Hemmings H C, Greengard P
J Neurosci. 1986 May;6(5):1469-81. doi: 10.1523/JNEUROSCI.06-05-01469.1986.
Rabbit antisera and mouse monoclonal antibodies have been prepared to bovine DARPP-32 (dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein, Mr = 32,000), and used to study its regional, tissue, and phylogenetic distributions. The antibodies, none of which distinguished between dephospho-DARPP-32 and phospho-DARPP-32, were characterized and used to develop a sensitive and specific radioimmunoassay for DARPP-32. The radioimmunoassay, in conjunction with immunolabeling of SDS/PAGE transfers and immunoprecipitation of phosphorylated tissue extracts, was used to measure immunoreactive DARPP-32 in microdissected regions of rat CNS, in peripheral nervous and non-nervous tissues, and in CNS tissue from various animal species. The distribution of DARPP-32 was generally consistent with the interpretation that it is localized primarily to dopaminoceptive cells that possess dopamine-sensitive adenylate cyclase (D-1 dopamine receptors coupled to adenylate cyclase). Within the rat CNS, DARPP-32 was most highly concentrated in the basal ganglia. DARPP-32 was present in neostriatum from all six mammalian species tested (mouse, rat, guinea pig, rabbit, cow, and rhesus monkey) at concentrations of from 96 to 144 pmol/mg total protein, which constituted from 0.22 to 0.32% of the total protein. DARPP-32 was also identified at low levels in several peripheral tissues, including choroid plexus, parathyroid cells, adrenal chromaffin cells, posterior pituitary gland, pineal gland, and superior cervical sympathetic ganglion. A phylogenetic survey was carried out of proteins immunologically related to DARPP-32 in nervous tissue from nonmammalian species. DARPP-32-like proteins were identified in dopaminoceptive brain regions from representative members of the amniote vertebrate classes (birds and reptiles), while none was identified in dopaminoceptive brain regions from representative members of the anamniote vertebrate classes (bony fishes and amphibians) or in nervous tissue from representative members of several invertebrate classes.
已制备出针对牛DARPP - 32（多巴胺和3':5'-单磷酸腺苷调节的磷蛋白，分子量 = 32,000）的兔抗血清和小鼠单克隆抗体，并用于研究其区域、组织和系统发育分布。这些抗体均无法区分去磷酸化的DARPP - 32和磷酸化的DARPP - 32，对其进行了特性鉴定，并用于开发一种灵敏且特异的DARPP - 32放射免疫测定法。该放射免疫测定法结合SDS/PAGE转印的免疫标记和磷酸化组织提取物的免疫沉淀，用于测量大鼠中枢神经系统微切割区域、外周神经和非神经组织以及来自各种动物物种的中枢神经系统组织中的免疫反应性DARPP - 32。DARPP - 32的分布总体上与以下解释一致，即它主要定位于具有多巴胺敏感腺苷酸环化酶（与腺苷酸环化酶偶联的D - 1多巴胺受体）的多巴胺感受细胞。在大鼠中枢神经系统内，DARPP - 32高度集中于基底神经节。在所测试的所有六种哺乳动物物种（小鼠、大鼠、豚鼠、兔、牛和恒河猴）的新纹状体中均存在DARPP - 32，浓度为96至144 pmol/mg总蛋白，占总蛋白的0.22%至0.32%。在包括脉络丛、甲状旁腺细胞、肾上腺嗜铬细胞、垂体后叶、松果体和颈上神经节在内的几种外周组织中也检测到低水平的DARPP - 32。对非哺乳动物物种神经组织中与DARPP - 32免疫相关的蛋白质进行了系统发育调查。在羊膜脊椎动物类（鸟类和爬行动物）的代表性成员的多巴胺感受脑区中鉴定出了DARPP - 32样蛋白，而在无羊膜脊椎动物类（硬骨鱼类和两栖动物）的代表性成员的多巴胺感受脑区或几种无脊椎动物类的代表性成员的神经组织中未鉴定出。
