Ottenweller J E, Hedge G A
Endocrinology. 1982 Aug;111(2):515-21. doi: 10.1210/endo-111-2-515.
A method for extracting and assaying TSH-like immunoreactivity (TSH-LI) in regions of the central nervous systems (CNS) of individual rats was developed and validated. Serial dilutions of tissue extracts paralleled dilutions of purified rat TSH standard in the RIA, and tissue TSH-LI comigrated electrophoretically with TSH standard. TSH-LI was measured in the pituitary (PIT), the medial basal hypothalamus (MBH), the remainder of the hypothalamus (HYPO), and the cerebral cortex (CC). Female rats were maintained on a normal 12-h light, 12-h dark cycle (LD; onset of light, 0730 h) or an inverted 12-h light, 12-h dark cycle (DL; onset of light, 1930 h) for 3 weeks and then killed at 4-h intervals throughout the day. Another group of rats on the LD photoperiod were either thyroidectomized or sham thyroidectomized and killed 3 weeks later at 1000 h. Diurnal variations of TSH-LI were present in the PIT, MBH, HYPO, and CC on the LD photoperiod (P less than 0.05). The acrophases of MBH, HYPO, and CC TSH-LI diurnal variations occurred soon after the onset of darkness, whereas the PIT TSH-LI variation peaked soon after the onset of light. Inverting the photoperiod inverted the PIT TSH-LI diurnal variation, but central nervous system TSH-LI variations were not phase shifted after 3 weeks on the inverted photoperiod. Thyroidectomy elevated plasma and MBH TSH-LI levels (P less than 0.05), but PIT, HYPO, and CC TSH-LI levels were not significantly different from those in sham-operated controls. These results indicated that brain TSH concentrations could change under different physiological states independently of changes in PIT or plasma TSH. Although brain TSH exhibited diurnal variations, the data suggested that the phase of these variations was not set by the light-dark cycle.
开发并验证了一种在个体大鼠中枢神经系统(CNS)区域提取和测定促甲状腺激素样免疫反应性(TSH-LI)的方法。组织提取物的系列稀释与放射免疫分析(RIA)中纯化大鼠促甲状腺激素标准品的稀释平行,并且组织TSH-LI与促甲状腺激素标准品在电泳中迁移一致。在垂体(PIT)、内侧基底下丘脑(MBH)、下丘脑其余部分(HYPO)和大脑皮层(CC)中测量TSH-LI。雌性大鼠维持在正常的12小时光照、12小时黑暗周期(LD;光照开始时间为0730时)或倒置的12小时光照、12小时黑暗周期(DL;光照开始时间为1930时)3周,然后在一天中每隔4小时处死。另一组处于LD光周期的大鼠进行甲状腺切除或假甲状腺切除,并在3周后的1000时处死。在LD光周期下,PIT、MBH、HYPO和CC中TSH-LI存在昼夜变化(P小于0.05)。MBH、HYPO和CC中TSH-LI昼夜变化的高峰相位在黑暗开始后不久出现;而PIT中TSH-LI变化在光照开始后不久达到峰值。颠倒光周期可使PIT中TSH-LI的昼夜变化颠倒,但在颠倒光周期3周后,中枢神经系统TSH-LI的变化未发生相位改变。甲状腺切除可提高血浆和MBH中TSH-LI水平(P小于0.05),但PIT、HYPO和CC中TSH-LI水平与假手术对照组无显著差异。这些结果表明,脑内促甲状腺激素浓度可在不同生理状态下独立于垂体或血浆促甲状腺激素的变化而改变。尽管脑内促甲状腺激素呈现昼夜变化,但数据表明这些变化的相位并非由明暗周期设定。