Tsujino T, Kose A, Saito N, Tanaka C
Department of Pharmacology, Kobe University School of Medicine, Japan.
J Neurosci. 1990 Mar;10(3):870-84. doi: 10.1523/JNEUROSCI.10-03-00870.1990.
We have localized the beta I-, beta II-, and gamma-subspecies of protein kinase C in cerebral neocortex with light and electron microscopic immunocytochemistry using a monoclonal antibody against gamma-PKC and polyclonal antisera to beta I- or beta II-PKC-specific oligopeptides. The gamma-PKC-immunopositive cell bodies were seen mostly in layers II, V, and VI, and the vast majority of them were pyramidal cells. The beta II-PKC immunoreactive cell bodies were observed in layers II, III, V, and VI, and most of them seemed to be pyramidal cells. Both gamma- and beta II-PKC were colocalized in some pyramidal cells in layers II, V, and VI. The small number of beta I-PKC immunoreactive cell bodies were observed in the neocortex, and many of them were nonpyramidal cells. About 80% of the beta I-PKC-immunoreactive cells were shown to be GABAergic neurons. The gamma-PKC-immunopositive neuropils were observed in layers I, II, V, and VI, while beta II-PKC-immunoreactive neuropils were seen in layers I-III, V, and VI. The distribution of each subspecies is much the same throughout all regions of the neocortex, although with different intensities of immunoreactivity. electron microscopic studies revealed that, in the perikarya, gamma-PKC was distributed throughout the cytoplasm, beta I-PKC was just adjacent to the plasma membrane, and beta II-PKC was located around the Golgi complex. The immunoreactivity of these 3 subspecies was also seen in dendrites and axons, but no immunoreactivity of these subspecies was found in the presynaptic terminals in the present study. The discrete cellular and intracellular distributions of protein kinase C subspecies imply that each subspecies has a specific role in neuronal activity in the cerebral neocortex.
我们利用针对γ-蛋白激酶C的单克隆抗体以及针对βI-或βII-蛋白激酶C特异性寡肽的多克隆抗血清,通过光镜和电镜免疫细胞化学方法,对大脑新皮质中蛋白激酶C的βI-、βII-和γ-亚型进行了定位。γ-蛋白激酶C免疫阳性细胞体主要见于II、V和VI层,其中绝大多数为锥体细胞。βII-蛋白激酶C免疫反应性细胞体在II、III、V和VI层中可见,其中大多数似乎是锥体细胞。γ-和βII-蛋白激酶C在II、V和VI层的一些锥体细胞中共定位。在新皮质中观察到少量βI-蛋白激酶C免疫反应性细胞体,其中许多是非锥体细胞。约80%的βI-蛋白激酶C免疫反应性细胞被证明是γ-氨基丁酸能神经元。γ-蛋白激酶C免疫阳性神经纤维网见于I、II、V和VI层,而βII-蛋白激酶C免疫反应性神经纤维网见于I-III、V和VI层。尽管免疫反应强度不同,但每个亚型在新皮质的所有区域分布大致相同。电镜研究显示,在胞体中,γ-蛋白激酶C分布于整个细胞质中,βI-蛋白激酶C紧邻质膜,βII-蛋白激酶C位于高尔基体复合体周围。这三种亚型的免疫反应性在树突和轴突中也可见,但在本研究中,这些亚型在突触前终末未发现免疫反应性。蛋白激酶C亚型在细胞和细胞内的离散分布表明,每个亚型在大脑新皮质的神经元活动中具有特定作用。