Vaughan Cheryl H, Zarebidaki Eleen, Ehlen J Christopher, Bartness Timothy J
Department of Biology, Neuroscience Institute and Center for Obesity Reversal, Georgia State University, Atlanta, Georgia, USA.
Department of Biology, Neuroscience Institute and Center for Obesity Reversal, Georgia State University, Atlanta, Georgia, USA; Department of Neurobiology, Neuroscience Institute, Morehouse School of Medicine, Atlanta, Georgia, USA.
Methods Enzymol. 2014;537:199-225. doi: 10.1016/B978-0-12-411619-1.00011-2.
Here, we provide a detailed account of how to denervate white and brown adipose tissue (WAT and BAT) and how to measure sympathetic nervous system (SNS) activity to these and other tissues neurochemically. The brain controls many of the functions of WAT and BAT via the SNS innervation of the tissues, especially lipolysis and thermogenesis, respectively. There is no clearly demonstrated parasympathetic innervation of WAT or the major interscapular BAT (IBAT) depot. WAT and BAT communicate with the brain neurally via sensory nerves. We detail the surgical denervation (eliminating both innervations) of several WAT pads and IBAT. We also detail more selective chemical denervation of the SNS innervation via intra-WAT/IBAT 6-hydroxy-dopamine (a catecholaminergic neurotoxin) injections and selective chemical sensory denervation via intra-WAT/IBAT capsaicin (a sensory nerve neurotoxin) injections. Verifications of the denervations are provided (HPLC-EC detection for SNS, ELIA for calcitonin gene-related peptide (proven sensory nerve marker)). Finally, assessment of the SNS drive to WAT/BAT or other tissues is described using the alpha-methyl-para-tyrosine method combined with HPLC-EC, a direct neurochemical measure of SNS activity. These methods have proven useful for us and for other investigators interested in innervation of adipose tissues. The chemical denervation approach has been extended to nonadipose tissues as well.
在此,我们详细介绍了如何去神经支配白色和棕色脂肪组织(WAT和BAT),以及如何通过神经化学方法测量交感神经系统(SNS)对这些组织及其他组织的活性。大脑通过对这些组织的SNS神经支配来控制WAT和BAT的许多功能,特别是分别控制脂肪分解和产热。目前尚无明确证据表明WAT或主要肩胛间BAT(IBAT)储存部位存在副交感神经支配。WAT和BAT通过感觉神经与大脑进行神经通讯。我们详细介绍了对几个WAT垫和IBAT进行手术去神经支配(消除两种神经支配)的方法。我们还详细介绍了通过向WAT/IBAT内注射6-羟基多巴胺(一种儿茶酚胺能神经毒素)对SNS神经支配进行更具选择性的化学去神经支配,以及通过向WAT/IBAT内注射辣椒素(一种感觉神经毒素)进行选择性化学感觉去神经支配的方法。提供了去神经支配的验证方法(用于SNS的高效液相色谱-电化学检测,用于降钙素基因相关肽(已证实的感觉神经标志物)的酶联免疫吸附测定)。最后,描述了使用α-甲基-对-酪氨酸方法结合高效液相色谱-电化学检测对WAT/BAT或其他组织的SNS驱动进行评估,这是一种直接测量SNS活性的神经化学方法。这些方法已被证明对我们以及其他对脂肪组织神经支配感兴趣的研究人员很有用。化学去神经支配方法也已扩展到非脂肪组织。