Perfringolysin O as a useful tool to study human sperm physiology

Capsule:
Perfringolysin O selectively permeabilizes the plasma membrane of human spermatozoa and can be used to assess changes in the amount and distribution of the active cholesterol fraction present in this membrane.

Authors:
Cristián A. Pocognoni, M.S., Gerardo A. De Blas, Ph.D., Alejandro P. Heuck, Ph.D., Silvia A. Belmonte, Ph.D., Luis S. Mayorga, Ph.D.

Volume 99, Issue 1, Pages 99-106.e2, January 2013

Abstract:

Objective:
To evaluate perfringolysin O, a cholesterol-dependent pore-forming cytolysin, as a tool to study several aspects of human sperm physiology.

Design:
Prospective study.

Setting:
Basic research laboratory.

Patient(s):
Human semen samples with normal parameters obtained from healthy donors.

Interventions:
Interaction of recombinant perfringolysin O with human spermatozoa.

Main Outcome Measures:
Assessment of perfringolysin O binding to spermatozoa. Tests for acrosome and plasma membrane integrity, and acrosomal exocytosis assays.

Results:
Perfringolysin O associated to human spermatozoa at 4ºC. The binding was sensitive to changes in cholesterol concentrations and distribution occurring in the plasma membrane of these cells during capacitation. When perfringolysin O-treated sperm were incubated at 37ºC, the plasma membrane became permeable whereas the acrosome membrane remained intact. Permeabilized spermatozoa were able to respond to exocytic stimuli. The process was inhibited by proteins that interfere with membrane fusion, indicating that large molecules, including antibodies, were able to permeate into the spermatozoa.

Conclusions:
PFO is a useful probe to assess changes in the amount and distribution of the active sterol fraction present in the sperm plasma membrane. The toxin can be used for the efficient and selective permeabilization of this membrane rendering a flexible experimental model suitable for studying molecular processes occurring in the sperm cytoplasm.

  • NicoGarrido

    Congratulations Dr Pocognoni for your nice work

    under the experimental viewpoint, the design is impeccable.
    I was wondering if you ahve any information of the use of PFO as a sperm quality marker in ART. TO this end, have you established any relationship between reproductive results and samples PFO binding.
    Moreover, given the relevance of cholesterol on sperm cryosurvival, have you related PFO binding levels with sperm cryosurvival rates?
    I think this could be a nice and interesting line
    all the best!

    • Dear Dr Garrido, thanks for your interest in our work and sorry for the delay.
      Sincerly we haven´t tried PFO as quality marker in ART because we do basic science.
      Regarding to sperm cryosurvival we haven´t tested that. I guess PFO binding patterns could be definitely proportional to the sperm cryosurvival rates and could be check for this purpose.
      If you have another question be free to ask to us.

      Best regards.

      Cristian Pocognoni

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