Extracellular vesicle distribution and localization in skeletal muscle at rest and following disuse atrophy.

IF 5.3 2区医学 Q2 CELL BIOLOGY

Skeletal Muscle Pub Date : 2023-03-10 DOI:10.1186/s13395-023-00315-1

Ahmed Ismaeel, Douglas W Van Pelt, Zachary R Hettinger, Xu Fu, Christopher I Richards, Timothy A Butterfield, Jonathan J Petrocelli, Ivan J Vechetti, Amy L Confides, Micah J Drummond, Esther E Dupont-Versteegden

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引用次数: 0

Abstract

Background: Skeletal muscle (SkM) is a large, secretory organ that produces and releases myokines that can have autocrine, paracrine, and endocrine effects. Whether extracellular vesicles (EVs) also play a role in the SkM adaptive response and ability to communicate with other tissues is not well understood. The purpose of this study was to investigate EV biogenesis factors, marker expression, and localization across cell types in the skeletal muscle. We also aimed to investigate whether EV concentrations are altered by disuse atrophy.

Methods: To identify the potential markers of SkM-derived EVs, EVs were isolated from rat serum using density gradient ultracentrifugation, followed by fluorescence correlation spectroscopy measurements or qPCR. Single-cell RNA sequencing (scRNA-seq) data from rat SkM were analyzed to assess the EV biogenesis factor expression, and cellular localization of tetraspanins was investigated by immunohistochemistry. Finally, to assess the effects of mechanical unloading on EV expression in vivo, EV concentrations were measured in the serum by nanoparticle tracking analysis in both a rat and human model of disuse.

Results: In this study, we show that the widely used markers of SkM-derived EVs, α-sarcoglycan and miR-1, are undetectable in serum EVs. We also found that EV biogenesis factors, including the tetraspanins CD63, CD9, and CD81, are expressed by a variety of cell types in SkM. SkM sections showed very low detection of CD63, CD9, and CD81 in myofibers and instead accumulation within the interstitial space. Furthermore, although there were no differences in serum EV concentrations following hindlimb suspension in rats, serum EV concentrations were elevated in human subjects after bed rest.

Conclusions: Our findings provide insight into the distribution and localization of EVs in SkM and demonstrate the importance of methodological guidelines in SkM EV research.

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静止时和废用性萎缩后骨骼肌中细胞外囊泡的分布和定位。

背景：骨骼肌（SkM）是一个大型分泌器官，可产生和释放具有自分泌、旁分泌和内分泌效应的肌动素。至于细胞外囊泡（EVs）是否也在骨骼肌适应性反应和与其他组织交流的能力中发挥作用，目前还不十分清楚。本研究的目的是调查骨骼肌中不同细胞类型的 EV 生物发生因子、标记物表达和定位情况。我们还旨在研究 EV 浓度是否会因废用性萎缩而改变：为了确定SkM衍生EV的潜在标记物，我们使用密度梯度超速离心法从大鼠血清中分离出EV，然后进行荧光相关光谱测量或qPCR。分析了大鼠SkM的单细胞RNA测序（scRNA-seq）数据，以评估EV生物发生因子的表达，并通过免疫组化研究了四泛素的细胞定位。最后，为了评估机械卸载对体内 EV 表达的影响，我们通过纳米粒子追踪分析法测量了大鼠和人类废用模型血清中的 EV 浓度：结果：在这项研究中，我们发现在血清 EVs 中检测不到广泛使用的 SkM 衍生 EVs 标记--α-肉毒杆菌素和 miR-1。我们还发现，EV 生物发生因子，包括四泛蛋白 CD63、CD9 和 CD81，在 SkM 中由多种细胞类型表达。SkM切片显示，肌纤维中CD63、CD9和CD81的检出率很低，而在间隙中则有积聚。此外，虽然大鼠后肢悬吊后血清中的 EV 浓度没有差异，但人类卧床休息后血清中的 EV 浓度却升高了：我们的研究结果让我们深入了解了EV在颅内肌中的分布和定位，并证明了颅内肌EV研究方法指南的重要性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Skeletal Muscle CELL BIOLOGY-

CiteScore

9.10

自引率

0.00%

发文量

审稿时长

12 weeks

期刊介绍： The only open access journal in its field, Skeletal Muscle publishes novel, cutting-edge research and technological advancements that investigate the molecular mechanisms underlying the biology of skeletal muscle. Reflecting the breadth of research in this area, the journal welcomes manuscripts about the development, metabolism, the regulation of mass and function, aging, degeneration, dystrophy and regeneration of skeletal muscle, with an emphasis on understanding adult skeletal muscle, its maintenance, and its interactions with non-muscle cell types and regulatory modulators. Main areas of interest include: -differentiation of skeletal muscle- atrophy and hypertrophy of skeletal muscle- aging of skeletal muscle- regeneration and degeneration of skeletal muscle- biology of satellite and satellite-like cells- dystrophic degeneration of skeletal muscle- energy and glucose homeostasis in skeletal muscle- non-dystrophic genetic diseases of skeletal muscle, such as Spinal Muscular Atrophy and myopathies- maintenance of neuromuscular junctions- roles of ryanodine receptors and calcium signaling in skeletal muscle- roles of nuclear receptors in skeletal muscle- roles of GPCRs and GPCR signaling in skeletal muscle- other relevant aspects of skeletal muscle biology. In addition, articles on translational clinical studies that address molecular and cellular mechanisms of skeletal muscle will be published. Case reports are also encouraged for submission. Skeletal Muscle reflects the breadth of research on skeletal muscle and bridges gaps between diverse areas of science for example cardiac cell biology and neurobiology, which share common features with respect to cell differentiation, excitatory membranes, cell-cell communication, and maintenance. Suitable articles are model and mechanism-driven, and apply statistical principles where appropriate; purely descriptive studies are of lesser interest.