Hendry S H, Jones E G, DeFelipe J, Schmechel D, Brandon C, Emson P C
Proc Natl Acad Sci U S A. 1984 Oct;81(20):6526-30. doi: 10.1073/pnas.81.20.6526.
Neurons in the cat and monkey cerebral cortex were stained immunocytochemically for glutamic acid decarboxylase (GluDCase; L-glutamate 1-carboxy-lyase, EC 4.1.1.15), somatostatin (SRIF), neuropeptide Y (NPY), and cholecystokinin octapeptide (CCK). In all areas of cortex examined (somatic sensory, motor, parietal and visual areas), neurons displaying immunoreactivity for each of these molecules were nonpyramidal cells. Co-localization of GluDCase immunoreactivity with peptide immunoreactivity in the same cells was demonstrated by (i) the antibody elution method, staining the same cells by immunofluorescence, first for a peptide and then for GluDCase; (ii) double staining of the same sections with sheep anti-GluDCase and rabbit anti-peptide antisera, the bound antibodies being localized by rhodamine-conjugated donkey anti-sheep and fluorescein-conjugated swine anti-rabbit secondary antisera. With both procedures, cell bodies immunoreactive for GluDCase and for each of the peptides were found in all areas of cortex examined. With double labeling on single sections, it was found that all CCK-, SRIF-, and NPY-immunoreactive cells in cat cortex and 90%-95% in monkey cortex are also GluDCase positive. Many more cells, however, are immunoreactive for GluDCase alone. GluDCase was co-localized with CCK, SRIF, or NPY not only in cell somata, but also in small punctate structures, which are likely to be axon terminals. From the data gained in previous electron microscopic studies, we postulate that neurons displaying GluDCase- and CCK-like immunoreactivity are a class separate from those displaying GluDCase- and SRIF-like immunoreactivity. NPY, however, is co-localized with SRIF immunoreactivity. These results imply that classes of cortical interneuron contain a conventional neurotransmitter (gamma-aminobutyric acid) and a neuromodulator (one of the peptides).
用免疫细胞化学方法对猫和猴大脑皮质中的神经元进行谷氨酸脱羧酶（GluDCase；L-谷氨酸1-羧基裂解酶，EC 4.1.1.15）、生长抑素（SRIF）、神经肽Y（NPY）和八肽胆囊收缩素（CCK）染色。在所检查的皮质所有区域（躯体感觉区、运动区、顶叶和视觉区），显示对这些分子中每一种有免疫反应性的神经元均为非锥体细胞。通过以下方法证明了同一细胞中GluDCase免疫反应性与肽免疫反应性的共定位：（i）抗体洗脱法，先用免疫荧光对同一细胞染色，首先针对一种肽，然后针对GluDCase；（ii）用羊抗GluDCase和兔抗肽抗血清对同一切片进行双重染色，结合的抗体用罗丹明偶联的驴抗羊和荧光素偶联的猪抗兔二抗进行定位。采用这两种方法，在所检查的皮质所有区域均发现了对GluDCase和每种肽有免疫反应性的细胞体。在单一切片上进行双重标记时，发现猫皮质中所有CCK、SRIF和NPY免疫反应性细胞以及猴皮质中90%-95%的此类细胞也呈GluDCase阳性。然而，更多的细胞仅对GluDCase有免疫反应性。GluDCase不仅与CCK、SRIF或NPY在细胞体中共定位，还与小的点状结构共定位，这些结构可能是轴突终末。根据先前电子显微镜研究获得的数据，我们推测显示GluDCase样和CCK样免疫反应性的神经元与显示GluDCase样和SRIF样免疫反应性的神经元是不同的类别。然而，NPY与SRIF免疫反应性共定位。这些结果表明，皮质中间神经元类别包含一种传统神经递质（γ-氨基丁酸）和一种神经调质（其中一种肽）。
