Peroxisomes are intracellular organelles responsible for crucial lipid metabolism including β-oxidation of very long chain fatty acids, metabolism of phytanic acid, plasmalogen synthesis, DHA synthesis, and hydrogen peroxide decomposition. Peroxisomes also seem to play important roles in spermatogenesis, as various grade male infertility has been frequently observed in knockout mice lacking genes related to peroxisomes. In this paper, we discuss the role of peroxisomes in spermatogenesis based on the localization of peroxisomal proteins in the testis and the analysis of knockout mice with genes involved in peroxisomal lipid metabolism.