Summary
This paper describes methods for the cytochemical demonstration of non-specific esterase and acid phosphatase activities in spermatozoa of the mouse. The acetate and phosphate esters of 1-naphthol were used as substrates, and hexazonium pararosanilin was used as coupler in both techniques. Specificity was assured by the use of appropriate positive and negative controls. Best results were obtained with unfixed smears which had been stored at room temperature for a few days prior to use. Chemical fixation, especially with formol-calcium solutions, is not recommended for use with rodent spermatozoa. According to our investigations the murine acrosome contains very low levels of non-specific esterase and acid phosphatase which are not amenable to detection by the “standard” methods employing short incubations and/or with material fixed in a formalin-containing fixative.
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These studies have been supported by funds from the office of General Research, The University of Georgia.
The authors thank Dr. J. Travis for gifts of trypsin inhibitors, Dr. H. A. Kent for the provision of Syrian hamsters, and Dr. W. J. Humphreys, Director, Electron Microscopy Laboratory, The University of Georgia, for use of photographic facilities. The senior author is indebted to Drs. Ellen M. Rasch and L. Ornstein for helpful discussions and advice concerning the development of the techniques described in this paper.
Dr. Unnithan was the recipient of a postdoctoral award from the University of Georgia.
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Bryan, J.H.D., Unnithan, R.R. Cytochemical localization of non-specific esterase and acid phosphatase in spermatozoa of the mouse (Mus musculus). Histochemie 33, 169–180 (1972). https://doi.org/10.1007/BF00305744
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DOI: https://doi.org/10.1007/BF00305744