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Differential immunostaining patterns of transient receptor potential (TRP) ion channels in the rat nodose ganglion
Journal of Anatomy ( IF 1.8 ) Pub Date : 2022-04-09 , DOI: 10.1111/joa.13656 Safdar Jawaid 1 , Amanda I Herring 1 , Paulina M Getsy 2 , Stephen J Lewis 2 , Michiko Watanabe 1 , Hana Kolesova 3
Journal of Anatomy ( IF 1.8 ) Pub Date : 2022-04-09 , DOI: 10.1111/joa.13656 Safdar Jawaid 1 , Amanda I Herring 1 , Paulina M Getsy 2 , Stephen J Lewis 2 , Michiko Watanabe 1 , Hana Kolesova 3
Affiliation
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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+/Ca2+-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+/Ca2+-across the membranes of cell bodies and axons of sensory afferents, efferent (motor) fibers coursing through the ganglia, and in vascular smooth muscle.
中文翻译：
大鼠结状神经节瞬时受体电位（TRP）离子通道的差异免疫染色模式
迷走神经传入调节许多生理功能，包括动脉血压、心率、呼吸和伤害感受。迷走神经传入细胞体位于下迷走神经（结状）神经节中，通过各种方式刺激它们被认为是调节心肺反应和控制疼痛感的一种方法。最近，通过暴露于红外光刺激结节被认为是引发反应的精确方法。这些反应可能涉及温度敏感膜结合通道的活动。虽然已经发表了追踪这些瞬时受体电位离子通道 (TRP) 表达的论文，但仍需要进一步的研究来确定结状神经节中内源性 TRP 蛋白的原位表达，以充分了解它们的表达模式、亚细胞位置、以及在该动物模型中的功能。 TRP离子通道是Na + /Ca 2+通道的超家族，其成员对温度和/或机械敏感，因此代表一组将被红外光直接或间接激活的潜在蛋白质。在这里，我们报告了来自幼年雄性斯普拉格-道利大鼠的结状神经节的六个 TRP 通道：TRPV1、TRPV4、TRPM3、TRPM8、TRPA1 和 TRPC1 的空间定位。使用冷冻切片上带有荧光标签的免疫组织学检测通道，并使用共聚焦显微镜成像。所有六个 TRP 通道均在神经元细胞体中检测到不同强度的水平，并且在轴突纤维和血管中也检测到一些通道。 TRP 受体的分布、表达模式和亚细胞表达/定位各不相同。 更具体地说，在迷走神经传入细胞体中发现了 TRPV1、TRPV4、TRPA1、TRPM8、TRPC1 和 TRPM3，从神经元到神经元具有广泛的免疫染色强度。 TRPV1、TRPV4 和 TRPA1 的免疫染色显示为分散在细胞体细胞质中的细颗粒。在轴突纤维的子集中，强烈的 TRPV1 免疫染色也很明显。 TRPM8 和 TRPC1 在粗颗粒中表达，表明其亚细胞区室与 TRPV1 不同。 TRPM3 的定位与其他 TRP 通道显着不同，其免疫染色模式位于细胞体子集的外围，而在大部分细胞质内检测到散射或未检测到免疫染色。 TRPV4和TRPC1也在血管壁上表达。所有六个 TRP 通道（代表四个亚族）都存在于结状神经节中的发现为旨在了解这些通道在迷走神经传入纤维内的感觉传递以及结状神经节暴露于红外线引起的反应中的作用的研究提供了基础光和其他刺激。根据 TRP 通道的位置和功能，它们可以调节 Na + /Ca 2+ - 穿过细胞体膜和感觉传入轴突、流过神经节的传出（运动）纤维以及血管平滑肌的流量。肌肉。
更新日期：2022-04-09
中文翻译：
大鼠结状神经节瞬时受体电位（TRP）离子通道的差异免疫染色模式
迷走神经传入调节许多生理功能，包括动脉血压、心率、呼吸和伤害感受。迷走神经传入细胞体位于下迷走神经（结状）神经节中，通过各种方式刺激它们被认为是调节心肺反应和控制疼痛感的一种方法。最近，通过暴露于红外光刺激结节被认为是引发反应的精确方法。这些反应可能涉及温度敏感膜结合通道的活动。虽然已经发表了追踪这些瞬时受体电位离子通道 (TRP) 表达的论文，但仍需要进一步的研究来确定结状神经节中内源性 TRP 蛋白的原位表达，以充分了解它们的表达模式、亚细胞位置、以及在该动物模型中的功能。 TRP离子通道是Na + /Ca 2+通道的超家族，其成员对温度和/或机械敏感，因此代表一组将被红外光直接或间接激活的潜在蛋白质。在这里，我们报告了来自幼年雄性斯普拉格-道利大鼠的结状神经节的六个 TRP 通道：TRPV1、TRPV4、TRPM3、TRPM8、TRPA1 和 TRPC1 的空间定位。使用冷冻切片上带有荧光标签的免疫组织学检测通道，并使用共聚焦显微镜成像。所有六个 TRP 通道均在神经元细胞体中检测到不同强度的水平，并且在轴突纤维和血管中也检测到一些通道。 TRP 受体的分布、表达模式和亚细胞表达/定位各不相同。 更具体地说，在迷走神经传入细胞体中发现了 TRPV1、TRPV4、TRPA1、TRPM8、TRPC1 和 TRPM3，从神经元到神经元具有广泛的免疫染色强度。 TRPV1、TRPV4 和 TRPA1 的免疫染色显示为分散在细胞体细胞质中的细颗粒。在轴突纤维的子集中，强烈的 TRPV1 免疫染色也很明显。 TRPM8 和 TRPC1 在粗颗粒中表达，表明其亚细胞区室与 TRPV1 不同。 TRPM3 的定位与其他 TRP 通道显着不同，其免疫染色模式位于细胞体子集的外围，而在大部分细胞质内检测到散射或未检测到免疫染色。 TRPV4和TRPC1也在血管壁上表达。所有六个 TRP 通道（代表四个亚族）都存在于结状神经节中的发现为旨在了解这些通道在迷走神经传入纤维内的感觉传递以及结状神经节暴露于红外线引起的反应中的作用的研究提供了基础光和其他刺激。根据 TRP 通道的位置和功能，它们可以调节 Na + /Ca 2+ - 穿过细胞体膜和感觉传入轴突、流过神经节的传出（运动）纤维以及血管平滑肌的流量。肌肉。
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