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Advantages of using the CRISPR/Cas9 system of genome editing to investigate male reproductive mechanisms using mouse models

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Asian Journal of Andrology 2015年第4期17卷 623-627页

作者： Samantha AM Young R John Aitken masahito ikawa Priority Research Centre in Reproductive Science Discipline of Biological Sciences Faculty of Scienceand IT University of Newcastle Callaghan NSWAustralia Animal Resource Centre for InfectiousDiseases Research Institute for Microbial DiseasesOsaka University Suita Osaka Japan.Correspondence: Prof. M Ikawa

Gene disruption technology has long been beneficial for the study of male reproductive biology. However, because of the time and cost involved, this technology was not a viable method except in specialist laboratories. The advent of the CRISPR/Cas9 system ofgene disruption has ushered in a new era of genetic investigation. Now, it is possible to generate gene-disrupted mouse models in very tittle time and at very little cost. 1his Highlight article discusses the application of this technology to study the genetics of male fertility and looks at some of the future uses of this system that could be used to reveal the essential and nonessential genetic components of male reproductive mechanisms.

关键词： Advantages investigate mechanisms

CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice

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Asian Journal of Andrology 2022年第3期24卷 266-272页

作者： Yuki Oyama Haruhiko Miyata Keisuke Shimada Yoshitaka Fujihara Keizo Tokuhiro Thomas X Garcia Martin M Matzuk masahito ikawa Graduate School of Pharmaceutical Sciences Osaka UniversitySuitaOsaka 565-0871Japan Department of Experimental Genome Research Research Institute for Microbial DiseasesOsaka UniversitySuitaOsaka 565-0871Japan Department of Bioscience and Genetics National Cerebral and Cardiovascular CenterSuitaOsaka 564-8565Japan Department of Genome Editing Institute of Biomedical ScienceKansai Medical UniversityHirakataOsaka 573-1191Japan Center for Drug Discovery Baylor College of MedicineHoustonTX 77030USA Department of Pathology and Immunology Baylor College of MedicineHoustonTX 77030USA Department of Biology and Biotechnology University of Houston-Clear LakeHoustonTX 77058USA The Institute of Medical Science The University of TokyoMinato-kuTokyo 108-8639Japan

Gene expression analyses suggest that more than 1000–2000 genes are expressed predominantly in mouse and human *** functional analyses of hundreds of these genes have been performed,there are still many testis-enriched genes whose functions remain *** gene function using knockout(KO)mice is a powerful tool to discern if the gene of interest is essential for sperm formation,function,and male fertility in *** this study,we generated KO mice for 12 testis-enriched genes,1700057G04Rik,4921539E11Rik,4930558C23Rik,Cby2,Ldhal6b,Rasef,Slc25a2,Slc25a41,Smim8,Smim9,Tmem210,and Tomm20l,using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)*** designed two gRNAs for each gene to excise almost all the protein-coding regions to ensure that the deletions in these genes result in a null *** tests of KO mice reveal that these 12 genes are not essential for male fertility,at least when individually ablated,and not together with other potentially compensatory paralogous *** results could prevent other laboratories from expending duplicative effort generating KO mice,for which no apparent phenotype exists.

关键词： CRISPR/Cas9 knockout mice male infertility spermatozoa testis

Ig SF11 regulates osteoclast differentiation through association with the scaffold protein PSD-95

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Bone Research 2020年第1期8卷 97-106页

作者： Hyunsoo Kim Noriko Takegahara Matthew CWalsh Sarah AMiddleton Jiyeon Yu Jumpei Shirakawa Jun Ueda Yoshitaka Fujihara masahito ikawa Masaru Ishii Junhyong Kim Yongwon Choi Department of Pathology and Laboratory Medicine University of Pennsylvania Perelman School of MedicinePhiladelphiaPA 19104USA Department of Biology Department of Computer and Information ScienceSchool of Arts and SciencesProgram in Single Cell BiologyUniversity of PennsylvaniaPhiladelphiaPA 19104USA Research Institute for Microbial Diseases Osaka UniversitySuitaOsaka 565-0871Japan Department of Immunology and Cell Biology Graduate School of Medicine and Frontier BiosciencesOsaka UniversitySuitaOsaka 565-0871Japan

Osteoclasts are multinucleated, giant cells derived from myeloid progenitors. While receptor activator of NF-κB ligand(RANKL)stimulation is the primary driver of osteoclast differentiation, additional signaling further contributes to osteoclast ***, we demonstrate that immunoglobulin superfamily member 11(Ig SF11), whose expression increases during osteoclast differentiation, regulates osteoclast differentiation through interaction with postsynaptic density protein 95(PSD-95), a scaffold protein with multiple protein interaction domains. Ig SF11 deficiency in vivo results in impaired osteoclast differentiation and bone resorption but no observed defect in bone formation. Consequently, Ig SF11-deficient mice exhibit increased bone *** in vitro osteoclast culture systems, we show that Ig SF11 functions through homophilic interactions. Additionally, we demonstrate that impaired osteoclast differentiation in Ig SF11-deficient cells is rescued by full-length Ig SF11 and that the Ig SF11-PSD-95 interaction requires the 75 C-terminal amino acids of Ig SF11. Our findings reveal a critical role for Ig SF11 during osteoclast differentiation and suggest a role for Ig SF11 in a receptor-and signal transduction molecule-containing protein complex.

关键词： SF11 osteoclast impaired

Cooperation-based sperm clusters mediate sperm oviduct entry and fertilization

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Protein & Cell 2021年第10期12卷 810-817页

作者： Yongcun Qu Qi Chen Shanshan Guo Chiyuan Ma Yonggang Lu Junchao Shi Shichao Liu Tong Zhou Taichi Noda Jingjing Qian Liwen Zhang Xili Zhu Xiaohua Lei Yujing Cao Wei Li Wei Li Nicolas Plachta Martin M.Matzuk masahito ikawa Enkui Duan Ying Zhang Hongmei Wang State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Division of Biomedical Sciences School of MedicineUniversity of CaliforniaRiversideCA 92521USA Research Institute for Microbial Diseases Osaka UniversitySuitaOsakaJapan Department of Physiology and Cell Biology Reno School of MedicineUniversity of NevadaRenoNVUSA Institute of Molecular and Cell Biology A*STARSingaporeSin gapore Center for Drug Discovery Department of Pathology and ImmunologyBaylor College of MedicineHoustonTX 77030USA Division of Biology and Biological Engineering California Institute of TechnologyPasadenaCAUSA Institute for Stem Cell and Regeneration Chinese Academy of SciencesBeijing 100101China The Key Laboratory of Cell Proliferation and Regulation Biology Ministry of EducationInstitute of Cell BiologyBeijing Normal UniversityBeijing 100875China

Dear Editor,Sperm cooperation has been observed in multiple species(Pizzari and Foster,2008),yet its existenee and benefit for reproductive success in mammals remains ***,combining tissue-clearing with deep three-dimensional(3D)imagi ng,we dem on strate that postcopulatory mouse sperm congregate into unidirectional sperm cooperative clusters at the utero-tubal junction(UTJ),a key physical barrier for passage into the oviduct.

关键词： passage media Foster

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PNAS:科学家发现新的精子运动调节器

现代生物医学进展 2017年第27期17卷 I0001-I0002页

作者： Castaneda, Julio M. Hua, Rong Miyata, Haruhiko Oji, Asami Guo, Yueshuai Cheng, Yiwei Zhou, Tao Guo, Xuejiang Cui, Yiqiang Shen, Bin Wang, Zibin Hu, Zhibin Zhou, Zuomin Sha, Jiahao Prunskaite-Hyyrylainen, Renata Yu, Zhifeng Ramirez-Solis, Ramiro ikawa, masahito Matzuk, Martin M. Liu, Mingxi Baylor Coll Med Dept Pathol & Immunol Houston TX 77030 USA Osaka Univ Res Inst Microbial Dis Suita Osaka 5650871 Japan Nanjing Med Univ State Key Lab Reprod Med Nanjing 210029 Jiangsu Peoples R China Nanjing Med Univ Dept Histol & Embryol Nanjing 210029 Jiangsu Peoples R China Osaka Univ Grad Sch Pharmaceut Sci Suita Osaka 5650871 Japan Nanjing Med Univ Anim Core Facil Nanjing 210029 Jiangsu Peoples R China Baylor Coll Med Ctr Reprod Med Houston TX 77030 USA Univ Oulu Fac Biochem & Mol Med FI-90014 Oulu Finland Baylor Coll Med Ctr Drug Discovery Houston TX 77030 USA Wellcome Trust Sanger Inst Hinxton CB10 1SA England Univ Tokyo Inst Med Sci Minato Ku Tokyo 1088639 Japan Baylor Coll Med Dept Mol & Cellular Biol Houston TX 77030 USA Baylor Coll Med Dept Mol & Human Genet Houston TX 77030 USA Baylor Coll Med Dept Pharmacol Houston TX 77030 USA

南京医科大学生殖医学国家重点实验室、基础医学院组织胚胎学系刘明兮课题组与贝勒医学院***课题组、大阪大学masahitoikawa课题组等共同解析了一个精子运动调节基因TCTE1，结果发表于《美国科学院院刊》（PNAs）。

关键词：精子运动调节器科学家国家重点实验室组织胚胎学课题组生殖医学医科大学

River system in Japan from a landscape ecological aspect

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Journal of Environmental Sciences 2003年第2期15卷 160-166页

作者： Nobukazu NAKAGOSHI masahito INOUE Graduate School for International Development and Cooperation Hiroshima UniversityGraduate School for International Development and Cooperation Hiroshima University 1-5-1 Kagamiyama Higashi-Hiroshima 739-8529 Japan1-5-1 Kagamiyama Higashi-Hiroshima 739-8529 Japan

The objective of this study was to elucidate characteristics associated with rivers by classification of major rivers in Japan into several types based on riparian conditions. As the results of principal component analysis (PCA) with use of parameters reflecting forms and artificial alterations of respective rivers, four major components such as “comprehensive riparian size', “intactness of water front', “continuity of streams' and “simplicity of landform in river basin' were extracted. Subsequently, cluster analysis was performed based on principal component scores, leading to successful classification of major rivers into 6 types. These findings disclosed that (1) the extracted principal components provide effective viewpoint for classification of rivers; (2) distribution of respective classes indicates area properties; and (3) the employed quantitative procedures were found effective for classification of major rivers.

关键词： river system landscape ecological aspect Japan

Intranasal administration of stem cell-derived exosomes for central nervous system diseases

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Neural Regeneration Research 2024年第6期19卷 1249-1255页

作者： Shuho Gotoh masahito Kawabori Miki Fujimura Department of Neurosurgery Hokkaido University Graduate School of MedicineHokkaidoJapan

Exosomes,lipid bilayer-enclosed small cellular vesicles,are actively secreted by various cells and play crucial roles in intercellular *** nanosized vesicles transport internalized proteins,mRNA,miRNA,and other bioactive *** findings have provided compelling evidence that exosomes derived from stem cells hold great promise as a therapeutic modality for central nervous system *** exosomes exhibit multifaceted properties including antiapoptotic,anti-inflammatory,neurogenic,and vasculogenic ***,exosomes offer several advantages over stem cell therapy,such as high preservation capacity,low immunogenicity,the ability to traverse the blood-brain barrier,and the potential for drug ***,researchers have turned their attention to exosomes as a novel therapeutic ***,akin to the limitations of stem cell treatment,the limited accumulation of exosomes in the injured brain poses a challenge to their clinical *** overcome this hurdle,intranasal administration has emerged as a non-invasive and efficacious route for delivering drugs to the central nervous *** exploiting the olfactory and trigeminal nerve axons,this approach enables the direct transport of therapeutics to the brain while bypassing the blood-brain ***,exosomes,owing to their small size,can readily access the nerve pathways using this *** a result,intranasal administration has gained increasing recognition as an optimal therapeutic strategy for exosomebased *** this comprehensive review,we aim to provide an overview of both basic and clinical research studies investigating the intranasal administration of exosomes for the treatment of central nervous system ***,we elucidate the underlying therapeutic mechanisms and offer insights into the prospect of this approach.

关键词： central nervous system disease exosome extracellular vesicle intranasal administration stem cell

Chemoprevention of obesity-related liver carcinogenesis by using pharmaceutical and nutraceutical agents

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World Journal of Gastroenterology 2016年第1期22卷 394-406页

作者： Hiroyasu Sakai Yohei Shirakami masahito Shimizu Department of Gastroenterology/Internal Medicine Gifu University Graduate School of Medicine

Obesity and its related metabolic disorders are serious health problems worldwide,and lead to various healthrelated complications,including *** human cancers,hepatocellular carcinoma(HCC)is one of the most common malignancies affected by ***,obesity and its related disorders might be a key target for the prevention of ***,new research indicates that the molecular abnormalities associated with obesity,including insulin resistance/hyperinsulinemia,chronic inflammation,adipokine imbalance,and oxidative stress,are possible molecular mechanisms underlying the pathogenesis of obesityrelated *** tea catechins and branched-chain amino acids,both of which are classified as nutraceutical agents,have been reported to prevent obesity-related HCC development by improving metabolic *** administration of acyclic retinoid,a pharmaceutical agent,reduced the incidence of HCC in obese and diabetic mice,and was also associated with improvements in insulin resistance and chronic *** this article,we review the detailed molecular mechanisms that link obesity to the development of HCC in obese *** also summarize recent evidence from experimental and clinical studies using either nutraceutical or pharmaceutical agents,and suggest that nutraceutical and pharmaceutical approaches targeting metabolic abnormalities might be a promising strategy to prevent the development of obesity-related HCC.

关键词： Hepatocellular carcinoma Obesity Greentea catechins Branched-chain amino acids Acyclicretinoid

Non-Kramers doublet ground state in a quaternary cubic compound PrRu_(2)In_(2)Zn_(18) investigated by ultrasonic measurements

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Chinese Physics B 2024年第6期33卷 405-411页

作者：张化远 Kazuhei Wakiya Mitsuteru Nakamura masahito Yoshizawa Yoshiki Nakanish Graduate School of Science and Engineering Iwate UniversityMorioka 020-8551Japan

We performed ultrasonic measurements on a quaternary cubic compound PrRu_(2)In_(2)Zn_(18) to explore the ground state properties derived from non-Kramers Γ_(3) doublet of Pr^(3+).PrRu_(2)In_(2)Zn_(18) is a quaternary derivative of the ternary compound PrRu_(2)Zn_(20) that exhibits a structural phase transition at T_S=138 *** PrRu_(2)In_(2)Zn_(18),the Zn atoms at the 16c site in PrRu_(2)Zn_(20) are selectively replaced by In atoms.A monotonic increase was observed in the temperature dependence of elastic constants C_L=(C_(11)+2C_(12)+4C_(44))/3 and C_(T)=(C_(11)-C_(12)+C_(44))/3 in the temperature range around T_(S) to which an elastic softening was observed in(C_(11)-C_(12))/2 for PrRu_(2)Zn_(20).The disappearance of the softening indicates that the structural transition in PrRu_(2)Zn_(20) is suppressed by the substitution of Zn ions by In ones with a larger ionic ***,the C_(T) of PrRu_(2)In_(2)Zn_(18) exhibits a precursor Curie-type elastic softening toward low temperatures being responsible for the non-Kramers Γ_(3) ground *** discuss the ground state and the evolution of the elastic properties of the different single-crystal samples of PrRu_(2)In_(2)Zn_(18) grown under different conditions.

关键词： ultrasonic measurements non-Kramers doublet structural phase transition crystalline electric field effect