Lactate is provided to spermatogenic cells by Sertoli cells as an energy substrate and its transport is regulated by H+-monocarboxylate co-transporters (MCTs). In the case of several cell types it is known that MCT1 is associated with basigin and MCT2 with embigin. Here we demonstrate co-localization and co-immunoprecipitation of basigin with both MCT1 and MCT2 in sperm, whereas no interaction with embigin was detectable. An investigation of the functional activity of MCT proteins revealed that it was mainly the application of L-lactate which resulted in a decrease in pHi. The pHi changes were blocked with α-cyano-4-OH cinnamate and the preference for L-lactate—as opposed to D-Lactate—was demonstrated by the determination of ATP after exposure to both lactate isomers. We propose that basigin interacts with MCT1 and MCT2 to locate them properly in the membrane of spermatogenic cells and that this may enable sperm to utilize lactate as an energy substrate contributing to cell survival. J. Cell. Physiol. 227: 2154–2162, 2012. © 2011 Wiley Periodicals, Inc.