Department of Biology, University of North Florida, Jacksonville, FL 32224, USA.
J Comp Physiol B. 2012 Feb;182(2):209-16. doi: 10.1007/s00360-011-0621-z. Epub 2011 Oct 9.
Purified epithelial brush border membrane vesicles (BBMV) were produced from the hepatopancreas of the Atlantic White shrimp, Litopeneaus setiferus, using standard methods originally developed for mammalian tissues and previously applied to other crustacean and echinoderm epithelia. These vesicles were used to study the cation dependency of sugar and amino acid transport across luminal membranes of hepatopancreatic epithelial cells. (3)H-D: -glucose uptake by BBMV against transient sugar concentration gradients occurred when either transmembrane sodium or potassium gradients were the only driving forces for sugar accumulation, suggesting the presence of a possible coupled transport system capable of using either cation. (3)H-L: -histidine transport was only stimulated by a transmembrane potassium gradient, while (3)H-L: -leucine uptake was enhanced by either a sodium or potassium gradient. These responses suggest the possible presence of a potassium-dependent transporter that accommodates either amino acid and a sodium-dependent system restricted only to L: -leucine. Uptake of (3)H-L: -leucine was significantly stimulated (P < 0.05) by several metallic cations (e.g., Zn(2+), Cu(2+), Mn(2+), Cd(2+), or Co(2+)) at external pH values of 7.0 or 5.0 (internal pH 7.0), suggesting a potential synergistic role of the cations in the transmembrane transfer of amino acids. (3)H-L: -histidine influxes (15 suptakes) were hyperbolic functions of external [zinc] or [manganese], following Michaelis-Menten kinetics. The apparent affinity constant (e.g., K (m)) for manganese was an order of magnitude smaller (K (m) = 0.22 μM Mn) than that for zinc (K (m) = 1.80 μM Zn), while no significant difference (P > 0.05) occurred between their maximal transport velocities (e.g., J (max)). These results suggest that a number of cation-dependent nutrient transport systems occur on the shrimp brush border membrane and aid in the absorption of these important dietary elements.
纯化的上皮刷状缘膜泡(BBMV)是使用最初为哺乳动物组织开发并以前应用于其他甲壳类动物和棘皮动物上皮的标准方法,从美洲白对虾(Litopeneaus setiferus)的肝胰腺中产生的。这些囊泡用于研究糖和氨基酸通过肝胰腺上皮细胞腔膜的阳离子依赖性转运。(3)H-D:-葡萄糖摄取通过 BBMV 发生在瞬态糖浓度梯度时,无论是跨膜钠或钾梯度是糖积累的唯一驱动力,表明存在一种可能的偶联转运系统,能够使用任何阳离子。(3)H-L:-组氨酸转运仅受跨膜钾梯度刺激,而(3)H-L:-亮氨酸摄取受钠或钾梯度增强。这些反应表明可能存在一种钾依赖性转运体,可容纳任何氨基酸和仅限于 L:-亮氨酸的钠依赖性系统。(3)H-L:-亮氨酸摄取在外部 pH 值为 7.0 或 5.0 时(内部 pH 值为 7.0)被几种金属阳离子(例如 Zn(2+),Cu(2+),Mn(2+),Cd(2+)或 Co(2+))显著刺激(P <0.05),表明阳离子在氨基酸的跨膜转运中可能具有协同作用。(3)H-L:-组氨酸内流(15 次摄取)是外部[锌]或[锰]的双曲线函数,遵循米氏动力学。锰的表观亲和力常数(例如 K(m))为 0.22 μM Mn)比锌(K(m)= 1.80 μM Zn)小一个数量级,而它们的最大转运速度(例如 J(max))之间没有显着差异(P> 0.05)。这些结果表明,虾刷状缘膜上存在许多阳离子依赖性营养转运系统,有助于吸收这些重要的膳食元素。