The blood-testis barrier is believed to be constituted by tight junctions between Sertoli cells in seminiferous tubules and possibly by myoid cells that encircle these tubules. We now show that testis microvessels are endowed with several markers of barrier properties of brain microvessels, such as the glucose transporter, P-glycoprotein, and gamma-glutamyl transpeptidase. Quantitative EM studies show that the endothelium in testis, as in brain, is continuous and has long junctional profiles and few vesicles. However, a small proportion of testis capillaries have expansions in their junctional clefts suggestive of patent paracellular channels, which may explain their higher permeability. Because barrier features are thought to be induced and/or maintained in brain microvessels by astrocytes, we assessed whether astrocyte-like cells exist in the testis. We found that the intertubular Leydig cells, adjacent to microvessels, express the astrocyte markers: glial fibrillary acidic protein, glutamine synthetase, and S-100 protein. We suggest that the testis endothelium contributes to the blood-testis barrier and that these endothelial barrier features are influenced by Leydig cells. We believe that the endothelial and the epithelial (Sertoli) components of the blood-testis barrier are "in series" and complement each other in achieving a stable milieu for spermatogenesis.