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睾丸细胞外囊泡货物的 repertoire 及其在与精子发生相关的隔间通讯中的作用。

The repertoire of testicular extracellular vesicle cargoes and their involvement in inter-compartmental communication associated with spermatogenesis.

机构信息

School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, SAR, China.

Dr. Li Dak-Sum Research Centre, University of Hong Kong, Hong Kong, SAR, China.

出版信息

BMC Biol. 2022 Mar 29;20(1):78. doi: 10.1186/s12915-022-01268-5.

DOI:10.1186/s12915-022-01268-5
PMID:35351114
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8966158/
Abstract

BACKGROUND

Spermatogenesis is regulated by a complex network of intercellular communication processes. Extracellular vesicles (EVs) are one of the important mediators in intercellular communication. Previous reports have demonstrated the involvement of EVs from the epididymis and prostate in sperm maturation and function. However, the presence of EVs in the testis and their potential involvement in spermatogenesis has not been explored. Here, we have established a testis dissociation protocol that allows the isolation and characterization of testicular EVs.

RESULTS

We show that testicular EVs are specifically and efficiently taken up by somatic cells and germ cells, including the spermatozoa in the interstitial space and the seminiferous tubule compartments. We profiled the proteome of testicular EVs and probed the cell types that release them, revealing the potential contributions from the Leydig cells and testicular macrophages. Moreover, we sequenced the small RNA cargoes of testicular EVs and identified sets of small non-coding RNAs that were overlooked in the testis transcriptome. Selected miRNA candidates in testicular EVs were found in sperm RNA payload and demonstrated specific resistance towards ribonuclease A independent of the vesicle membrane. Small molecule inhibition of EV secretion perturbed spermatogenesis via inter-compartmental communication.

CONCLUSIONS

Together, our study provides a valuable resource on the repertoire of cargoes carried by testicular EVs and uncovers a physiological function of testicular EVs in inter-compartmental communication associated to spermatogenesis.

摘要

背景

精子发生受细胞间复杂通讯过程网络调控。细胞外囊泡（EVs）是细胞间通讯的重要介质之一。先前的报告表明，附睾和前列腺中的 EVs 参与了精子成熟和功能。然而，睾丸中 EVs 的存在及其在精子发生中的潜在作用尚未被探索。在这里，我们建立了一种睾丸分离方案，可用于分离和鉴定睾丸 EVs。

结果

我们表明，睾丸 EVs 被体细胞和生殖细胞（包括间质空间中的精子和生精小管隔室中的精子）特异性且有效地摄取。我们对睾丸 EVs 的蛋白质组进行了分析，并研究了释放它们的细胞类型，揭示了 Leydig 细胞和睾丸巨噬细胞的潜在贡献。此外，我们对睾丸 EVs 的小 RNA 货物进行了测序，并鉴定了在睾丸转录组中被忽视的小非编码 RNA 集。在睾丸 EVs 中发现的选定 miRNA 候选物存在于精子 RNA 有效负载中，并表现出对核糖核酸酶 A 的特异性抗性，而与囊泡膜无关。小分子抑制 EV 分泌通过隔室间通讯干扰精子发生。

结论

总之，我们的研究提供了有关睾丸 EVs 携带的货物谱的有价值的资源，并揭示了睾丸 EVs 在与精子发生相关的隔室间通讯中的生理功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/8b0c88986195/12915_2022_1268_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/13aff9eb0603/12915_2022_1268_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/89b2183d1c39/12915_2022_1268_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/bb204d2cff81/12915_2022_1268_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/8b0c88986195/12915_2022_1268_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/13aff9eb0603/12915_2022_1268_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/89b2183d1c39/12915_2022_1268_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/bb204d2cff81/12915_2022_1268_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9a/8966158/8b0c88986195/12915_2022_1268_Fig4_HTML.jpg

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