Bossu J L, Dupont J L, Feltz A
Laboratoire d'Etude des Régulations Physiologiques, l'Université Louis Pasteur, Strasbourg, France.
Neuroscience. 1989;30(3):605-17. doi: 10.1016/0306-4522(89)90155-3.
Calcium permeabilities were examined in large cerebellar neurons maintained in culture, and morphologically identified as Purkinje cells. When cells were supplied with a Dulbecco Minimum Eagle's Medium with 10% horse serum added (5-10 days), somatic recordings revealed complex spikes and these were shown to be generated by Na and Ca components, the Na one being tetrodotoxin-sensitive. At the dendritic level, Ca currents were better resolved than at the soma. In dendrites, Ca entry was shown to occur through at least two distinct currents. The first was a low-threshold transient current (elicited above -60 mV from a holding potential of -80 mV) which was reduced by almost 30% by 50 microM cadmium. The second was a high-threshold current (above -20 mV) which gave rise to (1) a transient component exhibiting a steady-state inactivation and so requiring holding potentials at -80 mV, and (2) a sustained component. Both components were suppressed by 50 microns cadmium. We measured a total Ca current at the dendritic level reaching values of up to 1 nA. In another culture medium (Leibovitz medium) known to allow expression of three types of calcium currents in nodose cells we observed the development of the dendritic tree of Purkinje cells but with no simultaneous expression of the high-threshold Ca current.
对培养的大的小脑神经元进行钙通透性检测,这些神经元在形态上被鉴定为浦肯野细胞。当细胞用添加了10%马血清的杜氏改良伊格尔培养基培养(5 - 10天)时,体细胞记录显示有复合动作电位,且这些电位由钠和钙成分产生,其中钠成分对河豚毒素敏感。在树突水平,钙电流比在胞体处分辨得更好。在树突中,钙内流显示至少通过两种不同的电流发生。第一种是低阈值瞬态电流(从 - 80 mV的钳制电位在 - 60 mV以上激发),50 microM镉可使其降低近30%。第二种是高阈值电流(在 - 20 mV以上),它产生(1)一个表现出稳态失活的瞬态成分,因此需要 - 80 mV的钳制电位,以及(2)一个持续成分。两种成分都被50微米镉抑制。我们在树突水平测量到总钙电流值高达1 nA。在另一种已知能使结节细胞表达三种钙电流的培养基(莱博维茨培养基)中,我们观察到浦肯野细胞树突的发育,但没有同时表达高阈值钙电流。