Heat shock factor 5 is essential for spermatogenesis in mice: Detected by a new monoclonal antibody

Document Type : Original Article


1 Department of Cell and Molecular Biology, School of Biological Sciences, Kharazmi University, Tehran, Iran

2 Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

3 Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran


Objective(s): Here, we examined the function of our produced monoclonal antibody (mAb10C3) to recognize one of the most important members of the HEAT shock factor family, Hsf5, in embryonic development and in spermatogenic cells of adult mouse testis.
Materials and Methods: The targeting effects of mAb10C3 were investigated by immunohistochemistry analysis in the different phases of the embryo and in the adult testis tissue sections.
Results: The results of immunohistochemistry staining on the mouse embryos by the supernatant of hybridoma clone that produced mAb10C3, in the early and late phases (E7.5 and E14.5) of embryonic development, indicated that mAb10C3 could only detect Hsf5 in E7.5 and it did not have any targeting activity in the late phase of development. Therefore, we showed that the hsf5 gene has expressed in early mouse embryonic development. On the other hand, mAb10C3 could detect Hsf5 in spermatogonia and spermatocytes of adult testis in comparison with a known anti-Hsf5 antibody (ab98939) and an anti-PCNA antibody as a marker of spermatogonia cells.
Conclusion: Taken together, these data indicated that generated anti-testis mAb10C3 was generated against anti-testis proteins, specifically to target Hsf5, and can be useful as a scientific tool to investigate the critical genes in the development and spermatogenesis.


1.Naidu SD, Dinkova‐Kostova AT. Regulation of the mammalian heat shock factor 1. FEBS J 2017; 284:1606-1627.
2.Fung PC, Kong RK. The heat shock protein story-from taking mTORC1, 2 and heat shock protein inhibitors as therapeutic measures for treating cancers to development of cancer vaccines. Journal of Cancer Ther 2017; 8:962-1029.
3.Sarge KD. Male germ cell-specific alteration in temperature set point of the cellular stress response. J Biol Chem 1995; 270:18745-18748.
4.Abane R and Mezger V. Roles of heat shock factors in gametogenesis and development. FEBS J 2010; 277:4150-4172.
5.Gomez-Pastor R, Burchfiel ET, Thiele DJ. Regulation of heat shock transcription factors and their roles in physiology and disease. Nat Rev Mol Cell Biol 2018; 19:4.
6.Saju JM, Hossain MS, Liew WC, Pradhan A, Thevasagayam NM, Tan LSE, et al. Heat shock factor 5 is essential for Spermatogenesis in Zebrafish. Cell Rep 2018; 25:3252-3261.
7.Sikes RS, Gannon WL. Guidelines of the American Society of mammologists  for the use of wild mammals in research. J Mammal 2011; 92:235-253.
8.Vanage G, Lu YA, Tam JP, Koide SS. Infertility induced in rats by immunization with synthetic peptide segments of a sperm protein. Biochem Biophys Res Commun 1992; 183:538-543.
9.Sakakibara K, Sato KI, Iwasaki T, Kitamura K, Fukami Y. Generation of an antibody specific to Xenopus fertilized eggs by subtractive immunization. Genes Cells 2005; 10:345-356.
10.Hemati A, Azarnia M, Modarressi MM, Rahimi A. Obtaining and characterization of anti-testis monoclonal antibodies: Invaluable tools toward the identification of testis antigens involved in fertilization. Hum antibodies 2018; 26: 209-218.
11.Ditzel H, Erb K, Borup-Christensen P, Nielsen B, Jensenius JC. Evaluation of procedures for the fixation and processing of human tissue for immunohistochemical analysis of human monoclonal antibodies. Hum Antibodies 1991; 2:135-141.
12.Jassim A, Festenstein H. Immunological and morphological characterisation of nucleated cells other than sperm in semen of oligospermic donors. J Reprod Immunol 1987; 11: 77-89.
13.Eggers S, Ohnesorg T, Sinclair A. Genetic regulation of mammalian gonad development. "Nat. Rev. Endocrinol. 2014; 10:673.
14.Wilhelm D, Palmer S, Koopman P. Sex determination and gonadal development in mammals. Physiol Rev 2007; 87:1-28.
15.Biason-Lauber A. Control of sex development. Best Pract Res Clin Endocrinol Metab 2010; 24:163-186.
16.Leal MC, Cardoso ER, No´ brega RH, Batlouni SR, Bogerd J, Franc LR, et al. Histological and stereological evaluation of zebrafish (Danio rerio) spermatogenesis with an emphasis on spermatogonial generations. Biol Reprod 2009; 81:177-187.
17.Chowdhury AK and Steinberger E. Early changes in the germinal epithelium of rat testes following exposure to heat. J Reprod Fert 1970; 22: 205-212.
18.Mieusset R and Bujan, L. Testicular heating and its possible contributions to male infertility: a review. Int J Androl 1995; 18:169-184.
19.Fujimoto M and Nakai A. The heat shock factor family and adaptation to proteotoxic stress. FEBS J 2010; 277:4112-4125.
20.Bjo ̈rk JK and Sistonen L. Regulation of the members of the mammalian heat shock factor family. FEBS J 2010; 277:4126-4139.
21.Yamamoto N, Takemori Y, Sakurai M, Sugiyama K, Sakurai H. Differential recognition of heat shock elements by members of the heat shock transcription factor family. FEBS J 2009; 276: 1962-1974.
22.Morimoto RI. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones and negative regulators. Genes Dev 1998; 12: 3788-3796.
23.Park CJ and Seo YS. Heat shock proteins: A review of the molecular chaperones for plant immunity. Plant Pathol J 2015; 31:323-333.
24.MacPhee DJ. The Role of Heat Shock Proteins in Reproductive System Development and Function. Springer; 2017.
25.Jarvis S, Elliott DJ, Morgan D, Winston R, Readhead C. Molecular markers for the assessment of postnatal male germ cell development in the mouse. Fertil Steril 2004; 82:S6.