Testicular Diseases / Q000378:metabolism*; D013737:Testis / Q000098:blood supply* / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*
D025801:Ubiquitin; D043222:Ubiquitin Thiolesterase; C496976:Uchl1 protein, mouse