Neurological phenotype and synaptic function in mice lacking the CaV1.3 α subunit of neuronal L-type voltage-dependent Ca2+ channels

doi:10.1016/S0306-4522(03)00329-4

Neuroscience

Volume 120, Issue 2, 22 August 2003, Pages 435-442

https://doi.org/10.1016/S0306-4522(03)00329-4 Get rights and content

Abstract

Neuronal L-type calcium channels have been implicated in pain perception and neuronal synaptic plasticity. To investigate this we have examined the effect of disrupting the gene encoding the Ca_V1.3 (α1D) α subunit of L-type Ca²⁺ channels on neurological function, acute nociceptive behavior, and hippocampal synaptic function in mice. Ca_V1.3 α1 subunit knockout (Ca_V1.3α1^−/−) mice had relatively normal neurological function with the exception of reduced auditory evoked behavioral responses and lower body weight. Baseline thermal and mechanical thresholds were unaltered in these animals. Ca_V1.3α1^−/− mice were also examined for differences in N-methyl-d-aspartate (NMDA) receptor-dependent (100 Hz tetanization for 1 s) and NMDA receptor-independent (200 Hz in 100 μM DL-2-amino-5-phosphopentanoic acid) long-term potentiation within the CA1 region of the hippocampus. Both NMDA receptor-dependent and NMDA receptor-independent forms of long-term potentiation were expressed normally. Radioligand binding studies revealed that the density of (+)[³H]isradipine binding sites in brain homogenates was reduced by 20–25% in Ca_V1.3α1^−/− mice, without any detectable change in Ca_V1.2 (α1C) protein levels as detected using Western blot analysis.

Taken together these data indicate that following loss of Ca_V1.3α1 subunit expression there is sufficient residual activity of other Ca²⁺ channel subtypes to support NMDA receptor-independent long-term potentiation and some forms of sensory behavior/function.

Section snippets

Mice

All experiments were performed using male, age-matched Ca_v1.3α1^−/− and WT mice (Platzer et al., 2000). Animals were backcrossed for five generations into a C57Bl/6N genetic background. All experiments conformed to ethical guidelines for investigation of experimental pain in conscious animals (Zimmermann, 1983) and were carried out in accordance with the UK Animals (Scientific Procedures) Act 1986 and its associated guidelines. All efforts were made to minimize the number of animals and their

Histological and neurological evaluation

Detailed histological analysis of brains by light microscopy from 14 Ca_V1.3α1^−/− mice (for details see Experimental Procedures section) revealed that Ca_V1.3α1 deficiency did not lead to detectable changes in brain structure, or to a detectable loss of neurones in defined brain regions, such as hippocampus CA1 region, dentate gyrus, primary visual cortex VA1, and cerebellar cortex. The only exception was the degeneration of the cochlear inner hair cells and the auditory nerve. However, we found

Discussion

This study tested the hypotheses that Ca_V1.3α1-mediated L-type Ca²⁺ currents may be involved in the perception of acute pain, and in synaptic plasticity within the CA1 region of the hippocampus, using a strain of mice that were deficient in expression of Ca_V1.3α1 subunits (Platzer et al., 2000). While Ca_V1.3α1^−/− mice had significantly lower body weight, and did not respond to auditory stimuli, there were no overt differences in acute pain perception or hippocampal synaptic plasticity, compared

Acknowledgements

This work was supported in part by the European Commission UPRM-CT-2000-00082 and the Austrian Science Fund P-14820.

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Effective analgesic treatment for neuropathic pain remains an unmet need, so previous evidence that epidermal growth factor receptor inhibitors (EGFRIs) provide unexpected rapid pain relief in a clinical setting points to a novel therapeutic opportunity. The present study utilises rodent models to address the cellular and molecular basis for the findings, focusing on primary sensory neurons because clinical pain relief is provided not only by small molecule EGFRIs, but also by the anti-EGFR antibodies cetuximab and panitumumab, which are unlikely to access the central nervous system in therapeutic concentrations. We report robust, rapid and dose-dependent analgesic effects of EGFRIs in two neuropathic pain models, matched by evidence with highly selective antibodies that expression of the EGFR (ErbB1 protein) is limited to small nociceptive afferent neurons. As other ErbB family members can heterodimerise with ErbB1, we investigated their distribution, showing consistent co-expression of ErbB2 but not ErbB3 or ErbB4, with ErbB1 in cell bodies of nociceptors, as well as providing evidence for direct molecular interaction of ErbB1 with ErbB2 in situ. Co-administration of selective ErbB1 and ErbB2 inhibitors produced clear evidence of greater-than-additive, synergistic analgesia; highlighting the prospect of a unique new combination therapy in which enhanced efficacy could be accompanied by minimisation of side-effects. Peripheral (intraplantar) administration of EGF elicited hypersensitivity only following nerve injury and this was reversed by local co-administration of selective inhibitors of either ErbB1 or ErbB2. Investigating how ErbB1 is activated in neuropathic pain, we found evidence for a role of Src tyrosine kinase, which can be activated by signals from inflammatory mediators, chemokines and cytokines during neuroinflammation. Considering downstream consequences of ErbB1 activation in neuropathic pain, we found direct recruitment to ErbB1 of an adapter for PI 3-kinase and Akt signalling together with clear Akt activation and robust analgesia from selective Akt inhibitors. The known Akt target and regulator of vesicular trafficking, AS160 was strongly phosphorylated at a perinuclear location during neuropathic pain in an ErbB1-, ErbB2- and Akt-dependent manner, corresponding to clustering and translocation of an AS160-partner, the vesicular chaperone, LRP1. Exploring whether neuronal ion channels that could contribute to hyperexcitability might be transported by this vesicular trafficking pathway we were able to identify Na_v1.9, (Na_v1.8) and Ca_v1.2 moving towards the plasma membrane or into proximal axonal locations – a process prevented by ErbB1 or Akt inhibitors. Overall these findings newly reveal both upstream and downstream signals to explain how ErbB1 can act as a signalling hub in neuropathic pain models and identify the trafficking of key ion channels to neuronal subcellular locations likely to contribute to hyperexcitability. The new concept of combined treatment with ErbB1 plus ErbB2 blockers is mechanistically validated as a promising strategy for the relief of neuropathic pain.
Calcium Signaling in Neurons and Glial Cells: Role of Cav1 channels
2019, Neuroscience
Calcium (Ca²⁺) is an essential component in intracellular signaling of brain cells, and its control mechanisms are of great interest in biological systems. Ca²⁺ can signal differently in neurons and glial cells using the same intracellular pathways or cell membrane structural components. These types of machinery are responsible for entry, permanence, and removal of Ca²⁺ from the cellular environment and are of vital importance for brain homeostasis. This review highlights the importance of Ca²⁺ in neuronal and glial cell physiology as well as aspects of learning, memory, and Alzheimer's disease, focusing on the involvement of L-type voltage-gated Ca²⁺ channels.
Ca <sup>2+</sup> -Binding Protein 1 Regulates Hippocampal-dependent Memory and Synaptic Plasticity
2018, Neuroscience
Citation Excerpt :
CaBP1 also prolongs Ca2+ currents mediated by Cav1.2 L-type channels (Zhou et al., 2004; Zhou et al., 2005; Tippens and Lee, 2007). Cav1.2 is thought to be the dominant L-type calcium channel in the brain (Clark et al., 2003), and mediates somatodendritic Ca2+ signals coupled to gene transcription (Ma et al., 2013), LTP and spatial memory (Moosmang et al., 2005), and neurogenesis (De Jesus-Cortes et al., 2016; Temme et al., 2016; Volkening et al., 2017). While presynaptic Cav2.1 Ca2+ signals would be potentiated in C-KO mice, postsynaptic Cav1 currents would be more transient.
Ca²⁺-binding protein 1 (CaBP1) is a Ca²⁺-sensing protein similar to calmodulin that potently regulates voltage-gated Ca²⁺ channels. Unlike calmodulin, however, CaBP1 is mainly expressed in neuronal cell-types and enriched in the hippocampus, where its function is unknown. Here, we investigated the role of CaBP1 in hippocampal-dependent behaviors using mice lacking expression of CaBP1 (C-KO). By western blot, the largest CaBP1 splice variant, caldendrin, was detected in hippocampal lysates from wild-type (WT) but not C-KO mice. Compared to WT mice, C-KO mice exhibited mild deficits in spatial learning and memory in both the Barnes maze and in Morris water maze reversal learning. In contextual but not cued fear-conditioning assays, C-KO mice showed greater freezing responses than WT mice. In addition, the number of adult-born neurons in the hippocampus of C-KO mice was ∼40% of that in WT mice, as measured by bromodeoxyuridine labeling. Moreover, hippocampal long-term potentiation was significantly reduced in C-KO mice. We conclude that CaBP1 is required for cellular mechanisms underlying optimal encoding of hippocampal-dependent spatial and fear-related memories.
The L-type calcium channel Cav1.3 is required for proper hippocampal neurogenesis and cognitive functions
2015, Cell Calcium
Citation Excerpt :
These mice are congenitally deaf, and they show resting bradycardia and arrhythmia, which disappear during physical activity [44]. However, apart from that, Cav1.3 knockout does not compromise motor functions, spontaneous activity, food/water intake or acute pain, and on a molecular level, does not lead to compensatory up-regulation of Cav1.2 mRNA and protein in the brain [2,68]. The observed impairments in neurogenesis and memory functions in Cav1.3−/− mice could also be partly mediated by Cav1.3 deletion in cells in other brain regions than in the hippocampus.
L-type voltage gated Ca²⁺ channels (LTCCs) are widely expressed within different brain regions including the hippocampus. The isoforms Cav1.2 and Cav1.3 have been shown to be involved in hippocampus-dependent learning and memory, cognitive functions that require proper hippocampal neurogenesis. In vitro, functional LTCCs are expressed on neuronal progenitor cells, where they promote neuronal differentiation. Expression of LTCCs on neural stem and progenitor cells within the neurogenic regions in the adult brain in vivo has not been examined so far, and a contribution of the individual isoforms Cav1.2 and Cav1.3 to adult neurogenesis remained to be clarified. To reveal the role of these channels we first evaluated the expression patterns of Cav1.2 and Cav1.3 in the hippocampal dentate gyrus and the subventricular zone (SVZ) in adult (2- and 3-month old) and middle-aged (15-month old) mice on mRNA and protein levels. We performed immunohistological analysis of hippocampal neurogenesis in adult and middle-aged Cav1.3^−/− mice and finally addressed the importance of Cav1.3 for hippocampal function by evaluating spatial memory and depression-like behavior in adult Cav1.3^−/− mice. Our results showed Cav1.2 and Cav1.3 expression at different stages of neuronal differentiation. While Cav1.2 was primarily restricted to mature NeuN⁺ granular neurons, Cav1.3 was expressed in Nestin⁺ neural stem cells and in mature NeuN⁺ granular neurons. Adult and middle-aged Cav1.3^−/− mice showed severe impairments in dentate gyrus neurogenesis, with significantly smaller dentate gyrus volume, reduced survival of newly generated cells, and reduced neuronal differentiation. Further, Cav1.3^−/− mice showed impairment in the hippocampus dependent object location memory test, implicating Cav1.3 as an essential element for hippocampus-associated cognitive functions. Thus, modulation of LTCC activities may have a crucial impact on neurogenic responses and cognition, which should be considered for future therapeutic administration of LTCCs modulators.
The role of metaplasticity mechanisms in regulating memory destabilization and reconsolidation
2012, Neuroscience and Biobehavioral Reviews
Citation Excerpt :
Together these could be mechanisms by which strong or old hippocampus-dependent memories might appear resistant to destabilization under some conditions, but we expect that many other mechanisms must also be involved. To reconcile this model with the evidence indicating that calcium influx should suppress cell firing via reduced sAHP (and other components of excitability), it is possible that sAHP is controlled by the sensitive CaV1.3 (Gamelli et al., 2011; Lima and Marrion, 2007), but the much more prevalent CaV1.2 (Clark et al., 2003; Hell et al., 1993) could be primarily responsible for mediating plasticity (i.e. Clark et al., 2003; Moosmang et al., 2005). Indeed, CaV1.2 has specifically been observed to form complexes with the activity-regulating AKAP, PKA, and β2-adrenergic proteins (Davare et al., 2001; Oliveria et al., 2007).
Memory allows organisms to predict future events based on prior experiences. This requires encoded information to persist once important predictors are extracted, while also being modifiable in response to changes within the environment. Memory reconsolidation may allow stored information to be modified in response to related experience. However, there are many boundary conditions beyond which reconsolidation may not occur. One interpretation of these findings is that the event triggering memory retrieval must contain new information about a familiar stimulus in order to induce reconsolidation. Presently, the mechanisms that affect the likelihood of reconsolidation occurring under these conditions are not well understood. Here we speculate on a number of systems that may play a role in protecting memory from being destabilized during retrieval. We conclude that few memories may enter a state in which they cannot be modified. Rather, metaplasticity mechanisms may serve to alter the specific reactivation cues necessary to destabilize a memory. This might imply that destabilization mechanisms can differ depending on learning conditions.
Enhanced long-term potentiation and impaired learning in mice lacking alternative exon 33 of Ca<inf>V</inf>1.2 calcium channel
2022, Translational Psychiatry

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¹: These authors contributed equally to this work.

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Neuroscience

Abstract

Section snippets

Mice

Histological and neurological evaluation

Discussion

Acknowledgements

References (23)

Nomenclature of voltage-gated Ca²⁺ channels

Neuron

Assay for Ca²⁺ channels

Methods Enzymol

Up-regulation of alpha1D Ca²⁺ channel subunit mRNA expression in the hippocampus of aged F344 rats

Neurobiol Aging

Molecular cloning of multiple subtypes of a novel rat brain isoform of the alpha 1 subunit of the voltage-dependent Ca²⁺ channel

Neuron

Down-regulation of L-type calcium channels in inflamed circular smooth muscle cells of the canine colon

Gastroenterology

Is mechanism-based pain treatment attainable? clinical trial issues

Pain

Will ion-channel blockers be useful for management of nonneuropathic pain?

Pain

Increased messenger RNA expression of the 695 and 751 amino acid isoforms of the beta-amyloid protein precursor in the thalamus of 17-year-old cynomolgus (Macaca fascicularis) monkeys

Neuroscience

Structure and functional expression of alpha 1, alpha 2, and beta subunits of a novel human neuronal Ca²⁺ channel subtype

Neuron

Ethical guidelines for investigations of experimental pain in conscious animals

Pain

Somatic colocalisation of rat SK1 and D class (Ca_V1.3) L-type Ca²⁺ channels in rat CA1 hippocampal pyramidal neurons

J Neurosci

Cited by (64)

ErbB1-dependent signalling and vesicular trafficking in primary afferent nociceptors associated with hypersensitivity in neuropathic pain

Calcium Signaling in Neurons and Glial Cells: Role of Cav1 channels

Ca <sup>2+</sup> -Binding Protein 1 Regulates Hippocampal-dependent Memory and Synaptic Plasticity

The L-type calcium channel Cav1.3 is required for proper hippocampal neurogenesis and cognitive functions

The role of metaplasticity mechanisms in regulating memory destabilization and reconsolidation

Enhanced long-term potentiation and impaired learning in mice lacking alternative exon 33 of Ca<inf>V</inf>1.2 calcium channel

Neurological phenotype and synaptic function in mice lacking the CaV1.3 α subunit of neuronal L-type voltage-dependent Ca2+ channels

Abstract

Section snippets

Mice

Histological and neurological evaluation

Discussion

Acknowledgements

Neuron

Methods Enzymol

Neurobiol Aging

Neuron

Gastroenterology

Pain

Pain

Neuroscience

Neuron

Pain

Somatic colocalisation of rat SK1 and D class (CaV1.3) L-type Ca2+ channels in rat CA1 hippocampal pyramidal neurons

J Neurosci

Neurological phenotype and synaptic function in mice lacking the Ca_V1.3 α subunit of neuronal L-type voltage-dependent Ca²⁺ channels

Somatic colocalisation of rat SK1 and D class (Ca_V1.3) L-type Ca²⁺ channels in rat CA1 hippocampal pyramidal neurons