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首页> 美国卫生研究院文献>Journal of the Endocrine Society >SAT-601 Development of a Protocol for Stellate and Celiac Ganglia Dissection for Characterization of Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) Protein and Receptor Expression in Male and Female Mice Following Cold Acclimation
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SAT-601 Development of a Protocol for Stellate and Celiac Ganglia Dissection for Characterization of Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) Protein and Receptor Expression in Male and Female Mice Following Cold Acclimation

机译：SAT-601在冷适应后在冷驯化后SAT-601开发出星状腺苷酸和腹腔神经节目分析用于表征垂体腺苷酸环酶活化多肽（PACAP）蛋白和受体表达后冷置换后的雌性小鼠

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Pituitary adenylate cyclase-activating polypeptide (PACAP) is being studied to understand the endocrine regulation of energy balance and has been shown to be important in the regulation of the stress response (1,2). Specifically, PACAP has been shown to regulate thermogenesis, an energy burning process regulated by the sympathetic nervous system that contributes to achieving energy homeostasis in response to cold stress and overfeeding. PACAP is expressed in the sympathetic nervous system and is required at the adrenomedullary synapse to maintain epinephrine secretion from the adrenal medulla in response to physiological stress (3). Across the branches of the sympathetic nervous system, PACAP receptor expression is most well characterized in the superior cervical ganglia (SCG) (4). However, a detailed characterization of PACAP and its receptors has not been performed in ganglia whose postganglionic fibres innervate adipose tissues (stellate and celiac ganglia) in response to thermogenic stress. We hypothesized that PACAP is produced by preganglionic neurons innervating the stellate and celiac ganglia, and act on PACAP receptors expressed on the post-ganglionic neurons, and this expression will be upregulated in response to chronic cold stress. Due to their small and amorphous shape, we have developed a protocol to reliably isolate the stellate and celiac ganglia and validate their identity through the presence of tyrosine hydroxylase mRNA, using adrenal and SCG samples as positive controls. PACAP receptor expression (VPAC1, VPAC2, PAC1) was examined in the ganglia utilizing real-time PCR, and PACAP protein was visualized in the ganglia of transgenic mice that express eGFP under the control of the PACAP promoter (PACAP-eGFP mice) (5). This research demonstrates the expression of PACAP receptors in ganglia whose postganglionic fibres innervate adipose tissue, enhancing our understanding of PACAP’s role in the SNS, and its contribution to the regulation of adaptive thermogenesis. References: (1) Gray et al., Pacap: Regulator of the stress response. In: Fink G, ed. Stress: Physiology, biochemistry, and pathology. 2019:279-291. (2) Mustafa, Adv Pharmacol. San Diego, Calif:445-457. (3) Eiden et al., Pflungers Arch. 2018 Jan;470(1):79-88. (4) Braas et al., J Biol Chem. 1999 Sep 24;274(39):27702-27710. (5) Condro et al., J Comp Neurol. 2016 Dec 15; 524(18):3827-3848.
机译：垂体腺苷酸环化酶激活多肽（PACAP），正在研究以了解能量平衡的内分泌调节，并已被证明是在应激反应（1,2）的调节是重要的。具体而言，PACAP已显示调节热，由交感神经系统调节的能量燃烧过程有助于响应于冷应力和过度供料达到能量平衡。 PACAP在交感神经系统表达，并需要在肾上腺髓质突触维持从肾上腺髓质分泌肾上腺素响应于生理应力（3）。横跨交感神经系统的分支，PACAP受体表达最充分表征在颈上神经节（SCG）（4）。然而，PACAP及其受体的详细表征尚未在神经节，其节后纤维支配响应于产热应力脂肪组织（星状和腹腔神经节）进行。我们假设，PACAP由节前神经元支配星状和腹腔神经节产生的，并在对后神经节神经元中表达的PACAP受体起作用，并且该表达将响应于慢性冷应激上调。由于其小和无定形，我们已开发的协议，可靠地隔离星状和腹腔神经节和验证通过酪氨酸的存在下自己的身份羟化酶的mRNA，使用肾上腺和SCG样品作为阳性对照。 PACAP受体表达（VPAC1，VPAC2，PAC1）检查在利用实时PCR的神经节，和PACAP蛋白在转基因小鼠的表达eGFP的所述PACAP启动子（PACAP-EGFP小鼠）的控制下的神经节显现（5 ）。这项研究表明PACAP受体在神经节，其节后纤维支配脂肪组织中的表达，增强了我们的PACAP的在SNS的作用的认识，以及其适应性产热的调节作用。参考文献：（1）Gray等人，PACAP：应激反应的调节剂。在：芬克G，编压力：生理学，生物化学，病理检查。 2019：279-291。 （2）的Mustafa，进阶药理学。圣迭戈，加利福尼亚州：445-457。 （3）营电等，Pflungers拱。 2018扬; 470（1）：79-88。 （4）Braas等，生物化学杂志。 1999年09月24日; 274（39）：27702-27710。 （5）孔德罗等人，J小样神经病学。 2016年12月15; 524（18）：3827-3848。

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