Characterization and proteomics of chicken seminal plasma extracellular vesicles

Author(s):  
Xuliang Luo ◽  
Yan Guo ◽  
Ying Huang ◽  
Manman Cheng ◽  
Xiaohui Wu ◽  
...  
Author(s):  
Ana Carolina Pedrosa ◽  
Mariana A. Torres ◽  
Diego V. Alkmin ◽  
Jorge E.P. Pinzon ◽  
Simone Maria M.K. Martins ◽  
...  

2021 ◽  
Vol 10 (4) ◽  
pp. 5-13
Author(s):  
A.P. Sysoeva ◽  
◽  
N.P. Makarova ◽  
E.E. Kraevaya ◽  
◽  
...  

For a long time, the role of seminal plasma during human fertilization remained underestimated. Numerous studies related to the development of different methods for human embryo in vitro cultivation were gener-ally concerned with the quality of male and female gametes. However, in recent years, the development of Omix technologies provided a new insight into great seminal plasma influence on the morphofunctional characteristics of spermatozoa. This is especially true for the regulatory function of extracellular vesicles secreted by male reproductive tract cells. In this work, we attempted to analyze current data on the influence of extracellular seminal plasma vesicles on the morphofunctional characteristics of spermatozoa to solve male infertility topical issues. The review includes studies by foreign and Russian research groups that werу conducted within the past 5 years and found in PubMed, Google Scholar, and Cochrane Library databases. Very few studies demonstrate that seminal plasma vesicles act as functional regulators of male fertility and their dysfunction may lead to infertility. The use of seminal plasma extracellular vesicles in clinical practice may significantly increase the success of IVF programs, especially in impaired spermatogenesis. Keywords: extracellular vesicles, exosomes, biomarkers, seminal plasma, spermatozoa, assisted reproductive technology, cell biology, morphology


Author(s):  
Yaqun Ding ◽  
Ning Ding ◽  
Yu Zhang ◽  
Shenmin Xie ◽  
Mengna Huang ◽  
...  

Seminal plasma contains a large number of extracellular vesicles (EVs). However, the roles of these EVs and their interactions with sperm are not clear. To identify the important molecules affecting sperm motility in EVs, we analyzed RNA from seminal plasma EVs of boars with different sperm motility using whole-transcriptome sequencing and proteomic analysis. In total, 7 miRNAs, 67 lncRNAs, 126 mRNAs and 76 proteins were differentially expressed between the two groups. We observed that EV-miR-222 can obviously improve sperm motility. In addition, the results suggested that miR-222 was transferred into sperm by the EVs and that miR-222 affected sperm apoptosis by inhibiting the expression of EGFR, BCL2L11, BAX, CYCs, CASP9 and CASP3. The results of electron microscopy also showed that overexpression of miR-222 in EVs could reduce sperm apoptosis. The study of the whole transcriptomes and proteomes of EVs in boar semen revealed some miRNAs may play an important role in these EVs interactions with Duroc sperm, and the findings suggest that the release of miR-222 by semen EVs is an important mechanism by which sperm viability is maintained and sperm apoptosis is reduced. Our studies provide a new insight of miR-222 in EVs regulation for sperm motility and sperm apoptosis.


Reproduction ◽  
2021 ◽  
Vol 161 (5) ◽  
pp. 489-498 ◽  
Author(s):  
Luiz Cordeiro ◽  
Hsiu-Lien Herbie Lin ◽  
Anaïs Vitorino Carvalho ◽  
Isabelle Grasseau ◽  
Rustem Uzbekov ◽  
...  

Male subfertility causes are very varied and sometimes related to post-gonadic maturation disruption, involving seminal plasma constituents. Among them, extracellular vesicles are involved in key exchanges with sperm in mammals. However, in birds, the existence of seminal extracellular vesicles is still debated. The aim of the present work was first to clarify the putative presence of extracellular vesicles in the seminal plasma of chickens, secondly to characterize their size and protein markers in animals showing different fertility, and finally to make preliminary evaluations of their interactions with sperm. We successfully isolated extracellular vesicles from seminal plasma of males showing the highest differences in semen quality and fertility by using ultracentrifugation protocol (pool of 3 ejaculates/rooster, n =3/condition). Size characterization performed by electron microscopy revealed a high proportion of small extracellular vesicles (probably exosomes) in chicken seminal plasma. Smaller extracellular vesicles appeared more abundant in fertile than in subfertile roosters, with a mean diameter of 65.12 and 77.18 nm, respectively. Different protein markers of extracellular vesicles were found by western blotting (n = 6/condition). Among them, HSP90A was significantly more abundant in fertile than in subfertile males. In co-incubation experiments (n = 3/condition), extracellular vesicles enriched seminal fractions of fertile males showed a higher capacity to be incorporated into fertile than into subfertile sperm. Sperm viability and motility were impacted by the presence of extracellular vesicles from fertile males. In conclusion, we successfully demonstrated the presence of extracellular vesicles in chicken seminal plasma, with differential size, protein markers and putative incorporation capacity according to male fertility status.


Proteomes ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 17 ◽  
Author(s):  
Skalnikova ◽  
Bohuslavova ◽  
Turnovcova ◽  
Juhasova ◽  
Juhas ◽  
...  

Extracellular vesicles (EVs) are a highly attractive subject of biomedical research as possible carriers of nucleic acid and protein biomarkers. EVs released to body fluids enable indirect access to inner organs by so-called “liquid biopsies”. Obtaining a high-quality EV sample with minimum contaminants is crucial for proteomic analyses using LC–MS/MS or other techniques. However, the EV content in various body fluids largely differs, which may hamper subsequent analyses. Here, we present a comparison of extracellular vesicle yields from blood plasma, cerebrospinal fluid, and seminal plasma using an experimental pig model. Pigs are widely used in biomedical research as large animal models with anatomy and physiology close to those of humans and enable studies (e.g., of the nervous system) that are unfeasible in humans. EVs were isolated from body fluids by differential centrifugation followed by ultracentrifugation. EVs were characterized according to protein yields and to the quality of the isolated vesicles (e.g., size distribution, morphology, positivity for exosome markers). In our experimental setting, substantial differences in EV amounts were identified among body fluids, with the seminal plasma being the richest EV source. The yields of pellet proteins from ultracentrifugation of 1 mL of porcine body fluids may help to estimate body fluid input volumes to obtain sufficient samples for subsequent proteomic analyses.


2020 ◽  
Vol 21 (21) ◽  
pp. 7957
Author(s):  
Xiaogang Zhang ◽  
Harmjan R. Vos ◽  
Weiyang Tao ◽  
Willem Stoorvogel

Body fluids contain many populations of extracellular vesicles (EV) that differ in size, cellular origin, molecular composition, and biological activities. EV in seminal plasma are in majority originating from prostate epithelial cells, and hence are also referred to as prostasomes. Nevertheless, EV are also contributed by other accessory sex glands, as well as by the testis and epididymis. In a previous study, we isolated EV from seminal plasma of vasectomized men, thereby excluding contributions from the testis and epididymis, and identified two distinct EV populations with diameters of 50 and 100 nm, respectively. In the current study, we comprehensively analyzed the protein composition of these two EV populations using quantitative Liquid Chromatography-Mass Spectrometry (LC-MS/MS). In total 1558 proteins were identified. Of these, ≈45% was found only in the isolated 100 nm EV, 1% only in the isolated 50 nm EV, and 54% in both 100 nm and 50 nm EV. Gene ontology (GO) enrichment analysis suggest that both originate from the prostate, but with distinct biogenesis pathways. Finally, nine proteins, including KLK3, KLK2, MSMB, NEFH, PSCA, PABPC1, TGM4, ALOX15B, and ANO7, with known prostate specific expression and alternate expression levels in prostate cancer tissue were identified. These data have potential for the discovery of EV associated prostate cancer biomarkers in blood.


2021 ◽  
Vol 161 ◽  
pp. 26-40
Author(s):  
Maíra Bianchi Rodrigues Alves ◽  
Rubens Paes de Arruda ◽  
Leonardo Batissaco ◽  
Laura Nataly Garcia-Oliveros ◽  
Vitor Hugo Guilger Gonzaga ◽  
...  

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