Immunohistochemical localization of proteins involved in exocytosis of glutamate from P2X3 purinoceptor-expressing subserosal afferent nerve endings in the rat gastric antrum

IF 2.3 4区生物学 Q4 CELL BIOLOGY Acta histochemica Pub Date : 2024-05-01 DOI:10.1016/j.acthis.2024.152170

Masato Hirakawa , Takuya Yokoyama , Sayed Sharif Abdali , Yoshio Yamamoto , Tomoyuki Saino

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Abstract

We previously reported the presence of P2X3 purinoceptors (P2X3)-expressing subserosal afferent nerve endings consisting of net- and basket-like nerve endings in the rat gastric antrum. These nerve endings may morphologically be vagal mechanoreceptors activated by antral peristalsis. The present study investigated immunoreactivities for vesicular glutamate transporter (VGLUT) 1 and VGLUT2 as well as exocytosis-related proteins, i.e., core components of the SNARE complex (SNAP25, Stx1, and VAMP2) and synaptotagmin-1 (Syt1), in whole-mount preparations of the rat gastric antrum using double immunofluorescence. VGLUT1 immunoreactivity was not detected, whereas VGLUT2 immunoreactivity was observed in P2X3-immunoreactive subserosal nerve endings composed of both net- and basket-like endings. In net-like nerve endings, intense VGLUT2 immunoreactivity was localized in polygonal bulges of reticular nerve fibers and peripheral axon terminals. Furthermore, intense immunoreactivities for SNAP25, Stx1, and VAMP2 were localized in net-like nerve endings. Intense immunoreactivities for VAMP2 and Syt1 were observed in VGLUT2-immunoreactive net-like nerve endings. In basket-like nerve endings, VGLUT2 immunoreactivity was localized in pleomorphic terminal structures and small bulges surrounding the subserosal ganglion, whereas immunoreactivities for SNAP25, Stx1, and VAMP2 were weak in these nerve endings. VGLUT2-immunoreactive basket-like nerve endings were weakly immunoreactive for VAMP2 and Syt1. These results suggest that subserosal afferent nerve endings release glutamate by exocytosis mainly from net-like nerve endings to modulate their mechanoreceptor function.

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参与大鼠胃窦中表达 P2X3 嘌呤受体的浆膜下传入神经末梢谷氨酸外排的蛋白质的免疫组织化学定位。

我们以前曾报道过，在大鼠胃窦中存在由网状和篮状神经末梢组成的表达 P2X3 嘌呤受体（P2X3）的黏膜下传入神经末梢。这些神经末梢在形态上可能是由前胃蠕动激活的迷走机械感受器。本研究采用双重免疫荧光法研究了大鼠胃窦全贴片制备物中谷氨酸囊泡转运体（VGLUT）1 和 VGLUT2 以及外吞相关蛋白（即 SNARE 复合物的核心成分（SNAP25、Stx1 和 VAMP2）和 synaptotagmin-1 (Syt1)）的免疫活性。未检测到 VGLUT1 免疫反应，而在由网状末梢和篮状末梢组成的具有 P2X3 免疫反应的粘膜下神经末梢中观察到了 VGLUT2 免疫反应。在网状神经末梢中，网状神经纤维的多角形隆起和外周轴突末端具有强烈的 VGLUT2 免疫活性。此外，在网状神经末梢中，SNAP25、Stx1 和 VAMP2 也有强烈的免疫反应。在VGLUT2-免疫反应的网状神经末梢中观察到了VAMP2和Syt1的强免疫反应。在篮状神经末梢中，VGLUT2 免疫活性定位于多形性末端结构和围绕巩膜下神经节的小隆起中，而 SNAP25、Stx1 和 VAMP2 在这些神经末梢中的免疫活性较弱。VGLUT2免疫反应的篮状神经末梢对VAMP2和Syt1的免疫反应较弱。这些结果表明，骨膜下传入神经末梢主要通过网状神经末梢的外泌作用释放谷氨酸，以调节其机械感受器功能。

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来源期刊

Acta histochemica 生物-细胞生物学

CiteScore

4.60

自引率

4.00%

发文量

107

审稿时长

23 days

期刊介绍： Acta histochemica, a journal of structural biochemistry of cells and tissues, publishes original research articles, short communications, reviews, letters to the editor, meeting reports and abstracts of meetings. The aim of the journal is to provide a forum for the cytochemical and histochemical research community in the life sciences, including cell biology, biotechnology, neurobiology, immunobiology, pathology, pharmacology, botany, zoology and environmental and toxicological research. The journal focuses on new developments in cytochemistry and histochemistry and their applications. Manuscripts reporting on studies of living cells and tissues are particularly welcome. Understanding the complexity of cells and tissues, i.e. their biocomplexity and biodiversity, is a major goal of the journal and reports on this topic are especially encouraged. Original research articles, short communications and reviews that report on new developments in cytochemistry and histochemistry are welcomed, especially when molecular biology is combined with the use of advanced microscopical techniques including image analysis and cytometry. Letters to the editor should comment or interpret previously published articles in the journal to trigger scientific discussions. Meeting reports are considered to be very important publications in the journal because they are excellent opportunities to present state-of-the-art overviews of fields in research where the developments are fast and hard to follow. Authors of meeting reports should consult the editors before writing a report. The editorial policy of the editors and the editorial board is rapid publication. Once a manuscript is received by one of the editors, an editorial decision about acceptance, revision or rejection will be taken within a month. It is the aim of the publishers to have a manuscript published within three months after the manuscript has been accepted