Ro Seungil, Hwang Sung Jin, Muto Melodie, Jewett William Keith, Spencer Nick J
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, 89557, USA.
Am J Physiol Gastrointest Liver Physiol. 2006 Apr;290(4):G710-8. doi: 10.1152/ajpgi.00420.2005. Epub 2005 Dec 8.
It has been assumed that in piebald lethal mice that develop megacolon, impaired colonic motor activity is restricted to the aganglionic distal colon. Peristaltic mechanical recordings, immunohistochemistry, and quantitative PCR were used to investigate whether regions of the colon, other than the aganglionic segment, may also show anatomical modifications and dysfunctional colonic motor activity. Contrary to expectations, colonic migrating motor complexes (MMCs) were absent along the whole colon of piebald lethal homozygote mice and severely impaired in heterozygote siblings. Aganglionosis was detected not only in the distal colon of piebald homozygote lethal mice (mean length: 20.4 +/- 2.1 mm) but also surprisingly in their heterozygote siblings (mean length: 12.4 +/- 1.1 mm). Unlike homozygote lethal mice, piebald heterozygotes showed no signs of megacolon. Interestingly, mRNA expression for PGP 9.5 was also dramatically reduced (by 71-99%) throughout the entire small and large bowel in both homozygote lethal and heterozygous littermates (by 67-87%). Histochemical staining confirmed a significant reduction in myenteric ganglia along the whole colon. In summary, the piebald mutation in homozygote lethal and heterozygote siblings is associated with dramatic reductions in myenteric ganglia throughout the entire colon and not limited to the distal colon as originally thought. Functionally, this results in an absence or severe impairment of colonic MMC activity in both piebald homozygote lethal and heterozygote siblings, respectively. The observation that piebald heterozygotes have an aganglionic distal colon (mean length: 12 mm) but live a normal murine life span without megacolon suggests that aganglionosis >12 mm and the complete absence of colonic MMCs may be required before any symptoms of megacolon arise.
人们一直认为,在发生巨结肠的花斑致死小鼠中,结肠运动活动受损仅限于无神经节的远端结肠。采用蠕动机械记录、免疫组织化学和定量PCR来研究除无神经节段之外的结肠区域是否也会出现解剖学改变和结肠运动活动功能障碍。与预期相反,花斑致死纯合子小鼠的整个结肠均未出现结肠移行性运动复合波(MMC),杂合子同胞小鼠的MMC则严重受损。不仅在花斑纯合子致死小鼠的远端结肠(平均长度:20.4±2.1毫米)检测到无神经节症,令人惊讶的是,在其杂合子同胞小鼠中(平均长度:12.4±1.1毫米)也检测到了。与纯合子致死小鼠不同,花斑杂合子未表现出巨结肠的迹象。有趣的是,在纯合子致死和杂合子同窝小鼠的整个小肠和大肠中,PGP 9.5的mRNA表达也显著降低(降低了71 - 99%)(杂合子降低了67 - 87%)。组织化学染色证实整个结肠的肌间神经节显著减少。总之,纯合子致死和杂合子同胞中的花斑突变与整个结肠肌间神经节的显著减少有关,而不仅限于最初认为的远端结肠。在功能上,这分别导致花斑纯合子致死和杂合子同胞的结肠MMC活动缺失或严重受损。花斑杂合子有一个无神经节的远端结肠(平均长度:12毫米),但能正常度过小鼠寿命且无巨结肠,这一观察结果表明,在出现任何巨结肠症状之前,可能需要无神经节症超过12毫米且完全没有结肠MMC。