Department of Biological Sciences, University of Manitoba, Winnipeg, MB, Canada.
Comp Biochem Physiol A Mol Integr Physiol. 2012 Jan;161(1):27-35. doi: 10.1016/j.cbpa.2011.08.017. Epub 2011 Sep 3.
The present study has examined the role of the colon in regulating ammonia and urea nitrogen balance in two species of chondrichthyans, the ratfish, Hydrolagus colliei (a holocephalan) and the spiny dogfish, Squalus acanthias (an elasmobranch). Stripped colonic tissue from both the dogfish and ratfish was mounted in an Ussing chamber and in both species bi-directional urea flux was found to be negligible. Urea uptake by the mucosa and serosa of the isolated colonic epithelium through accumulation of (14)C-urea was determined to be 2.8 and 6.2 fold greater in the mucosa of the dogfish compared to the serosa of the dogfish and the mucosa of the ratfish respectively. Furthermore, there was no difference between serosal and mucosal accumulation of (14)C-urea in the ratfish. Through the addition of 2mM NH(4)Cl to the mucosal side of each preparation the potential for ammonia flux was also examined. This was again found to be negligible in both species suggesting that the colon is an extremely tight epithelium to the movement of both urea and ammonia. Plasma, chyme and bile fluid samples were also taken from the agastric ratfish and were compared with solute concentrations of equivalent body fluids in the dogfish. Finally molecular analysis revealed expression of 3 isoforms of the urea transport protein (UT) and an ammonia transport protein (Rhbg) in the gill, intestine, kidney and colon of the ratfish. Partial nucleotide sequences of the UT-1, 2 and 3 isoforms in the ratfish had 95, 95 and 92% identity to the equivalent UT isoforms recently identified in another holocephalan, the elephantfish, Callorhinchus milii. Finally, the nucleotide sequence of the Rhbg identified in the ratfish had 73% identity to the Rhbg protein recently identified in the little skate, Leucoraja erinacea.
本研究考察了结肠在调节两种软骨鱼类(圆口鱼,Hydrolagus colliei(全头鱼)和尖齿鲨,Squalus acanthias(软骨鱼))氨和尿素氮平衡中的作用。从鲨鱼和鼠鱼的剥离结肠组织都被安装在 Ussing 室中,在这两种物种中,双向尿素通量都被发现可以忽略不计。通过积累(14)C-尿素,确定分离的结肠上皮黏膜和浆膜对(14)C-尿素的摄取分别比鲨鱼的浆膜和鼠鱼的黏膜高 2.8 和 6.2 倍。此外,在鼠鱼中,浆膜和黏膜对(14)C-尿素的积累没有差异。通过向每个制剂的黏膜侧添加 2mM NH(4)Cl,还检查了氨通量的潜力。这在两种物种中都被发现可以忽略不计,这表明结肠是一种对尿素和氨运动极其紧密的上皮组织。还从无胃鼠鱼中采集了血浆、食糜和胆汁液样本,并将其与鲨鱼中相当的体液的溶质浓度进行了比较。最后,分子分析表明,在鼠鱼的鳃、肠、肾和结肠中表达了 3 种尿素转运蛋白(UT)和 1 种氨转运蛋白(Rhbg)同工型。鼠鱼的 UT-1、2 和 3 同工型的部分核苷酸序列与最近在另一种全头鱼,象鼻鱼 Callorhinchus milii 中鉴定的等效 UT 同工型具有 95、95 和 92%的同一性。最后,在鼠鱼中鉴定的 Rhbg 的核苷酸序列与最近在小鲨鱼,Leucoraja erinacea 中鉴定的 Rhbg 蛋白具有 73%的同一性。