Blockade of ion channel expression in xenopus oocytes with complementary DNA probes to Na+ and K+ channel mRNAs

I. Lotan; A. Volterra; P. Dash; S. A. Siegelbaum; P. Goelet

doi:10.1016/0896-6273(88)90153-5

Blockade of ion channel expression in xenopus oocytes with complementary DNA probes to Na⁺ and K⁺ channel mRNAs

I. Lotan, A. Volterra, P. Dash, S. A. Siegelbaum, P. Goelet

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

Ionic currents were recorded from Xenopus oocytes injected with RNA isolated from chick or mouse brain. Three currents were studied: a rapid tetrodotoxin-sensitive Na⁺ current (I_Na), an early outward K⁺ current sensitive to 4-aminopyridine (IA), and an inward current activated by the excitatory amino acid receptor agonist kainate. Oligonucleotides (60-80 bases long complementary to rat brain Na+ channel sequences were prehybridized to chick brain RNA. These DNA sequences, upon injection into oocytes, specifically inhibited expression of IN, relative to IA and the kainate-induced current in a dose-dependent manner. By contrast, prehybridization of oligonucleotides complementary to sequences either from the Drosophila Shaker locus (which codes for an early K⁺ current in Drosophila muscle) or from a homologous clone from mouse brain did not block the expression of the early outward K⁺ current induced in the oocytes by mRNA from chick or mouse brain. This method provides a convenient means for testing the functional role of cloned DNA species.

Original language	English (US)
Pages (from-to)	963-971
Number of pages	9
Journal	Neuron
Volume	1
Issue number	10
DOIs	https://doi.org/10.1016/0896-6273(88)90153-5
State	Published - Dec 1988
Externally published	Yes

ASJC Scopus subject areas

General Neuroscience

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10.1016/0896-6273(88)90153-5

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title = "Blockade of ion channel expression in xenopus oocytes with complementary DNA probes to Na+ and K+ channel mRNAs",

abstract = "Ionic currents were recorded from Xenopus oocytes injected with RNA isolated from chick or mouse brain. Three currents were studied: a rapid tetrodotoxin-sensitive Na+ current (INa), an early outward K+ current sensitive to 4-aminopyridine (IA), and an inward current activated by the excitatory amino acid receptor agonist kainate. Oligonucleotides (60-80 bases long complementary to rat brain Na+ channel sequences were prehybridized to chick brain RNA. These DNA sequences, upon injection into oocytes, specifically inhibited expression of IN, relative to IA and the kainate-induced current in a dose-dependent manner. By contrast, prehybridization of oligonucleotides complementary to sequences either from the Drosophila Shaker locus (which codes for an early K+ current in Drosophila muscle) or from a homologous clone from mouse brain did not block the expression of the early outward K+ current induced in the oocytes by mRNA from chick or mouse brain. This method provides a convenient means for testing the functional role of cloned DNA species.",

author = "I. Lotan and A. Volterra and P. Dash and Siegelbaum, {S. A.} and P. Goelet",

year = "1988",

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T1 - Blockade of ion channel expression in xenopus oocytes with complementary DNA probes to Na+ and K+ channel mRNAs

AU - Lotan, I.

AU - Volterra, A.

AU - Dash, P.

AU - Siegelbaum, S. A.

AU - Goelet, P.

PY - 1988/12

Y1 - 1988/12

N2 - Ionic currents were recorded from Xenopus oocytes injected with RNA isolated from chick or mouse brain. Three currents were studied: a rapid tetrodotoxin-sensitive Na+ current (INa), an early outward K+ current sensitive to 4-aminopyridine (IA), and an inward current activated by the excitatory amino acid receptor agonist kainate. Oligonucleotides (60-80 bases long complementary to rat brain Na+ channel sequences were prehybridized to chick brain RNA. These DNA sequences, upon injection into oocytes, specifically inhibited expression of IN, relative to IA and the kainate-induced current in a dose-dependent manner. By contrast, prehybridization of oligonucleotides complementary to sequences either from the Drosophila Shaker locus (which codes for an early K+ current in Drosophila muscle) or from a homologous clone from mouse brain did not block the expression of the early outward K+ current induced in the oocytes by mRNA from chick or mouse brain. This method provides a convenient means for testing the functional role of cloned DNA species.

AB - Ionic currents were recorded from Xenopus oocytes injected with RNA isolated from chick or mouse brain. Three currents were studied: a rapid tetrodotoxin-sensitive Na+ current (INa), an early outward K+ current sensitive to 4-aminopyridine (IA), and an inward current activated by the excitatory amino acid receptor agonist kainate. Oligonucleotides (60-80 bases long complementary to rat brain Na+ channel sequences were prehybridized to chick brain RNA. These DNA sequences, upon injection into oocytes, specifically inhibited expression of IN, relative to IA and the kainate-induced current in a dose-dependent manner. By contrast, prehybridization of oligonucleotides complementary to sequences either from the Drosophila Shaker locus (which codes for an early K+ current in Drosophila muscle) or from a homologous clone from mouse brain did not block the expression of the early outward K+ current induced in the oocytes by mRNA from chick or mouse brain. This method provides a convenient means for testing the functional role of cloned DNA species.

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Blockade of ion channel expression in xenopus oocytes with complementary DNA probes to Na⁺ and K⁺ channel mRNAs

Abstract

ASJC Scopus subject areas

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