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Decreased Expression of KIFC1 in Human Testes with Globozoospermic Defects.

ABSTRACT: Globozoospermia is a rare (prevalence of <0.1%) but severe male infertility condition. In our previous study, we found that robust KIFC1 immunostaining was detected in the human elongating/elongated spermatids during human acrosomogenesis. However, the relationship between the decreased expression of KIFC1 and human globozoospermia remains largely unknown. Testicular biopsies of 30 globozoospermia and 30 obstructive azoospermia patients who underwent infertility evaluation and treatment were utilized in this study. Reverse transcriptase polymerase chain reaction (RT-PCR), Western blots, immunohistochemistry, an in vivo model, and intratesticular injection of small inhibitory RNA (siRNA) against the Kifc1 gene were employed, and sperm abnormalities were evaluated by hematoxylin and eosin (H&E) staining and immunocytochemistry. We revealed that the testicular level of KIFC1 mRNA in globozoospermia was significantly reduced compared with that in obstructive azoospermia, and the KIFC1 protein was barely detectable in testicular specimens in 30% (9 of 30) of patients with globozoospermia. Furthermore, knockdown of the Kifc1 gene in mice increased the percentage of sperm with globozoospermic defects (26.5%). Decreased KIFC1 expression was mainly observed in the testes of patients with globozoospermia at the spermatid stage, which may be useful for counseling and management of such patients.

SUBMITTER: Zhi E

PROVIDER: S-EPMC5083914 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Deletions of chromosome 1p36 are associated with a high incidence of congenital heart defects (CHDs). The arginine-glutamic acid dipeptide repeats gene (RERE) is located in a critical region for CHD on chromosome 1p36 and encodes a cardiac-expressed nuclear receptor co-regulator. Mutations affecting RERE cause atrial and ventricular septal defects (VSDs) in humans, and RERE-deficient mice also develop VSDs. During cardiac development, mesenchymal cells destined to form part of the atrioventricular (AV) septum are generated when endocardial cells in the AV canal undergo epithelial-to-mesenchymal transition (EMT) and migrate into the space between the endocardium and the myocardium. These newly generated mesenchymal cells then proliferate to fill the developing AV endocardial cushions. Here, we demonstrate that RERE-deficient mouse embryos have reduced numbers of mesenchymal cells in their AV endocardial cushions owing to decreased levels of EMT and mesenchymal cell proliferation. In the endocardium, RERE colocalizes with GATA4, a transcription factor required for normal levels of EMT and mesenchymal cell proliferation. Using a combination of in vivo and in vitro studies, we show that Rere and Gata4 interact genetically in the development of CHDs, RERE positively regulates transcription from the Gata4 promoter and GATA4 levels are reduced in the AV canals of RERE-deficient embryos. Tissue-specific ablation of Rere in the endocardium leads to hypocellularity of the AV endocardial cushions, defective EMT and VSDs, but does not result in decreased GATA4 expression. We conclude that RERE functions in the AV canal to positively regulate the expression of GATA4, and that deficiency of RERE leads to the development of VSDs through its effects on EMT and mesenchymal cell proliferation. However, the cell-autonomous role of RERE in promoting EMT in the endocardium must be mediated by its effects on the expression of proteins other than GATA4.This article has an associated First Person interview with the first author of the paper.

Project description:Purpose: Kinesin-14 KIFC1 proteins are highly expressed in testes and function in male spermatogenesis, especially acrosome biogenesis. However, the mechanisms of KIFC1 during acrosomal vesicles transportation and vesicle fusion are still unknown. This study revealed the in vivo functions of KIFC1 proteins in mouse spermiogenesis using two KIFC1 specific inhibitors, AZ82 and CW069. Methods: Testicular mRNA profiles of male mice PBS (Control), AZ82 and CW069 injected groups were progressed with deep sequencing, in triplicate through Illumina sequencing platform (HiSeqTM 2500). Genes with adjusted P-value < 0.05 and foldChange > 1.2 were defined as differentially expressed genes (DEGs) and subjected to the following GO and KEGG enrichment. qRT-PCR was performed to validate several essential genes using QuantStudio 5 (Thermo Fisher) and SYBR Green assays. Results: We have mapped over 40 million sequence reads to the mice genome in each specimen. Only a few genes showed significant differences in our gene expression profile of KIFC1 inhibition groups compared with the control. The transcriptome of testes in the AZ82 treatment group reflected the defects in protein maturation. CW069 activated apparent drug responses. In results, AZ82 and CW069 reduced cell viability with an enhanced apoptosis signal and excessive oxidation. Comparing the GO enrichment data, we confirmed the role of KIFC1 in actin filament organization and intercellular adhesion. KEGG enrichment showed a series of lipid, organic acid and steroid metabolism disorders after KIFC1 inhibition, resulting in fatty acid degradation and lipid oxidation. Conclusions: Our study confirmed that KIFC1 functioned in the microfilament organization and proacrosomal vesicle transportation. The acrosin could not accumulate on the nuclear surface with fatty acid degradation. The RNA-seq results were consistent with our in vivo experiments which proved that KIFC1 plays an essential role in mouse acrosome biogenesis.

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Decreased Expression of KIFC1 in Human Testes with Globozoospermic Defects.

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