Ono J K
Neuroscience. 1986 Aug;18(4):957-74. doi: 10.1016/0306-4522(86)90111-9.
Immunohistochemical procedures were applied to wholemounts of the central nervous system and posterior intestine of the mollusc, Aplysia californica, to facilitate localization of cells that were immunoreactive to several antisera recognizing various epitopes of the peptides cholecystokinin and gastrin. Only antisera that recognized the carboxyl terminal sequence common to cholecystokinin and gastrin reacted with the Aplysia tissues tested. Intracellular electrophysiological studies of identified postsynaptic targets of immunoreactive neurons in the cerebral ganglia indicated that mammalian forms of gastrin 1-17, several cholecystokinin fragments, and the related peptide, amphibian caerulein, did not mimick the synaptic response mediated by the immunoreactive presynaptic neurons. Combinations of electrophysiological, immunohistochemical, and biochemical studies of several neurons in the buccal ganglia indicated that neurons B7 and B13 were immunoreactive to antisera against cholecystokinin and gastrin and that neuron B13 also contained a concentration of the neurotransmitter acetylcholine as high as in the identified cholinergic buccal neurons, B4 and B5. Several differences in the immunoreactivity of the various antisera were observed. Only one of the antisera was effective in staining neurons in the abdominal ganglia and another antiserum stained subsets of neurons that were immunoreactive to most of the other antisera recognizing the carboxyl terminus common to cholecystokinin and gastrin. The giant serotoninergic metacerebral neurons in Aplysia were not immunoreactive to the cholecystokinin/gastrin antisera even though it has been reported that the homologous neurons in a pulmonate mollusc contain cholecystokinin-like immunoreactivity. These studies demonstrated that there are many neurons with cholecystokinin/gastrin-like immunoreactivity in the Aplysia central and peripheral nervous system and suggested that the peptide may differ from vertebrate forms of cholecystokinin and gastrin. The identification of immunoreactive neurons with known postsynaptic target neurons and buccal neurons with acetylcholine co-localized with a cholecystokinin/gastrin-like peptide will facilitate elucidation of the functions of peptides in the nervous system since the Aplysia preparation is well known to be amenable to multidisciplinary studies.
免疫组织化学方法应用于海兔（Aplysia californica）的中枢神经系统和后肠整体标本，以利于定位对几种抗血清呈免疫反应的细胞，这些抗血清可识别胆囊收缩素和胃泌素肽的各种表位。只有识别胆囊收缩素和胃泌素共同的羧基末端序列的抗血清与所测试的海兔组织发生反应。对脑神经节中免疫反应性神经元的已鉴定突触后靶标的细胞内电生理研究表明，胃泌素1 - 17的哺乳动物形式、几种胆囊收缩素片段以及相关肽两栖蛙皮素，均不能模拟免疫反应性突触前神经元介导的突触反应。对颊神经节中几个神经元进行的电生理、免疫组织化学和生化研究的组合表明，神经元B7和B13对针对胆囊收缩素和胃泌素的抗血清呈免疫反应，并且神经元B13中神经递质乙酰胆碱的浓度与已鉴定的胆碱能颊神经元B4和B5中的浓度一样高。观察到各种抗血清在免疫反应性方面存在若干差异。只有一种抗血清能有效染色腹神经节中的神经元，另一种抗血清则对大多数识别胆囊收缩素和胃泌素共同羧基末端的其他抗血清呈免疫反应的神经元亚群进行染色。海兔中的巨大5-羟色胺能大脑中神经元对胆囊收缩素/胃泌素抗血清无免疫反应，尽管有报道称肺螺类软体动物中的同源神经元含有胆囊收缩素样免疫反应性。这些研究表明，海兔中枢和外周神经系统中有许多具有胆囊收缩素/胃泌素样免疫反应性的神经元，并提示该肽可能与脊椎动物形式的胆囊收缩素和胃泌素不同。由于海兔标本众所周知适合进行多学科研究，因此鉴定具有已知突触后靶标的免疫反应性神经元以及乙酰胆碱与胆囊收缩素/胃泌素样肽共定位的颊神经元，将有助于阐明神经系统中肽的功能。
