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二肽基肽酶10(DPP10)在大鼠脑内的免疫组化定位支持神经元中存在钾通道蛋白4(Kv4)/钾通道相互作用蛋白(KChIP)/二肽基肽酶样蛋白(DPPL)三元复合物。

Immunohistochemical localization of DPP10 in rat brain supports the existence of a Kv4/KChIP/DPPL ternary complex in neurons.

作者信息

Wang Wan-Chen, Cheng Chau-Fu, Tsaur Meei-Ling

机构信息

Institute of Neuroscience, Brain Research Center, National Yang-Ming University, Taipei 112, Taiwan.

出版信息

J Comp Neurol. 2015 Mar 1;523(4):608-28. doi: 10.1002/cne.23698. Epub 2014 Dec 19.

Abstract

Subthreshold A-type K(+) currents (ISA s) have been recorded from the cell bodies of hippocampal and neocortical interneurons as well as neocortical pyramidal neurons. Kv4 channels are responsible for the somatodendritic ISA s. It has been proposed that neuronal Kv4 channels are ternary complexes including pore-forming Kv4 subunits, K(+) channel-interacting proteins (KChIPs), and dipeptidyl peptidase-like proteins (DPPLs). However, colocalization evidence was still lacking. The distribution of DPP10 mRNA in rodent brain has been reported but its protein localization remains unknown. In this study, we generated a DPP10 antibody to label DPP10 protein in adult rat brain by immunohistochemistry. Absent from glia, DPP10 proteins appear mainly in the cell bodies of DPP10(+) neurons, not only at the plasma membrane but also in the cytoplasm. At least 6.4% of inhibitory interneurons in the hippocampus coexpressed Kv4.3, KChIP1, and DPP10, with the highest density in the CA1 strata alveus/oriens/pyramidale and the dentate hilus. Colocalization of Kv4.3/KChIP1/DPP10 was also detected in at least 6.9% of inhibitory interneurons scattered throughout the neocortex. Both hippocampal and neocortical Kv4.3/KChIP1/DPP10(+) inhibitory interneurons expressed parvalbumin or somatostatin, but not calbindin or calretinin. Furthermore, we found colocalization of Kv4.2/Kv4.3/KChIP3/DPP10 in neocortical layer 5 pyramidal neurons and olfactory bulb mitral cells. Together, although DPP10 is also expressed in some brain neurons lacking Kv4 (such as parvalbumin- and somatostatin-positive Golgi cells in the cerebellum), colocalization of DPP10 with Kv4 and KChIP at the plasma membrane of ISA -expressing neuron somata supports the existence of Kv4/KChIP/DPPL ternary complex in vivo.

摘要

在海马体和新皮质中间神经元以及新皮质锥体神经元的细胞体中已记录到阈下A 型钾电流(ISA s)。Kv4通道负责树突-胞体的ISA s。有人提出神经元Kv4通道是三元复合物,包括形成孔道的Kv4亚基、钾通道相互作用蛋白(KChIPs)和二肽基肽酶样蛋白(DPPLs)。然而,仍缺乏共定位证据。已报道了DPP10 mRNA在啮齿动物脑中的分布,但其蛋白定位仍不清楚。在本研究中,我们制备了一种DPP10抗体,通过免疫组织化学标记成年大鼠脑中的DPP10蛋白。DPP10蛋白不存在于神经胶质细胞中,主要出现在DPP10(+)神经元的细胞体中,不仅在质膜上,也在细胞质中。海马体中至少6.4%的抑制性中间神经元共表达Kv4.3、KChIP1和DPP10,在CA1层的齿状回/原层/锥体层和齿状回门区密度最高。在散布于整个新皮质的至少6.9%的抑制性中间神经元中也检测到Kv4.3/KChIP1/DPP10的共定位。海马体和新皮质的Kv4.3/KChIP1/DPP10(+)抑制性中间神经元均表达小白蛋白或生长抑素,但不表达钙结合蛋白或钙视网膜蛋白。此外,我们在新皮质第5层锥体神经元和嗅球 mitral细胞中发现了Kv4.2/Kv4.3/KChIP3/DPP10的共定位。总之,尽管DPP10也在一些缺乏Kv4的脑神经元中表达(如小脑中表达小白蛋白和生长抑素的高尔基细胞),但在表达ISA的神经元胞体质膜上DPP10与Kv4和KChIP的共定位支持了体内Kv4/KChIP/DPPL三元复合物的存在。

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