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Hormonal stimulation of carp is accompanied with changes of seminal plasma and spermatozoa proteome

ABSTRACT: Hormonal stimulation in common carp is a routine practice to enhance sperm production and control gamete maturation. The mechanism regulating functional changes of semen composition upon hormone treatment is unknown at present. The aim of the study was to compare proteome of carp spermatozoa and seminal plasma from control (PBS, n=10) and hormonally stimulated males (D-Ala6,Pro9-NEt)-mGnRH+metoclopramide, n=10). Comparative proteomics based on two-dimensional differential in-gel electrophoresis was employed to quantitatively analyze the spermatozoa and seminal plasma proteins changed in abundance due to hormonal stimulation. Changes in phosphorylation and oxidation level of spermatozoa proteins were also analyzed. The proteomic results were validated by western blot, enzymatic assay and mRNA expression analysis.In total we identified 62 spots (38 up and 24 down regulated) corresponding to 48 proteins with differential abundance in spermatozoa and 36 spots (23 up and 13 down regulated) corresponding to 23 proteins differentially abundant in seminal plasma upon hormone treatment (P<0.05; fold change 1.2). The majority of spermatozoa proteins changed in abundance as well as phosphorylation and carbonylation level were involved in cillium assembly and organization, energy metabolism, protein folding, and signal transduction while differentially abundant seminal plasma proteins were associated with immune and stress response, lipid metabolism and tissue remodelling. Our results strongly suggest that hormonal treatment is not only related to hydration of testis but also induces modification of spermatozoa proteins and synthesis of seminal plasma proteins. ROS signaling and cAMP/PKA and calcium/calmodulin pathways are involved in sperm response to hormonal stimulation.

INSTRUMENT(S): autoflex

ORGANISM(S): Cyprinus Carpio (common Carp)

SUBMITTER: Mariola Dietrich

LAB HEAD: Mariola Dietrich

PROVIDER: PXD011049 | Pride | 2021-04-20

REPOSITORIES: Pride

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Project description:Male factors account for approximately 40% of all infertility. Conventional semen analysis focus on seminal physicochemical property and spermatozoa morphology. They yield no information concerning the functional competence of the spermatozoa(Petrunkina et al., 2007). Owning to the essential role of proteins in the reproductive process, comprehensive and systematic identification of proteome is critical to gain new insights into spermatogenesis and infertility. Recent advances in mass spectrometry have allowed the identification of hundreds to thousands of protein in spermatozoa(Oliva et al., 2008; Amaral et al., 2014; Codina et al., 2015). In major domestic mammalian species, global spermatozoa proteomic profiles have been described in rodents(Baker et al., 2008a; Baker et al., 2008b) and bovine(Peddinti et al., 2008). Meanwhile, lacking of new protein synthesis in spermatozoa supports the current view that the regulation of sperm maturation is controlled by exogenous proteins(O'Rand et al., 2011) (e.g., during epididymal transit). Among the seminal plasma proteins, The human seminal plasma has been comprehensively described (Pilch and Mann, 2006; Batruch et al., 2011; Milardi et al., 2012; Milardi et al., 2013; Sharma et al., 2013). In domestic animal species, some studies performing a systematic analysis on using high throughput proteomics have been performed(Kelly et al., 2006; Moura et al., 2007; Souza et al., 2012; Druart et al., 2013; Soleilhavoup et al., 2014). Buffalos (Bubalus bubalis) are adapted to hot–humid tropical climatic conditions, but have low reproductive efficiency. The low sperm motility maybe contributed to protein components variation of spermatozoa and seminal plasma. The composition of buffalo spermatozoa and buffalo seminal plasma (BSP) remains unknown. Proteomic study would be beneficial to the elucidation of the roles of sperm and BSP proteins in regulation of maturation, motility and fertilization. As such, the aim of the current study was to extensively characterize the differentially expressed protein of buffalo spermatozoa and seminal plasma using a comparative proteomics.

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Hormonal stimulation of carp is accompanied with changes of seminal plasma and spermatozoa proteome

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