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在生殖细胞中缺失的雄性小鼠精子发生的转基因拯救

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

作者信息

Biswas Barnali, Batista Frank, Akintayo Ayodele, Aguilan Jennifer, Stanley Pamela

机构信息

Department of Cell Biology, Albert Einstein College of Medicine, New York, NY, United States.

Laboratory for Macromolecular Analysis and Proteomics, Department of Pathology, Albert Einstein College of Medicine, New York, NY, United States.

出版信息

Front Cell Dev Biol. 2020 Apr 2;8:212. doi: 10.3389/fcell.2020.00212. eCollection 2020.

Abstract

MGAT1 and complex N-glycans are required for spermatogenesis and fertility. Conditional deletion of in spermatogonia ( 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 cKO males that also expressed a transgene under the control of a germ cell-specific promoter - for spermatogonia, for spermatocytes and for spermatids. Males expressing each transgene were fertile, and both males and females transmitted each transgene. When --HA was expressed in 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 expressed in spermatocytes, nor the - transgene expressed in spermatids rescued spermatogenesis or fertility in cKO males. Therefore, MGAT1 must be expressed in spermatogonia for spermatogenesis to proceed normally.

摘要

MGAT1和复合N-聚糖是精子发生和生育所必需的。在精原细胞中条件性敲除(cKO)会导致ERK1/2信号传导减少以及多核生殖细胞(MNC)的形成。在这里我们表明,对从固定睾丸切片中释放的N-聚糖进行糖组学分析,并通过基质辅助激光解吸电离成像质谱(MALDI-IMS)进行分析,基于MGAT1的低聚甘露糖基底物的积累,揭示了所有生殖细胞中MGAT1活性的丧失。为了确定在哪种类型的生殖细胞中MGAT1对精子发生至关重要,我们构建了cKO雄性小鼠,它们还在生殖细胞特异性启动子的控制下表达转基因——用于精原细胞的、用于精母细胞的和用于精子细胞的。表达每种转基因的雄性小鼠都具有生育能力,并且雄性和雌性都能传递每种转基因。当在cKO雄性小鼠中表达--HA时,基于睾丸切片的形态、基底膜上N-聚糖的组成、凝集素组织化学、MALDI-IMS和生育能力,精子发生得以挽救。相比之下,在精母细胞中表达的和在精子细胞中表达的转基因都不能挽救cKO雄性小鼠的精子发生或生育能力。因此,MGAT1必须在精原细胞中表达,精子发生才能正常进行。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/baf8/7142241/52fb32637187/fcell-08-00212-g001.jpg

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