A5019005784- Sperm and Testicular Function
- Reproductive Biology and Fertility
- CRISPR and Genetic Engineering
- Genetic and Clinical Aspects of Sex Determination and Chromosomal Abnormalities
- DNA Repair Mechanisms
- RNA and protein synthesis mechanisms
- Reproductive biology and impacts on aquatic species
- Pluripotent Stem Cells Research
- Cancer-related Molecular Pathways
- Reproductive System and Pregnancy
- Ovarian cancer diagnosis and treatment
- Cancer Research and Treatments
- Animal Genetics and Reproduction
- Renal and related cancers
- Epigenetics and DNA Methylation
- Genetics, Aging, and Longevity in Model Organisms
- Microtubule and mitosis dynamics
- Ubiquitin and proteasome pathways
- Neuroendocrine regulation and behavior
- Hormonal and reproductive studies
- Genetic and Kidney Cyst Diseases
- Ovarian function and disorders
- Toxoplasma gondii Research Studies
- Cancer, Hypoxia, and Metabolism
- RNA regulation and disease
Osaka University
2016-2025
Kumamoto University
2021-2025
Rockefeller University
2020
National Institutes of Health
2020
Yale University
2020
Harvard University
2020
Brigham and Women's Hospital
2020
Federation of American Societies for Experimental Biology
2020
Kobe University
2010-2015
Nara Medical University
2010-2012
The RNA-guided endonuclease Cas9 cleaves its target DNA and is a powerful genome-editing tool. However, the widely used Streptococcus pyogenes enzyme (SpCas9) requires an NGG protospacer adjacent motif (PAM) for recognition, thereby restricting targetable genomic loci. Here, we report rationally engineered SpCas9 variant (SpCas9-NG) that can recognize relaxed NG PAMs. crystal structure revealed loss of base-specific interaction with third nucleobase compensated by newly introduced...
Significance The sperm−oocyte fusion step is important to transport the male genome into oocytes. So far, IZUMO1 and FIMP have been identified as fusion-related proteins in spermatozoa, but molecular mechanisms underpinning all of required for this essential process remain unclear. In study, using CRISPR-Cas9−mediated gene knockouts mice, we discover that sperm SOF1, TMEM95, SPACA6 are fertility. As these genes conserved among mammals including human, they may explain not only also...
Gene-expression analysis studies from Schultz et al. estimate that more than 2,300 genes in the mouse genome are expressed predominantly male germ line. As of their 2003 publication [Schultz N, Hamra FK, Garbers DL (2003) Proc Natl Acad Sci USA 100(21):12201-12206], functions majority these testis-enriched during spermatogenesis and fertilization were largely unknown. Since study by al., functional hundreds reproductive-tract-enriched have been performed, but there remain many for which...
Sperm-oocyte fusion is a critical event in mammalian fertilization, categorized by three indispensable proteins. Sperm membrane protein IZUMO1 and its counterpart oocyte JUNO make complex allowing sperm to interact with the oocyte, subsequent sperm-oocyte fusion. Oocyte tetraspanin CD9 also contributes However, process cannot be explained solely these essential factors. In this study, we focused on analyzing testis-specific gene 4930451I11Rik generated mutant mice using CRISPR/Cas9 system....
Abstract Targeted gene disrupted mice can be efficiently generated by expressing a single guide RNA (sgRNA)/CAS9 complex in the zygote. However, limited success of complicated genome editing, such as large deletions, point mutations, and knockins, remains to improved. Further, mosaicism founder generations complicates genotypic phenotypic analyses these animals. Here we show that deletions with two sgRNAs well dsDNA-mediated mutations are efficient mouse embryonic stem cells (ESCs). The...
Abstract More than 1000 genes are predicted to be predominantly expressed in mouse testis, yet many of them remain unstudied terms their roles spermatogenesis and sperm function essentiality male reproduction. Since individually indispensable factors can provide important implications for the diagnosis genetically related idiopathic infertility may serve as candidate targets development nonhormonal contraceptives, our laboratories continuously analyze functions testis-enriched vivo by...
Local control of sperm maturation Newly produced spermatozoa within the testis do not have fertilizing ability but become fully functional when they mature in epididymis. The development epididymis itself is dependent on testicular factors arriving via luminal flow. Improper signaling between and hypothesized to result male infertility. Kiyozumi et al. identified NELL2 as a protein that binds its receptor, ROS1, surface induces epididymal differentiation (see Perspective by Lord Oatley). In...
Abstract The process of sperm-egg fusion is critical for successful fertilization, yet the underlying mechanisms that regulate these steps have remained unclear in vertebrates. Here, we show both mouse and zebrafish DCST1 DCST2 are necessary sperm to fertilize egg, similar their orthologs SPE-42 SPE-49 C. elegans Sneaky D. melanogaster . Mouse Dcst1 Dcst2 single knockout (KO) able undergo acrosome reaction normal relocalization IZUMO1, an essential factor fusion, equatorial segment. While KO...
Spermatozoa need to undergo an exocytotic event called the acrosome reaction before fusing with eggs. Although calcium ion (Ca 2+ ) is essential for reaction, its molecular mechanism remains unknown. Ferlin a single transmembrane protein multiple Ca -binding C2 domains, and there are six ferlins, dysferlin (DYSF), otoferlin (OTOF), myoferlin (MYOF), fer-1–like 4 (FER1L4), FER1L5, FER1L6, in mammals. Dysf , Otof Myof knockout mice have been generated, each mouse line exhibited membrane fusion...
The recombinant clustered regulatory interspaced short palindromic repeats ( CRISPR )/Cas system has opened a new era for mammalian genome editing. Here, we constructed pX330 plasmids expressing humanized Cas9 (hCas9) and single guide RNA s (sg s) against mouse genes validated them both in vitro vivo . When randomly chose 291 target sequences within protein coding regions of 73 genes, an average number off‐target candidates (exact match 13 nucleotides from 3′ NGG) found by Bowtie software...
Significance While the emergence of gene modification technologies has produced major discoveries in biomedical sciences, recent development CRISPR/Cas9 system dramatically altered trajectory phenotypic analysis animal models. In this study, we identified male-specific clusters ( Cst and Pate ) family genes Gdpd Lypd found specific members to be required for male fertility, especially sperm fertilizing ability. Our findings support important roles these proteins function could used develop...
The mammalian spermatozoa produced in the testis require functional maturation epididymis for their full competence. Epididymal sperm is regulated by lumicrine signalling pathways which testis-derived secreted signals relocate to lumen and promote differentiation. However, detailed mechanisms of regulation are unclear. Herein, we demonstrate that a small protein, NELL2-interacting cofactor (NICOL), plays crucial role mice. NICOL expressed male reproductive organs, including testis, forms...
Spermatozoa have a streamlined shape to swim through the oviduct fertilize oocytes. To become svelte spermatozoa, spermatid cytoplasm must be eliminated in several steps including sperm release, which is part of spermiation. Although this process has been well observed, molecular mechanisms that underlie it remain unclear. In male germ cells, there are membraneless organelles called nuage, observed by electron microscopy various forms dense material. Reticulated body (RB) and chromatoid...
NFκB is the key modulator in inflammatory disorders. However, regulators that activate, fine-tune or shut off activity conditions are poorly understood. In this study, we aim to investigate roles NFκB-specific long non-coding RNAs (lncRNAs) play regulating networks.Using first genetic-screen identify lncRNAs, performed RNA-seq from p65-/- and Ikkβ-/- mouse embryonic fibroblasts report identification of an evolutionary conserved lncRNA designated mNAIL (mice) hNAIL (human). upregulated human...
Abstract Background Spermatozoa become competent for fertilization during transit through the epididymis. As spermatozoa from proximal caudal epididymis can fertilize eggs, proteins caput and corpus are required sperm maturation. Objectives Microarray analysis identified that more than 17,000 genes expressed in epididymis; however, few of these demonstrate expression restricted to To analyze epididymis‐enriched gene function vivo, we generated knockout ( KO ) mutations nine abundantly region...
Abstract Spermatozoa are produced in the testis but gain their fertilizing ability during epididymal migration. This necessary step sperm maturation includes posttranslational modification of membrane proteins that protein processing by proteases. However, molecular mechanism underpinning this remains unknown. In study, we focused on transmembrane serine protease 12 (Tmprss12). Based multi-tissue expression analysis PCR, Tmprss12 was specifically expressed testis, and its started day 10...
Spermatogenesis is a complex process of sperm generation, including mitosis, meiosis, and spermiogenesis. During spermiogenesis, histones in post-meiotic spermatids are removed from chromatin replaced by protamines. Although histone-to-protamine exchange important for nuclear condensation, the underlying regulatory mechanism still poorly understood. Here, we identify PHD finger protein 7 (PHF7) as an E3 ubiquitin ligase histone H3K14 spermatids. Generation Phf7-deficient mice Phf7 C160A...
Abstract The clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system is a research hotspot in gene therapy. However, the widely used Streptococcus pyogenes Cas9 (WT-SpCas9) requires an NGG protospacer adjacent motif (PAM) for target recognition, thereby restricting targetable disease mutations. To address this issue, we recently reported engineered SpCas9 nuclease variant (SpCas9-NG) recognizing NGN PAMs. Here, as feasibility study, report SpCas9-NG-mediated repair of...
The acrosome is a cap-shaped, Golgi-derived membranous organelle that located over the anterior of sperm nucleus and highly conserved throughout evolution. Although morphological changes during biogenesis in spermatogenesis have been well described, molecular mechanism underlying this process still largely unknown. Family with sequence similarity 71, member F1 F2 (FAM71F1 FAM71F2) are testis-enriched proteins contain RAB2B-binding domain, small GTPase involved vesicle transport membrane...
Sperm acrosomal membrane proteins, such as Izumo sperm–egg fusion 1 (IZUMO1) and sperm acrosome-associated 6 (SPACA6), play essential roles in mammalian gamete binding or fusion. How their biosynthesis is regulated during spermiogenesis has largely remained elusive. Here, we show that 1700029I15Rik knockout male mice are severely subfertile spermatozoa do not fuse with eggs. a type-II transmembrane protein expressed early round spermatids but mature spermatozoa. It interacts proteins...
Abstract The effect on the reproductive system and fertility of living in a space environment remains unclear. Here, we caged 12 male mice under artificial gravity (≈1 gravity) (AG) or microgravity (MG) International Space Station (ISS) for 35 days, characterized organs (testes, epididymides, accessory glands) after their return to earth. Mice earth during days served as “ground” control (GC). Only decrease gland weight was detected AG MG males; however, none showed any overt microscopic...
Cilia and flagella are ancient structures that achieve controlled motor functions through the coordinated interaction based on microtubules some attached projections. Radial spokes (RSs) facilitate beating motion of these organelles by mediating signal transduction between dyneins a central pair (CP) singlet microtubules. RS complex isolation from Chlamydomonas axonemes enabled detection 23 radial spoke proteins (RSP1-RSP23), although roles remain unknown. Recently, RSP15 has been reported...