Methods for assessing rat sperm motility

doi:10.1016/0890-6238(92)90183-T

Reproductive Toxicology

Volume 6, Issue 3, 1992, Pages 267-273

https://doi.org/10.1016/0890-6238(92)90183-T Get rights and content

Abstract

Computer-assisted sperm analysis (CASA) systems are becoming more widely used. With this spread to technology come more data from toxicology studies, designed to determine if treatment with putative toxicants affects sperm motion parameters. While these CASA methods provide us with more ways to evaluate toxicity and thus perhaps increase our chances of successfully protecting human health, there is also a greater likelihood that different laboratories will use different methods of collecting data on sperm motility. Different systems used with different methods in different laboratories will inevitably generate data that are difficult to compare. In a prospective attempt to address this issue of comparability and limit the problems, a group of individuals using CASA systems to analyze rat sperm motility convened to discuss methodologic issues, share data, and try to reach a consensus about methods for performing these studies. This article shares those meetings and data in the hope that common methods will enhance interlaboratory comparisons.

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Citation Excerpt :
Sperm from the cauda epididymis were collected, and motility was evaluated by Computer Assisted Sperm Analysis (Hamilton Thorne Research, Beverly, USA) at 37 °C in medium 199 (Sigma Aldrich Chemical Co.) [44,45]; morphology was evaluated with eosin staining. Spermatozoan number (from cauda epididymis) and sperm head count (from total testis lysate) were evaluated by light microscopy [46]. Daily sperm production (DSP) was calculated by dividing the total number of spermatids/sperm per gram testis by 6.1 days, the lifetime of step 19 spermatids in the seminiferous tubules [47].
Reproductive functions are controlled by a finely tuned balance between estrogens and androgens. To further characterize the gonadal pathways leading to hormonal balance disruption by atrazine, vinclozolin, methoxychlor, and bisphenol A in rat, we investigated their effects in male and female young adult animals. Specifically, we assessed reproductive tract alterations, sex hormone balance in serum and gonads, tissue dosimetry, and mRNA expression. Remarkably, we observed different aromatase regulation profiles between animals with similar estrogen-to-androgen ratios but with different chemical treatments. For example, increased estrogen-to-androgen ratios in atrazine-treated females could be partly linked to aromatase upregulation, while in methoxychlor- and bisphenol A-treated females, peripheral mechanisms such as conjugation/deconjugation processes might be more likely to elevate estrogen levels. In vinclozolin-treated animals, the decreased estrogen-to-androgen ratios reported might be due to an increase of peripheral (adrenal) steroidogenesis. Thus, measurement of many endpoints is necessary for good risk assessment.
Measuring mouse sperm parameters using a particle counter and sperm quality analyzer: A simple and inexpensive method
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This study examined a method for analyzing the count, motility, and morphology of mouse epididymal sperm, optimizing the diluent, incubation time, sample concentration, and temperature, using a particle counter (CDA-500) to count and size sperm and a sperm quality analyzer (SQA-IIC) to measure sperm motility, quantified as the sperm motility index (SMI). The optimal conditions consisted of a 30-min incubation in D-MEM (Dulbecco's modified Eagle's medium; considering cost and availability) at 37 °C, with 5 × 10⁶ cells mL⁻¹ in the original solution. Furthermore, the influence of formalin fixation, and the correlation between the automated counter and a manual method were investigated. The sample fixation had no marked effect on the sperm count or morphology assessment. A linear correlation was observed between the manual and automated methods (y = 0.920x + 0.276; r² = 0.571; p < 0.001; range: (3–6) × 10⁶). The suitability of the proposed method was confirmed using spermatozoa prepared from mice treated with the reproductive toxin diethylstilbestrol (DES). Using sperm from the cauda epididymidis on one side per mouse, we confirmed that measurement of these sperm parameters using the two devices was simple, rapid, inexpensive, and reproducible.
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Sperm quality of the sea urchin, Anthocidaris crassispina, after exposure to environmentally realistic UV-B irradiances, was assessed by changes in sperm motility (measured by the computer-assisted sperm analysis (CASA) system), and related to subsequent fertilization success. Percentage motile sperm of A. crassispina declined significantly after exposure to a UV-B dose of 16.2 $kJ m^{−2}$ , while sperm motion velocity as measured by curvilinear velocity (VCL), straight line velocity (VSL), and average path velocity (VAP) showed significant reduction after exposure to a UV-B dose of 5.4 $kJ m^{−2}$ . A parallel study showed that fertilization success was significantly reduced after sperm were exposed to UV-B doses $⩾5.4 kJ m^{−2}$ . Notably, the four sperm motility parameters were strongly correlated with fertilization success (P<0.001), followed the increasing order: VSL $(r=0.8)<% motile sperm (r=0.804)< VCL (r=0.912)< VAP (r=0.928)$ . Fertilization success is best predicted by VAP using the exponential model: y=8.678+90.202/[1+exp(82.83−x)/10.27)] (r²=0.95). Thus, impairment of sperm motility of sea urchin, as measured by the CASA method, can be used to predict reproductive success and ecological effects.
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^☆: Disclaimer: The research described in this article has been reviewed viewed by the Health Effects Laboratory, US Environment Protection Agency, and approved for publication. Approval does not signify that the contents necessarily reflect the views and policies of the Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use.

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Special Consensus PapersMethods for assessing rat sperm motility☆

Abstract

Fundam Appl Toxicol

Fertil Steril

Fertil Steril

Fertil Steril

Fertil Steril

Comut Meth Prog Biomed

Fundam Appl Toxicol

Fertil Steril

Fertil Steril

Fundam Appl Toxicol

Reprod Toxicol

Reprod Toxicol

Reprod Toxicol

fertil Steril

Fertil Steril

A reexamination of motility patterns of rat epididymal spermatozoa

Anat Rec

Rat sperm are mechanically immobilized in the cauda epididymis by “immobilin”, a high molecular weight glycoprotein

Biol Reprod

Special Consensus Papers
Methods for assessing rat sperm motility☆