GSE195744
Mouse testicular gene expression profile after KIFC1 inhibition
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. Overall design: Mouse testicular mRNA profiles of PBS, AZ82 and CW069 injection groups.