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Induction of voltage-dependent sodium channels by in vitro differentiation of human retinoblastoma cells.

Abstract：

:1. Neuronlike differentiation of Y-79 retinoblastoma was chemically induced in vitro, by plating the cells onto a poly-D-lysine and laminin substrate. The changes in voltage-dependent conductances after 48-72 h were examined with the whole-cell tight-seal and the perforated-patch recording techniques. 2. Although outward currents carried by potassium ions appeared qualitatively similar before and after differentiation, the depolarization-activated transient inward current displayed a pronounced acceleration of its activation and inactivation kinetics. 3. After differentiation, both the threshold of activation and the steady-state inactivation curve of the inward current are displaced in the negative direction by approximately 10-20 mV as compared with untreated cells. The current attains its peak amplitude in approximately 1 ms at maximum activating voltages, and decays within 3 ms. In contrast, in undifferentiated cells these values are on the order of 6 and 60 ms, respectively. The time to recover from inactivation is also shortened 20-fold in differentiated cells. 4. Unlike the mixed conductance of undifferentiated cells, which requires extracellular calcium, the inward current of the neuronlike differentiated cells is insensitive to manipulations of external calcium. Instead, it can be completely abolished in a reversible way by sodium removal or by micromolar concentrations of tetrodotoxin (TTX) in the bathing solution. As such, it resembles in all salient respects the voltage-dependent sodium conductance of nerve cells. 5. The fast sodium current expressed after neuronal differentiation is not the result of a progressive enhancement of an existing conductance, because no such component is discernible in undifferentiated cells. Moreover, recordings performed in cells at early stages of differentiation also failed to reveal the coexistence of the immature and the differentiated inward currents. 6. A possible account of the present observations is that the native inward current of undifferentiated Y-79 cells may correspond to a precursor form of the mature channel, and the observed developmental changes induced by chemical differentiation could be a consequence of progressive modification of the original channels, rather than expression of a separate class of proteins.

journal_name

J Neurophysiol

journal_title

Journal of neurophysiology

authors

del Pilar Gomez M,Waloga G,Nasi E

doi

10.1152/jn.1993.70.4.1487

subject

Has Abstract

pub_date

1993-10-01 00:00:00

pages

1487-96

issue

eissn

0022-3077

issn

1522-1598

journal_volume

pub_type

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Induction of voltage-dependent sodium channels by in vitro differentiation of human retinoblastoma cells.