The influence of germ cells (greater than 85% pachytene spermatocytes) on vectorial secretion of androgen binding protein (ABP) and transferrin by immature rat Sertoli cells was investigated using two-compartment culture chambers. The ratio of ABP secreted into the outer and inner compartment in control cultures of Sertoli cells alone was 1.9, and was not influenced by either FSH or testosterone. Co-culture of Sertoli cells in direct contact with germ cells in the presence of FSH decreased this ratio, the decrease being most pronounced (0.7) after 2 days of co-culture. This effect was not observed if the germ cells were not in direct contact with Sertoli cell monolayers. The outer to inner compartment ratio of transferrin in Sertoli cell-alone cultures was 1.6 and, in contrast to ABP, was not significantly influenced by the addition of germ cells, even in the presence of FSH. It is concluded that in immature rat Sertoli cells the polarity of ABP secretion, but not that of transferrin, may be regulated by pachytene spermatocytes (and possibly other germ cells), and that this process is FSH-dependent.