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Volume 1, Issue 2, Number 2, April-June 2000


Evaluation of Rat Sperm Motility by CASA


Mohammad Mehdi Akhondi Corresponding Author
Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran

Hari Moore
University of Sheffield, Sheffield, United Kingdom

Abstract
Introduction: When mammalian spermatozoa first enter the epididymis from testis they
display little or no progressive motility. On transit along the epididymis, spermatozoa
gradually acquire the potential for full progressive motility. This process which begins
after sperm leave the testis, is called sperm maturation and requires specific factors from
the epididymal epithelium. The aim of this study was to investigate the use of
computer-assisted sperm analysis for the objective determination of rat sperm motility.
Having validated this system, it would be used to assess in vitro and in vivo sperm
maturation.Material and methods: Sperm samples from various regions of the epididymis and
ejaculates, were prepared. Primary cultures of the epithelium from the rat epididymis
were established. A novel computer-aided sperm analysis was used to analyze and
characterize the motility of the rat spermatozoa during in vitro and in vivo maturation.
Sperm motility analysis also examined during co-culture. Image analyses of sperm
movement from video recording tapes were also examined.
Results: This study used a novel CASA system and image analysis to monitor the
changes in motility as spermatozoa undergo maturation in the rat epididymis, during
ejaculation and during co-incubation with epididymal epithelial cultures and medium
preparations. It is interesting to note that changes in epididymal sperm motility were
occurred in vivo and in vitro. The co-incubation of immature rat spermatozoa from
the caput epididymis with epithelial cultures led to significant changes in progressive
sperm motility parameters associated with maturation.Conclusion: This study indicate the CASA and other image analysis techniques, were
able to characterize in detail the objective determination and changes in motility
parameters of rat spermatozoa. Validation of this system provides the best facility to
assess rat sperm maturation in vitro.




Keywords: rat spermatozoa, CASA system, image analyzer, epididymal sperm maturation, in vitro sperm maturation


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