Paton Julian F R, Machado Benedito H, Moraes Davi J A, Zoccal Daniel B, Abdala Ana P, Smith Jeffrey C, Antunes Vagner R, Murphy David, Dutschmann Mathias, Dhingra Rishi R, McAllen Robin, Pickering Anthony E, Wilson Richard J A, Day Trevor A, Barioni Nicole O, Allen Andrew M, Menuet Clément, Donnelly Joseph, Felippe Igor, St-John Walter M
Manaaki Manawa - The Centre for Heart Research, Faculty of Medical & Health Science, University of Auckland, Grafton, Auckland, New Zealand.
Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
J Physiol. 2022 May;600(9):2049-2075. doi: 10.1113/JP281953. Epub 2022 Apr 7.
Twenty-five years ago, a new physiological preparation called the working heart-brainstem preparation (WHBP) was introduced with the claim it would provide a new platform allowing studies not possible before in cardiovascular, neuroendocrine, autonomic and respiratory research. Herein, we review some of the progress made with the WHBP, some advantages and disadvantages along with potential future applications, and provide photographs and technical drawings of all the customised equipment used for the preparation. Using mice or rats, the WHBP is an in situ experimental model that is perfused via an extracorporeal circuit benefitting from unprecedented surgical access, mechanical stability of the brain for whole cell recording and an uncompromised use of pharmacological agents akin to in vitro approaches. The preparation has revealed novel mechanistic insights into, for example, the generation of distinct respiratory rhythms, the neurogenesis of sympathetic activity, coupling between respiration and the heart and circulation, hypothalamic and spinal control mechanisms, and peripheral and central chemoreceptor mechanisms. Insights have been gleaned into diseases such as hypertension, heart failure and sleep apnoea. Findings from the in situ preparation have been ratified in conscious in vivo animals and when tested have translated to humans. We conclude by discussing potential future applications of the WHBP including two-photon imaging of peripheral and central nervous systems and adoption of pharmacogenetic tools that will improve our understanding of physiological mechanisms and reveal novel mechanisms that may guide new treatment strategies for cardiorespiratory diseases.
25年前，一种名为工作心脏-脑干制备（WHBP）的新的生理学制备方法被引入，据称它将提供一个新的平台，使心血管、神经内分泌、自主神经和呼吸研究中以前无法进行的研究成为可能。在此，我们回顾了WHBP所取得的一些进展、一些优缺点以及潜在的未来应用，并提供了用于该制备的所有定制设备的照片和技术图纸。使用小鼠或大鼠，WHBP是一种原位实验模型，通过体外循环进行灌注，受益于前所未有的手术通路、用于全细胞记录的大脑机械稳定性以及类似于体外方法的药理学试剂的无阻碍使用。该制备方法揭示了一些新的机制见解，例如不同呼吸节律的产生、交感神经活动的神经发生、呼吸与心脏和循环之间的耦合、下丘脑和脊髓控制机制以及外周和中枢化学感受机制。对高血压、心力衰竭和睡眠呼吸暂停等疾病也有了深入了解。原位制备的研究结果已在清醒的体内动物中得到验证，并在进行测试时转化到了人类身上。我们通过讨论WHBP潜在的未来应用来结束本文，包括外周和中枢神经系统的双光子成像以及采用药物遗传学工具，这将提高我们对生理机制的理解，并揭示可能指导心肺疾病新治疗策略的新机制。
