Elfers Kristin, Sehnert Alina Sophia, Wagner Alexander, Zwirner Ulrich, Linge Helena, Kulik Ulf, Poehnert Daniel, Winny Markus, Gundert Benjamin, Aselmann Heiko, Mazzuoli-Weber Gemma
Institute for Physiology and Cell Biology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Center for Systems Neuroscience (ZSN), Hannover, Germany.
Gastro Hep Adv. 2024 Aug 24;4(1):100537. doi: 10.1016/j.gastha.2024.08.016. eCollection 2025.
The enteric nervous system independently controls gastrointestinal function including motility, which is primarily mediated by the myenteric plexus, therefore also playing a crucial role in functional intestinal disorders. Live recordings from human myenteric neurons proved to be challenging due to technical difficulties. Using the neuroimaging technique, we are able to record human colonic myenteric neuronal activity and investigate their functional properties in a large cohort of patients.
Activity from myenteric neurons in wholemount preparations of different sampling sites of fresh, human colonic tissue was recorded using neuroimaging with the voltage sensitive dye 1-(3-sulfanatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl]vinyl]pyridinium betaine. Neuronal responses were analyzed following stimulation with nicotine and serotonin (5-HT) for differences based on the donor's age, the disorder indicative for surgery and the colonic region. Immunohistochemistry was performed to calculate the total neuronal numbers.
Stimulation with nicotine and 5-HT elicited reproducible action potential discharge in a proportion of human myenteric neurons. The responses to 5-HT were significantly greater in tissues from older patients and from those with inflammatory disorders, while neuronal activity to nicotinergic stimulation was comparable in all patients. Neuronal numbers declined with rising patient's age and was highest in the sigmoid colon.
Neuroimaging with 1-(3-sulfanatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl]vinyl]pyridinium betaine was successfully adapted to record reproducible responses from human colonic myenteric neurons upon pharmacological stimulation. Evidence exists for an impact of age and inflammation on the serotonergic neuronal signaling and for differences in neuronal numbers in the distinct colonic regions as well as a neuronal decrease with age.
肠神经系统独立控制包括蠕动在内的胃肠功能，蠕动主要由肌间神经丛介导，因此在功能性肠道疾病中也起着关键作用。由于技术困难，对人类肌间神经进行实时记录具有挑战性。利用神经成像技术，我们能够记录大量患者的人类结肠肌间神经活动并研究其功能特性。
使用对电压敏感的染料1-(3-磺丙基)-4-[β-[2-(二正辛基氨基)-6-萘基]乙烯基]吡啶鎓甜菜碱，通过神经成像记录新鲜人类结肠组织不同采样部位整装标本中肌间神经的活动。在用尼古丁和5-羟色胺（5-HT）刺激后，根据供体年龄、手术指征疾病和结肠区域分析神经元反应的差异。进行免疫组织化学以计算神经元总数。
尼古丁和5-HT刺激在一定比例的人类肌间神经元中引发了可重复的动作电位发放。老年患者和患有炎症性疾病患者的组织对5-HT的反应明显更大，而所有患者对烟碱能刺激的神经元活动相当。神经元数量随患者年龄增长而减少，在乙状结肠中最高。
1-(3-磺丙基)-4-[β-[2-(二正辛基氨基)-6-萘基]乙烯基]吡啶鎓甜菜碱神经成像已成功应用于记录人类结肠肌间神经元在药理学刺激下的可重复反应。有证据表明年龄和炎症对5-羟色胺能神经元信号传导有影响，不同结肠区域的神经元数量存在差异，且神经元数量随年龄减少。
