Kubota Y, Hattori R, Yui Y
Laboratory for Neural Systems, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Brain Res. 1994 Jun 27;649(1-2):159-73. doi: 10.1016/0006-8993(94)91060-x.
GABAergic interneurons in the rat frontal cortex were subdivided on the basis of immunoreactivity for calcium binding proteins, neuropeptides and nitric oxide synthase, using double immunofluorescence and mirror image immunohistochemical methods. The results indicate that in this region of the neocortex there are at least three distinct subpopulations of local circuit neurons. The first subgroup consists of parvalbumin-immunoreactive cells. Those do not contain neuropeptide, calretinin or nitric oxide synthase immunoreactivity. A substantial number of parvalbumin-immunoreactive cells in layer II/III were also immunoreactive for calbindin D28k. The second subgroup consists of cells immunoreactive for calretinin. Most were usually immunoreactive for vasoactive intestinal polypeptide as well, but a few cells in layer II/III were immunoreactive for one or the other only. Calretinin-immunoreactive cells do not colocalize parvalbumin, somatostatin or nitric oxide synthase, and only a few colocalize calbindin D28k. The third subgroup consists of cells most of which contain somatostatin, and is entirely separate from the parvalbumin- and calretinin-immunoreactive populations. There was substantial colocalization of somatostatin and calbindin D28k and of somatostatin and neuropeptide Y. Some somatostatin-immunoreactive cells showed nitric oxide synthase immunoreactivity. All of the populations of immunoreactive cells examined in the present study also showed GABA immunoreactivity. About 10% of calbindin D28k-immunoreactive cells and all of those strongly stained for calbindin D28k in layer II/III showed GABA immunoreactivity. Most calbindin D28k-positive cells in deep layers also showed GABA immunoreactivity. These results support that almost all calbindin D28k-immunoreactive non-pyramidal cells are probably GABAergic.
利用双重免疫荧光和镜像免疫组织化学方法，根据钙结合蛋白、神经肽和一氧化氮合酶的免疫反应性，对大鼠额叶皮质中的γ-氨基丁酸能中间神经元进行了细分。结果表明，在新皮质的这一区域，至少存在三种不同的局部回路神经元亚群。第一亚群由小白蛋白免疫反应性细胞组成。这些细胞不含有神经肽、钙视网膜蛋白或一氧化氮合酶免疫反应性。在第II/III层中，大量小白蛋白免疫反应性细胞也对钙结合蛋白D28k呈免疫反应性。第二亚群由对钙视网膜蛋白呈免疫反应性的细胞组成。大多数细胞通常也对血管活性肠肽呈免疫反应性，但第II/III层中的少数细胞仅对其中一种呈免疫反应性。钙视网膜蛋白免疫反应性细胞不与小白蛋白、生长抑素或一氧化氮合酶共定位，只有少数与钙结合蛋白D28k共定位。第三亚群由大多数含有生长抑素的细胞组成，并且与小白蛋白和钙视网膜蛋白免疫反应性群体完全分开。生长抑素与钙结合蛋白D28k以及生长抑素与神经肽Y之间存在大量共定位。一些生长抑素免疫反应性细胞显示出一氧化氮合酶免疫反应性。在本研究中检测的所有免疫反应性细胞群体也显示出γ-氨基丁酸免疫反应性。在第II/III层中，约10%的钙结合蛋白D28k免疫反应性细胞以及所有对钙结合蛋白D28k强染色的细胞显示出γ-氨基丁酸免疫反应性。深层中的大多数钙结合蛋白D28k阳性细胞也显示出γ-氨基丁酸免疫反应性。这些结果支持几乎所有钙结合蛋白D28k免疫反应性非锥体细胞可能都是γ-氨基丁酸能的。
