The C-terminal kinesin motor KIFC1 may participate in nuclear reshaping and flagellum formation during spermiogenesis of Larimichthys crocea

Ding

et al. 2019

Genes

Larimichthys crocea is an economically important marine fish in China. To date, the molecular mechanisms underlying testicular development and spermatogenesis in L. crocea have not been thoroughly elucidated. In this study, we conducted a comparative transcriptome analysis between testes (TES) and pooled multiple tissues (PMT) (liver, spleen, heart, and kidney) from six male individuals. More than 54 million clean reads were yielded from TES and PMT libraries. After mapping to the draft genome of L. crocea, we acquired 25,787 genes from the transcriptome dataset. Expression analyses identified a total of 3853 differentially expressed genes (DEGs), including 2194 testes-biased genes (highly expressed in the TES) and 1659 somatic-biased genes (highly expressed in the PMT). The dataset was further annotated by blasting with multi-databases. Functional genes and enrichment pathways involved in spermatogenesis and testicular development were analyzed, such as the neuroactive ligand-receptor interaction pathway, gonadotropin-releasing hormone (GnRH) and mitogen-activated protein kinase (MAPK) signaling pathways, cell cycle pathway, and dynein, kinesin, myosin, actin, heat shock protein (hsp), synaptonemal complex protein 2 (sycp2), doublesex-and mab-3-related transcription factor 1 (dmrt1), spermatogenesis-associated genes (spata), DEAD-Box Helicases (ddx), tudor domain-containing protein (tdrd), and piwi genes. The candidate genes identified by this study lay the foundation for further studies into the molecular mechanisms underlying testicular development and spermatogenesis in L. crocea.Genes 2019, 10, 958 2 of 22 remains for somatic growth; these changes eventually reduce the culture benefit of L. crocea [3] and hinder the conservation of its germplasm resources. Therefore, the molecular mechanisms underlying reproduction regulation must be investigated, and the genes involved in gametogenesis and gonadal development must be identified, to provide a theoretical reference for further studies on the above problems in L. crocea.Directly correlated with sexual maturation and reproduction, in various animal species, including L. crocea, the testis is an essential basic component of the animal reproductive system and responsible for the production of male gametes via spermatogenesis [4]. During the process of spermatogenesis, diploid spermatogonia slowly evolve into many highly specialized spermatozoa through mitosis, meiosis, and spermiogenesis. Stringent spatial and temporal expression of genes during both transcriptional and translational processes are fundamental to ensure the precise processes of spermatogenesis [5]. Previous studies have focused on the reproductive and developmental biology of L. crocea based on anatomical/histological aspects [6,7], whereas few studies have focused on genes related to spermatogenesis and testicular development. To the best of our knowledge, only a small number of reproduction-related genes have been identified in L. crocea [8][9][10][11]. To further comprehensively explore the mol...

mentioning

confidence: 99%

Transcriptome Sequencing Reveals the Traits of Spermatogenesis and Testicular Development in Large Yellow Croaker (Larimichthys crocea)

Luo

Ding

et al. 2019

Genes

“…RNA was extracted from testes in O. Bidens. RNA extraction was performed as previously described [56]. First, tissue samples were mixed with TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) and homogenized.…”

Section: Full-length Cdna Cloning Of Ob-phbmentioning

confidence: 99%

“…The samples used for WB were the same as described in qPCR. Protein extraction assay was performed based on Zhang's research [56]. First, RIPA (Solarbio, Shanghai, China) was used to extract the total protein from each tissue, with PMSF as a protease inhibitor.…”

Section: Western Blottingmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Mitochondrial Prohibitin in Opsariichthys bidens and Its Potential Functions in Spermatogenesis

Wang

et al. 2022

IJMS

Spermatogenesis is the intricate and coordinated process by which spermatogonia develop into haploid differentiated spermatozoa. Mitochondria are essential for spermatogenesis, and prohibitin (PHB) is closely associated with mitochondrial structure and function during spermatogenesis. Although PHB has been implicated in spermatogenesis in some taxa, its roles in Opsariichthys bidens have not been determined. In this study, the expression patterns and potential functions of PHB in spermatogenesis in O. bidens were characterized using histological microscopic observations, PCR cloning, real-time quantitative PCR (qPCR), Western blotting (WB) and immunofluorescence (IF). The full-length cDNA of Ob-phb was 1500 bp encoding 271 amino acids. A sequence alignment demonstrated that the PHB protein is conserved among different animals. qPCR revealed that phb mRNA is widely distributed in O. bidens and highly expressed in the testes at stages IV and V. WB revealed that Ob-PHB is located in the mitochondria of testes. IF revealed the colocalization of PHB signals and mitochondria. Signals were detected around nuclei in spermatogonia and spermatocytes, gradually moving to the tail region during spermiogenesis, and finally aggregating in the midpiece. These results indicate that Ob-PHB was expressed in the mitochondria during spermatogenesis. In addition, this study proposed Ob-PHB may participate in the degradation of mitochondria and cell differentiation during spermatogenesis.

“…L. crocea undergoes spermatid remodeling, which involves nuclear reshaping, tail formation, and excess cytoplasm removal, during spermiogenesis. It has been used as a model for studying spermiogenesis in sh (Luo et al 2019;Mu et al 2019;Zhang et al 2017). This study aimed to investigate the functions of KIF17 in spermatid remodeling during L. crocea spermiogenesis to contribute to improved understanding of the mechanism underlying sh spermiogenesis and to provide a basis for further research on sh reproduction and development.…”

Section: Introductionmentioning

confidence: 99%

The Potential Function of KIF17 in Large Yellow Croaker (Larimichthys crocea) Spermatid Remodeling: Molecular Characterization and Expression Pattern During Spermiogenesis

Wang

Liu

et al. 2021

Preprint

Self Cite

KIF17, which belongs to the kinesin-2 protein family, plays an indispensable role in mammalian spermiogenesis. However, the role of KIF17 in fish spermatid remodeling during spermiogenesis remains poorly understood. Therefore, we aimed to study the role of KIF17 in spermatid remodeling during Larimichthys crocea (L. crocea) spermiogenesis. The kif17 cDNA sequence, 3247 bp in length, was cloned from L. crocea testis, which consisted of a 347 bp 5ʹ-untranslated region (UTR), 413 bp 3ʹ -UTR, and 2487 bp open reading frame. Bioinformatic analyses revealed that KIF17 obtained from L. crocea (Lc-KIF17) exhibited a high sequence identity compared with those from other teleosts and possessed the structural features of other kinesin-2 proteins. Based on structural similarity, we speculate that the role of Lc-KIF17 may be similar to that of KIF17 in other animals. Lc-kif17 mRNA was diffusely expressed in L. crocea tissues and was highly expressed in the testis, especially at stage IV testicular development. Immunofluorescence analysis revealed that Lc-KIF17 signals colocalized with β-tubulin signals and migrated from the perinuclear cytoplasm to the side of the nucleus where the tail forms during spermiogenesis. These findings revealed that KIF17 may be involved in L. crocea spermiogenesis. In particular, KIF17 may participate in spermatid remodeling by interacting with perinuclear microtubules during L. crocea spermiogenesis. Collectively, this study contributes to an improved understanding of the mechanism underlying L. crocea spermiogenesis and provides a basis for further research on L. crocea reproduction and development.