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      RSPH6A is required for sperm flagellum formation and male fertility in mice

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          Abstract

          <p id="d636751e275">The flagellum is an evolutionarily conserved appendage used for sensing and locomotion. Its backbone is the axoneme and a component of the axoneme is the radial spoke (RS), a protein complex implicated in flagellar motility regulation. Numerous diseases occur if the axoneme is improperly formed, such as primary ciliary dyskinesia (PCD) and infertility. Radial spoke head 6 homolog A (RSPH6A) is an ortholog of <i>Chlamydomonas</i> RSP6 in the RS head and is evolutionarily conserved. While some RS head proteins have been linked to PCD, little is known about RSPH6A. Here, we show that mouse RSPH6A is testis-enriched and localized in the flagellum. <i>Rsph6a</i> knockout (KO) male mice are infertile as a result of their short immotile spermatozoa. Observation of the KO testis indicates that the axoneme can elongate but is disrupted before accessory structures are formed. Manchette removal is also impaired in the KO testis. Further, RSPH9, another radial spoke protein, disappeared in the <i>Rsph6a</i> KO flagella. These data indicate that RSPH6A is essential for sperm flagellar assembly and male fertility in mice. </p><p id="d636751e286"> <a data-untrusted="" href="http://dx.doi.org/10.1242/jcs.225920" id="d636751e288" target="xrefwindow">This article has an associated First Person interview with the first author of the paper. </a> </p><p class="first" id="d636751e292"> <b>Summary:</b> Removal of RSPH6A via CRISPR/Cas9 results in short-tailed immotile spermatozoa and loss of fertilizing ability in male mice. </p>

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          Most cited references34

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          Is Open Access

          SMART: recent updates, new developments and status in 2015

          SMART (Simple Modular Architecture Research Tool) is a web resource (http://smart.embl.de/) providing simple identification and extensive annotation of protein domains and the exploration of protein domain architectures. In the current version, SMART contains manually curated models for more than 1200 protein domains, with ∼200 new models since our last update article. The underlying protein databases were synchronized with UniProt, Ensembl and STRING, bringing the total number of annotated domains and other protein features above 100 million. SMART's ‘Genomic’ mode, which annotates proteins from completely sequenced genomes was greatly expanded and now includes 2031 species, compared to 1133 in the previous release. SMART analysis results pages have been completely redesigned and include links to several new information sources. A new, vector-based display engine has been developed for protein schematics in SMART, which can also be exported as high-resolution bitmap images for easy inclusion into other documents. Taxonomic tree displays in SMART have been significantly improved, and can be easily navigated using the integrated search engine.
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            Efficient selection for high-expression transfectants with a novel eukaryotic vector

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              Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization

              A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).
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                Author and article information

                Journal
                Journal of Cell Science
                J Cell Sci
                The Company of Biologists
                0021-9533
                1477-9137
                October 11 2018
                October 01 2018
                October 01 2018
                September 05 2018
                : 131
                : 19
                : jcs221648
                Article
                10.1242/jcs.221648
                6198453
                30185526
                7eff31c3-ada4-4b21-85a3-ce6eb9f40843
                © 2018

                http://www.biologists.com/user-licence-1-1

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