CHOLINOCEPTIVE NEURONS IN THE RETINA

乙酰胆碱(ACh)是脊椎动物视网膜中的一种递质，它影响 许多神经节细胞的反应特性。八个阿尔法和三个贝塔 神经型烟碱型乙酰胆碱受体(NAChRs)的亚基已经被 克隆了它们，并针对它们产生了抗体。这些亚单位可以 组合以形成大量的nAChR亚型，每个亚型的特征是 独特的亚基组成和生理和药理特性。 具体的目标是基于这样的假设：1)不同的视网膜 细胞类型表达nAChR亚型的不同组合，2) 不同的nAChR亚型在视网膜中起着不同的功能作用。 1.抗血清生产：合成抗血清的生产 与鸡和哺乳动物α基因独特序列相对应的多肽 贝塔亚基将继续存在。然后，这些文件将用于 具体目标2、3和5中描述的研究。 2.禽和鸡nAChR表达细胞的特性 哺乳动物视网膜：针对α和β亚基的抗体和抗血清 将用于确定nAChR在鸡、大鼠、 和兔眼视网膜。NAChr表达细胞的鉴定将基于 它们的神经递质、酶或其他分子标志物的含量 通过传统的双标记研究揭示。空间关系 表达nAChR的细胞的树突与 其他神经元将以同样的方式进行研究。 3.视网膜nAChR亚基组成和丰度的测定 亚型：免疫沉淀实验使用相同的抗体和 上述抗血清将被用来确定亚基组成 鸡和兔视网膜nAChR，并量化它们的相对丰度。 4.胆碱能无长突细胞亚群的特征 兔视网膜胆碱能无长突细胞至少由两个 基于nAChR和GABA表达的亚群。这些亚群 将根据它们的密度、分布和 这些细胞的树突与这些细胞的树突的空间关系 其他视网膜神经元。 5.哺乳动物视网膜发育过程中的胆碱能回路：亚单位 特定的抗体和抗血清将与 用于确定胆碱模式的市售抗血清 乙酰基转移酶、GABA和nAChR在出生后视网膜中的表达 已知年龄的雪貂。

Acetylcholine (ACh) is a transmitter in the vertebrate retina that affects the response properties of many ganglion cells. Eight alpha and three beta subunits of neuronal nicotinic acetylcholine receptors (nAChRs) have been cloned, and antibodies have been raised against them. These subunits can combine to form a large number of nAChR subtypes, each characterized by a unique subunit composition and physiological and pharmacological profile. The specific aims are based on the hypothesis that 1) different retinal cell types express different combinations of nAChR subtypes, and that 2) different nAChR subtypes serve different functional roles in the retina. 1. Antisera production: The production of antisera against synthetic peptides corresponding to unique sequences of chick and mammalian alpha and beta subunits will be continued. These will then be used for use in the studies described in Specific Aims 2, 3, and 5. 2. Characterization of the nAChR-expressing cells in the avian and mammalian retina: Antibodies and antisera against alpha and beta subunits will be used to determine the nAChR expression pattern in the chick, rat, and rabbit retina. nAChr-expressing cells will be identified based upon their content of neurotransmitters, enzymes, or other molecular markers as revealed by conventional double label studies. The spatial relationships between the dendrites of the nAChR-expressing cells and the processes of other neurons will be studied in the same fashion. 3. Determination of the subunit composition and abundance of retinal nAChR subtypes: Immune precipitation experiments using the same antibodies and antisera as above will be used to determine the subunit composition of chick and rabbit retinal nAChR and to quantify their relative abundance. 4. Characterization of subpopulations of cholinergic amacrine cells: The cholinergic amacrine cells of the rabbit retina comprise at least two subpopulations based upon nAChR and GABA expression. These subpopulations will be characterized in terms of their density, distribution, and the spatial relationships between the dendrites of these cells and those of other retinal neurons. 5. Cholinergic circuitry in the developing mammalian retina: Subunit specific antibodies and antisera will be used in conjunction with commercially available antisera to determine the pattern of choline acetyltransferase, GABA, and nAChR expression in retinas from postnatal ferrets of known ages.