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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 短暂麻醉，如暴露于高水平的二氧化碳后，在断头之前被认为是一个更人道的替代啮齿动物安乐死，相比单独使用断头。对血浆中某些应激激素（如皮质酮和促肾上腺皮质激素（ACTH））水平的研究支持在内分泌和分子研究中使用这种安乐死方法。在当前研究中，将大鼠短暂暴露于充满二氧化碳的室中，直至横卧（20-25 s），并立即通过断头处死，收集躯干血液;将结果与通过断头处死但未暴露于二氧化碳的大鼠进行比较。用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（ir）。虽然ACTH-IR水平保持稳定后，短暂暴露于二氧化碳（根据其他研究人员的结果），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-质谱法的证实，表明血浆AVP-ir水平快速增加的观察结果不是由于放射免疫测定中使用的抗体的非特异性识别。同样，使用毛细管液相色谱-质谱法，我们观察到血浆催产素水平迅速增加后，二氧化碳暴露。这些令人惊讶的发现对于涉及斩首前短暂接触二氧化碳的研究以及因二氧化碳诱导窒息而实施安乐死的研究的设计和解释具有重要意义。描述这项工作的手稿已出版（300。 B。Reed，J. Varon，B.T. Chait，M. J. Kreek“二氧化碳诱导的麻醉导致血浆血管加压素水平迅速增加”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.