Epididymal Sperm Count

Yefan Wang1

1 TherImmune Research Corporation, Gaithersburg, Maryland
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 16.6
DOI:  10.1002/0471140856.tx1606s14
Online Posting Date:  February, 2003
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Abstract

Making Epididymal Sperm Counts (Yefan Wang, TherImmune Research Corporation, Gaithersberg, Maryland). Epididymal sperm counts are a widely used, simple and sensitive method of assessing the effects of male reproductive toxicants on the epididymal and/or testicular site of action. After careful dissection of the tissues and further processing, the sperm suspensions are counted using a hemacytometer and analyzed for effect.

     
 
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Table of Contents

  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1:

  Materials
  • PBS ( appendix 2A)
  • Necropsied male rats or mice, treated and control, euthanized
  • 15 × 60–mm (rats) or 15 × 30–mm (mice) and 1.5 × 2.0–cm petri dishes
  • Slide warmer
  • 13 × 100–mm test tubes and appropriate rack
  • Microscope: 10× and 40× objectives and 10× or 15× eyepieces
  • Necropsy board
  • 0.1‐mm‐deep improved Neubauer hemacytometer and coverslips
  • No. 11 scalpel blades
  • 60°C water bath
  • Gauze
  • Hand tally counter
  • Data collection system (i.e., hand‐generated data form or computer‐generated data collection system)
  • Additional reagents and equipment for using a hemacytometer ( appendix 3B)
NOTE: All protocols using live animals must first be reviewed and approved by an Institutional Animal Care and Use Committee (IACUC) and must follow officially approved procedures for the care and use of laboratory animals.
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Figures

Videos

Literature Cited

Literature Cited
   Hamilton Thorne Research 1989. HTM‐IVOS Toxicology Application Version 10.8 Operation Manual. Hamilton Thorne Biosciences, Beverly, Mass.
   Kempinas, W.G. and Lamano‐Carvalho, T.L. 1988. A method for estimating the concentration of spermatozoa in the rat cauda epididymis. Lab. Anim. 22:154‐156.
   Klassen, C.D. 1996. Fifth Casarett & Doull's Toxicology: The Basic Science of Poisons, Fifth Edition. McGraw‐Hill, New York.
   Saksena, S.K., Lau, I‐F., and Chang, M‐C. 1979. Age dependent changes in the sperm population and fertility in the male rat. Exp. Aging Res. 5:373‐381.
   Ratnasooriya, W.D. and Wadsworth, R.M. 1987. Effect of mating on sperm distribution in the reproductive tract of the male rat. Gamete Res. 17:261‐266.
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