Nerve growth factor (NGF), which is required for the survival and differentiation of the nervous system, has been proved to play important roles in male reproductive physiology. Several studies have focused on the roles of NGF in the testes. However, no study has reported on the mechanism of paracrine and autocrine actions of NGF in Sertoli cells. This study showed that NGF stimulated mitochondrial activity and biogenesis in TM4 Sertoli cells. Moreover, our results demonstrated that peroxisome proliferator-activated receptor-gamma coactivator-1α is a possible downstream key target of the NGF signalling pathway. In a 3-nitropropionic acid cell model, NGF treatment attenuated mitochondrial activity defect and depolarisation. This NGF-triggered signalling may help in discovering new therapeutic targets for certain male infertility disorders.