Effect of heat shock transcription factor 5(Hsf5) knockdown on heat shock family in mouse Leydig cells and Sertoli cells

doi:10.16352/j.issn.1001-6325.2023.04.0626

Abstract

Abstract: Objective To investigate the effect of knockdown heat shock transcription factor 5(Hsf5) on the expression of heat shock family in mouse Leydig cells (TM3) and Sertoli cells(TM4). Methods Mouse TM3 and TM4 cells were cultured and randomly divided into control group and Hsf5 knockdown group. The expression of heat shock transcription factor(HSF) and heat shock protein(HSP) mRNA in the two groups of cells was detected by real-time quantitative PCR. According to the results of RT-qPCR, HSPs with significant differences in the two kinds of cells were selected (P＜0.01). Western blot were used to detect the expression of HSPA2, HSPA5, HSP90ab1 in TM3 cells and HSPA2,HSP90aa1 in TM4 cells. Results Hsf5 knockdown model was successfully constructed by RNA interference. After knockdown of Hsf5, the mRNA expression of Hspa2,Hspa5,Hsp90ab1 in TM3 cells and Hsf2,Hspa2, Hsp90aa1, Hsp90ab1, Hspd1 in TM4 cells was all significantly down-regulated(P＜0.05). In Hsf5 knockdown group, the expression of HSPA2,HSPA5 proteins in TM3 cells and HSPA2 protein in TM4 cells were significantly down-regulated (P＜0.05). Conclusions HSF5 is potentially involved in spermatogenesis by regulating the expression of HSPA2 and HSPA5 in mouse Leydig cells and Sertoli cells.

Key words: heat shock transcription factor 5, heat shock transcription factors, heat shock proteins, spermatogenesis

CLC Number:

R363.1+6

ZHONG Lin, ZHONG Hongli, DAI Yujie, ZHANG Qinghua. Effect of heat shock transcription factor 5(Hsf5) knockdown on heat shock family in mouse Leydig cells and Sertoli cells[J]. Basic & Clinical Medicine, 2023, 43(4): 626-631.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/10.16352/j.issn.1001-6325.2023.04.0626

http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/Y2023/V43/I4/626

References

[1]Agarwal A, Baskaran S, Parekh N, et al. Male infertility[J]. Lancet, 2021,397:319-333.
[2]Widlak W, Vydra N. The role of heat shock factors in mammalian spermatogenesis[J]. Adv Anat Embryol Cell Biol, 2017,222:45-65.
[3]Abane R, Mezger V. Roles of heat shock factors in gametogenesis and development[J]. FEBS J, 2010,277:4150-4172.
[4]Saju JM, Hossain MS, Liew WC, et al. Heat shock factor 5 is essential for spermatogenesis in zebrafish[J]. Cell Rep, 2018,25:3252-3261.
[5]Radons J. The human HSP70 family of chaperones: where do we stand?[J]. Cell Stress Chaperones, 2016,21:379-404.
[6]Nowicka-Bauer K, Malcher A, Wloczkowska O, et al. Evaluation of seminal plasma HSPA2 protein as a biomarker of human spermatogenesis status[J]. Reprod Biol, 2022,22:100597.doi:10.1016/j.repbio.2021.100597.
[7]Lobo V, Parte P. Membrane-bound glucose regulated protein 78 interacts with alpha-2-macroglobulin to promote actin reorganization in sperm during epididymal maturation[J]. Mol Hum Reprod, 2019,25:137-155.
[8]Kajiwara C, Kondo S, Uda S, et al. Spermatogenesis arrest caused by conditional deletion of Hsp90alpha in adult mice[J]. Biol Open, 2012,1:977-982.
[9]Saribek B, Jin Y, Saigo M, et al. HSP90beta is involved in signaling prolactin-induced apoptosis in newt testis[J]. Biochem Biophys Res Commun, 2006,349:1190-1197.
[10]Asquith KL, Harman AJ, McLaughlin EA, et al. Localization and significance of molecular chaperones, heat shock protein 1, and tumor rejection antigen gp96 in the male reproductive tract and during capacitation and acrosome reaction[J]. Biol Reprod, 2005,72:328-337.
[11]钟洪莉,肖城良,史祥睿,等. 精子发生调控因子重组人HSF5的原核表达、活性检测与结构预测[J]. 基础医学与临床, 2021. 41:1323-1328.
[12]钟洪莉, 钟琳, 代宇婕, 等. 热休克转录因子5在小鼠睾丸不同细胞中的表达及其功能探索[J]. 免疫学杂志, 2022,38:890-895.
[13]Redgrove KA, Nixon B, Baker MA, et al. The molecular chaperone HSPA2 plays a key role in regulating the expression of sperm surface receptors that mediate sperm-egg recognition.[J]. PloS one, 2012,7:e50851.doi:10.1371/journal.pone.0050851.
[14]Dix DJ, Allen JW, Collins BW, et al. Targeted gene disruption of Hsp70-2 results in failed meiosis, germ cell apoptosis, and male infertility[J]. Proc Natl Acad Sci U S A, 1996,93:3264-3268.
[15]Lachance C, Bailey JL, Leclerc P. Expression of Hsp60 and Grp78 in the human endometrium and oviduct, and their effect on sperm functions[J]. Hum Reprod, 2007,22:2606-2614.
[16]Abdou HS, Villeneuve G, Tremblay JJ. The calcium signaling pathway regulates leydig cell steroidogenesis through a transcriptional cascade involving the nuclear receptor NR4A1 and the steroidogenic acute regulatory protein[J]. Endocrinology, 2013,154:511-520.
[17]Guzel E, Basar M, Ocak N, et al. Bidirectional interac-tion between unfolded-protein-response key protein HSPA5 and estrogen signaling in human endometrium[J]. Biol Reprod, 2011,85:121-127.