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MGAT1 and complex N-glycans are required for spermatogenesis and fertility. Conditional deletion of  Mgat1  in spermatogonia ( Mgat1  cKO) causes reduced ERK1/2 signaling and the formation of multinucleated germ cells (MNC). Here we show that glycomics analysis of N-glycans released from fixed testis sections and analyzed by MALDI imaging mass spectrometry (MALDI-IMS) revealed a loss of MGAT1 activity in all germ cells based on the accumulation of the oligomannosyl substrate of MGAT1. To determine in which type of germ cell MGAT1 is essential for spermatogenesis, we generated  Mgat1  cKO males that also expressed a  Mgat1-HA  transgene under the control of a germ cell-specific promoter –  Stra8  for spermatogonia,  Ldhc  for spermatocytes and  Prm1  for spermatids. Males expressing each  Mgat1-HA  transgene were fertile, and both males and females transmitted each transgene. When  Stra8 - Mgat1 -HA was expressed in  Mgat1  cKO males, spermatogenesis was rescued based on the morphology of testis sections, the complement of N-glycans on basigin, lectin histochemistry, MALDI-IMS, and fertility. By contrast, neither  Ldhc-Mgat1-HA  expressed in spermatocytes, nor the  Prm1 - Mgat1-HA  transgene expressed in spermatids rescued spermatogenesis or fertility in  Mgat1  cKO males. Therefore, MGAT1 must be expressed in spermatogonia for spermatogenesis to proceed normally.

Transgenic Rescue of Spermatogenesis in Males With Mgat1 Deleted in Germ Cells