Sui G P, Wu C, Fry C H
Institute of Urology and Nephrology, London, United Kingdom.
J Urol. 2004 Feb;171(2 Pt 1):938-43. doi: 10.1097/01.ju.0000108120.28291.eb.
We measured the membrane electrical characteristics as well as the response to adenosine triphosphate of cells isolated from the suburothelial layer of the bladder.
Suburothelial cells were isolated from biopsy samples of human bladder by collagenase disruption. Electrophysiological measurements were done under current and voltage clamp to record membrane potential and ionic currents using patch pipettes with a K+ based filling solution. Intracellular [Ca2+] was measured with Fura-2.
Cells were different from epithelial cells by their spindle-shaped appearance with projections at either end. The cells stained for vimentin but epithelial and smooth muscle cells did not. The cells had small membrane capacitance (27 +/- 16 pF) and a specific membrane resistance of 90 +/- 48 x 10(9) Omega cm2. Average membrane potential was -63 +/- 14 mV but cells showed spontaneous spikes or random fluctuations of membrane potential. A small net inward current was superimposed by a larger outward current. Inward current was attenuated by the removal of extracellular Ca. Outward current showed large spontaneous fluctuations and was greatly decreased by 30 mM tetraethyl ammonium chloride. Adenosine triphosphate (30 to 100 microM) elicited an inward current of about 50 pA and large intracellular Ca2+ transients.
These cells are electrically active which, in conjunction with the previous observation of connexin 43 labeling, suggests that they could act as an electrical network. A quantitative model of voltage distribution in such a network after the generation of inward current suggests that individual cells could not act as pacemakers, but rather a group of simultaneously activated cells could exert a peripheral excitatory effect that would amplify the magnitude of the original response. The implications of this in terms of bladder sensation are discussed.
我们测量了从膀胱黏膜下层分离出的细胞的膜电特性以及对三磷酸腺苷的反应。
通过胶原酶消化从人膀胱活检样本中分离出黏膜下层细胞。使用基于K⁺的填充溶液的膜片吸管,在电流钳和电压钳下进行电生理测量,以记录膜电位和离子电流。用Fura - 2测量细胞内[Ca²⁺]。
这些细胞与上皮细胞不同,呈纺锤形,两端有突起。这些细胞波形蛋白染色阳性,而上皮细胞和平滑肌细胞则不着色。这些细胞具有小的膜电容(27±16 pF)和90±48×10⁹Ω·cm²的比膜电阻。平均膜电位为 - 63±14 mV,但细胞表现出自发的尖峰或膜电位的随机波动。一个小的内向净电流叠加在一个较大的外向电流上。去除细胞外Ca可使内向电流减弱。外向电流表现出大的自发波动,并且在30 mM四乙铵氯化物作用下大大降低。三磷酸腺苷(30至100 μM)引起约50 pA的内向电流和大的细胞内Ca²⁺瞬变。
这些细胞具有电活性,结合先前连接蛋白43标记的观察结果,表明它们可能构成一个电网络。内向电流产生后这种网络中电压分布的定量模型表明,单个细胞不能作为起搏器,而是一组同时被激活的细胞可发挥外周兴奋作用,放大原始反应幅度。讨论了其在膀胱感觉方面的意义。