Mashburn Kendall L, Atkinson Shannon
University of Alaska, Fairbanks and Alaska SeaLife Center, P.O. Box 1329, 301 Railway Avenue, Seward, AL 99664, USA.
Gen Comp Endocrinol. 2004 May 1;136(3):371-81. doi: 10.1016/j.ygcen.2004.01.016.
Fecal corticosterone concentrations, measured via radioimmunoassay (RIA), were validated as a method to monitor adrenal function in Steller sea lion physiology. Quantification of adrenal response to an acute stressor and relevance of data produced by developed methodologies was determined through physiological challenge with exogenous administration of adrenocorticotropic hormone (ACTH) to captive adult, reproductively intact, Steller sea lions of both sexes (n=3, 1 male, 2 female) during seasonal molt. Following ACTH administration, serial blood and fecal samples were collected and analyzed by RIA to determine adrenal response. Storage regimens and weather exposure were examined to establish external impact on fecal corticosterone concentrations. High-pressure liquid chromatography (HPLC) of both serum and feces of Steller sea lions was employed to explore potential gender-based differences extant in either sample media. ACTH challenges produced >3-fold increases in serum cortisol concentrations which were reflected in >18-fold increases in fecal corticosterone concentrations post-injection at 3.25 and 32 h, respectively, and fecal corticosterone concentrations returned to baseline 52 h post-injection. Neither outdoor exposure to weather nor variation in duration and temperature of freezer storage impacted fecal corticosterone concentrations. HPLC of individual fecal samples produced eluate immunoreactivity profiles that differed consistently with both sex and age class. Techniques developed herein effectively detected physiologically relevant corticosterone data in Steller sea lion feces, unaffected by conditions likely to be encountered with field collection samples. Additionally, results quantify an acute response to ACTH and provide methodology for examining chronically heightened adrenal activity in Steller sea lions.
通过放射免疫分析法(RIA)测量的粪便皮质酮浓度,被验证为一种监测海獭生理中肾上腺功能的方法。通过对圈养的成年、生殖功能正常、处于季节性换毛期的雌雄海獭(n = 3,1只雄性,2只雌性)进行外源性促肾上腺皮质激素(ACTH)的生理刺激,确定肾上腺对急性应激源的反应量化以及所开发方法产生的数据的相关性。在注射ACTH后,采集系列血液和粪便样本,并通过RIA进行分析以确定肾上腺反应。检查储存方案和天气暴露情况,以确定对粪便皮质酮浓度的外部影响。采用高效液相色谱法(HPLC)分析海獭的血清和粪便,以探索两种样本介质中存在的潜在性别差异。ACTH刺激使血清皮质醇浓度分别在注射后3.25小时和32小时增加了3倍以上,这反映在粪便皮质酮浓度分别增加了18倍以上,并且粪便皮质酮浓度在注射后52小时恢复到基线水平。户外天气暴露以及冷冻储存时间和温度的变化均未影响粪便皮质酮浓度。对单个粪便样本进行的HPLC产生的洗脱液免疫反应谱在性别和年龄组之间始终存在差异。本文开发的技术有效地检测了海獭粪便中与生理相关的皮质酮数据,不受野外采集样本可能遇到的条件的影响。此外,结果量化了对ACTH的急性反应,并提供了检查海獭慢性肾上腺活动增强的方法。