Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice
Kelch-like 1 (KLHL1) is a neuronal actin-binding protein that modulates voltage-gated CaV2.1 (P/Q-type) and CaV3.2 (α1H T-type) calcium channels; KLHL1 knockdown experiments (KD) cause down-regulation of both channel types and altered synaptic properties in cultured rat hippocampal neurons (Perissin...
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doaj-1f4d51d0bbea480d847d13029f7caa382020-11-25T00:19:57ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022015-01-01810.3389/fncel.2014.00444124485Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout micePaula Patricia Perissinotti0Elizabeth G. Ethington1Erik eAlmazan2Elizabeth eMartínez-Hernández3Jennifer eKalil4Michael D. Koob5Erika S. Piedras-Renteria6Erika S. Piedras-Renteria7Loyola University Chicago, Stritch School of MedicineLoyola University Chicago, Stritch School of MedicineLoyola University Chicago, Stritch School of MedicineLoyola University Chicago, Stritch School of MedicineLoyola University Chicago, Stritch School of MedicineUniversity of MinnesotaLoyola University Chicago, Stritch School of MedicineLoyola University ChicagoKelch-like 1 (KLHL1) is a neuronal actin-binding protein that modulates voltage-gated CaV2.1 (P/Q-type) and CaV3.2 (α1H T-type) calcium channels; KLHL1 knockdown experiments (KD) cause down-regulation of both channel types and altered synaptic properties in cultured rat hippocampal neurons (Perissinotti et al., 2014). Here, we studied the effect of ablation of KLHL1 on calcium channel function and synaptic properties in cultured hippocampal neurons from KLHL1 knockout (KO) mice. Western blot data showed the P/Q-type channel α1A subunit was less abundant in KO hippocampus compared to wildtype (WT); and PQ-type calcium currents were smaller in KO neurons than WT during early days in vitro, although this decrease was compensated for at late stages by increases in L-type calcium current. In contrast, T-type currents did not change in culture. However, biophysical properties and western blot analysis revealed a differential contribution of T-type channel isoforms in the KO, with CaV3.2 α1H subunit being down-regulated and CaV3.1 α1G up-regulated. Synapsin I levels were reduced in the KO hippocampus; cultured neurons displayed a concomitant reduction in synapsin I puncta and decreased miniature excitatory postsynaptic current (mEPSC) frequency. In summary, genetic ablation of the calcium channel modulator resulted in compensatory mechanisms to maintain calcium current homeostasis in hippocampal KO neurons; however, synaptic alterations resulted in a reduction of excitatory synapse number, causing an imbalance of the excitatory-inhibitory synaptic input ratio favoring inhibition.http://journal.frontiersin.org/Journal/10.3389/fncel.2014.00444/fullactin-binding proteinsHVASCA8Synapsin IKelch-like 1 proteinT-type voltage-gated calcium channels |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Paula Patricia Perissinotti Elizabeth G. Ethington Erik eAlmazan Elizabeth eMartínez-Hernández Jennifer eKalil Michael D. Koob Erika S. Piedras-Renteria Erika S. Piedras-Renteria |
spellingShingle |
Paula Patricia Perissinotti Elizabeth G. Ethington Erik eAlmazan Elizabeth eMartínez-Hernández Jennifer eKalil Michael D. Koob Erika S. Piedras-Renteria Erika S. Piedras-Renteria Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice Frontiers in Cellular Neuroscience actin-binding proteins HVA SCA8 Synapsin I Kelch-like 1 protein T-type voltage-gated calcium channels |
author_facet |
Paula Patricia Perissinotti Elizabeth G. Ethington Erik eAlmazan Elizabeth eMartínez-Hernández Jennifer eKalil Michael D. Koob Erika S. Piedras-Renteria Erika S. Piedras-Renteria |
author_sort |
Paula Patricia Perissinotti |
title |
Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice |
title_short |
Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice |
title_full |
Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice |
title_fullStr |
Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice |
title_full_unstemmed |
Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice |
title_sort |
calcium current homeostasis and synaptic deficits in hippocampal neurons from kelch-like 1 knockout mice |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular Neuroscience |
issn |
1662-5102 |
publishDate |
2015-01-01 |
description |
Kelch-like 1 (KLHL1) is a neuronal actin-binding protein that modulates voltage-gated CaV2.1 (P/Q-type) and CaV3.2 (α1H T-type) calcium channels; KLHL1 knockdown experiments (KD) cause down-regulation of both channel types and altered synaptic properties in cultured rat hippocampal neurons (Perissinotti et al., 2014). Here, we studied the effect of ablation of KLHL1 on calcium channel function and synaptic properties in cultured hippocampal neurons from KLHL1 knockout (KO) mice. Western blot data showed the P/Q-type channel α1A subunit was less abundant in KO hippocampus compared to wildtype (WT); and PQ-type calcium currents were smaller in KO neurons than WT during early days in vitro, although this decrease was compensated for at late stages by increases in L-type calcium current. In contrast, T-type currents did not change in culture. However, biophysical properties and western blot analysis revealed a differential contribution of T-type channel isoforms in the KO, with CaV3.2 α1H subunit being down-regulated and CaV3.1 α1G up-regulated. Synapsin I levels were reduced in the KO hippocampus; cultured neurons displayed a concomitant reduction in synapsin I puncta and decreased miniature excitatory postsynaptic current (mEPSC) frequency. In summary, genetic ablation of the calcium channel modulator resulted in compensatory mechanisms to maintain calcium current homeostasis in hippocampal KO neurons; however, synaptic alterations resulted in a reduction of excitatory synapse number, causing an imbalance of the excitatory-inhibitory synaptic input ratio favoring inhibition. |
topic |
actin-binding proteins HVA SCA8 Synapsin I Kelch-like 1 protein T-type voltage-gated calcium channels |
url |
http://journal.frontiersin.org/Journal/10.3389/fncel.2014.00444/full |
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