F Douglas Stephens Surgical Research Laboratory, Murdoch Childrens Research Institute, Parkville, Victoria, Australia.
Cell Tissue Res. 2010 Jul;341(1):33-48. doi: 10.1007/s00441-010-0981-9. Epub 2010 May 21.
Cholinergic nerves are identified by labelling molecules in the ACh synthesis, release and destruction pathway. Recently, antibodies against another molecule in this pathway have been developed. Choline reuptake at the synapse occurs via the high-affinity choline transporter (CHT1). CHT1 immunoreactivity is present in cholinergic nerve fibres containing vesicular acetylcholine transporter (VAChT) in the human and rat central nervous system and rat enteric nervous system. We have examined whether CHT1 immunoreactivity is present in nerve fibres in human intestine and whether it is colocalised with markers of cholinergic, tachykinergic or nitrergic circuitry. Human ileum and colon were fixed, sectioned and processed for fluorescence immunohistochemistry with antibodies against CHT1, class III beta-tubulin (TUJ1), synaptophysin, common choline acetyl-transferase (cChAT), VAChT, nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP). CHT1 immunoreactivity was present in many nerve fibres in the circular and longitudinal muscle, myenteric and submucosal ganglia, submucosa and mucosa in human colon and ileum and colocalised with immunoreactivity for TUJ1 and synaptophysin confirming its presence in nerve fibres. In nerve fibres in myenteric ganglia and muscle, CHT1 immunoreactivity colocalised with immunoreactivity for VAChT and cChAT. Some colocalisation occurred with SP immunoreactivity, but little with immunoreactivity for VIP or NOS. In the mucosa, CHT1 immunoreactivity colocalised with that for VIP and SP in nerve fibres and was also present in vascular nerve fibres in the submucosa and on epithelial cells on the luminal border of crypts. The colocalisation of CHT1 immunoreactivity with VAChT immunoreactivity in cholinergic enteric nerves in the human bowel thus suggests that CHT1 represents another marker of cholinergic nerves.
胆碱能神经通过标记 ACh 合成、释放和破坏途径中的分子来识别。最近，已经开发出针对该途径中另一种分子的抗体。突触处的胆碱再摄取是通过高亲和力胆碱转运体 (CHT1) 进行的。CHT1 免疫反应性存在于包含人、大鼠中枢神经系统和大鼠肠神经系统中囊泡乙酰胆碱转运体 (VAChT) 的胆碱能神经纤维中。我们已经检查了 CHT1 免疫反应性是否存在于人类肠内的神经纤维中，以及它是否与胆碱能、速激肽能或氮能电路的标志物共定位。固定、切片并用人 CHT1、III 类β-微管蛋白 (TUJ1)、突触素、胆碱乙酰转移酶 (cChAT)、VAChT、一氧化氮合酶 (NOS)、P 物质 (SP) 和血管活性肠肽 (VIP) 的抗体进行荧光免疫组织化学处理。CHT1 免疫反应性存在于人结肠和回肠的环形和纵行肌、肌间和黏膜下神经节、黏膜下和黏膜中的许多神经纤维中，并与 TUJ1 和突触素的免疫反应性共定位，证实其存在于神经纤维中。在肌间神经节和肌肉中的神经纤维中，CHT1 免疫反应性与 VAChT 和 cChAT 的免疫反应性共定位。与 SP 免疫反应性有一些共定位，但与 VIP 或 NOS 的免疫反应性很少。在黏膜中，CHT1 免疫反应性与 VIP 和 SP 在神经纤维中的免疫反应性共定位，并且也存在于黏膜下的血管神经纤维和隐窝腔上皮细胞的腔侧边界上。因此，CHT1 与 VAChT 免疫反应性在人类肠道中的胆碱能肠神经中的共定位表明 CHT1 代表了另一种胆碱能神经标志物。
