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雏鸡降钙素分泌细胞中的电压门控钠电流和钾电流及其变化

Voltage-gated sodium and potassium currents and their variation in calcitonin-secreting cells of the chick.

作者信息

Kawa K

机构信息

Department of Pharmacology, Gunma University School of Medicine, Maebashi, Japan.

出版信息

J Physiol. 1988 May;399:93-113. doi: 10.1113/jphysiol.1988.sp017070.

Abstract
  1. The electrical properties of dissociated ultimobranchial cells from chick embryos (18-20 days after fertilization) were studied using whole-cell patch electrodes. Antibodies for immunohistological identification of calcitonin-secreting cells in the preparation were obtained by immunizing rabbits with a conjugated analogue of eel calcitonin. 2. In a proportion of cells, spike-like action potentials were generated in response to depolarization when cells were immersed in standard saline containing 140 mM-Na+ but no Ca2+. When the membrane potential was shifted from a holding potential (-83 - -103 mV) to a test depolarization (-50 mV or more positive) under voltage-clamp conditions, a transient inward current was produced which was followed by a slowly developing outward current. 3. The inward current was identified as a Na+-carried current, since (1) the kinetics of the current seemed fast and the amplitude consistently depended on the holding potential, (2) replacement of external Na+ with choline ions reversibly abolished the current, and (3) external application of tetrodotoxin (1 microM) abolished the current completely. The cells in which inward currents were detected showed intense to intermediate degrees of staining with anti-calcitonin antibodies. 4. In some other cells, no regenerative potentials were evoked even with intense depolarization, but a delayed decrease in membrane depolarization during the current pulse was observed. Voltage-clamp experiments in these cells revealed the existence of slowly developing outward currents, and the cells showed an intermediate degree of antibody staining. 5. The outward currents in both types of cells were selectively diminished in the presence of K+ channel blockers such as tetraethylammonium (1-10 mM) or 4-aminopyridine (1 mM). When the pipette contained 120 mM-CsCl, none of the dissociated cells exhibited any appreciable outward currents. Thus, the outward currents were most likely to be membrane potential-dependent K+ currents. The potential dependency of activation and inactivation of the currents were consistent with those of delayed K+ rectifier. 6. In the remaining cells, only passive responses of membrane potentials were observed with current injection. No discernible voltage-dependent inward or outward currents were detected under voltage-clamp conditions. Although these cells had a similar appearance to the two types of cells previously mentioned under phase-contrast microscopy, none of them showed significant antibody staining. These cells were presumed to represent non-secretory or supporting cells within the gland.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 使用全细胞膜片电极研究了来自鸡胚(受精后18 - 20天)的分离鳃后体细胞的电特性。通过用鳗鱼降钙素的偶联类似物免疫兔子,获得了用于免疫组织化学鉴定制剂中降钙素分泌细胞的抗体。2. 在一定比例的细胞中,当细胞浸入含有140 mM - Na⁺但无Ca²⁺的标准盐溶液中时,去极化会产生尖峰状动作电位。当在电压钳制条件下将膜电位从保持电位(-83 - -103 mV)转变为测试去极化(-50 mV或更正)时,会产生一个短暂的内向电流,随后是一个缓慢发展的外向电流。3. 内向电流被确定为Na⁺携带的电流,因为(1)电流动力学似乎很快,且幅度始终取决于保持电位;(2)用胆碱离子替代外部Na⁺可使电流可逆地消失;(3)外部施加河豚毒素(1 μM)可使电流完全消失。检测到内向电流的细胞用抗降钙素抗体染色呈强至中度。4. 在其他一些细胞中,即使强烈去极化也不会诱发再生电位,但在电流脉冲期间观察到膜去极化延迟下降。这些细胞的电压钳实验显示存在缓慢发展的外向电流,且细胞用抗体染色呈中度。5. 在存在钾通道阻滞剂如四乙铵(1 - 10 mM)或4 - 氨基吡啶(1 mM)的情况下,两种类型细胞中的外向电流均选择性降低。当移液管中含有120 mM - CsCl时,分离的细胞均未表现出任何明显的外向电流。因此,外向电流很可能是膜电位依赖性钾电流。电流激活和失活的电位依赖性与延迟钾整流器的一致。6. 在其余细胞中,电流注入时仅观察到膜电位的被动反应。在电压钳制条件下未检测到可辨别的电压依赖性内向或外向电流。尽管这些细胞在相差显微镜下与上述两种类型的细胞外观相似,但它们均未显示出明显的抗体染色。这些细胞被推测代表腺体内的非分泌或支持细胞。(摘要截短至400字)
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/987f/1191654/d6eae743fb57/jphysiol00510-0107-a.jpg

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