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Transcriptome analysis of the testes of male chickens with high and low sperm motility.

ABSTRACT: The reproductive performance of chicken breeders has significant economic importance in the poultry industry, and sperm motility is an indicator of reproductive performance. This study performed RNA-seq of the testes of Gushi chicken roosters with high and low sperm motility and identified differentially expressed RNAs involved in sperm motility. RNA-seq analysis showed that 73 and 67 differentially expressed mRNAs were up- and downregulated, and 47 and 56 differentially expressed long non-coding RNAs were up- and downregulated, respectively. The genes related to sperm motility and spermatogenesis included KIFC1, KCNK2, and REC8. Functional enrichment analysis revealed that the pathways related to sperm motility included oxidative phosphorylation and glycine, serine, and threonine metabolism. In addition, the MSTRG.15920.1-REC8-MSTRG.11860.2-VWC2 pathway may regulate sperm motility. This study helped elucidate the molecular genetic mechanism of sperm motility in chicken.

SUBMITTER: Du Z

PROVIDER: S-EPMC9554828 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Transcriptome analysis of the testes of male chickens with high and low sperm motility.

Du Zhenwei Z Li WenTing W Liu Cong C Wang ChunXiu C Wang DongXue D Zhu ShuaiPeng S Kang XiangTao X Jiang RuiRui R Deng LeKun L Li DongHua D Sun GuiRong G

Poultry science 20220914 12

The reproductive performance of chicken breeders has significant economic importance in the poultry industry, and sperm motility is an indicator of reproductive performance. This study performed RNA-seq of the testes of Gushi chicken roosters with high and low sperm motility and identified differentially expressed RNAs involved in sperm motility. RNA-seq analysis showed that 73 and 67 differentially expressed mRNAs were up- and downregulated, and 47 and 56 differentially expressed long non-codin ...[more]

PMID: 36215742

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Project description:Molecular mechanisms underlying sperm motility have not been fully explained, particularly in chickens. The objective was to identify seminal plasma proteins associated with chicken sperm motility by comparing the seminal plasma proteomic profile of roosters with low sperm motility (LSM, n = 4) and high sperm motility (HSM, n = 4). Using a label-free MS-based method, a total of 522 seminal plasma proteins were identified, including 386 (∼74%) previously reported and 136 novel ones. A total of 70 differentially abundant proteins were defined, including 48 more-abundant, 15 less-abundant, and seven proteins unique to the LSM group (specific proteins). Key secretory proteins like less-abundant adhesion G-protein coupled receptor G2 (ADGRG2) and more-abundant serine peptidase inhibitor Kazal-type 2 (SPINK2) in the LSM suggested that the corresponding secretory tissues played a crucial role in maintaining sperm motility. Majority (80%) of the more-abundant and five specific proteins were annotated to the cytoplasmic domain which might be a result of higher plasma membrane damage and acrosome dysfunction in LSM. Additionally, more-abundant mitochondrial proteins were detected in LSM seminal plasma associated with lower spermatozoa mitochondrial membrane potential (ΔΨm) and ATP concentrations. Further studies showed that the spermatozoa might be suffering from oxidative stress, as the amount of spermatozoa reactive oxygen species (ROS) were largely enhanced, seminal malondialdehyde (MDA) concentrations were increased, and the seminal plasma total antioxidant capacity (T-AOC) were decreased. Our study provides an additional catalogue of chicken seminal plasma proteome and supports the idea that seminal plasma could be as an indicator of spermatozoa physiology. More-abundant of acrosome, mitochondria and sperm cytoskeleton proteins in the seminal plasma could be a marker of sperm dysfunction and loss of motility. The degeneration of spermatozoa caused by the reduced seminal T-AOC and enhanced oxidative stress might be potential determinants of low sperm motility. These results could extend our understanding of sperm motility and sperm physiology regulation.

Project description:BackgroundExosomes are nanosized membranous vesicles secreted by various types of cells, which facilitate intercellular communication by transporting bioactive compounds. Exosomes are abundant in biological fluids including semen, and their protein composition and the potential of seminal plasma exosomes (SPEs) as fertility biomarkers were elucidated in humans, however, little information is available regarding buffalo (Bubalus bubalis). Here, we examined protein correlation between spermatozoa, seminal plasma (SP), and SPEs, and we compared and analyzed protein differences between high-motility (H-motility) and low-motility (L-motility) SPEs in buffalo.ResultsSPEs were concentrated and purified by ultracentrifugation combined with sucrose density gradient centrifugation, followed by verification using western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Protein composition in spermatozoa, SP and SPEs, and protein difference in H- and L-motility SPEs were identified by LC-MS/MS proteomic analysis and were functionally analyzed through comprehensive bioinformatics. Many SPEs proteins originated from spermatozoa and SP, and nearly one third were also present in spermatozoa and SP. A series of proteins associated with reproductive processes including sperm capacitation, spermatid differentiation, fertilization, sperm-egg recognition, membrane fusion, and acrosome reaction were integrated in a functional network. Comparative proteomic analyses showed 119 down-regulated and 41 up-regulated proteins in L-motility SPEs, compared with H-motility SPEs. Gene Ontology (GO) enrichment of differentially expressed proteins (DEPs) showed that most differential proteins were located in sperm and vesicles, with activities of hydrolase and metalloproteinase, and were involved in sperm-egg recognition, fertilization, single fertilization, and sperm-zona pellucida binding processes, etc. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that differential proteins were mainly involved in the PPRP signaling pathway, calcium signaling pathway, and cAMP signaling pathway, among others. Furthermore, 6 proteins associated with reproduction were validated by parallel reaction monitoring analysis.ConclusionThis study provides a comprehensive description of the seminal plasma exosome proteome and may be of use for further screening of biomarkers associated with male infertility.

Dataset Information

Transcriptome analysis of the testes of male chickens with high and low sperm motility.

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Transcriptome analysis of the testes of male chickens with high and low sperm motility.

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