Haruhiko Miyata's research while affiliated with Osaka University and other places

... Given that many lncRNAs are expressed in a tissue-specific manner, with the testis showing the greatest number of tissue-specific lncRNAs in mammals, many unannotated CDSs likely remain to be identified in lncRNAs of the testis (38). We performed a comprehensive screening of testis-specific lncRNAs with the PhyloCSF algorithm, and thereby found that the mouse lncRNA locus Gm9999 gives rise to two transcripts with different transcription start sites, each of which is translated to produce a polypeptide (35). We named these two polypeptides Kastor and Polluks with reference ...

... The strong transcriptional effect of MORC proteins in early male gametogenesis is reminiscent of the known role of some animal MORC family members in the male germline. [59][60][61][62] However, plant MORC proteins are not required for male fertility, and we did not observe any pollen development defect in the morc hextuple mutant, thus leaving open the question of whether Arabidopsis MORCs play a role in pollen biology. ...

... To date, 45 human kinesin superfamily (KIF) proteins have been identified and shown to exhibit a diverse range of functions. Of these, KIF9 is a highly conserved kinesin that is enriched in the testes of mice and has been shown to interact with hydrocephalus-inducing protein homolog (HYDIN) and to localize to the axonemal CP (17,18). Deletion of murine Kif9 impairs flagellar motility of sperm, resulting in male infertility (18). ...

... Ablation of PPP3CC, PPP3R2 or SPATA33 result in the decreased flexibility of sperm flagellar midpiece, which causes the impairment of sperm penetration through egg zona pellucida and male infertility (25,26). Interestingly, while in vitro supplementation of culture medium with the calcineurin inhibitor FK506 or cyclosporin does not alter sperm flagellar flexibility, the in vivo treatment with these inhibitors makes animals male infertile and the sperm flagellum of treated animals becomes rigid (25). ...

... Consistent with a possible role for crotonylation in the histone-to-protamine exchange, CDYL (chromodomain Y-like protein, an eraser of crotonylation) knockout mice exhibit reduced levels of chromatin-bound transition proteins, sperm motility defects, and decreased fertility (58). Given the general enrichment in spermatids for modifications canonically associated with transcriptional activation in somatic cells, together with the well-documented pervasive transcription observed in round spermatids (59)(60)(61) and the lack of reported phenotypes for many spermatid-specific expressed genes, this begs the question of whether the physical process of gene transcription may be important in nucleosome destabilization and subsequent exchange-a hypothesis that will need to be evaluated in future studies (62)(63)(64)(65). ...

... In addition, ZFP541 KO mice were generated as a model in which differentiation stops early in meiosis. Tbc1d21 KO mice were used as a model of teratozoospermia and were generated using the methodology previously described in Ref. 19 Kit w /Kit wv , Zfp541 KO, and Kctd19 KO mice underwent MRI imaging at 4 weeks of age because their testes atrophy considerably with age. Tbc1d21 KO mice were imaged at 40 weeks of age. ...

... Karyopherin alpha (KPNA) are nuclear transporters (NTRs) that consist of a cluster of basic amino acids, which selectively through the nuclear pore complex (NPC) (Hazawa et al., 2020;Miyamoto et al., 2020). NPC is composed of 30 nucleoporin (NUP) proteins, which is the sole channel between the nucleus and the cytoplasm (Hazawa et al., 2020). ...

... Sperm Motility Analysis. Sperm motility was analyzed as described previously (5,37). Spermatozoa obtained from the cauda epididymis were incubated in a 100 μl drop of Toyoda, Yokoyama, and Hoshi (TYH) medium (38). ...

... GPI-APs were HA-tagged by cloning their coding cDNA amplified from the human cDNA library or from Invitrogen plasmids, as described (60), into the mammalian expression vector (pME-puro) and co-transfected into the PIGG-KO cells with PIGG cDNA or the empty vector as control. The expression levels of the GPI-APs were preprint (which was not certified by peer review) is the author/funder. ...

... Histology Testes were fixed in Bouin's Fixative and stained with periodic acid-Schiff's reagent (PAS, Sigma Aldrich) as described in Ref. [40]. The sections were observed and imaged using DM LB2 (Leica) and Zeiss Axio Imager motorized histology (Zeiss) microscopes. ...