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新生大鼠单个视网膜神经节细胞中的钙通道

Calcium channels in solitary retinal ganglion cells from post-natal rat.

作者信息

Karschin A, Lipton S A

机构信息

Department of Neurology, Children's Hospital-G4, Boston, MA.

出版信息

J Physiol. 1989 Nov;418:379-96. doi: 10.1113/jphysiol.1989.sp017847.

Abstract
  1. Calcium currents from identified, post-natal retinal ganglion cell neurones from rat were studied with whole-cell and single-channel patch-clamp techniques. Na+ and K+ currents were suppressed with pharmacological agents, allowing isolation of current carried by either 10 mM-Ca2+ or Ba2- during whole-cell recordings. For cell-attached patch recordings, the recording pipette contained 96-110 mM-BaCl2 while the bath solution consisted of isotonic potassium aspartate in order to zero the neuronal membrane potential. 2. A transient component, present in approximately one-third of the whole-cell recordings resembles closely the T-type calcium current observed previously in other tissues. This component activates at low voltages (-40 to -50 mV from holding potentials negative to -80 mV), inactivates with a time constant of 10-30 ms at 35 degrees C, and is carried equally well by Ba2+ or Ca2+. In single-channel recordings small (8 pS) channels are observed whose aggregate microscopic kinetics correspond well to the macroscopic current obtained during whole-cell measurements. 3. During whole-cell recordings, a more prolonged component activates in all retinal ganglion cells at -40 to -20 mV from a holding potential of -90 mV. This component is substantially larger when equimolar Ba2+ replaces Ca2+ as the charge carrier, and is sensitive to the dihydropyridine agonist Bay K8644 (5 microM) and antagonists nifedipine (1-10 microM) and nimodipine (1-10 microM). Thus, the dihydropyridine pharmacology of this prolonged component resembles that of the L-type calcium current found in dorsal root ganglion neurones and in heart cells. Also reminiscent of the L-current, the prolonged component in this preparation is less inactivated at depolarized holding potentials (-60 to -40 mV) than the transient component. In cell-attached recordings, large (20 pS) channels are observed with activation properties similar to those of the prolonged portion of the whole-cell current. 4. omega-Conotoxin fraction GVIA (omega-CgTX VIA), a peptide from the venom of the snail Conus geographus, produces a readily reversible blockade of all components of the calcium current in these central mammalian neurones. This finding is in contrast to that of other preparations in which this toxin is responsible for an ephemeral block of T-current but a long-lasting block of other components of calcium current. 5. In summary, at least two components of calcium current with discrete underlying unitary events are present in post-natal retinal ganglion cells from rat. One component closely resembles the T or transient current observed in other cell types.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用全细胞膜片钳技术和单通道膜片钳技术,对出生后大鼠视网膜神经节细胞神经元的钙电流进行了研究。用药物抑制钠电流和钾电流,以便在全细胞记录过程中分离出由10 mM - Ca2+或Ba2+携带的电流。对于细胞贴附式膜片钳记录,记录微电极中含有96 - 110 mM - BaCl2,而浴液由等渗天冬氨酸钾组成,以使神经元膜电位归零。2. 在大约三分之一的全细胞记录中出现的一个瞬态成分,与先前在其他组织中观察到的T型钙电流非常相似。该成分在低电压下(相对于 - 80 mV的钳制电位为 - 40至 - 50 mV)激活,在35℃时以10 - 30 ms的时间常数失活,并且由Ba2+或Ca2+携带的情况相同。在单通道记录中,观察到小的(8 pS)通道,其总的微观动力学与全细胞测量中获得的宏观电流很好地对应。3. 在全细胞记录过程中,一个持续时间更长的成分在所有视网膜神经节细胞中,从 - 90 mV的钳制电位在 - 40至 - 20 mV时激活。当等摩尔的Ba2+替代Ca2+作为电荷载体时,该成分显著更大,并且对二氢吡啶激动剂Bay K8644(5 microM)以及拮抗剂硝苯地平(1 - 10 microM)和尼莫地平(1 - 10 microM)敏感。因此,这个持续时间更长的成分的二氢吡啶药理学与在背根神经节神经元和心脏细胞中发现的L型钙电流相似。同样让人联想到L电流的是,该制剂中的持续时间更长的成分在去极化钳制电位( - 60至 - 40 mV)下比瞬态成分失活程度更小。在细胞贴附式记录中,观察到具有与全细胞电流持续时间更长部分相似激活特性的大的(20 pS)通道。4. ω - 芋螺毒素GVIA(ω - CgTX VIA),一种来自芋螺毒液的肽,对这些中枢哺乳动物神经元的钙电流的所有成分产生易于逆转的阻断。这一发现与其他制剂的情况形成对比,在其他制剂中该毒素导致T电流的短暂阻断,但对钙电流的其他成分产生持久阻断。5. 总之,出生后大鼠视网膜神经节细胞中存在至少两种具有离散潜在单一事件的钙电流成分。一种成分与在其他细胞类型中观察到的T或瞬态电流非常相似。(摘要截短至400字)

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