Dataset Information

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

ABSTRACT: 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.

SUBMITTER: Biswas B

PROVIDER: S-EPMC7142241 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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

Biswas Barnali B Batista Frank F Akintayo Ayodele A Aguilan Jennifer J Stanley Pamela P

Frontiers in cell and developmental biology 20200402

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. ...[more]

PMID: 32300591

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Project description:In mammals, Sirt1, a member of the sirtuin family of proteins, functions as a nicotinamide adenine dinucleotide-dependent protein deactylase, and has important physiological roles, including the regulation of glucose metabolism, cell survival, and mitochondrial respiration. The initial investigations of Sirt1 deficient mice have revealed a phenotype that includes a reduced lifespan, small size, and an increased frequency of abnormal sperm. We have now performed a detailed analysis of the molecular and functional effects of Sirt1 deficiency in the germ line of Sirt1 knock-out (-/-) mice. We find that Sirt1 deficiency markedly attenuates spermatogenesis, but not oogenesis. Numbers of mature sperm and spermatogenic precursors, as early as d15.5 of development, are significantly reduced ( approximately 2-10-fold less; P</=0.004) in numbers in Sirt1-/- mice, whereas Sirt1 deficiency did not effect the efficiency oocyte production following superovulation of female mice. Furthermore, the proportion of mature sperm with elevated DNA damage ( approximately 7.5% of total epididymal sperm; P = 0.02) was significantly increased in adult Sirt1-/- males. Analysis of global gene expression by microarray analysis in Sirt1 deficient testis revealed dysregulated expression of 85 genes, which were enriched (P<0.05) for genes involved in spermatogenesis and protein sumoylation. To assess the function of Sirt1 deficient germ cells, we compared the efficiency of generating embryos and viable offspring in in vitro fertilization (IVF) experiments using gametes from Sirt1-/- and sibling Sirt1+/- mice. While viable animals were derived in both Sirt1-/- X wild type and Sirt1-/- X Sirt1-/- crosses, the efficiency of producing both 2-cell zygotes and viable offspring was diminished when IVF was performed with Sirt1-/- sperm and/or oocytes. Together, these data support an important role for Sirt1 in spermatogenesis, including spermatogenic stem cells, as well as germ cell function.

Dataset Information

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

Publications

Transgenic Rescue of Spermatogenesis in Males With <i>Mgat1</i> Deleted in Germ Cells.

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