Comparison of streptozotocin-induced diabetic and insulin resistant effects on spermatogenesis with proliferating cell nuclear antigen (PCNA) immunostaining of adult rat testis

Abstract

Male reproductive function is one of the mammalian systems that are impaired by diabetes mellitus and on the average, it will affect many more men prior to and during their reproductive years. The present study aimed to compare proliferating cell nuclear antigen (PCNA) immunostaining effects at stage VII of the spermatogenic cycle in streptozotocin-induced and insulin resistant diabetic rat testis. Male adult Sprague-Dawley rats (120-140 g) were randomly divided into 3 groups. Group 1: control group; fed on normal rat pellets. Group 2: streptozotocin diabetic group; received a single dose IP injection of streptozotocin 45 mg/kg BW in Na+ citrate buffer pH 4.5. Group 3: insulin resistant diabetic group; fed ad libitum on a special diet containing 25% fructose W/W. Following hyperglycaemia confirmation, animals were perfused with 4% Paraformaldehyde (PFA). Testes were isolated, and fixed in 4% PFA overnight, embedded in paraffin, 5μm thick sections were made and mounted on poly-L-lysine coated slides. Immunohistochemical staining was carried out on the positively charged slides using the PCNA as primary antibody.The results of this study showed that diabetes mellitus induced by streptozotocin injection and chronic fructose consumption through insulin resistance significantly (P<0.05) reduced PCNA index, mean seminiferous tubular diameter and testicular diameter. It could also be inferred from the results that diabetes mellitus and insulin resistance may necessarily not impair differentiation of primordial germ cells (gonocytes) into Type A and Type B spermatogonia. J. Exp. Clin. Med., 2012; 29:209-214