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Deficient spermiogenesis in mice lacking Rlim.

ABSTRACT: The X-linked gene Rlim plays major roles in female mouse development and reproduction, where it is crucial for the maintenance of imprinted X chromosome inactivation in extraembryonic tissues of embryos. However, while females carrying a systemic Rlim knockout (KO) die around implantation, male Rlim KO mice appear healthy and are fertile. Here, we report an important role for Rlim in testis where it is highly expressed in post-meiotic round spermatids as well as in Sertoli cells. Systemic deletion of the Rlim gene results in lower numbers of mature sperm that contains excess cytoplasm, leading to decreased sperm motility and in vitro fertilization rates. Targeting the conditional Rlim cKO specifically to the spermatogenic cell lineage largely recapitulates this phenotype. These results reveal functions of Rlim in male reproduction specifically in round spermatids during spermiogenesis.

SUBMITTER: Wang F

PROVIDER: S-EPMC7935487 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Deficient spermiogenesis in mice lacking Rlim.

Wang Feng F Gervasi Maria Gracia MG Bošković Ana A Sun Fengyun F Rinaldi Vera D VD Yu Jun J Wallingford Mary C MC Tourzani Darya A DA Mager Jesse J Zhu Lihua Julie LJ Rando Oliver J OJ Visconti Pablo E PE Strittmatter Lara L Bach Ingolf I

eLife 20210223

The X-linked gene Rlim plays major roles in female mouse development and reproduction, where it is crucial for the maintenance of imprinted X chromosome inactivation in extraembryonic tissues of embryos. However, while females carrying a systemic Rlim knockout (KO) die around implantation, male Rlim KO mice appear healthy and are fertile. Here, we report an important role for Rlim in testis where it is highly expressed in post-meiotic round spermatids as well as in Se ...[more]

PMID: 33620316

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Project description:BackgroundLubricin, a mucinous glycoprotein secreted by synoviocytes and chondrocytes plays an important role in reducing the coefficient of friction in mammalian joints. Elevated cartilage surface friction is thought to cause chondrocyte loss; however, its quantification and methodological approaches have not been reported. We adapted a stereological method and incorporated vital cell staining to assess cellular loss in superficial and upper intermediate zones in lubricin deficient mouse cartilage.MethodsThe femoral condyle cartilage of the intact knees from lubricin wild type (Prg4 (+/+)), heterozygote (Prg4 (+/-)), and knockout (Prg4 (-/-)) mice was imaged using fluorescein diacetate (FDA), propidium iodide (PI), and Hoechst staining, and confocal microscopy. Three dimensional reconstructions of confocal images to a depth of 14 ?m were analyzed using Matlab to determine the volume fraction occupied by chondrocytes in cartilage of both medial and lateral femoral condyles. Living chondrocyte volume fraction was defined as FDA stained chondrocyte volume/total volume of superficial?+?upper intermediate zone. Living and dead (total) chondrocyte volume fraction was defined as FDA?+?PI stained chondrocyte volume/total volume of superficial?+?upper intermediate zone. MicroCT provided an orthogonal measure of cartilage thickness. Immunohistology for activated caspase-3 and TUNEL staining were performed to evaluate the presence of apoptotic chondrocytes in Prg4 mutant mice.ResultsLiving chondrocyte volume fraction of the medial femoral condyle was significantly lower in Prg4 (-/-) mice compared to Prg4 (+/+) (p?=?0.002) and Prg4 (+/-) (p?=?0.002) littermates. There was no significant difference in medial condyle chondrocyte volume fraction between Prg4 (+/+) and Prg4 (+/-) mice (p?=?0.82). No significant differences were observed for the chondrocyte volume fraction for the lateral condyle (p?>?0.26). Cartilage thickness increased in the medial condyle for Prg4 (-/-) mice compared to Prg4 (+/+) (p?=?0.02) and Prg4 (+/-) (p?=?0.03) littermates, and the lateral condyle for Prg4 (-/-) mice compared to Prg4 (+/+) (p?<?0.0001) and Prg4 (+/-) (p?<?0.0001) littermates, indicating that a multi-dimensional increase in cartilage volume did not artifactually lower the chondrocyte volume fraction in the medial condyle. Significantly higher number of caspase-3 positive cells were observed in the superficial and upper intermediate zone cartilage of the medial femoral condyle of Prg4 (-/-) mice compared to Prg4 (+/+) (p?=?0.01) and Prg4 (+/-) (p?=?0.04) littermates, and the lateral femoral condyle of Prg4 (-/-) mice compared to Prg4 (+/+) (p?=?0.02) and Prg4 (+/-) (p?=?0.02) littermates. There were no significant differences in TUNEL staining among different Prg4 genotypes in both condyles (p?>?0.05 for all comparisons).ConclusionsIncreased Caspase-3 activation is observed in Prg4 deficient mice compared to Prg4 sufficient littermates. Absence of Prg4 induces loss of chondrocytes in the superficial and upper intermediate zone of mouse cartilage that is quantifiable by a novel image processing technique.

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Deficient spermiogenesis in mice lacking Rlim.

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Deficient spermiogenesis in mice lacking <i>Rlim</i>.

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