CO2-INDUCED ANESTHESIA RESULTS IN RAPID INCREASE IN PLASMA LEVELS OF VASOPRESSIN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 与单独使用断头相比，短暂的麻醉，如暴露在高水平的二氧化碳中，在斩首之前被认为是更人道的啮齿动物安乐死选择。对血浆中某些应激激素水平的研究，如皮质酮和促肾上腺皮质激素(ACTH)，已经支持在内分泌学和分子研究中使用这种安乐死方法。在目前的研究中，大鼠被短暂地暴露在充满二氧化碳的房间中，直到卧倒(20-25岁，S)，然后立即断头处死，收集躯干血；研究结果与没有暴露在二氧化碳中的断头处死大鼠进行了比较。放射免疫法测定血浆精氨酸加压素(AVP)和促肾上腺皮质激素(ACTH)免疫反应性。虽然ACTH-ir水平在短暂接触二氧化碳后保持稳定(根据其他研究人员的结果)，但AVP-ir水平增加了一个数量级以上。这些结果得到了定量毛细管液相色谱-质谱仪的证实，表明这种血浆AVP-ir水平的快速升高不是由于放射免疫分析中使用的抗体的非特异性识别所致。同样，使用毛细管-液-质联用技术，我们观察到二氧化碳暴露后血浆催产素水平迅速升高。这些令人惊讶的发现对于设计和解释涉及斩首前短暂接触二氧化碳的研究以及那些由二氧化碳诱导窒息导致的安乐死的研究具有重要意义。描述这项工作的手稿已经出版(300。B.Reed，J.Varon，B.T.Chait，M.J.Kreek<二氧化碳诱导的麻醉导致血浆中加压素水平的快速增加>，内分泌学150(2009)2934-9。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Brief anesthesia, such as following exposure to high levels of carbon dioxide, prior to decapitation is considered a more humane alternative for the euthanasia of rodents, compared to use of the decapitation alone. Studies of the levels of certain stress hormones in plasma such as corticosterone and adrenocorticotropic hormone (ACTH) have supported the use of this method of euthanasia in endocrinological and molecular studies. In the current study, rats were briefly exposed to a chamber filled with carbon dioxide until recumbent (20-25 s), and immediately sacrificed via decapitation, and trunk blood collected; findings were compared to rats sacrificed via decapitation with no exposure to carbon dioxide. Radioimmunassays were used to measure arginine vasopressin (AVP) and ACTH immunoreactivity (ir) in plasma. While ACTH-ir levels remained steady following brief exposure to carbon dioxide (in accordance with results of other investigators), AVP-ir levels were increased by more than an order of magnitude. These results were confirmed by quantitative capillary-liquid chromatography-mass spectrometry, indicating this observation of rapid increase in plasma AVP-ir levels is not due to nonspecific recognition by the antibody used in the radioimmunoassay. Likewise, using capillary-liquid chromatography-mass spectrometry, we observed a rapid increase in plasma oxytocin levels following carbon dioxide exposure. These surprising findings have important implications for the design and interpretation of studies involving brief carbon dioxide exposure prior to decapitation, as well as those with euthanasia resulting from carbon dioxide-induced asphyxiation. A manuscript describing this work has been published (300. B. Reed, J. Varon, B.T. Chait, M.J. Kreek "Carbon Dioxide-Induced Anesthesia Results in a Rapid Increase in Plasma Levels of Vasopressin" Endocrinology 150 (2009) 2934-9.