Freeman T C, Bentsen B S, Thwaites D T, Simmons N L
Sánger Centre, Hinxton, Cambridge, UK.
Pflugers Arch. 1995 Jul;430(3):394-400. doi: 10.1007/BF00373915.
In order to examine the intestinal expression of the recently cloned H+/di-tripeptide transporter (PepT1), oligonucleotide probes were synthesized and their specificity confirmed by Northern blot analysis of rabbit jejunal RNA. In situ hybridization studies, using these probes, show that PepT1 is expressed all along the small intestine and at a very much reduced level in the colon. In contrast, PepT1 mRNA was not detected in the stomach, sacculus rotundus or caecum. Microscopic examination of tissue sections showed PepT1 expression to be restricted to intestinal epithelium with no detectable expression in the lamina propria, muscularis mucosae, muscularis or serosa. The accumulation of PepT1 mRNA along the crypt-villus axis was also investigated. In all regions of the small intestine (in duodenum, jejunum and ileum), PepT1 mRNA was undetectable in deeper epithelial cells of the crypts. Expression was first detectable at or near the crypt-villus junction, the amount of PepT1 mRNA increasing rapidly in the lower villus to a maximum approximately 100-200 microns from this point. Along the length of the small intestine PepT1 mRNA was most abundant in duodenal and jejunal enterocytes, with lower levels in the ileal epithelium. PepT1 expression is greatly depressed in the follicle-associated epithelium of the Peyer's patch relative to both interfollicular and adjacent "normal" villi. These data are discussed in the context of the known physiological role of PepT1 in the gastrointestinal tract.
为了检测最近克隆的H⁺/二肽 - 三肽转运体(PepT1)的肠道表达情况,合成了寡核苷酸探针,并通过兔空肠RNA的Northern印迹分析证实了其特异性。使用这些探针进行的原位杂交研究表明,PepT1在整个小肠均有表达,而在结肠中的表达水平则大幅降低。相比之下,在胃、圆小囊或盲肠中未检测到PepT1 mRNA。组织切片的显微镜检查显示,PepT1的表达仅限于肠上皮,在固有层、黏膜肌层、肌层或浆膜中未检测到表达。还研究了PepT1 mRNA沿隐窝 - 绒毛轴的积累情况。在小肠的所有区域(十二指肠、空肠和回肠),隐窝较深的上皮细胞中未检测到PepT1 mRNA。在隐窝 - 绒毛交界处或其附近首次可检测到表达,PepT1 mRNA的量在绒毛下部迅速增加,从该点起约100 - 200微米处达到最大值。在小肠全长中,PepT1 mRNA在十二指肠和空肠肠上皮细胞中最为丰富,在回肠上皮中的水平较低。相对于滤泡间和相邻的“正常”绒毛,PepT1在派尔集合淋巴结的滤泡相关上皮中的表达大大降低。将结合PepT1在胃肠道中已知的生理作用来讨论这些数据。