ANALYSIS OF ENDOGENOUS CORTISOL CONCENTRATIONS IN THE HAIR OF RHESUS MACAQUES

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。下丘脑-垂体-肾上腺皮质（HPA）系统活动的短期变化通常通过测量血浆、唾液、尿液或粪便中的糖皮质激素或代谢物浓度来评估。然而，目前还没有确定该系统长期（即周或月）活动的方法。在这里，我们描述的发展和验证一个简单的程序测量皮质醇浓度在恒河猴的头发。该程序包括对头发进行两次简短的异丙醇洗涤以去除表面污染物，随后将洗涤和干燥的头发粉末化，24小时甲醇提取，然后蒸发溶剂并在实验缓冲液中重构提取物，最后通过敏感和特异性酶免疫分析法（EIA）分析提取的皮质醇。我们的结果证实了皮质醇的特异性，表明头发的近端和远端部分在皮质醇浓度上没有差异。这种新方法对于评估相对较长时间内非人类灵长类动物（以及其他物种）的基线HPA活性，以及监测可能与各种实验操作相关的慢性应激，应该是有价值的

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. Short-term changes in activity of the hypothalamic-pituitary-adrenocortical (HPA) system are routinely assessed by measuring glucocorticoid or metabolite concentrations in plasma, saliva, urine, or feces. However, there are no current methods for determining long-term (i.e., weeks or months) activity of this system. Herein we describe the development and validation of a simple procedure for measuring cortisol concentrations in the hair of rhesus macaques. This procedure involves two brief isopropanol washes of the hair strands to remove surface contaminants, subsequent powdering of the washed and dried hair, a 24-h methanol extraction followed by evaporation of the solvent and reconstitution of the extract in assay buffer, and finally analysis of the extracted cortisol by a sensitive and specific enzyme immunoassay (EIA). Our results confirm the specificity of the procedure for cortisol, show that proximal and distal segments of hair do not differ in their cortisol concentration. This new procedure should be valuable for assessing baseline HPA activity in non-human primates (and, with appropriate validation, in other species as well) over relatively long periods of time, and also for monitoring chronic stress that might be associated with various experimental manipulation