Alves P, Soveral G, Macey R I, Moura T F
Dep. Química, Centro de Química Fina e Biotecnologia, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2825 Monte de Caparica, Portugal.
J Membr Biol. 1999 Sep 15;171(2):177-82. doi: 10.1007/s002329900569.
Brush border membrane vesicles, BBMV, from eel intestinal cells or kidney proximal tubule cells were prepared in a low osmolarity cellobiose buffer. The osmotic water permeability coefficient P(f) for eel vesicles was not affected by pCMBS and was measured at 1.6 x 10(-3) cm sec(-1) at 23 degrees C, a value lower than 3.6 x 10(-3) cm sec(-1) exhibited by the kidney vesicles and similar to published values for lipid bilayers. An activation energy E(a) of 14.7 Kcal mol(-1) for water transport was obtained for eel intestine, contrasting with 4.8 Kcal mol(-1) determined for rabbit kidney proximal tubule vesicles using the same method of analysis. The high value of E(a), as well as the low P(f) for the eel intestine is compatible with the absence of water channels in these membrane vesicles and is consistent with the view that water permeates by dissolution and diffusion in the membrane. Further, the initial transient observed in the osmotic response of kidney vesicles, which is presumed to reflect the inhibition of water channels by membrane stress, could not be observed in the eel intestinal vesicles. The P(f) dependence on the tonicity of the osmotic shock, described for kidney vesicles and related to the dissipation of pressure and stress at low tonicity shocks, was not seen with eel vesicles. These results indicate that the membranes from two volume transporter epithelia have different mechanisms of water permeation. Presumably the functional water channels observed in kidney vesicles are not present in eel intestine vesicles. The elastic modulus of the membrane was estimated by analysis of swelling kinetics of eel vesicles following hypotonic shock. The value obtained, 0.79 x 10(-3) N cm(-1), compares favorably with the corresponding value, 0. 87 x 10(-3) N cm(-1), estimated from measurements at osmotic equilibrium.
用低渗纤维二糖缓冲液制备了来自鳗鱼肠道细胞或肾近端小管细胞的刷状缘膜囊泡(BBMV)。鳗鱼囊泡的渗透水渗透系数P(f)不受对氯汞苯甲酸(pCMBS)影响,在23℃下测得为1.6×10⁻³ cm·sec⁻¹,该值低于肾囊泡显示的3.6×10⁻³ cm·sec⁻¹,与已发表的脂质双层的值相似。用相同分析方法测得鳗鱼肠道水转运的活化能E(a)为14.7千卡·mol⁻¹,相比之下,兔肾近端小管囊泡的该值为4.8千卡·mol⁻¹。鳗鱼肠道E(a)的高值以及低P(f)值与这些膜囊泡中不存在水通道相符,并且与水通过在膜中溶解和扩散进行渗透的观点一致。此外,在肾囊泡渗透反应中观察到的初始瞬变现象(据推测反映了膜应激对水通道的抑制)在鳗鱼肠道囊泡中未观察到。肾囊泡中描述的P(f)对渗透压休克张力的依赖性以及与低张力休克时压力和应激消散相关的现象,在鳗鱼囊泡中未见到。这些结果表明,来自两种容积转运上皮的膜具有不同的水渗透机制。推测在肾囊泡中观察到的功能性水通道在鳗鱼肠道囊泡中不存在。通过分析低渗休克后鳗鱼囊泡的肿胀动力学来估计膜的弹性模量。得到的值为0.79×10⁻³ N·cm⁻¹,与在渗透平衡时测量估计的相应值0.87×10⁻³ N·cm⁻¹相比很有利。