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如何研究肠神经系统:在体内和体外分析运动障碍的各种方法。

How to approach the ENS: various ways to analyse motility disorders in situ and in vitro.

作者信息

Schäfer K-H, Hagl C I, Wink E, Holland-Cunz S, Klotz M, Rauch U, Waag K-L

机构信息

Department of Paediatric Surgery, Clinical Hospital Mannheim, University Heidelberg, Theodor-Kutzer-Ufer 1-3, 68135 Mannheim, Germany.

出版信息

Eur J Pediatr Surg. 2003 Jun;13(3):163-9. doi: 10.1055/s-2003-41266.

Abstract

Motility disorders of the human intestine are so variable that they cannot be diagnosed by just one technique. Their aetiology is obviously so varied that they have to be approached with a broad range of technical methods. These reach from the simple haematoxylin-stained section to the isolation of stem or precursor cells. In this study, various methods to investigate the enteric nervous system and its surrounding tissue are demonstrated. While sections from paraffin-embedded material or cryostat sections provide only a two-dimensional perspective of the ENS, the whole-mount method yields three-dimensional perspectives of large areas of the gut wall. The three-dimensional impression can even be enhanced by electron microscopy of the isolated ENS. Dynamical aspects of ENS development can be tackled by in vitro studies. The myenteric plexus can be isolated and cultivated under the influence of the microenvironment (protein extracts). Although the postnatal myenteric plexus is not fully developed, the choice of embryological neuronal cells seems to be more effective for certain approaches. They can be isolated from the embryonic mouse gut and cultivated under the influence of various factors. This method seems to us a valuable tool for the investigation of the aetiology of motility disorders, although only a "complete" approach which considers all available methods will yield at the end a clear understanding which might lead to new therapeutical concepts.

摘要

人类肠道的运动障碍具有很大的变异性,以至于无法仅通过一种技术进行诊断。其病因显然多种多样,因此必须采用广泛的技术方法来研究。这些方法从简单的苏木精染色切片到干细胞或前体细胞的分离。在本研究中,展示了多种研究肠神经系统及其周围组织的方法。虽然石蜡包埋材料的切片或低温恒温器切片仅提供肠神经系统的二维视角,但整装法可呈现肠壁大面积区域的三维视角。通过对分离出的肠神经系统进行电子显微镜观察,三维印象甚至可以得到增强。肠神经系统发育的动态方面可以通过体外研究来解决。肌间神经丛可以在微环境(蛋白质提取物)的影响下进行分离和培养。虽然出生后的肌间神经丛尚未完全发育,但对于某些方法而言,选择胚胎神经元细胞似乎更有效。它们可以从胚胎小鼠肠道中分离出来,并在各种因素的影响下进行培养。在我们看来,这种方法是研究运动障碍病因的一种有价值的工具,尽管只有采用考虑所有可用方法的“完整”方法,最终才能获得清晰的认识,进而可能产生新的治疗理念。

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