| Summary: | 碩士 === 國立臺灣大學 === 生物科技研究所 === 107 === Proper spermatogenesis is critical to maintain the long-term male fertility and the health status of offspring. DNA methyltransferase 3-like (DNMT3L) mutation leads to spermatogenesis arrest and failure to repress transposons elements (TE), eventually cause male infertility. DNMT3L is an epigenetic modulator well known for facilitating DNA methylation during embryonic male germ cell development. The expression and function of postnatal DNMT3L are scarcely described. In this study, I identified a novel isoform, DNMT3L-at (“at” refers to adult testis), highly enriched in post-meiotic germ cells. I also demonstrated its surprising cytoplasmic localization, in contrast to the known epigenomic modulation property for canonical DNMT3L-s (“s” refers to stem cells). I deduced Dnmt3l-at promoter and transcriptional activity from various omics profiles of different male germ cell developmental stages. In addition, I clarified the DNMT3L-at postnatal expression and cytoplasmic localization by immunohistochemistry (IHC) of seminiferous tubules from different ages, and Western bolt from cytoplasm and nucleus fractioned protein samples. Furthermore, combination of various protein co-localization analysis results, I discovered possible interaction between DNMT3L-PLZF, and PLZF-piRNA pathway components. These data implied a novel function of DNMT3L-at beyond epigenetic regulation. Functional mutagenesis is being designed to precisely eliminate DNMT3L-at without interfering DNMT3L-s, in order to test the significance of DNMT3L-at in regulating meiosis.
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