Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus

Voltage-dependent Ca2+ channels are a major pathway for Ca2+ entry in neurons. We have studied the electrophysiological, pharmacological, and molecular properties of voltage-gated Ca2+ channels in motoneurons of the rat facial nucleus in slices of the brainstem. Most facial motoneurons express both...

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Autores principales:	Plant, T.D., Schirra, C., Katz, E., Uchitel, O.D., Konnerth, A.
Formato:	JOUR
Materias:	α1-subunit ω- conotoxin ω-agatoxin-GVIA ω-agatoxin-IVA ω-conotoxin-MVIIC Calcium channel Calcium current Facial nucleus Motoneuron Single-cell RT- PCR calcium calcium channel dihydropyridine omega conotoxin alpha chain animal cell animal experiment animal tissue article channel gating facial nerve nucleus gene amplification membrane conductance membrane electrophysiology motoneuron muscle reinnervation nerve ending nerve fiber transection neurotransmitter release newborn nonhuman priority journal rat reverse transcription polymerase chain reaction Animals Animals, Newborn Brain Stem Cadmium Calcium Channel Blockers Calcium Channels Calcium Channels, L-Type Dihydropyridines DNA, Complementary Facial Nerve Ion Channel Gating Motor Neurons Nickel Nitrendipine omega-Agatoxin IVA omega-Conotoxin GVIA omega-Conotoxins Patch-Clamp Techniques Peptides Rats Rats, Wistar Receptors, Cholinergic Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Spider Venoms
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_02706474_v18_n23_p9573_Plant
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_02706474_v18_n23_p9573_Plant2023-10-03T15:14:33Z Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus Plant, T.D. Schirra, C. Katz, E. Uchitel, O.D. Konnerth, A. α1-subunit ω- conotoxin ω-agatoxin-GVIA ω-agatoxin-IVA ω-conotoxin-MVIIC Calcium channel Calcium current Facial nucleus Motoneuron Single-cell RT- PCR calcium calcium channel dihydropyridine omega conotoxin alpha chain animal cell animal experiment animal tissue article channel gating facial nerve nucleus gene amplification membrane conductance membrane electrophysiology motoneuron muscle reinnervation nerve ending nerve fiber transection neurotransmitter release newborn nonhuman priority journal rat reverse transcription polymerase chain reaction Animals Animals, Newborn Brain Stem Cadmium Calcium Channel Blockers Calcium Channels Calcium Channels, L-Type Dihydropyridines DNA, Complementary Facial Nerve Ion Channel Gating Motor Neurons Nickel Nitrendipine omega-Agatoxin IVA omega-Conotoxin GVIA omega-Conotoxins Patch-Clamp Techniques Peptides Rats Rats, Wistar Receptors, Cholinergic Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Spider Venoms Voltage-dependent Ca2+ channels are a major pathway for Ca2+ entry in neurons. We have studied the electrophysiological, pharmacological, and molecular properties of voltage-gated Ca2+ channels in motoneurons of the rat facial nucleus in slices of the brainstem. Most facial motoneurons express both low voltage-activated (LVA) and high voltage-activated (HVA) Ca2+ channel currents. The HVA current is composed of a number of pharmacologically separable components, including 30% of N-type and ~5% of L-type. Despite the dominating role of P-type Ca2+ channels in transmitter release at facial motoneuron terminals described in previous studies, these channels were not present in the cell body. Remarkably, most of the HVA current was carried through a new type of Ca2+ channel that is resistant to toxin and dihydropyridine block but distinct from the R-type currents described in other neurons. Using reverse transcription followed by PCR amplification (RT-PCR) with a powerful set of primers designed to amplify all HVA subtypes of the α1-subunit, we identified a highly heterogeneous expression pattern of Ca2+ channel α1-subunit mRNA in individual neurons consistent with the Ca2+ current components found in the cell bodies and axon terminals. We detected mRNA for α(1A) in 86% of neurons, α(1B) in 59%, α(1C) in 18%, (α(1D) in 18%, and α(1E) in 59%. Either α(1A) or α(1B) mRNAs (or both) were present in all neurons, together with various other α1-subunit mRNAs. The most frequently occurring combination was α(1A) with α(1B) and α(1E). Taken together, these results demonstrate that the Ca2+ channel pattern found in facial motoneurons is highly distinct from that found in other brainstem motoneurons. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_02706474_v18_n23_p9573_Plant
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	α1-subunit ω- conotoxin ω-agatoxin-GVIA ω-agatoxin-IVA ω-conotoxin-MVIIC Calcium channel Calcium current Facial nucleus Motoneuron Single-cell RT- PCR calcium calcium channel dihydropyridine omega conotoxin alpha chain animal cell animal experiment animal tissue article channel gating facial nerve nucleus gene amplification membrane conductance membrane electrophysiology motoneuron muscle reinnervation nerve ending nerve fiber transection neurotransmitter release newborn nonhuman priority journal rat reverse transcription polymerase chain reaction Animals Animals, Newborn Brain Stem Cadmium Calcium Channel Blockers Calcium Channels Calcium Channels, L-Type Dihydropyridines DNA, Complementary Facial Nerve Ion Channel Gating Motor Neurons Nickel Nitrendipine omega-Agatoxin IVA omega-Conotoxin GVIA omega-Conotoxins Patch-Clamp Techniques Peptides Rats Rats, Wistar Receptors, Cholinergic Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Spider Venoms
spellingShingle	α1-subunit ω- conotoxin ω-agatoxin-GVIA ω-agatoxin-IVA ω-conotoxin-MVIIC Calcium channel Calcium current Facial nucleus Motoneuron Single-cell RT- PCR calcium calcium channel dihydropyridine omega conotoxin alpha chain animal cell animal experiment animal tissue article channel gating facial nerve nucleus gene amplification membrane conductance membrane electrophysiology motoneuron muscle reinnervation nerve ending nerve fiber transection neurotransmitter release newborn nonhuman priority journal rat reverse transcription polymerase chain reaction Animals Animals, Newborn Brain Stem Cadmium Calcium Channel Blockers Calcium Channels Calcium Channels, L-Type Dihydropyridines DNA, Complementary Facial Nerve Ion Channel Gating Motor Neurons Nickel Nitrendipine omega-Agatoxin IVA omega-Conotoxin GVIA omega-Conotoxins Patch-Clamp Techniques Peptides Rats Rats, Wistar Receptors, Cholinergic Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Spider Venoms Plant, T.D. Schirra, C. Katz, E. Uchitel, O.D. Konnerth, A. Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
topic_facet	α1-subunit ω- conotoxin ω-agatoxin-GVIA ω-agatoxin-IVA ω-conotoxin-MVIIC Calcium channel Calcium current Facial nucleus Motoneuron Single-cell RT- PCR calcium calcium channel dihydropyridine omega conotoxin alpha chain animal cell animal experiment animal tissue article channel gating facial nerve nucleus gene amplification membrane conductance membrane electrophysiology motoneuron muscle reinnervation nerve ending nerve fiber transection neurotransmitter release newborn nonhuman priority journal rat reverse transcription polymerase chain reaction Animals Animals, Newborn Brain Stem Cadmium Calcium Channel Blockers Calcium Channels Calcium Channels, L-Type Dihydropyridines DNA, Complementary Facial Nerve Ion Channel Gating Motor Neurons Nickel Nitrendipine omega-Agatoxin IVA omega-Conotoxin GVIA omega-Conotoxins Patch-Clamp Techniques Peptides Rats Rats, Wistar Receptors, Cholinergic Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Spider Venoms
description	Voltage-dependent Ca2+ channels are a major pathway for Ca2+ entry in neurons. We have studied the electrophysiological, pharmacological, and molecular properties of voltage-gated Ca2+ channels in motoneurons of the rat facial nucleus in slices of the brainstem. Most facial motoneurons express both low voltage-activated (LVA) and high voltage-activated (HVA) Ca2+ channel currents. The HVA current is composed of a number of pharmacologically separable components, including 30% of N-type and ~5% of L-type. Despite the dominating role of P-type Ca2+ channels in transmitter release at facial motoneuron terminals described in previous studies, these channels were not present in the cell body. Remarkably, most of the HVA current was carried through a new type of Ca2+ channel that is resistant to toxin and dihydropyridine block but distinct from the R-type currents described in other neurons. Using reverse transcription followed by PCR amplification (RT-PCR) with a powerful set of primers designed to amplify all HVA subtypes of the α1-subunit, we identified a highly heterogeneous expression pattern of Ca2+ channel α1-subunit mRNA in individual neurons consistent with the Ca2+ current components found in the cell bodies and axon terminals. We detected mRNA for α(1A) in 86% of neurons, α(1B) in 59%, α(1C) in 18%, (α(1D) in 18%, and α(1E) in 59%. Either α(1A) or α(1B) mRNAs (or both) were present in all neurons, together with various other α1-subunit mRNAs. The most frequently occurring combination was α(1A) with α(1B) and α(1E). Taken together, these results demonstrate that the Ca2+ channel pattern found in facial motoneurons is highly distinct from that found in other brainstem motoneurons.
format	JOUR
author	Plant, T.D. Schirra, C. Katz, E. Uchitel, O.D. Konnerth, A.
author_facet	Plant, T.D. Schirra, C. Katz, E. Uchitel, O.D. Konnerth, A.
author_sort	Plant, T.D.
title	Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
title_short	Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
title_full	Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
title_fullStr	Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
title_full_unstemmed	Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus
title_sort	single-cell rt-pcr and functional characterization of ca2+ channels in motoneurons of the rat facial nucleus
url	http://hdl.handle.net/20.500.12110/paper_02706474_v18_n23_p9573_Plant
work_keys_str_mv	AT planttd singlecellrtpcrandfunctionalcharacterizationofca2channelsinmotoneuronsoftheratfacialnucleus AT schirrac singlecellrtpcrandfunctionalcharacterizationofca2channelsinmotoneuronsoftheratfacialnucleus AT katze singlecellrtpcrandfunctionalcharacterizationofca2channelsinmotoneuronsoftheratfacialnucleus AT uchitelod singlecellrtpcrandfunctionalcharacterizationofca2channelsinmotoneuronsoftheratfacialnucleus AT konnertha singlecellrtpcrandfunctionalcharacterizationofca2channelsinmotoneuronsoftheratfacialnucleus
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Single-cell RT-PCR and functional characterization of Ca2+ channels in motoneurons of the rat facial nucleus

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