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Abstract
This study was performed to understand the regional distribution of superoxide anion radicals and hydrogen peroxide within the spermatozoa of mice during both normal and altered situations of epididymal maturation. The intracellular O2- levels were probed employing dichlorodihydrofluorescein diacetate (DDF) as a reporter. The testicular spermatozoa from normal animals showed strong regional heterogeneity in the DDF fluorescence patterns over the various domains. Vasectomy resulted in strong inhibition in the O2- levels of spermatozoa at all stages of maturation. Interestingly, the fluorescein diacetate staining pattern was strong in the head of spermatozoa from the testis, caput, corpus, and cauda region. Further, there was a progressive reduction in the fluorescence in the head region in the spermatozoa toward the cauda region. The midpiece and tail showed moderate fluorescence, which also diminished as the spermatozoa matured. The spermatozoa from the vas deferens exhibited a weak fluorescence over the head domain, with the other domain staining extremely weak. Here again, vasectomy introduced considerable loss in the fluorescence intensity. The implications of programmed production of reactive oxygen species in specific domains of the spermatozoa during various stages of development are discussed.