Ionic currents of cultured olfactory receptor neurons from antennae of male manduca sexta

Frank Zufall, Monika Stengl, Christian Franke, John G Hildebrand, Hanns Hatt

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Whole-cell and single-channel voltage-clamp techniques were used to identify and characterize the ionic currents of insect olfactory receptor neurons (ORNs) in vitro. The cells were isolated from the antennae of male Manduca sexta pupae at stages 3-5 of adult development and maintained in primary cell culture. After 2-3 weeks in vitro, the presumptive ORNs had resting potentials of -62 ± 12 mV (n = 18) and expressed at least 1 type of Na+ channel and at least 3 types of K+ channels. Na+ currents, recorded in the whole-cell mode, were reversibly blocked by 0.1 μM tetrodotoxin. The predominant type of K+ channel observed was a voltage-activated K+ channel (γ = 30 pS) with characteristics similar to those of the delayed rectifier. The activity of the 30-pS K+ channel could be inhibited by the application of nucleotides to the cytoplasmic face of inside-out patches of membrane. The nucleotides had relative potencies as follows: ATP > cGMP > cAMP, with an inhibition constant for ATP of K1 = 0.18 mM. Raising the intracellular Ca2+ concentration from 0.1 to 5 μM induced the opening of a Ca2+- activated K+ channel (γ = 66 pS at 0 mV) that had a low voltage sensitivity. A third, transient type of K+ channel (γ = 12-18 pS) could be activated by depolarizing voltage steps from very negative resting potentials. Properties of this channel were similar to those of the "A-channel." These results support the conclusion that M. sexta ORNs differentiate in vitro and provide the basis for studying primary mechanisms of olfactory transduction.

Original languageEnglish (US)
Pages (from-to)956-965
Number of pages10
JournalJournal of Neuroscience
Volume11
Issue number4
StatePublished - 1991

Fingerprint

Olfactory Receptor Neurons
Manduca
Membrane Potentials
Nucleotides
Adenosine Triphosphate
Calcium-Activated Potassium Channels
Pupa
Primary Cell Culture
Tetrodotoxin
Patch-Clamp Techniques
Insects
Membranes
In Vitro Techniques

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Ionic currents of cultured olfactory receptor neurons from antennae of male manduca sexta. / Zufall, Frank; Stengl, Monika; Franke, Christian; Hildebrand, John G; Hatt, Hanns.

In: Journal of Neuroscience, Vol. 11, No. 4, 1991, p. 956-965.

Research output: Contribution to journalArticle

Zufall, Frank ; Stengl, Monika ; Franke, Christian ; Hildebrand, John G ; Hatt, Hanns. / Ionic currents of cultured olfactory receptor neurons from antennae of male manduca sexta. In: Journal of Neuroscience. 1991 ; Vol. 11, No. 4. pp. 956-965.
@article{f22f8587af2549848160b2f9a73aa767,
title = "Ionic currents of cultured olfactory receptor neurons from antennae of male manduca sexta",
abstract = "Whole-cell and single-channel voltage-clamp techniques were used to identify and characterize the ionic currents of insect olfactory receptor neurons (ORNs) in vitro. The cells were isolated from the antennae of male Manduca sexta pupae at stages 3-5 of adult development and maintained in primary cell culture. After 2-3 weeks in vitro, the presumptive ORNs had resting potentials of -62 ± 12 mV (n = 18) and expressed at least 1 type of Na+ channel and at least 3 types of K+ channels. Na+ currents, recorded in the whole-cell mode, were reversibly blocked by 0.1 μM tetrodotoxin. The predominant type of K+ channel observed was a voltage-activated K+ channel (γ = 30 pS) with characteristics similar to those of the delayed rectifier. The activity of the 30-pS K+ channel could be inhibited by the application of nucleotides to the cytoplasmic face of inside-out patches of membrane. The nucleotides had relative potencies as follows: ATP > cGMP > cAMP, with an inhibition constant for ATP of K1 = 0.18 mM. Raising the intracellular Ca2+ concentration from 0.1 to 5 μM induced the opening of a Ca2+- activated K+ channel (γ = 66 pS at 0 mV) that had a low voltage sensitivity. A third, transient type of K+ channel (γ = 12-18 pS) could be activated by depolarizing voltage steps from very negative resting potentials. Properties of this channel were similar to those of the {"}A-channel.{"} These results support the conclusion that M. sexta ORNs differentiate in vitro and provide the basis for studying primary mechanisms of olfactory transduction.",
author = "Frank Zufall and Monika Stengl and Christian Franke and Hildebrand, {John G} and Hanns Hatt",
year = "1991",
language = "English (US)",
volume = "11",
pages = "956--965",
journal = "Journal of Neuroscience",
issn = "0270-6474",
publisher = "Society for Neuroscience",
number = "4",

}

TY - JOUR

T1 - Ionic currents of cultured olfactory receptor neurons from antennae of male manduca sexta

AU - Zufall, Frank

AU - Stengl, Monika

AU - Franke, Christian

AU - Hildebrand, John G

AU - Hatt, Hanns

PY - 1991

Y1 - 1991

N2 - Whole-cell and single-channel voltage-clamp techniques were used to identify and characterize the ionic currents of insect olfactory receptor neurons (ORNs) in vitro. The cells were isolated from the antennae of male Manduca sexta pupae at stages 3-5 of adult development and maintained in primary cell culture. After 2-3 weeks in vitro, the presumptive ORNs had resting potentials of -62 ± 12 mV (n = 18) and expressed at least 1 type of Na+ channel and at least 3 types of K+ channels. Na+ currents, recorded in the whole-cell mode, were reversibly blocked by 0.1 μM tetrodotoxin. The predominant type of K+ channel observed was a voltage-activated K+ channel (γ = 30 pS) with characteristics similar to those of the delayed rectifier. The activity of the 30-pS K+ channel could be inhibited by the application of nucleotides to the cytoplasmic face of inside-out patches of membrane. The nucleotides had relative potencies as follows: ATP > cGMP > cAMP, with an inhibition constant for ATP of K1 = 0.18 mM. Raising the intracellular Ca2+ concentration from 0.1 to 5 μM induced the opening of a Ca2+- activated K+ channel (γ = 66 pS at 0 mV) that had a low voltage sensitivity. A third, transient type of K+ channel (γ = 12-18 pS) could be activated by depolarizing voltage steps from very negative resting potentials. Properties of this channel were similar to those of the "A-channel." These results support the conclusion that M. sexta ORNs differentiate in vitro and provide the basis for studying primary mechanisms of olfactory transduction.

AB - Whole-cell and single-channel voltage-clamp techniques were used to identify and characterize the ionic currents of insect olfactory receptor neurons (ORNs) in vitro. The cells were isolated from the antennae of male Manduca sexta pupae at stages 3-5 of adult development and maintained in primary cell culture. After 2-3 weeks in vitro, the presumptive ORNs had resting potentials of -62 ± 12 mV (n = 18) and expressed at least 1 type of Na+ channel and at least 3 types of K+ channels. Na+ currents, recorded in the whole-cell mode, were reversibly blocked by 0.1 μM tetrodotoxin. The predominant type of K+ channel observed was a voltage-activated K+ channel (γ = 30 pS) with characteristics similar to those of the delayed rectifier. The activity of the 30-pS K+ channel could be inhibited by the application of nucleotides to the cytoplasmic face of inside-out patches of membrane. The nucleotides had relative potencies as follows: ATP > cGMP > cAMP, with an inhibition constant for ATP of K1 = 0.18 mM. Raising the intracellular Ca2+ concentration from 0.1 to 5 μM induced the opening of a Ca2+- activated K+ channel (γ = 66 pS at 0 mV) that had a low voltage sensitivity. A third, transient type of K+ channel (γ = 12-18 pS) could be activated by depolarizing voltage steps from very negative resting potentials. Properties of this channel were similar to those of the "A-channel." These results support the conclusion that M. sexta ORNs differentiate in vitro and provide the basis for studying primary mechanisms of olfactory transduction.

UR - http://www.scopus.com/inward/record.url?scp=0025995411&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025995411&partnerID=8YFLogxK

M3 - Article

C2 - 1849175

AN - SCOPUS:0025995411

VL - 11

SP - 956

EP - 965

JO - Journal of Neuroscience

JF - Journal of Neuroscience

SN - 0270-6474

IS - 4

ER -