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大鼠结状神经节中瞬时受体电位(TRP)离子通道的免疫染色差异模式。

Differential immunostaining patterns of transient receptor potential (TRP) ion channels in the rat nodose ganglion.

机构信息

Divisions of Pediatric Cardiology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.

Pediatric Pulmonology, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH, USA.

出版信息

J Anat. 2022 Aug;241(2):230-244. doi: 10.1111/joa.13656. Epub 2022 Apr 9.

Abstract

Vagal afferents regulate numerous physiological functions including arterial blood pressure, heart rate, breathing, and nociception. Cell bodies of vagal afferents reside in the inferior vagal (nodose) ganglia and their stimulation by various means is being considered as a way to regulate cardiorespiratory responses and control pain sensations. Stimulation of the nodose by exposure to infrared light is recently being considered as a precise way to elicit responses. These responses would likely involve the activity of temperature-sensitive membrane-bound channels. While papers have been published to track the expression of these transient receptor potential ion channels (TRPs), further studies are warranted to determine the in situ expression of the endogenous TRP proteins in the nodose ganglia to fully understand their pattern of expression, subcellular locations, and functions in this animal model. TRP ion channels are a superfamily of Na /Ca -channels whose members are temperature- and/or mechano-sensitive and therefore represent a potential set of proteins that will be activated directly or indirectly by infrared light. Here, we report the spatial localization of six TRP channels, TRPV1, TRPV4, TRPM3, TRPM8, TRPA1, and TRPC1, from nodose ganglia taken from juvenile male Sprague-Dawley rats. The channels were detected using immunohistology with fluorescent tags on cryosections and imaged using confocal microscopy. All six TRP channels were detected with different levels of intensity in neuronal cell bodies and some were also detected in axonal fibers and blood vessels. The TRP receptors differed in their prevalence, in their patterns of expression, and in subcellular expression/localization. More specifically, TRPV1, TRPV4, TRPA1, TRPM8, TRPC1, and TRPM3 were found in vagal afferent cell bodies with a wide range of immunostaining intensity from neuron to neuron. Immunostaining for TRPV1, TRPV4, and TRPA1 appeared as fine particles scattered throughout the cytoplasm of the cell body. Intense TRPV1 immunostaining was also evident in a subset of axonal fibers. TRPM8 and TRPC1 were expressed in courser particles suggesting different subcellular compartments than for TRPV1. The localization of TRPM3 differed markedly from the other TRP channels with an immunostaining pattern that was localized to the periphery of a subset of cell bodies, whereas a scattering or no immunostaining was detected within the bulk of the cytoplasm. TRPV4 and TRPC1 were also expressed on the walls of blood vessels. The finding that all six TRP channels (representing four subfamilies) were present in the nodose ganglia provides the basis for studies designed to understand the roles of these channels in sensory transmission within vagal afferent fibers and in the responses elicited by exposure of nodose ganglia to infrared light and other stimuli. Depending on the location and functionality of the TRP channels, they may regulate the flux of Na /Ca -across the membranes of cell bodies and axons of sensory afferents, efferent (motor) fibers coursing through the ganglia, and in vascular smooth muscle.

摘要

迷走神经传入纤维调节许多生理功能,包括动脉血压、心率、呼吸和痛觉。迷走神经传入纤维的细胞体位于下迷走(结状)神经节,通过各种手段刺激它们被认为是调节心肺反应和控制疼痛感觉的一种方式。最近,人们认为通过暴露在红外光下刺激结状神经节是一种精确的诱发反应的方法。这些反应可能涉及到温度敏感的膜结合通道的活性。虽然已经发表了一些论文来跟踪这些瞬时受体电位离子通道(TRP)的表达,但进一步的研究是必要的,以确定结状神经节中内源性 TRP 蛋白的原位表达,以充分了解它们在这个动物模型中的表达模式、亚细胞定位和功能。TRP 离子通道是一个 Na+/Ca2+-通道的超家族,其成员对温度和/或机械敏感,因此代表了一组潜在的蛋白质,它们将直接或间接通过红外光激活。在这里,我们报告了从幼年雄性 Sprague-Dawley 大鼠的结状神经节中分离出的六种 TRP 通道,即 TRPV1、TRPV4、TRPM3、TRPM8、TRPA1 和 TRPC1 的空间定位。使用带有荧光标签的冷冻切片免疫组织化学检测到这些通道,并使用共聚焦显微镜进行成像。在神经元细胞体中,用不同的强度检测到所有六种 TRP 通道,其中一些也在轴突纤维和血管中检测到。TRP 受体在其普遍性、表达模式和亚细胞表达/定位上存在差异。更具体地说,TRPV1、TRPV4、TRPA1、TRPM8、TRPC1 和 TRPM3 存在于迷走神经传入纤维的细胞体中,从神经元到神经元的免疫染色强度有很大差异。TRPV1、TRPV4 和 TRPA1 的免疫染色表现为散布在细胞体细胞质中的细颗粒。TRPV1 的强烈免疫染色也明显存在于一部分轴突纤维中。TRPM8 和 TRPC1 表达在较粗的颗粒中,表明与 TRPV1 不同的亚细胞隔室。TRPM3 的定位与其他 TRP 通道明显不同,其免疫染色模式定位于细胞体的一部分周围,而在细胞质的大部分中检测到散射或无免疫染色。TRPV4 和 TRPC1 也在血管壁上表达。在结状神经节中存在所有六种 TRP 通道(代表四个亚家族)的发现为研究这些通道在迷走传入纤维中的感觉传递以及结状神经节暴露于红外光和其他刺激物所引起的反应中的作用提供了基础。根据 TRP 通道的位置和功能,它们可能调节 Na+/Ca2+-穿过感觉传入纤维的细胞体和轴突的膜的流动,以及通过神经节的传出(运动)纤维,以及血管平滑肌。

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