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鼠全胃降钙素基因相关肽免疫反应性轴突的形态与分布

Organization and morphology of calcitonin gene-related peptide-immunoreactive axons in the whole mouse stomach.

机构信息

Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida, USA.

Auckland Bioengineering Institute, University of Auckland, Auckland, New Zealand.

出版信息

J Comp Neurol. 2023 Nov;531(16):1608-1632. doi: 10.1002/cne.25519. Epub 2023 Sep 11.

Abstract

Nociceptive afferent axons innervate the stomach and send signals to the brain and spinal cord. Peripheral nociceptive afferents can be detected with a variety of markers (e.g., substance P [SP] and calcitonin gene-related peptide [CGRP]). We recently examined the topographical organization and morphology of SP-immunoreactive (SP-IR) axons in the whole mouse stomach muscular layer. However, the distribution and morphological structure of CGRP-IR axons remain unclear. We used immunohistochemistry labeling and applied a combination of imaging techniques, including confocal and Zeiss Imager M2 microscopy, Neurolucida 360 tracing, and integration of axon tracing data into a 3D stomach scaffold to characterize CGRP-IR axons and terminals in the whole mouse stomach muscular layers. We found that: (1) CGRP-IR axons formed extensive terminal networks in both ventral and dorsal stomachs. (2) CGRP-IR axons densely innervated the blood vessels. (3) CGRP-IR axons ran in parallel with the longitudinal and circular muscles. Some axons ran at angles through the muscular layers. (4) They also formed varicose terminal contacts with individual myenteric ganglion neurons. (5) CGRP-IR occurred in DiI-labeled gastric-projecting neurons in the dorsal root and vagal nodose ganglia, indicating CGRP-IR axons were visceral afferent axons. (6) CGRP-IR axons did not colocalize with tyrosine hydroxylase or vesicular acetylcholine transporter axons in the stomach, indicating CGRP-IR axons were not visceral efferent axons. (7) CGRP-IR axons were traced and integrated into a 3D stomach scaffold. For the first time, we provided a topographical distribution map of CGRP-IR axon innervation of the whole stomach muscular layers at the cellular/axonal/varicosity scale.

摘要

伤害性传入轴突支配胃,并将信号发送到大脑和脊髓。外周伤害性传入可以用多种标志物(例如 P 物质[SP]和降钙素基因相关肽[CGRP])来检测。我们最近检查了整个小鼠胃肌层中 SP 免疫反应性(SP-IR)轴突的拓扑结构和形态。然而,CGRP-IR 轴突的分布和形态结构仍不清楚。我们使用免疫组织化学标记,并应用了包括共聚焦和蔡司 Imager M2 显微镜、Neurolucida 360 追踪、以及将轴突追踪数据整合到 3D 胃支架中的组合成像技术,以表征整个小鼠胃肌层中的 CGRP-IR 轴突和末梢。我们发现:(1)CGRP-IR 轴突在胃的腹侧和背侧形成广泛的末梢网络。(2)CGRP-IR 轴突密集地支配血管。(3)CGRP-IR 轴突与纵行和环形肌肉平行运行。一些轴突以一定角度穿过肌层。(4)它们还与单个肌间神经节神经元形成曲张终末接触。(5)CGRP-IR 存在于背根和迷走神经结状神经节中标记的胃投射神经元中,表明 CGRP-IR 轴突是内脏传入轴突。(6)CGRP-IR 轴突与胃中的酪氨酸羟化酶或囊泡乙酰胆碱转运体轴突不共定位,表明 CGRP-IR 轴突不是内脏传出轴突。(7)CGRP-IR 轴突被追踪并整合到 3D 胃支架中。我们首次提供了整个胃肌层 CGRP-IR 轴突支配的细胞/轴突/曲张体的拓扑分布图谱。

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