Flow cytometric evaluation of the effects of doxorubicin on rat spermatogenesis

Histopathologic examination of testicular tissue allows testicular impairment to be investigated. As an alternative to histopathology, flow cytometry (FCM) using a triple staining technique that combines DNA-ploidy with mitochondria stainability and vimentin immunostaining has also been utilized to evaluate testicular damage. In this article we evaluate the effects on spermatogenesis after acute exposure of rats to doxorubicin. Testicular cell suspensions of treated and control animals were analyzed by FCM. This allows several cell types to be identified and quantified, giving a control pattern. Deviations from this control pattern are considered as an indication of testicular damage. Doxorubicin produced a depletion of spermatogonia as early as 3 d after treatment. This effect could be followed through the temporal evolution of spermatogenesis. Comparable results were obtained by histopathology. The presented results show that FCM is a suitable and sensitive method for the detection of testicular damage. The advantages of FCM over other techniques include its rapidity and objectivity.