KATNB1 is a microtubule regulatory enzyme which complexes with enzymatic microtubule-severing proteins to regulate their severing activity. Previously, we have shown that Katnb1 mutation disrupts germ cell microtubule dynamics resulting in male sterility[1]. The enzyme(s) mediating KATNB1 function in this context has not been identified. This study sought to characterize the microtubule-severing enzyme KATNAL2 in spermatogenesis and determine if it could mediate KATNB1 testis functions. To study this, we used a Katnal2 point mutant and a Katnal2 KO mouse model. Both Katnal2 full ablation and loss of function resulted in male sterility. Spermatids in these mice exhibited defects in head shaping due to manchette dysregulation and acrosome detachment from the nucleus. Concomitantly, supernumerary centrioles were observed in the spermatids, followed by an absence of axoneme generation in later steps and disorganisation of the mitochondrial sheath. Sperm production rates and testis weight were normal in these mice, however; elongated spermatids were retained resulting in a 96% reduction in epididymal sperm content. Our analysis suggests this spermiation failure arises from an absence of tubulobulbar complex and residual body formation. Interestingly, ectoplasmic specialisation disassembly appeared normal. To ascertain if these spermatogenic abnormalities were germ cell autonomous effects or reliant on interactions between germ and Sertoli cells, Katnal2 germ cell specific KO mice were generated using a Stra8-Cre and found to phenocopy the Katnal2 mutant and KO mice. To determine if KATNAL2 and KATNB1 associate to form microtubule-severing complexes in the testis, in situ protein ligation and co-immunoprecipitation assays were performed. Consistent with similarities in sperm head shaping between the Katnal2 and Katnb1 mouse models, we successfully immunoprecipitated KATNAL2-KATNB1 complexes and observed that these complexes localized to the spermatid manchette. Collectively, our findings demonstrate that KATNAL2, likely under KATNB1 regulation, has multiple essential roles in regulating germ cell microtubule dynamics during spermatogenesis.