Cloning and characterization of a novel neuropeptide using TRH knockout mice.

据报道，TRH在大脑中具有几种神经生理作用。为了深入了解这些效应背后的分子机制，我们试图利用TRH敲除小鼠、减法cDNA分析和微阵列系统克隆TRH调控的cDNA。通过这些分析获得的野生型和TRH^<-/->小脑的100多个克隆进行了分析。其中，一个无性系添加甲状腺激素后，正常甲状腺TRH^<-/->小脑mDP1 mRNA水平较野生型显著降低。对下丘脑、纹状体、中脑海马、脑桥、小脑、皮质等7个脑区进行Northern blot分析发现，mDP1 mRNA在下丘脑和小脑显著表达。此外，饥饿72小时导致下丘脑和小脑mDP1 mRNA水平显著下降。这些结果表明mDP1可能是调节饱腹感行为的物质。

TRH has been reported to possess several neurophysiological actions in the brain. To gain insights into the molecular mechanisms underlying these effects, we attempted to clone a cDNA that was regulated by TRH using TRH knockout mice and subtractive cDNA analysis and a microarray system. Over 100 clones obtained by these analyses between the wild-type and TRH^<-/-> cerebellum were analyzed. Among them, a clone, mDP1 mRNA level in the euthyroid TRH^<-/-> cerebellum supplemented with thyroid hormone were significantly decreased compared with those in the wild-type. Northern blot analysis of seven brain regions including hypothalamus, striatum, midbrain hippocampus, pons, cerebellum, and cortex revealed that mDP1 mRNA were expressed significantly in the hypothalamus and cerebellum. Furthermore, starvation for 72 hours lead to a significant decrease in mDP1 mRNA level in the hypothalamus and cerebellum. These results suggested that mDP1 might be a substance for regulating satiety behavior.