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Molecular Cloning of MSRG-11 Gene Related to Apoptosis of Mouse Spermatogenic cells

Acta Biochimica et Biophysica Sinica 2005年第3期37卷 159-166页

作者： Yun DENG Dong-Song NIE Jian WANG Xiao-Jun TAN Zhao-Yan NIE Hong-Mei YANG Liang-Sha HU Guang-Xiu LU Human Reproductive and stem cell Engineering Institute,Central South University,Changsha 410078,China Human Reproductive and stem cell Engineering Institute Central South UniversityChangsha 410078 China

Beginning with a new contig of the expressed sequence tags(Mm.63892)obtained bycomparing testis libraries with other tissue and cell line libraries using the digital differential display program,we cloned a new gene which is related to the apoptosis of mouse spermatogenic cells using the Genscanprogram and polymerase chain reaction(PCR)*** sequence data have been submitted to theGenBank database under accession number *** full cDNA length is 1074 bp,and the gene with7 exons and 6 introns is located in mouse chromosome 1 *** protein is recognized as a new member ofcahnodulin(CaM)binding protein family because the sequence contains three short calmodulin-binding motifscontaining conserved Ile and Gln residues(IQ motif)and is considered to play a critical role in interactions ofIQ motif-containing proteins with CaM *** putative protein encoded by this gene has 192 aminoacid residues with a theoretical molecular mass of 23.7 kDa and a calculated isoelectric point of *** shares no significant homology with any known protein in ***-PCR and Northern blotanalyses revealed that 1.3 kb MSRG-11 transcript was strongly expressed in adult mouse testis but weaklyexpressed in the spleen and *** MSRG-11 gene was expressed at various levels,faintly at twoweeks postpartum and strongly from three weeks postpartum in adult *** green fluorescence pro-duced by pEGFP-C2/MSRG-11 was detected in the cytoplasm of COS7 cells 24 h ***3.1(-)/MSRG-11 plasmid was constructed and introduced into COS7 cells using Lipofectamine 2000transfection reagent(Invitrogen,Carlsbad,USA).MSRG-11 can accelerate COS7 cell apoptosis,whichsuggests that this gene may play an important role in the development of mouse testes and is a candidate geneof testis-specific *** on these observations,it was considered that we cloned a new gene whichprobably accelerates spermatogenetic cell apoptosis in mouse.

关键词： MSRG-11 gene testis-specific expression spermatogenetic cell apoptosis IQ motif

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Cloning,Characterization and Primary Function Study of a Novel Gene,Cymg1,Related to Family 2 Cystatins

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Acta Biochimica et Biophysica Sinica 2005年第1期37卷 11-18页

作者： Yang XIANG Dong-Song NIE Jian WANG Xiao-Jun TAN Yun DENG Shu-Wei LUO Guang-Xiu LU~* Human Reproductive and stem cell Engineering Institute,Central South University,Changsha 410078,China Human Reproductive and stem cell Engineering Institute Central South UniversityChangsha 410078 China

Cystatins are cysteine proteinase inhibitors,We found two expression sequence tags (ESTs),CA463109 and AV042522,from a mouse testis library using Digital differential display (DDD).By electricalhybridization,a novel gene,Cymgl(GenBank accession ***600990),which has a full length of 0.78 kb,and contains four exons and three introns,was cloned from a mouse testis eDNA *** gene is locatedin the 2G3 area of chromosome *** full eDNA encompasses the entire open reading frame,encoding 141amino acid *** protein has a cysteine protease inhibitor domain that is related to the family 2cystatins but lacks critical consensus sites important for cysteine protease *** characteristicsare seen in the CRES subfamily,which are related to the family 2 cystatins and are expressed specifically inthe male reproductive ***1 has a 44%(48/108)identity with mouse CRES and 30%(42/140)identity with mouse cystatin *** blot analysis showed that the Cymgl is specifically expressed inadult mouse *** location studies showed that the GFP-tagged CYMG 1 protein was localized in thecytoplasm of HeLa cells,lmmunohistochemistry revealed that the CYMG1 protein was expressed in mousetestes spermatogonium,spermatocytes,round spermatids,elongating spermatids and ***-PCRresults also showed that Cymgl was expressed in mouse testes and *** Cymgl expressionlevel varied in different developmental stages:it was low 1 week postpartum,steadily increased 2 to 5 weekspostpartum,and was highest 7 weeks *** expression level at 5 weeks postpartum was main-tained during 13 to 57 weeks *** Cymgl expression level in the testes over different develop-mental stages correlates with the mouse spermatogenesis and sexual maturation *** these indicatethat Cymgl might play an important role in mouse spermatogenesis and sexual maturation. Cystatins are cysteine proteinase inhibitors,We found two expression sequence tags(EST

关键词： cystatins gene cloning testis spermatogenesis sexual maturation

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Liquid biopsies for liquid tumors: emerging potential of circulating free nucleic acid evaluation for the management of hematologic malignancies

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Cancer Biology & Medicine 2016年第2期13卷 215-225页

作者： Jay Hocking Sridurga Mithraprabhu Anna Kalff Andrew Spencer Myeloma research Group Australian Center for Blood Diseases Monash University Malignant Haematology & stem cell Transplantation Service Alfred Hospital

Circulating free nucleic acids; cell free DNA and circulating micro-RNA, are found in the plasma of patients with hematologic and solid malignancies at levels higher than that of healthy individuals. In patients with hematologic malignancy cell free DNA reflects the underlying tumor mutational profile, whilst micro-RNAs reflect genetic interference mechanisms within a tumor and potentially the surrounding microenvironment and immune effector cells. These circulating nucleic acids offer a potentially simple, non-invasive, repeatable analysis that can aid in diagnosis, prognosis and therapeutic decisions in cancer treatment.

关键词： Circulating nucleic acids DNA miRNA hematologic malignancy biopsy

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PIWI proteins and their interactors in piRNA biogenesis,germline development and gene expression

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National Science Review 2014年第2期1卷 205-218页

作者： Hsueh-Yen Ku Haifan Lin Yale stem cell Center and Department of cell Biology Yale University School of Medicine

PIWI-interacting RNAs(piRNAs) are a complex class of small non-coding RNAs that are mostly 24–32nucleotides in length and composed of at least hundreds of thousands of species that speciically interact with the PIWI protein subfamily of the ARGONAUTE family. Recent studies revealed that PIWI proteins interact with a number of proteins, especially the TUDOR-domain-containing proteins, to regulate piRNA biogenesis and regulatory function. Current research also provides evidence that PIWI proteins and pi RNAs are not only crucial for transposon silencing in the germline, but also mediate novel mechanisms of epigenetic programming, DNA rearrangements, m RNA turnover, and translational control both in the germline and in the soma. hese new discoveries begin to reveal an exciting new dimension of gene regulation in the cell.

关键词： PIWI piRNA TDRD epigenetic RNA decay translational regulation

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A dysregulated calcium homeostasis as the earliest pathological sign in stem cell-derived Parkinson's disease neurons?

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Neural Regeneration research 2024年第7期19卷 1421-1422页

作者： Nilima Prakash Working Group Human stem cell research Department Hamm 2Hamm-Lippstadt University of Applied SciencesHammGermany

Parkinson’s disease(PD) is characterized by the slow and progressive demise of dopamine(DA)-synthesizing neurons in the substantia nigra pars compacta(SNc),a nucleus located in the human ventral *** death also affects other regions in the brain at later stages of *** concomitant lack of DA in the human forebrain(striatum) leads to the typical motor symptoms of this still uncurable neurodegenerative disorder.

关键词： homeostasis dopamine death

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In sight on multiple morphological abno rmalities of sperm flagella in male infertility:what is new?

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Asian Journal of Andrology 2020年第3期22卷 236-245页

作者： Wei-Li Wang Chao-Feng Tu Yue-Qiu Tan Institute of Reproductive and stem cell Engineering School of Basic Medical ScienceCentral South UniversityChangsha 410078China Reproductive and Genetic Hospital of CITIC-Xiangya Changsha 410078China National Engineering and research Center of Human stem cell Changsha 410078China

The syndrome of multiple morphological abnormalities of the sperm flagella(MMAF)is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities(absent,short,bent,coiled,and irregular flagella).MMAF was proposed in 2014 and has attracted increasing attention;however,it has not been clearly *** this review,we elucidate the definition of MMAF from a systematical view,the difference between MMAF and other conditions with asthenoteratozoospermia or asthenozoospermia(such as primary mitochondrial sheath defects and primary ciliary dyskinesia),the knowledge regarding its etiological mechanism and related genetic findings,and the clinical significance of MMAF for intracytoplasmic sperm injection and genetic coun sell *** review provides the basic kno wledge for MMAF and puts forward some suggestions for further investigations.

关键词： asthe no teratozoospermia disease-causing ge nes in tracytoplasmic sperm in jecti on male in fertility multiple morphological abnormalities of the sperm flagella

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A neurovascular unit-on-a-chip:culture and differentiation of human neural stem cells in a three-dimensional microfluidic environment

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Neural Regeneration research 2022年第10期17卷 2260-2266页

作者： Wen-Juan Wei Ya-Chen Wang Xin Guan Wei-Gong Chen Jing Liu stem cell Clinical research Center National Joint Engineering LaboratoryRegenerative Medicine Centerthe First Affiliated Hospital of Dalian Medical UniversityDalianLiaoning ProvinceChina Dalian Innovation Institute of stem cell and Precision Medicine DalianLiaoning ProvinceChina

Biological studies typically rely on a simple monolayer cell culture,which does not reflect the complex functional characteristics of human tissues and organs,or their real response to external *** technology has advantages of high-throughput screening,accurate control of the fluid velocity,low cell consumption,long-term culture,and high *** combining the multipotential differentiation of neural stem cells with high throughput and the integrated characteristics of microfluidic technology,an in vitro model of a functionalized neurovascular unit was established using human neural stem cell-derived neurons,astrocytes,oligodendrocytes,and a functional microvascular *** model comprises a multi-layer vertical neural module and vascular module,both of which were connected with a syringe *** provides controllable conditions for cell inoculation and nutrient supply,and simultaneously simulates the process of ischemic/hypoxic injury and the process of inflammatory factors in the circulatory system passing through the blood-brain barrier and then acting on the nerve tissue in the *** in vitro functionalized neurovascular unit model will be conducive to central nervous system disease research,drug screening,and new drug development.

关键词： (neural)differentiation astrocyte blood-brain barrier brain microvascular endothelial cells central nervous system microfluidics neural stem cells neuron neurovascular unit oligodendrocyte organ-on-a-chip

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Genome editing and animal models

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Chinese Science Bulletin 2014年第1期59卷 1-6页

作者： Ruby Yanru Chen-Tsai Ruhong Jiang Luping Zhuang Junfeng Wu Lingsong Li Jiarui Wu Department of Animal Disease Modeling Center for Stem Cell and NanoMedicine Shanghai Advanced Research InstituteChinese Academy of Sciences

Transgenic technology allows a gene of interest to be introduced into the genome of a laboratory animal,and provides an extremely powerful tool to dissect the molecular mechanisms of *** mouse models made by microinjection of DNA into zygotic pronuclei in particular have been widely used by the genetics community for 30 ***,it remains a rather crude approach:injected sequences randomly insert in multiple copies as concatamers,they can be mutagenic,and they have variable or silenced expression depending on the site of integration,a phenomenon called position *** a result,multiple lines are required in order to confirm appropriate transgene *** can be partially overcome by flanking transgenes with insulator sequences to protect the transgene from the influence of the surrounding regulatory ***(\300 kb)BACbased transgenic vectors have also been shown to be more resistant to position ***,animals carrying extra copies of fairly large regions of the genome could have unpredictable *** most effective method used to control for position effects is to target transgene insertion to specific genomic loci,the so-called targeted transgenesis;for instance,the fast,site-specific transgenic technology Targatt *** purpose of this review is to provide an overview on the current existing methods for making targeted transgenic mouse models.

关键词：动物模型基因组转基因小鼠模型转基因技术编辑位置效应显微注射 DNA导入

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Boosting the cellular potency of embryonic stem cells by spliceosome targeting

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Signal Transduction and Targeted Therapy 2021年第9期6卷 2613-2614页

作者： Wilfried A.Kues Friedrich-Loeffler-lnstitut Federal Research Institute for Animal HealthBiotechnologyStem Cell PhysiologyNeustadtGermany

In recent work published in cell,Shen et al.1 identified spliceosome inhibition in embryonic stem(ES)cells as a key mechanism for the transition from pluri-to *** inhibition,achieved by RNA interference or the chemical inhibitor pladienolide B,may gain widespread relevance to the culture of totipotent ES cells,in vitro differentiation of extra-embryonal tissue and organoids,translation to the maintenance of pluripotent cells of other mammal species,including humans,and a better molecular understanding of cellular potency in stem cells and cancer.

关键词： some culture

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Luteolin induces hippocampal neurogenesis in the Ts65Dn mouse model of Down syndrome

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Neural Regeneration research 2019年第4期14卷 613-620页

作者： Wen-Bo Zhou Zong-Ning Miao Bin Zhang Wei Long Fang-Xiu Zheng Jing Kong Bin Yu Changzhou Women and Children Health Hospital Affiliated to Nanjing Medical University The stem cell research Laboratory Wuxi Third People's Hospital

Studies have shown that the natural flavonoid luteolin has neurotrophic activity. In this study, we investigated the effect of luteolin in a mouse model of Down syndrome. Ts65 Dn mice, which are frequently used as a model of Down syndrome, were intraperitoneally injected with 10 mg/kg luteolin for 4 consecutive weeks starting at 12 weeks of age. The Morris water maze test was used to evaluate learning and memory abilities, and the novel object recognition test was used to assess recognition memory. Immunohistochemistry was performed for the neural stem cell marker nestin, the astrocyte marker glial fibrillary acidic protein, the immature neuron marker DCX, the mature neuron marker NeuN, and the cell proliferation marker Ki67 in the hippocampal dentate gyrus. Nissl staining was used to observe changes in morphology and to quantify cells in the dentate gyrus. Western blot assay was used to analyze the protein levels of brain-derived neurotrophic factor(BDNF) and phospho-extracellular signal-regulated kinase 1/2(p-ERK1/2) in the hippocampus. Luteolin improved learning and memory abilities as well as novel object recognition ability, and enhanced the proliferation of neurons in the hippocampal dentate gyrus. Furthermore, luteolin increased expression of nestin and glial fibrillary acidic protein, increased the number of DCX^+ neurons in the granular layer and NeuN^+ neurons in the subgranular region of the dentate gyrus, and increased the protein levels of BDNF and p-ERK1/2 in the hippocampus. Our findings show that luteolin improves behavioral performance and promotes hippocampal neurogenesis in Ts65 Dn mice. Moreover, these effects might be associated with the activation of the BDNF/ERK1/2 pathway.

关键词： nerve regeneration luteolin hippocampus Ts65Dn mice neurogenesis ERK BDNF nestin GFAP NeuN Down syndrome neural regeneration

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