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mouse testis RNA

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https://www.readbyqxmd.com/read/28298247/tissue-specific-inhibition-of-urokinase-type-plasminogen-activator-expression-in-the-testes-of-mice-by-inducible-lentiviral-rna-interference-causes-male-infertility
#1
Kai Zhao, Yan Liu, Zhe Xiong, Lian Hu, Cheng-Liang Xiong
Urokinase-type plasminogen activator (uPA) is involved in many physiological processes, including male infertility. To explore the effects of uPA in male reproduction, we constructed an inducible uPA short hairpin RNA (shRNA) system expressed by lentiviral vectors. After proving inhibition of uPA expression in the mouse Sertoli cell line TM4 by 1μgmL-1 doxycycline (Dox), two lentivirus (pLenti4-shRNA and pLenti6/TR) were co-microinjected into mouse testes to produce TetR&shuPA mice model. Though oral gavage by 0...
March 16, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28291811/systematic-identification-and-characterization-of-long-non-coding-rnas-in-mouse-mature-sperm
#2
Xiaoning Zhang, Fengxin Gao, Jianbo Fu, Peng Zhang, Yuqing Wang, Xuhui Zeng
Increasing studies have shown that mature spermatozoa contain many transcripts including mRNAs and miRNAs. However, the expression profile of long non-coding RNAs (lncRNAs) in mammalian sperm has not been systematically investigated. Here, we used highly purified RNA to investigate lncRNA expression profiles in mouse mature sperm by stranded-specific RNA-seq. We identified 20,907 known and 4,088 novel lncRNAs transcripts, and the existence of intact lncRNAs was confirmed by RT-PCR and fluorescence in situ hybridization on two representative lncRNAs...
2017: PloS One
https://www.readbyqxmd.com/read/28278534/transcription-and-splicing-factor-tdp-43-role-in-regulation-of-gene-expression-in-testis
#3
Prabhakara P Reddi
TDP-43 (TAR DNA binding Protein of 43 kD) is a transcription factor and RNA-binding protein with diverse functions. We cloned TDP-43 from the mouse testis in a screen for promoter-binding proteins and showed that it functions as a transcriptional repressor. TDP-43 plays a role in maintaining the precise pattern of spatiotemporal expression of the spermatid-specific Acrv1 gene during spermatogenesis by facilitating RNA polymerase II pausing at the promoter. We also showed that TDP-43 plays a partial role in preventing somatic cell expression of the Acrv1 gene by acting as an insulator-binding protein...
March 2017: Seminars in Reproductive Medicine
https://www.readbyqxmd.com/read/28214373/improved-detection-of-zika-virus-rna-in-human-and-animal-specimens-by-a-novel-highly-sensitive-and-specific-real-time-rt-pcr-assay-targeting-the-5-untranslated-region-of-zika-virus
#4
Jasper Fuk-Woo Chan, Cyril Chik-Yan Yip, Kah-Meng Tee, Zheng Zhu, Jessica Oi-Ling Tsang, Kenn Ka-Heng Chik, Terance Gi-Wai Tsang, Chris Chung-Sing Chan, Vincent Kwok-Man Poon, Siddharth Sridhar, Feifei Yin, Ivan Fan-Ngai Hung, Sandy Ka-Yee Chau, Anna Jinxia Zhang, Kwok-Hung Chan, Kwok-Yung Yuen
OBJECTIVE AND METHOD: We developed and evaluated five novel real-time RT-PCR assays targeting conserved regions in the 5'-untranslated region (5'-UTR), envelope (E'), non-structural protein 2A (NS2A), NS5, and 3'-UTR of the ZIKV genome. RESULTS: The ZIKV-5'-UTR assay exhibited the lowest in-vitro limit of detection (5-10 RNA copies/reaction and 3.0×10(-1) plaque-forming units/ml). Compared to the modified version of a widely adopted RT-PCR assay targeting the ZIKV-E gene, the ZIKV-5'-UTR assay showed better sensitivity in human clinical specimens, and representative mouse specimens, including many organs which are known to be involved in human ZIKV infection but difficult to obtain in clinical settings...
February 18, 2017: Tropical Medicine & International Health: TM & IH
https://www.readbyqxmd.com/read/28209515/cell-type-specific-role-of-the-rna-binding-protein-nono-in-the-dna-double-strand-break-response-in-the-mouse-testes
#5
Shuyi Li, Feng-Jue Shu, Zhentian Li, Lahcen Jaafar, Shourong Zhao, William S Dynan
The tandem RNA recognition motif protein, NONO, was previously identified as a candidate DNA double-strand break (DSB) repair factor in a biochemical screen for proteins with end-joining stimulatory activity. Subsequent work showed that NONO and its binding partner, SFPQ, have many of the properties expected for bona fide repair factors in cell-based assays. Their contribution to the DNA damage response in intact tissue in vivo has not, however, been demonstrated. Here we compare DNA damage sensitivity in the testes of wild-type mice versus mice bearing a null allele of the NONO homologue (Nono (gt))...
March 2017: DNA Repair
https://www.readbyqxmd.com/read/28196369/leveraging-online-resources-to-prioritize-candidate-genes-for-functional-analyses-using-the-fetal-testis-as-a-test-case
#6
Kathryn S McClelland, Humphrey H-C Yao
With each new microarray or RNA-seq experiment, massive quantities of transcriptomic information are generated with the purpose to produce a list of candidate genes for functional analyses. Yet an effective strategy remains elusive to prioritize the genes on these candidate lists. In this review, we outline a prioritizing strategy by taking a step back from the bench and leveraging the rich range of public databases. This in silico approach provides an economical, less biased, and more effective solution. We discuss the publicly available online resources that can be used to answer a range of questions about a gene...
2017: Sexual Development: Genetics, Molecular Biology, Evolution, Endocrinology, Embryology, and Pathology of Sex Determination and Differentiation
https://www.readbyqxmd.com/read/28191733/a-novel-dynamic-expression-of-vasa-in-male-germ-cells-during-spermatogenesis-in-the-chinese-soft-shell-turtle-pelidiscus-sinensis
#7
Wei Li, Piaoyi Zhang, Xuling Wu, Xinping Zhu, Hongyan Xu
vasa gene encodes a highly conserved DEAD-box RNA helicase, required for germ cell development across animal kingdom. Vasa mutations cause male infertility in mammals. It has been widely used as a biomarker for studying animal fertility or manipulating germ cells in organisms. However, in reptilians, the functions of vasa gene involved in germ cell differentiation are largely unclear; this hampers the development of biological techniques and the improvement of the productivity in these species. Here a vasa cDNA was isolated in Chinese soft-shell turtle and it predicts a protein of 691 amino acid residues, which is 72%, 69%, 58%, 59%, and 54-56% identical to its homolog from mouse, platypus, frog, chicken, and fish, respectively, and named as PsVasa...
February 13, 2017: Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution
https://www.readbyqxmd.com/read/28145452/testis-specific-transcriptional-regulators-selectively-occupy-boris-bound-ctcf-target-regions-in-mouse-male-germ-cells
#8
Samuel Rivero-Hinojosa, Sungyun Kang, Victor V Lobanenkov, Gabriel E Zentner
Despite sharing the same sequence specificity in vitro and in vivo, CCCTC-binding factor (CTCF) and its paralog brother of the regulator of imprinted sites (BORIS) are simultaneously expressed in germ cells. Recently, ChIP-seq analysis revealed two classes of CTCF/BORIS-bound regions: single CTCF target sites (1xCTSes) that are bound by CTCF alone (CTCF-only) or double CTCF target sites (2xCTSes) simultaneously bound by CTCF and BORIS (CTCF&BORIS) or BORIS alone (BORIS-only) in germ cells and in BORIS-positive somatic cancer cells...
February 1, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28096900/ms_histonedb-a-manually-curated-resource-for-proteomic-analysis-of-human-and-mouse-histones
#9
Sara El Kennani, Annie Adrait, Alexey K Shaytan, Saadi Khochbin, Christophe Bruley, Anna R Panchenko, David Landsman, Delphine Pflieger, Jérôme Govin
BACKGROUND: Histones and histone variants are essential components of the nuclear chromatin. While mass spectrometry has opened a large window to their characterization and functional studies, their identification from proteomic data remains challenging. Indeed, the current interpretation of mass spectrometry data relies on public databases which are either not exhaustive (Swiss-Prot) or contain many redundant entries (UniProtKB or NCBI). Currently, no protein database is ideally suited for the analysis of histones and the complex array of mammalian histone variants...
2017: Epigenetics & Chromatin
https://www.readbyqxmd.com/read/27992363/germ-cell-specific-sustained-activation-of-wnt-signalling-perturbs-spermatogenesis-in-aged-mice-possibly-through-non-coding-rnas
#10
Manish Kumar, Joshua Atkins, Murray Cairns, Ayesha Ali, Pradeep S Tanwar
Dysregulated Wnt signalling is associated with human infertility and testicular cancer. However, the role of Wnt signalling in male germ cells remains poorly understood. In this study, we first confirmed the activity of Wnt signalling in mouse, dog and human testes. To determine the physiological importance of the Wnt pathway, we developed a mouse model with germ cell-specific constitutive activation of βcatenin. In young mutants, similar to controls, germ cell development was normal. However, with age, mutant testes showed defective spermatogenesis, progressive germ cell loss, and flawed meiotic entry of spermatogonial cells...
December 15, 2016: Oncotarget
https://www.readbyqxmd.com/read/27941834/lin28a-promotes-self-renewal-and-proliferation-of-dairy-goat-spermatogonial-stem-cells-sscs-through-regulation-of-mtor-and-pi3k-akt
#11
Fanglin Ma, Zhe Zhou, Na Li, Liming Zheng, Chongyang Wu, Bowen Niu, Furong Tang, Xin He, Guangpeng Li, Jinlian Hua
Lin28a is a conserved RNA-binding protein that plays an important role in development, pluripotency, stemness maintenance, proliferation and self-renewal. Early studies showed that Lin28a serves as a marker of spermatogonial stem cells (SSCs) and promotes the proliferation capacity of mouse SSCs. However, there is little information about Lin28a in livestock SSCs. In this study, we cloned Capra hircus Lin28a CDS and found that it is evolutionarily conserved. Lin28a is widely expressed in different tissues of Capra hircus, but is expressed at a high level in the testis...
December 12, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27895724/expression-profile-of-spaca5-spaca5-in-spermatogenesis-and-transitional-cell-carcinoma-of-the-bladder
#12
Xiao-Yue Zhang, Qiu-Xia Yan, Xiao-Yan Guo, Cai-Rong Chen, Run-Qiang Chen, Zhi-Ming Cai, Ai-Fa Tang
The majority of bladder cancer-associated mortalities are due to transitional cell carcinoma (TCC), which is the most prevalent and chemoresistant malignancy of the bladder. Sperm acrosome associated 5 (SPACA5)/Spaca5 is a sperm acrosome-associated, c-type lysozyme-like protein that has been recently identified, and has been designated as an attractive candidate antigen for cancer testis. In the present study, the expression profile of SPACA5/Spaca5 was analyzed in spermatogenesis and TCC of the bladder using diverse molecular and cellular biology methods...
November 2016: Oncology Letters
https://www.readbyqxmd.com/read/27856695/multiplex-shrna-screening-of-germ-cell-development-by-in-vivo-transfection-of-mouse-testis
#13
Nicholas R Y Ho, Abul R Usmani, Yan Yin, Liang Ma, Donald F Conrad
Spermatozoa are one of the few mammalian cell types that cannot be fully derived in vitro, severely limiting the application of modern genomic techniques to study germ cell biology. The current gold standard approach of characterizing single-gene knockout mice is slow as generation of each mutant line can take 6-9 months. Here, we describe an in vivo approach to rapid functional screening of germline genes based on a new nonsurgical, nonviral in vivo transfection method to deliver nucleic acids into testicular germ cells...
January 5, 2017: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/27780883/human-umbilical-perivascular-cells-a-novel-source-of-mscs-to-support-testicular-niche-regeneration
#14
Leila Maghen, Ekaterina Shlush, Itai Gat, Melissa Filice, Tanya Barretto, Keith Jarvi, Kirk Lo, Andree S Gauthier-Fisher, Clifford L Librach
The expansion of functional testicular biopsy-derived human spermatogonial stem cells (hSSC) ex-vivo may enable the restoration of fertility in pre-pubertal males having undergone gonadotoxic therapies or men with severe male factor infertility. Various somatic cells are known to regulate SSC homeostasis and spermatogenesis in the developing and adult testis. Prior attempts to recapitulate this niche demonstrated the requirement of feeder cells, such as endogenous testicular somatic cells, for germ cell expansion ex-vivo...
October 25, 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/27734358/identification-of-mouse-pirna-pathway-components-using-anti-miwi2-antibodies
#15
Takamasa Hirano, Hidetoshi Hasuwa, Haruhiko Siomi
The mouse testis has served as a popular model system to study a wide range of biological processes, including germ cell development, meiosis, epigenetic changes of chromatin, transposon silencing, and small RNA-mediated epigenetic modifications. PIWI-interacting RNAs (piRNAs) are a class of small RNAs that are almost exclusively expressed in animal gonads. They repress transposons by forming effector complexes with PIWI proteins to maintain genome integrity of the germline. Here we describe detailed procedures of how to produce monoclonal antibodies against a mouse nuclear PIWI protein, MIWI2, which functions in de novo DNA methylation of target transposon loci...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27733379/tspan8-expression-distinguishes-spermatogonial-stem-cells-in-the-prepubertal-mouse-testis
#16
Kazadi Mutoji, Anukriti Singh, Thu Nguyen, Heidi Gildersleeve, Amy V Kaucher, Melissa J Oatley, Jon M Oatley, Ellen K Velte, Christopher B Geyer, Keren Cheng, John R McCarrey, Brian P Hermann
Precise separation of spermatogonial stem cells (SSCs) from progenitor spermatogonia that lack stem cell activity and are committed to differentiation remains a challenge. To distinguish between these spermatogonial subtypes, we identified genes that exhibited bimodal mRNA levels at the single-cell level among undifferentiated spermatogonia from Postnatal Day 6 mouse testes, including Tspan8, Epha2, and Pvr, each of which encode cell surface proteins useful for cell selection. Transplantation studies provided definitive evidence that a TSPAN8-high subpopulation is enriched for SSCs...
December 2016: Biology of Reproduction
https://www.readbyqxmd.com/read/27560004/expression-dynamics-relationships-and-transcriptional-regulations-of-diverse-transcripts-in-mouse-spermatogenic-cells
#17
Xiwen Lin, Miao Han, Lu Cheng, Jian Chen, Zhuqiang Zhang, Ting Shen, Min Wang, Bo Wen, Ting Ni, Chunsheng Han
Among all tissues of the metazoa, the transcritpome of testis displays the highest diversity and specificity. However, its composition and dynamics during spermatogenesis have not been fully understood. Here, we have identified 20,639 message RNAs (mRNAs), 7,168 long non-coding RNAs (lncRNAs) and 15,101 circular RNAs (circRNAs) in mouse spermatogenic cells, and found many of them were specifically expressed in testes. lncRNAs are significantly more testis-specific than mRNAs. At all stages, mRNAs are generally more abundant than lncRNAs, and linear transcripts are more abundant than circRNAs...
October 2, 2016: RNA Biology
https://www.readbyqxmd.com/read/27519264/regulatory-complexity-revealed-by-integrated-cytological-and-rna-seq-analyses-of-meiotic-substages-in-mouse-spermatocytes
#18
Robyn L Ball, Yasuhiro Fujiwara, Fengyun Sun, Jianjun Hu, Matthew A Hibbs, Mary Ann Handel, Gregory W Carter
BACKGROUND: The continuous and non-synchronous nature of postnatal male germ-cell development has impeded stage-specific resolution of molecular events of mammalian meiotic prophase in the testis. Here the juvenile onset of spermatogenesis in mice is analyzed by combining cytological and transcriptomic data in a novel computational analysis that allows decomposition of the transcriptional programs of spermatogonia and meiotic prophase substages. RESULTS: Germ cells from testes of individual mice were obtained at two-day intervals from 8 to 18 days post-partum (dpp), prepared as surface-spread chromatin and immunolabeled for meiotic stage-specific protein markers (STRA8, SYCP3, phosphorylated H2AFX, and HISTH1T)...
August 12, 2016: BMC Genomics
https://www.readbyqxmd.com/read/27486773/integrated-analysis-mirna-and-mrna-profiling-in-patients-with-severe-oligozoospermia-reveals-mir-34c-3p-downregulates-plcxd3-expression
#19
Zhiming Li, Zaozao Zheng, Jun Ruan, Zhi Li, Xuan Zhuang, Chi-Meng Tzeng
Our previous research suggested that an integrated analysis of microRNA (miRNA) and messenger RNA (mRNA) expression is helpful to explore miRNA-mRNA interactions and to uncover the molecular mechanisms of male infertility. In this study, microarrays were used to compare the differences in the miRNA and mRNA expression profiles in the testicular tissues of severe oligozoospermia (SO) patients with obstructive azoospermia (OA) controls with normal spermatogenesis. Four miRNAs (miR-1246, miR-375, miR-410, and miR-758) and six mRNAs (SLC1A3, PRKAR2B, HYDIN, WDR65, PRDX1, and ADATMS5) were selected to validate the microarray data using quantitative real-time PCR...
August 16, 2016: Oncotarget
https://www.readbyqxmd.com/read/27473657/mouse-d1pas1-a-dead-box-rna-helicase-is-required-for-the-completion-of-first-meiotic-prophase-in-male-germ-cells
#20
Hiroki Inoue, Narumi Ogonuki, Michiko Hirose, Yuki Hatanaka, Shogo Matoba, Shinichiro Chuma, Kimio Kobayashi, Shigeharu Wakana, Junko Noguchi, Kimiko Inoue, Kentaro Tanemura, Atsuo Ogura
D1Pas1 is a mouse autosomal DEAD-box RNA helicase expressed predominantly in the testis. To assess its possible function, we generated D1Pas1-deficient mice using embryonic stem cells with a targeted D1Pas1 allele. Deletion of D1Pas1 did not cause noticeable embryonic defects or death, indicating that D1Pas1 is not essential for embryogenesis. Whereas homozygous knockout female mice showed normal reproductive performance, homozygous knockout male mice were completely sterile. The seminiferous epithelium of D1Pas1-deficient males contained no spermatids or spermatozoa because of spermatogenic arrest at the late pachytene stage...
September 16, 2016: Biochemical and Biophysical Research Communications
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