Cold-induced RNA-binding protein (CIRBP) regulates the expression of Src-associated during mitosis of 68 kDa (Sam68) and extracellular signal-regulated kinases (ERK) during heat stress-induced testicular injury
Chengcheng Xu
A * , Dandan Ke A B * , Liping Zou A , Nianyu Li A , Yingying Wang A , Xiaorong Fan A , Changhong Zhu A C and Wei Xia A C D
+ Author Affiliations
- Author Affiliations
A Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
B Department of Obstetrics and Gynaecological Ultrasound Imaging, Renmin Hospital of Wuhan University, No. 99 Zhang Zhidong Road, No. 238 Jiefang Road, Wuhan 430060, China.
C Reproductive Medicine Centre, Tongji Medical College, Huazhong University of Science and Technology, No. 128 Sanyang Road, Wuhan 430010, China.
D Corresponding author. Email: tjxiawei@hust.edu.cn
Reproduction, Fertility and Development 32(18) 1357-1364 https://doi.org/10.1071/RD20253
Submitted: 26 September 2020 Accepted: 18 November 2020 Published: 14 December 2020
Abstract
In this study, the ability of cold-induced RNA-binding protein (CIRBP) to regulate the expression of Src-associated during mitosis of 68 kDa (Sam68) and extracellular signal-regulated kinases (ERK) in the mouse testis and mouse primary spermatocytes (GC-2spd cell line) before and after heat stress was examined to explore the molecular mechanism by which CIRBP decreases testicular injury. A mouse testicular hyperthermia model, a mouse primary spermatocyte hyperthermia model and a low CIRBP gene-expression cell model were constructed and their relevant parameters were analysed. The mRNA and protein levels of CIRBP and Sam68 were significantly decreased in the 3-h and 12-h testicular heat-stress groups, extracellular signal-regulated kinase 1/2 (ERK1/2) protein expression was not significantly affected but phospho-ERK1/2 protein levels were significantly decreased. GC-2spd cellular heat-stress results showed that the mRNA and protein concentrations of CIRBP and Sam68 were reduced 48 h after heat stress. In the low CIRBP gene-expression cell model, CIRBP protein expression was significantly decreased. Sam68 mRNA expression was significantly decreased only at the maximum transfection concentration of 50 nM and Sam68 protein expression was not significantly affected. These findings suggest that CIRBP may regulate the expression of Sam68 at the transcriptional level and the expression of phospho-ERK1/2 protein, both of which protect against heat-stress-induced testicular injury in mice.
Keywords: CIRBP, ERK1/2, heat stress, Sam68, spermatocytes, testis.
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