Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 4, pp 495–503 | Cite as

Progesterone requires heat shock protein 90 (HSP90) in human sperm to regulate motility and acrosome reaction

  • Vrushali Sagare-Patil
  • Rashmi Bhilawadikar
  • Mosami Galvankar
  • Kusum Zaveri
  • Indira Hinduja
  • Deepak Modi
Gamete Biology

Abstract

Purpose

The aims of this paper were to study whether heat shock protein 90 (HSP90) is a regulator of sperm functions and to determine its association with oligoasthenozoospermia.

Methods

The levels of HSP90 in sperm lysates were measured by ELISA. Localization of HSP90 and its isoforms was evaluated by immunofluorescence. Sperm motility and kinetics were assessed by computer-assisted sperm analysis. Acrosome reaction was determined by lectin staining.

Results

The levels of HSP90 were lower in oligoasthenozoospermic men and correlated positively with the number of motile spermatozoa. In capacitated human spermatozoa, HSP90α was mostly found in residual nuclear envelope, and the HSP90β isoform was higher in the flagella. Inhibition of HSP90 by geldanamycin or 17-AAG did not affect basal motility, but suppressed progesterone-mediated forward progressive motility, hyperactivation and acrosome reaction. Progesterone treatment dephosphorylated both HSP90α and HSP90β at Ser/Thr-Pro residues, but not Tyr residues.

Conclusion

HSP90 levels are downregulated in oligoasthenozoospermia, and its functional inhibition attenuates progesterone-mediated sperm motility and acrosome reaction.

Keywords

Heat shock protein HSP90 Human sperm Motility Acrosome reaction Progesterone Oligoasthenozoospermia Phosphorylation 

Notes

Acknowledgements

We are grateful to Dr. A S Sreedhar (Scientist, Centre for Cellular and Molecular Biology, Hyderabad, India) for generously sparing the geldanamycin and 17-AAG used in this study. The study (RA/382/06-2016) is funded by grants from the Department of Biotechnology, Innovative Young Biotechnologist Award (IYBA) and Indian Council of Medical Research (ICMR), Government of India, New Delhi, India. VSP is a recipient of a Senior Research Fellowship from ICMR. DM is a recipient of the IYBA grant.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Funding source

Indian Council of Medical Research, Department of Biotechnology, Government of India

Supplementary material

10815_2017_879_MOESM1_ESM.jpg (95 kb)
Fig. S1 Effect of HSP90 inhibitors on sperm motility kinetics. CASA was used to estimate the motility kinetics which include average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), lateral head displacement (ALH), beat cross frequency (BCF), linearity (Lin = VSL/VCL), and straightness (Str = VSL/VAP). a Effect of HSP90 inhibitors on basal motility, velocity and other sperm kinetics. b Capacitated spermatozoa were incubated with HSP90 inhibitors (geldanamycin and 17-AAG) with or without progesterone and kinematic parameters were assessed by CASA. In both the graphs, values on Y axis are mean + SE (n = 3 independent experiments) of fold change with respect to a control (untreated) sample. *Indicates value significantly different as compared to control, # indicates value is significantly different as compared to progesterone treatment (p < 0.05) (JPG 95 kb)

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Molecular and Cellular Biology LaboratoryNational Institute for Research in Reproductive Health (ICMR)MumbaiIndia
  2. 2.Hinduja IVF CentrePD Hinduja Hospital and Medical Research CenterMumbaiIndia

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