Gautvik K M, de Lecea L, Gautvik V T, Danielson P E, Tranque P, Dopazo A, Bloom F E, Sutcliffe J G
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8733-8. doi: 10.1073/pnas.93.16.8733.
We applied the directional tag PCR subtractive hybridization method to construct a rat hypothalamic cDNA library from which cerebellar and hippocampal sequences had been depleted, enriching 20-30-fold for sequences expressed selectively in the hypothalamus. We studied a sample of 94 clones selected for enrichment in the subtracted library. These clones corresponded to 43 distinct mRNA species, about half of which were novel. Thirty-eight of these 43 mRNAs (corresponding to 85 of the clones in the sample) exhibited enrichment in the hypothalamus; 23 were highly enriched. In situ hybridization studies revealed that one novel species was restricted to cells in a small bilaterally symmetric area of the paraventricular hypothalamus. Other novel mRNAs showed substantial enrichment in basal diencephalic structures, particularly the hypothalamus, without restriction to single hypothalamic nuclei. The data suggest that the hypothalamus utilizes at least two distinct strategies for employing its selectively expressed proteins. Secretory neuropeptides utilized for intercellular communication are produced by functionally discrete nuclei, while several other proteins are shared by structures that are unrelated in their physiological roles but may share biochemical systems.
我们应用定向标签PCR消减杂交方法构建了一个大鼠下丘脑cDNA文库，其中已去除小脑和海马序列，使在下丘脑中选择性表达的序列富集了20至30倍。我们研究了从消减文库中选择用于富集的94个克隆样本。这些克隆对应于43种不同的mRNA种类，其中约一半是新的。这43种mRNA中的38种（对应于样本中的85个克隆）在下丘脑中表现出富集；23种高度富集。原位杂交研究表明，一种新的mRNA种类仅限于下丘脑室旁核双侧对称的小区域中的细胞。其他新的mRNA在间脑基部结构，特别是下丘脑中显示出大量富集，但不限于单个下丘脑核。数据表明，下丘脑利用至少两种不同的策略来利用其选择性表达的蛋白质。用于细胞间通讯的分泌性神经肽由功能上离散的核产生，而其他几种蛋白质则由生理作用无关但可能共享生化系统的结构共享。
