Piezo2 在许旺细胞体积调节中的作用及其对神经营养释放调节的影响

IF 2.5 Q3 CELL BIOLOGY
Chawapun Suttinont, Katsuyuki Maeno, Mamiko Yano, Kaori Sato-Numata, Tomohiro Numata, Moe Tsutsumi
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引用次数: 0

摘要

背景/目的:触觉依赖于机械感受器和神经纤维,包括c纤维、Aβ纤维和Aδ纤维。许旺细胞(SC)在支持神经纤维方面起着至关重要的作用,非髓鞘化的许旺细胞包裹着 c 纤维,而髓鞘化的许旺细胞则包裹着 Aβ 和 Aδ 纤维。最近的研究揭示了皮肤感觉 SC 的新功能,强调了痛觉 SC 和 Meissner 冠状 SC 在痛觉和触觉中的参与。此外,之前与梅克尔细胞触觉敏感性相关的 Piezo2 也在 SCs 中被发现。本研究的目的是调查与SC机械敏感性和神经营养因子分泌释放过程有关的通道:方法:IFRS1 永生化 SCs 和人类原代 SCs 产生了两种不同亚型的 SCs:未分化 SCs 和已分化 SCs。定量 PCR 被用来评估分化标记和机械敏感通道的表达,包括 TRP 通道(TRPV4、TRPM7 和 TRPA1)和 Piezo 通道(Piezo1 和 Piezo2)。为了验证特定机械敏感通道的功能,在低渗条件下进行了 Ca2+ 成像和电子细胞大小实验,并使用了抑制剂和 siRNA。蛋白表达通过 Western 印迹和免疫染色法进行评估。此外,还进行了分泌组分析,以评估神经营养因子在低张刺激下的释放情况,并使用 ELISA 对具有代表性的营养因子 BDNF 进行了量化:结果:分化诱导增加了Piezo2 mRNA在IFRS1和人类原代SC中的表达水平。两种细胞类型对低渗溶液都有反应,分化后的SCs反应更明显。Gd3+和FM1-43能有效抑制低张性诱导的Ca2+瞬态,这与Piezo2通道有关。相反,Piezo1 和 TRPM7 的抑制剂(分别为 Dooku1 和 NS8593)则没有明显的影响。此外,在低渗刺激诱导细胞肿胀后,分化的SC中的Piezo2似乎参与了调节性体积下降(RVD)。Piezo2缺乏与RVD降低和细胞肿胀延长相关,通过上调内源性表达的Ca2+渗透性TRPV4的功能,导致神经营养因子BDNF的释放增加:我们的研究揭示了SCs的机械敏感性,并发现Piezo2通道与SCs神经营养因子的释放有关。这些结果表明,Piezo2 可能有助于 RVD,从而维持细胞稳态,也可能是神经营养因子释放的负调控因子。这些发现强调了进一步研究 Piezo2 在 SC 功能和神经营养调节中的作用的必要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Role of Piezo2 in Schwann Cell Volume Regulation and Its Impact on Neurotrophic Release Regulation.

Background/aims: Tactile perception relies on mechanoreceptors and nerve fibers, including c-fibers, Aβ-fibers and Aδ-fibers. Schwann cells (SCs) play a crucial role in supporting nerve fibers, with non-myelinating SCs enwrapping c-fibers and myelinating SCs ensheathing Aβ and Aδ fibers. Recent research has unveiled new functions for cutaneous sensory SCs, highlighting the involvement of nociceptive SCs in pain perception and Meissner corpuscle SCs in tactile sensation. Furthermore, Piezo2, previously associated with Merkel cell tactile sensitivity, has been identified in SCs. The goal of this study was to investigate the channels implicated in SC mechanosensitivity and the release process of neurotrophic factor secretion.

Methods: Immortalized IFRS1 SCs and human primary SCs generated two distinct subtypes of SCs: undifferentiated and differentiated SCs. Quantitative PCR was employed to evaluate the expression of differentiation markers and mechanosensitive channels, including TRP channels (TRPV4, TRPM7 and TRPA1) and Piezo channels (Piezo1 and Piezo2). To validate the functionality of specific mechanosensitive channels, Ca2+ imaging and electronic cell sizing experiments were conducted under hypotonic conditions, and inhibitors and siRNAs were used. Protein expression was assessed by Western blotting and immunostaining. Additionally, secretome analysis was performed to evaluate the release of neurotrophic factors in response to hypotonic stimulation, with BDNF, a representative trophic factor, quantified using ELISA.

Results: Induction of differentiation increased Piezo2 mRNA expression levels both in IFRS1 and in human primary SCs. Both cell types were responsive to hypotonic solutions, with differentiated SCs displaying a more pronounced response. Gd3+ and FM1-43 effectively inhibited hypotonicity-induced Ca2+ transients in differentiated SCs, implicating Piezo2 channels. Conversely, inhibitors of Piezo1 and TRPM7 (Dooku1 and NS8593, respectively) had no discernible impact. Moreover, Piezo2 in differentiated SCs appeared to participate in regulatory volume decreases (RVD) after cell swelling induced by hypotonic stimulation. A Piezo2 deficiency correlated with reduced RVD and prolonged cell swelling, leading to heightened release of the neurotrophic factor BDNF by upregulating the function of endogenously expressed Ca2+-permeable TRPV4.

Conclusion: Our study unveils the mechanosensitivity of SCs and implicates Piezo2 channels in the release of neurotrophic factors from SCs. These results suggest that Piezo2 may contribute to RVD, thereby maintaining cellular homeostasis, and may also serve as a negative regulator of neurotrophic factor release. These findings underscore the need for further investigation into the role of Piezo2 in SC function and neurotrophic regulation.

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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
86
审稿时长
1 months
期刊介绍: Cellular Physiology and Biochemistry is a multidisciplinary scientific forum dedicated to advancing the frontiers of basic cellular research. It addresses scientists from both the physiological and biochemical disciplines as well as related fields such as genetics, molecular biology, pathophysiology, pathobiochemistry and cellular toxicology & pharmacology. Original papers and reviews on the mechanisms of intracellular transmission, cellular metabolism, cell growth, differentiation and death, ion channels and carriers, and the maintenance, regulation and disturbances of cell volume are presented. Appearing monthly under peer review, Cellular Physiology and Biochemistry takes an active role in the concerted international effort to unravel the mechanisms of cellular function.
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