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Fig. 1 | Reproductive Biology and Endocrinology

Fig. 1

From: Flutamide induces alterations in the cell-cell junction ultrastructure and reduces the expression of Cx43 at the blood-testis barrier with no disturbance in the rat seminiferous tubule morphology

Fig. 1

a-f Testis morphology (a-b), apoptosis (c-d) and proliferation (e-f) in control and flutamide-treated rats. a-b Morphology of the testis. Representative section of control (a) and flutamide-treated (b) rats. H-E staining. Scale bars represent 15 μm. Note full spermatogenesis in seminiferous tubules (ST) of both control and flutamide-treated testes (a-b) and enlarged interstitial tissue (IT) after flutamide exposure (b). Representative semi-thin section of control (a’) and flutamide-treated rats (b’). Scale bars represent 15 μm. Both cross-sections (a’, b’) illustrate different cell types present during spermatogenesis. From the basal membrane to the lumen: spermatogonia (sg), spermatocytes (sp), round spermatids (r.s) and elongated spermatids (e.s) are visible. Sertoli cell (Sc) nucleus is situated close to the basal membrane, while Sertoli cell cytoplasm extends towards the lumen being in close contact with the other cell types. Note any loss of germ cells within the seminiferous epithelium (a’, b’). Abundant distribution of ectoplasmic specializations (arrows) close to the basal membrane of control testis (a’). Note the reduction of the cell surface occupied by the basal ectoplasmic specializations connecting adjoined Sertoli cells (arrow) following flutamide treatment (b’). C-F. TUNEL staining, caspase 3 protein, and proliferating nuclear antigen (PCNA) expression in testes of control and flutamide-treated rats. Representative TUNEL staining (arrow, c). Scale bars represent 15 μm. Immunoreactive cells were absent in negative control sections (c, top). Quantitative analysis of TUNEL-positive cells (d). Representative immunoblots for caspase 3 (e, top) and PCNA (f, top). The band at 32 kDa represents inactive proenzyme (procaspase 3) and the band at 17 kDa corresponds to active form (cleaved caspase 3). Actin was used as a loading control, and each set of shown actin immunoblots corresponds to the target protein that was investigated within a given panel. The relative level of cleaved caspase 3 (e, bottom) and PCNA (f, bottom) protein normalized against its corresponding β-actin. Data obtained from three separate analyses is expressed as mean ± SD. Control (n = 6) and flutamide (n = 6) group

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Reproductive Biology and Endocrinology

ISSN: 1477-7827

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