Home LiteratureArticle Details
PMID: 25541943 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Heat shock protein 90 has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-responsive sperm function in human sperm.

PloS one ·Vol. 9 ·No. 12 ·2014-00-00 ·Pages e115841

Li K, Xue Y, Chen A, Jiang Y, Xie H, Shi Q, Zhang S, Ni Y

Abstract

Heat shock protein 90 plays critical roles in client protein maturation, signal transduction, protein folding and degradation, and morphological evolution; however, its function in human sperm is not fully understood. Therefore, our objective in this study was to elucidate the mechanism by which heat shock protein 90 exerts its effects on human sperm function. By performing indirect immunofluorescence staining, we found that heat shock protein 90 was localized primarily in the neck, midpiece, and tail regions of human sperm, and that its expression increased with increasing incubation time under capacitation conditions. Geldanamycin, a specific inhibitor of heat shock protein 90, was shown to inhibit this increase in heat shock protein 90 expression in western blotting analyses. Using a multifunctional microplate reader to examine Fluo-3 AM-loaded sperm, we observed for the first time that inhibition of heat shock protein 90 by using geldanamycin significantly decreased intracellular calcium concentrations during capacitation. Moreover, western blot analysis showed that geldanamycin enhanced tyrosine phosphorylation of several proteins, including heat shock protein 90, in a dose-dependent manner. The effects of geldanamycin on human sperm function in the absence or presence of progesterone was evaluated by performing chlortetracycline staining and by using a computer-assisted sperm analyzer. We found that geldanamycin alone did not affect sperm capacitation, hyperactivation, and motility, but did so in the presence of progesterone. Taken together, these data suggest that heat shock protein 90, which increases in expression in human sperm during capacitation, has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-stimulated sperm function. In this study, we provide new insights into the roles of heat shock protein 90 in sperm function.

MeSH Terms
Adult Calcium/metabolism HSP90 Heat-Shock Proteins/analysis,metabolism Humans Male Phosphorylation Progesterone/metabolism Sperm Capacitation Sperm Motility Spermatozoa/cytology,metabolism
Chemicals
HSP90 Heat-Shock Proteins Progesterone Calcium
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Li Kun
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
Xue Yamei
Reproductive Medicine Center, Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, China.
Chen Aijun
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
Jiang Youfang
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
Xie Haifeng
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
Shi Qixian
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
Zhang Songying
Reproductive Medicine Center, Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, China.
Ni Ya
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013, China.
References (56)
56 references, click to expand
  1. Progesterone utilizes the PI3K-AKT pathway in human spermatozoa to regulate motility and hyperactivation but not acrosome reaction.
    Mol Cell Endocrinol. 2013 Jul 15;374(1-2):82-91 PMID: 23623968
  2. The role of molecular chaperones in spermatogenesis and the post-testicular maturation of mammalian spermatozoa.
    Hum Reprod Update. 2012 Jul;18(4):420-35 PMID: 22523110
  3. The decline of porcine sperm motility by geldanamycin, a specific inhibitor of heat-shock protein 90 (HSP90).
    Theriogenology. 2000 Mar 15;53(5):1177-84 PMID: 10798494
  4. Modulation of Akt kinase activity by binding to Hsp90.
    Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):10832-7 PMID: 10995457
  5. Secretory phospholipase A2 group IID Is involved in progesterone-induced acrosomal exocytosis of human spermatozoa.
    J Androl. 2012 Sep-Oct;33(5):975-83 PMID: 22240557
  6. Tyrosine phosphorylation of HSP-90 during mammalian sperm capacitation.
    Biol Reprod. 2003 Dec;69(6):1801-7 PMID: 12890735
  7. Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism.
    Asian J Androl. 2003 Mar;5(1):43-6 PMID: 12647002
  8. Hsp90 is involved in apoptosis of Candida albicans by regulating the calcineurin-caspase apoptotic pathway.
    PLoS One. 2012;7(9):e45109 PMID: 23028789
  9. Analysis of the mechanism by which calcium negatively regulates the tyrosine phosphorylation cascade associated with sperm capacitation.
    J Cell Sci. 2004 Jan 15;117(Pt 2):211-22 PMID: 14676274
  10. Inhibition of transforming activity of tyrosine kinase oncogenes by herbimycin A.
    Virology. 1988 May;164(1):294-8 PMID: 2452516
  11. Inhibition of sperm capacitation and fertilizing capacity by adjudin is mediated by chloride and its channels in humans.
    Hum Reprod. 2013 Jan;28(1):47-59 PMID: 23117128
  12. Bradykinin and ATP accelerate Ca(2+) efflux from rat sensory neurons via protein kinase C and the plasma membrane Ca(2+) pump isoform 4.
    Neuron. 2002 Jan 3;33(1):113-22 PMID: 11779484
  13. Penetration, adhesion, and fusion in mammalian sperm-egg interaction.
    Science. 2002 Jun 21;296(5576):2183-5 PMID: 12077404
  14. Different expression time of the 105-kDa protein and 90-kDa heat-shock protein in rat testis.
    FEBS Lett. 1991 Sep 2;289(1):110-2 PMID: 1893998
  15. HSP86 and HSP84 exhibit cellular specificity of expression and co-precipitate with an HSP70 family member in the murine testis.
    Dev Genet. 1993;14(2):119-26 PMID: 8482016
  16. Reactive oxygen species generation and human spermatozoa: the balance of benefit and risk.
    Bioessays. 1994 Apr;16(4):259-67 PMID: 8031303
  17. Exocytosis in spermatozoa in response to progesterone and zona pellucida.
    Science. 1994 Dec 2;266(5190):1578-81 PMID: 7985030
  18. Progesterone receptor structure and function altered by geldanamycin, an hsp90-binding agent.
    Mol Cell Biol. 1995 Dec;15(12):6804-12 PMID: 8524246
  19. Localization of heat shock proteins in mouse male germ cells: an immunoelectron microscopical study.
    Exp Cell Res. 1996 Nov 25;229(1):77-85 PMID: 8940251
  20. Amino acid neurotransmitter receptor/chloride channels of mammalian sperm and the acrosome reaction.
    Biol Reprod. 1997 Mar;56(3):569-74 PMID: 9046998
  21. Effects of gamma-aminobutyric acid on human sperm motility and hyperactivation.
    Mol Hum Reprod. 1996 Oct;2(10):733-8 PMID: 9239690
  22. Human sperm capacitation induced by biological fluids and progesterone, but not by NADH or NADPH, is associated with the production of superoxide anion.
    J Androl. 1998 Mar-Apr;19(2):215-25 PMID: 9570746
  23. Repression of heat shock transcription factor HSF1 activation by HSP90 (HSP90 complex) that forms a stress-sensitive complex with HSF1.
    Cell. 1998 Aug 21;94(4):471-80 PMID: 9727490
  24. Effect of seminal plasma on capacitation and hyperactivation in human spermatozoa.
    Hum Reprod. 1998 Aug;13(8):2139-46 PMID: 9756285
  25. Human sperm chemotaxis: is progesterone a chemoattractant?
    Biol Reprod. 1999 Jun;60(6):1314-9 PMID: 10330086
  26. Association of NASP with HSP90 in mouse spermatogenic cells: stimulation of ATPase activity and transport of linker histones into nuclei.
    J Biol Chem. 2005 Jan 28;280(4):2904-11 PMID: 15533935
  27. Reassessing the role of progesterone in fertilization--compartmentalized calcium signalling in human spermatozoa?
    Hum Reprod. 2005 Oct;20(10):2675-80 PMID: 15980011
  28. Whole-cell patch-clamp measurements of spermatozoa reveal an alkaline-activated Ca2+ channel.
    Nature. 2006 Feb 9;439(7077):737-40 PMID: 16467839
  29. Mammalian sperm translate nuclear-encoded proteins by mitochondrial-type ribosomes.
    Genes Dev. 2006 Feb 15;20(4):411-6 PMID: 16449571
  30. Geldanamycin prevents hemorrhage-induced ATP loss by overexpressing inducible HSP70 and activating pyruvate dehydrogenase.
    Am J Physiol Gastrointest Liver Physiol. 2006 Jul;291(1):G117-27 PMID: 16565416
  31. Hsp90 inhibition transiently activates Src kinase and promotes Src-dependent Akt and Erk activation.
    Proc Natl Acad Sci U S A. 2006 Jul 25;103(30):11318-22 PMID: 16844778
  32. Sperm competition enhances functional capacity of mammalian spermatozoa.
    Proc Natl Acad Sci U S A. 2006 Oct 10;103(41):15113-7 PMID: 16990431
  33. Acrosome reaction induced by recombinant human zona pellucida 3 peptides rhuZP3a22 approximately 176 and rhuZP3b177 approximately 348 and their mechanism.
    J Androl. 2007 May-Jun;28(3):381-8 PMID: 17192598
  34. Analysis of chaperone proteins associated with human spermatozoa during capacitation.
    Mol Hum Reprod. 2007 Sep;13(9):605-13 PMID: 17595329
  35. Geldanamycin augments nitric oxide production and promotes capacitation in boar spermatozoa.
    Anim Reprod Sci. 2008 Feb 1;104(1):56-68 PMID: 17280805
  36. Protein synthesis in sperm: dialog between mitochondria and cytoplasm.
    Mol Cell Endocrinol. 2008 Jan 30;282(1-2):45-55 PMID: 18226442
  37. Comparative immunolocalization of heat shock proteins (Hsp)-60, -70, -90 in boar, stallion, dog and cat spermatozoa.
    Reprod Domest Anim. 2008 Aug;43(4):385-92 PMID: 18226022
  38. Chloride Is essential for capacitation and for the capacitation-associated increase in tyrosine phosphorylation.
    J Biol Chem. 2008 Dec 19;283(51):35539-50 PMID: 18957426
  39. Role of translation by mitochondrial-type ribosomes during sperm capacitation: an analysis based on a proteomic approach.
    Proteomics. 2009 Mar;9(5):1385-99 PMID: 19253287
  40. Nongenomic activation of spermatozoa by steroid hormones: facts and fictions.
    Mol Cell Endocrinol. 2009 Sep 24;308(1-2):39-46 PMID: 19549590
  41. The effect of anti-eppin antibodies on ionophore A23187-induced calcium influx and acrosome reaction of human spermatozoa.
    Hum Reprod. 2010 Jan;25(1):29-36 PMID: 19801569
  42. CFTR is essential for sperm fertilizing capacity and is correlated with sperm quality in humans.
    Hum Reprod. 2010 Feb;25(2):317-27 PMID: 19923167
  43. Heat shock protein and heat shock factor expression in sperm: relation to oligozoospermia and varicocele.
    J Urol. 2010 Mar;183(3):1248-52 PMID: 20096881
  44. HSP90 at the hub of protein homeostasis: emerging mechanistic insights.
    Nat Rev Mol Cell Biol. 2010 Jul;11(7):515-28 PMID: 20531426
  45. The HSP90AA1 sperm content and the prediction of the boar ejaculate freezability.
    Theriogenology. 2010 Oct 1;74(6):940-50 PMID: 20580074
  46. Protein homeostasis and the phenotypic manifestation of genetic diversity: principles and mechanisms.
    Annu Rev Genet. 2010;44:189-216 PMID: 21047258
  47. The molecular chaperone Hsp90α is required for meiotic progression of spermatocytes beyond pachytene in the mouse.
    PLoS One. 2010;5(12):e15770 PMID: 21209834
  48. Hsp90β1 knockout targeted to male germline: a mouse model for globozoospermia.
    Fertil Steril. 2011 Mar 15;95(4):1475-7.e1-4 PMID: 21208614
  49. Progesterone activates the principal Ca2+ channel of human sperm.
    Nature. 2011 Mar 17;471(7338):387-91 PMID: 21412339
  50. Developmental expression of heat shock proteins 60, 70, 90, and A2 in rabbit testis.
    Cell Tissue Res. 2011 May;344(2):355-63 PMID: 21538228
  51. Ion channels, phosphorylation and mammalian sperm capacitation.
    Asian J Androl. 2011 May;13(3):395-405 PMID: 21540868
  52. Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm.
    J Androl. 2011 Jul-Aug;32(4):402-10 PMID: 21088232
  53. Mathematical modeling of calcium signaling during sperm hyperactivation.
    Mol Hum Reprod. 2011 Aug;17(8):500-10 PMID: 21606121
  54. Conventional progesterone receptors (PR) B and PRA are expressed in human spermatozoa and may be involved in the pathophysiology of varicocoele: a role for progesterone in metabolism.
    Int J Androl. 2011 Oct;34(5 Pt 1):430-45 PMID: 20946440
  55. Modelling the regulation of thermal adaptation in Candida albicans, a major fungal pathogen of humans.
    PLoS One. 2012;7(3):e32467 PMID: 22448221
  56. Substantial decrease of heat-shock protein 90 precedes the decline of sperm motility during cooling of boar spermatozoa.
    Theriogenology. 1999 Apr 1;51(5):1007-16 PMID: 10729022
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0115841
Published
2014-00-00
Epub
2014-00-26
Pages
e115841
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC4277372
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0115841 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2