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On stage single cell identification of rat spermatogenic cells
Author(s) -
Reyes Juan G,
Diaz Alvaro,
Osses Nelson,
Opazo Carlos,
Benos Dale J
Publication year - 1997
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/s0248-4900(99)80081-1
Subject(s) - biology , identification (biology) , stage (stratigraphy) , microbiology and biotechnology , cell , computational biology , genetics , ecology , paleontology
Summry— The study of spermatogenic cell physiology has been hindered by the absence of unbiased methods of identification of cells upon which single cell techniques are being applied. In this work, we have used histochemical techniques, digital videoimaging, quantification of chromatin‐bound DNA probes, and measurements of cell diameter to identify single spermatogenic cells at different periods of development. Our criteria of identification permit the definition of four developmental stages of spermatogenesis on which to perform single cell analyses: spermatogonia B/preleptotene spermatocytes, leptotene/zygotene spermatocytes, pachytene spermatocytes, and round spermatids. The use of voltage‐sensitive dyes and Ca 2+ ‐sensitive dyes does not interfere with the estimations of DNA content. The estimations of DNA content of spermatogenic cells can be performed both with near‐UV exciged dyes (H 33342 ) and long wavelength‐excited dyes (ethidium bromide), allowing the use of a wide range of physiological and immunocytochemical fluorescent probes to study the spermatogenic process.