Kriegsfeld Lance J, Korets Ruslan, Silver Rae
Department of Psychology, Columbia University New York, 1190 Amsterdam Avenue, New York, 10027, USA.
Eur J Neurosci. 2003 Jan;17(2):212-20. doi: 10.1046/j.1460-9568.2003.02431.x.
The circadian clock located in the suprachiasmatic nuclei (SCN) of the hypothalamus regulates daily temporal organization in behaviour and neuroendocrine function. The molecular basis for circadian rhythm generation is an interacting transcriptional/translational feedback loop comprised of several 'clock genes' and their respective protein products. Clock genes are expressed not only in the SCN but also in numerous other locations throughout the brain, including regions rich in neuroendocrine cells. In order to investigate whether neuroendocrine cells function as autonomous oscillators, we used female transgenic mice in which an unstable, degradable jellyfish green fluorescent protein (GFP) gene is driven by a mouse Period 1 (Per1) gene promoter. Mice were injected (s.c.) with fluorogold (FG) in order to label neuroendocrine cells and brain sections were double-labelled for either FG and Per1 mRNA (labelled by GFP immunostaining) or FG and PER1 protein using fluorescence immunocytochemistry. Mice were killed during either the day or night. Neuroendocrine cells contained Per1 mRNA and PER1 protein in several brain regions with the greatest proportion of double-labelled cells occurring in the arcuate nucleus (Arc). The number of neuroendocrine cells labelled was not affected by the stage of the estrous cycle. Fewer FG-labelled cells expressed Per1 message and protein during the night compared to the day. In the Arc, staining for tyrosine hydroxylase revealed that neuroendocrine cells expressing Per1 message and protein were dopaminergic. Together, these findings suggest that neuroendocrine cells contain the molecular machinery necessary to oscillate independently. It remains to be determined whether these cells actually function as autonomous oscillators or whether these rhythms are driven by signals from the SCN. These findings also indicate that the endocrine system represents an opportunity to study the interactions between central (SCN and neuroendocrine cells) and peripheral circadian (endocrine gland) oscillators.
位于下丘脑视交叉上核(SCN)的生物钟调节着行为和神经内分泌功能的日常时间组织。昼夜节律产生的分子基础是一个由几个“时钟基因”及其各自的蛋白质产物组成的相互作用的转录/翻译反馈环。时钟基因不仅在SCN中表达,还在大脑中的许多其他位置表达,包括富含神经内分泌细胞的区域。为了研究神经内分泌细胞是否作为自主振荡器发挥作用,我们使用了雌性转基因小鼠,其中不稳定的、可降解的水母绿色荧光蛋白(GFP)基因由小鼠周期1(Per1)基因启动子驱动。给小鼠皮下注射荧光金(FG)以标记神经内分泌细胞,并使用荧光免疫细胞化学对脑切片进行FG和Per1 mRNA(通过GFP免疫染色标记)或FG和PER1蛋白的双重标记。在白天或晚上处死小鼠。神经内分泌细胞在几个脑区含有Per1 mRNA和PER1蛋白,双标记细胞比例最高的是弓状核(Arc)。标记的神经内分泌细胞数量不受发情周期阶段的影响。与白天相比,夜间标记的FG细胞中表达Per1信息和蛋白的细胞较少。在Arc中,酪氨酸羟化酶染色显示,表达Per1信息和蛋白的神经内分泌细胞是多巴胺能的。总之,这些发现表明神经内分泌细胞含有独立振荡所需的分子机制。这些细胞是否实际作为自主振荡器发挥作用,或者这些节律是否由来自SCN的信号驱动,仍有待确定。这些发现还表明,内分泌系统为研究中枢(SCN和神经内分泌细胞)和外周昼夜节律(内分泌腺)振荡器之间的相互作用提供了一个机会。