Role Of L-type Calcium Channels Research Articles

Comparing tonic and phasic dendritic calcium in cholinergic pedunculopontine neurons and dopaminergic substantia nigra neurons.

Several brainstem nuclei degenerate in Parkinson's disease (PD). In addition to the well-characterized dopaminergic neurons of the substantia nigra pars compacta (SNc), the cholinergic neurons of the pedunculopontine nucleus (PPN) also degenerate in PD. One leading hypothesis of selective vulnerability is that pacemaking activity and the activation of low-threshold L-type calcium current are major contributors to tonic calcium load and cellular stress in SNc dopaminergic neurons. However, it is not yet clear whether the vulnerable PPN cholinergic neurons share this property. Therefore, we used two-photon dendritic calcium imaging and whole-cell electrophysiology to evaluate the role of L-type calcium channels in tonic and phasic dendritic calcium signals in PPN and SNc neurons. In addition, we investigated N- and P/Q-type calcium channel regulation of firing properties and dendritic calcium in PPN neurons. We found that blocking L-type channels reduces tonic firing rate and dendritic calcium levels in SNc neurons. By contrast, the tonic calcium load in PPN neurons did not depend on L-, N- or P/Q-type channels. However, we found that blocking either L-type (with nifedipine) or N- and P/Q-type (with omega-conotoxin MVIIC) channels reducesphasic calcium influx in PPN dendrites. Together, these findings show that L-type calcium channels play different roles in the activity of SNc and PPN neurons, and suggest that low-threshold L-type channels are not responsible for tonic calcium levels in PPN cholinergic neurons and are therefore not likely to be a source of selective vulnerability in these cells.

Role of L-Type Calcium Channels in the Increased Fatigue of Rat Soleus Muscle under Functional Unloading

Excessive long-term accumulation of calcium ions in the myoplasm of skeletal muscles can negatively affect mitochondria and lead to muscle dysfunction. The aim of our study was to identify the role of L-type calcium channels in the development of increased fatigue rat soleus muscle under functional unloading. Young male Wistar rats were divided into three groups of 8 animals each: the vivarium control group (C), the group subjected to hind limb unloading for 7 days (7HS) and the group with 7 days of hindlimb unloading with daily intraperitoneal injections of nifedipine (7 mg/kg body weight). The administration of nifidipine during hindlimb unloading prevented the upregulation of calcium-dependent phosphorylation of calcium-calmodulin kinase II (CaMK II), prevented the increase in fatigue and contributed to the preservation of mitochondrial proteins, DNA and mRNA expression of a number of genes that regulate mitochondrial biogenesis.

Role of L-Type Calcium Channels in Increased Fatigue of the Rat Soleus Muscle under Functional Unloading

Role of L-Type Calcium Channels in Increased Fatigue of the Rat Soleus Muscle under Functional Unloading

The inhibitory effect of escitalopram on mouse detrusor contractility: The role of L-type calcium channels

Selective serotonin reuptake inhibitors (SSRIs) are associated with urinary problems attributed to their central effects. ESC is a preferred SSRI and several case reports described that ESC is related to urinary retention. However, the direct effect of ESC on detrusor contractility is still not completely elucidated. Thus, we investigated the effect of ESC on detrusor contractility and mechanism(s) of its action in isolated mouse detrusor strips. Molecular docking and measurement of intracellular calcium were performed to determine the possible calcium channel blocking effect of ESC. The contractile responses to carbachol (CCh), KCl and electrical field stimulation of detrusor strips were significantly abolished by ESC (10 or 100 μM). ESC relaxed KCl-precontracted detrusor strips concentration-dependently, which was not affected by tetraethylammonium, glibenclamide, 4-aminopyridine, propranolol, L-NAME or methylene blue. ESC (10 or 100 μM) reduced both the CaCl2- and CCh-induced contractions under calcium-free conditions, indicating the role of calcium-involved mechanisms in ESC-mediated relaxation. Furthermore, ESC significantly decreased Bay K8644-induced contraction and the cytosolic calcium level in fura-2-loaded A7r5 cells. Molecular docking study also revealed the potential of ESC to bind L-type calcium (Cav1) channels. Our results demonstrate that ESC inhibits detrusor contractility via blocking Cav1 channels, which provides evidence for the direct effect of ESC on detrusor contractility and its mechanism.

Aging differentially affects LTCC function in hippocampal CA1 and piriform cortex pyramidal neurons.

Aging is associated with cognitive decline and memory loss in humans. In rats, aging-associated neuronal excitability changes and impairments in learning have been extensively studied in the hippocampus. Here, we investigated the roles of L-type calcium channels (LTCCs) in the rat piriform cortex (PC), in comparison with those of the hippocampus. We employed spatial and olfactory tasks that involve the hippocampus and PC. LTCC blocker nimodipine administration impaired spontaneous location recognition in adult rats (6-9months). However, the same blocker rescued the spatial learning deficiency in aged rats (19-23months). In an odor-associative learning task, infusions of nimodipine into either the PC or dorsal CA1 impaired the ability of adult rats to learn a positive odor association. Again, in contrast, nimodipine rescued odor associative learning in aged rats. Aged CA1 neurons had higher somatic expression of LTCC Cav1.2 subunits, exhibited larger afterhyperpolarization (AHP) and lower excitability compared with adult neurons. In contrast, PC neurons from aged rats showed higher excitability and no difference in AHP. Cav1.2 expression was similar in adult and aged PC somata, but relatively higher in PSD95- puncta in aged dendrites. Our data suggest unique features of aging-associated changes in LTCCs in the PC and hippocampus.

Activation of L-type calcium channels and attenuation of oxidative stress are involved in the improving effect of methyl jasmonate on learning and memory and its anxiolytic property in rats.

The present study was designed to evaluate the effect of plant bioactive compound methyl jasmonate on learning and memory, anxiety-like behaviors, and brain oxidative stress in rats. It has been indicated that methyl jasmonate stimulates calcium-binding protein expression and increases intracellular calcium (Ca2+). Therefore, we investigated the potential role of L-type calcium channel on methyl jasmonate effects. The animals were intracerebroventriculary (i.c.v.) injected with different doses of methyl jasmonate (0.5, 2.5, and 5 µg/rat). L-type calcium channel blocker (nifedipine 5 µg/rat, i.c.v.) was injected 30 min before methyl jasmonate (5 µg/rat). Shuttle box apparatus was used to evaluate passive avoidance memory. Anxiety-like behaviors were assessed by open field and elevated plus maze tests. Lastly, oxidative stress-related indices were assessed in hippocampus and prefrontal cortex. The data showed that methyl jasmonate dose-dependently could improve passive avoidance learning and memory and reduce anxiogenic behaviors. The methyl jasmonate effects were significantly prevented by nifedipine. Furthermore, central microinjection of methyl jasmonate significantly decreased hydrogen peroxide concentration, and increased reactive oxygen species scavenger activity (catalase and peroxide enzymes) in rats' hippocampus as well as prefrontal cortex. Indeed, the results indicated that the beneficial effects of methyl jasmonate on learning and memory and anxiety might be partly associated with L-type calcium channel and partly on the inhibition of oxidant indices.

The role of L-type calcium channels in neuronal excitability and aging

Over the last two decades there has been significant progress towards understanding the neural substrates that underlie age-related cognitive decline. Although many of the exact molecular and cellular mechanisms have yet to be fully understood, there is consensus that alterations in neuronal calcium homeostasis contribute to age-related deficits in learning and memory. Furthermore, it is thought that the age-related changes in calcium homeostasis are driven, at least in part, by changes in calcium channel expression. In this review, we focus on the role of a specific class of calcium channels: L-type voltage-gated calcium channels (LVGCCs). We provide the reader with a general introduction to voltage-gated calcium channels, followed by a more detailed description of LVGCCs and how they serve to regulate neuronal excitability via the post burst afterhyperpolarization (AHP). We conclude by reviewing studies that link the slow component of the AHP to learning and memory, and discuss how age-related increases in LVGCC expression may underlie cognitive decline by mediating a decrease in neuronal excitability.

L-type calcium channel contributions to intrinsic excitability and synaptic activity during basolateral amygdala postnatal development.

The amygdala contributes toward emotional processes such as fear, anxiety, and social cognition. Furthermore, evidence suggests that increased excitability of basolateral amygdala (BLA) principal neurons underlie certain neuropsychiatric disorders. Gain-of-function mutations in neuronal L-type calcium channels (LTCCs) are linked to neurodevelopmental diseases, including autism spectrum disorders (ASDs). While LTCCs are expressed throughout the BLA, direct evidence for increased LTCC activity affecting BLA excitability and potentially contributing to disease pathophysiology is lacking. In this study, we utilized a pharmacological approach to examine the contributions of LTCCs to BLA principal cell excitability and synaptic activity at immature (postnatal day 7, P7) and juvenile (P21) developmental stages. Acute upregulation of LTCC activity in brain slices by application of the agonist (S)-Bay K 8644 resulted in increased intrinsic excitability properties including firing frequency response, plateau potential, and spike-frequency adaptation selectively in P7 neurons. Contrastingly, for P21 neurons, the main effect of (S)-Bay K 8644 was to enhance burst firing. (S)-Bay K 8644 increased spontaneous inhibitory synaptic currents at both P7 and P21 but did not affect evoked synaptic currents at either stage. (S)-Bay K 8644 did not alter P7 spontaneous excitatory synaptic currents, although it increased current amplitude in P21 neurons. Overall, the results provide support for the notion that alteration of LTCC activity at specific periods of early brain development may lead to functional alterations to neuronal network activity and subsequently contribute to underlying mechanisms of amygdala-related neurological disorders.NEW & NOTEWORTHY The role of L-type calcium channels (LTCCs) in regulating neuronal electrophysiological properties during development remains unclear. We show that in basolateral amygdala principal neurons, an increase of LTCC activity alters both neuronal excitability and synaptic activity. The results also provide evidence for the distinct contributions of LTCCs at different stages of neurodevelopment and shed insight into our understanding of LTCC dysfunction in amygdala-related neurological disorders.

The role of l-type calcium channels in the mediation of fast oscillation of arterial blood pressure and renal blood flow in rats

The investigation of dynamic characteristics of blood pressure and renal blood flow provides detailed information about the fast regulatory mechanisms involved in arterial blood pressure (ABP) and renal blood flow (RBF) autoregulation. The aim of our study was to investigate the role of L-type Ca2+ channels in the mediation of fast oscillations of arterial blood pressure and renal blood flow in rats by means of spectral analysis. The experiments were performed on anesthetized male Wistar rats (n=7) at the age of 12-14 weeks. The ABP was measured directly in femoral artery; RBF was registered by perivascular ultrasonic Doppler probe (Transonic system) in control and experimental period. The spectrograms from APB and RBF were derived in Lab View 3.11 software. In experimental period the selective L-type Ca2+ channel blocker amlodipine besylate (AML) in dose 200 mkg.kg-1 bolus followed by 50 mkg.kg-1.h infusions was applied. Our results by using a spectral method of analysis of ABP and RBF confirmed involvement of L-type Ca2+ channels in the mediation of dynamic nature of myogenic vascular response. The L-type Ca2+ channels in different specific manner participate in the regulation of renal blood flow and arterial blood pressure.

Cellular Proliferative Domains: Barriers to Migration

An important aspect of the analysis of the proliferation of cell populations as composed of a set of intercommunicating logistic domains is the nature of the barriers between adjacent domains and the regulation of cellular migration between them. The nature of these inter-domain boundaries is discussed in relation to the central role of L-type calcium channels in the control of cell motility and it is proposed that domain boundaries are determined by proteoglycans which act as calcium chelators and hence regulate the local extracellular Ca⁺⁺ concentration. The selective migratory advantage gained by overexpression of calcium channels is briefly alluded to.

The Role of L-type Calcium Channels in Olfactory Learning and Its Modulation by Norepinephrine

L type calcium channels (LTCCs) are prevalent in different systems and hold immense importance for maintaining/performing selective functions. In the nervous system, CaV1.2 and CaV1.3 are emerging as critical modulators of neuronal functions. Although the general role of these calcium channels in modulating synaptic plasticity and memory has been explored, their role in olfactory learning is not well understood. In this review article we first discuss the role of LTCCs in olfactory learning especially focusing on early odor preference learning in neonate rodents, presenting evidence that while NMDARs initiate stimulus-specific learning, LTCCs promote protein-synthesis dependent long-term memory (LTM). Norepinephrine (NE) release from the locus coeruleus (LC) is essential for early olfactory learning, thus noradrenergic modulation of LTCC function and its implication in olfactory learning is discussed here. We then address the differential roles of LTCCs in adult learning and learning in aged animals.

The role of L-type calcium channels in mouse oocyte maturation, activation and early embryonic development

Calcium ion fluctuation is closely related to the transformation of cell cycle. However, little is known about the function of L-type calcium channel in mammalian oocyte and embryo development. We thus studied the roles of L-type calcium channel in mouse oocyte meiotic maturation, parthenogenetic activation and early embryonic development. We used the antagonist Amlodipine to block L-type calcium channel. Oocytes or zygotes were cultured to different time points with 0 μM, 10 μM, 30 μM and 50 μM Amlodipine. Then we checked the rate of first polar body extrusion, spindle formation, asymmetric division parthenogenetic activation and early embryo cleavage. The results showed that Amlodipine treatment did not affect germinal vesicle breakdown, but caused disruption of cytoskeleton organization, symmetric division, formation of mature oocytes with a large polar body, or reduced the first polar body extrusion, depending on its concentrations. Amlodipine treatment also resulted in decreased parthenogenetic activation and arrested early embryonic development. Overall, these data suggest that proper function of L-type calcium channel is critical for oocyte maturation, activation, and early embryonic development.

Roles of L-type calcium channels (CaV1.2) and the distal C-terminus (DCT) in differentiation and mineralization of rat dental apical papilla stem cells (rSCAPs)

ObjectiveVoltage-gated inward Ca2+ currents (ICa) are triggered by cell depolarization and commonly produce transient increases in the cytoplasmic free Ca2+ concentration. The CaV1.2 distal C-terminus is susceptible to proteolytic cleavage, which yields a truncated CaV1.2 subunit and a cleaved C-terminal fragment (CCt or DCT). Stem cells from the apical papilla (SCAPs) has a capacity for differentiation into the odontoblastic-like cells in vitro and dentin forming in vivo, which makes SCAPs advantages in tissue engineering and regenerative endodontic. The aim of this study was to investigate the effect of CaV1.2 and its distal C-terminal fragment in the odontoblastic differentiation of rat SCAPs (stem cells from the apical papilla). DesignIn this study, we generated stable CaV1.2 knockdown and DCT over-expressed rSCAPs using short hairpin RNA and DCT gene containing Lentivirus vectors, respectively. The transfected apical papilla cells were induced to differentiate into the odontoblast-like cells, and the expression of markers for odontoblastic differentiation were analyzed by alizarin red staining, Real-time Polymerase chain reaction (RT-PCR), and Western blot analysis. ResultsThe knockdown of CaV1.2 and excess expression of DCT both suppressed the expression of DSPP, ALP in mRNA level and the formation of calcium nodules. ConclusionsOur results suggest that CaV1.2 and DCT play important roles in the differentiation of rSCAPs, DCT might act as a transcription factor and regulate the differentiation of rSCAPs.

Tumor-selective killing and TRAIL sensitization by garlic organosulfur compounds in human melanoma cells: Role of l-type calcium channel

Tumor-selective killing and TRAIL sensitization by garlic organosulfur compounds in human melanoma cells: Role of l-type calcium channel

Effects of iron overload condition on liver toxicity and hepcidin/ferroportin expression in thalassemic mice

AimsAlthough iron-overload conditions can be found in β-thalassemic patients, resulting in cellular damage, particularly in the liver, the mechanism for this iron-mediated hepatic injury specifically in β-thalassemic (HT) mice is unclear. This study aimed to investigate the roles of L-type calcium channels (LTCC), T-type calcium channels (TTCC) and divalent metal transporter1 (DMT1) in iron-mediated hepatic injury in HT mice. Main methodsIron chelator deferoxamine (DFO), LTCC blocker, TTCC blocker and DMT1 blocker were used to determine the roles of these channels regarding liver iron accumulation, apoptosis and iron regulatory protein expression in HT mice. Key findingsTTCC and DMT1 blockers and DFO decreased liver iron and malondialdehyde (MDA) in HT mice indicating their antioxidant effects, whereas LTCC blocker produced no decrease in liver iron or MDA. However, only DFO decreased liver apoptosis through the reduced Bax/Bcl-2 ratio in wild type (WT) mice. The levels of iron regulatory hormone hepcidin were markedly higher in HT mice even before iron loading while ferroportin levels did not alter. Each of the pharmacological interventions increased ferroportin protein back to normal levels only in WT while HT mice showed no difference. SignificanceThalassemic mice have different hepcidin/ferroportin and apoptotic protein expression as a defense mechanism to iron-overload compared with those in WT mice. DFO was the most effective intervention in preventing liver apoptosis under iron-overload conditions in WT but did not have the same effect in HT mice.

Pharmacology of L-type Calcium Channels: Novel Drugs for Old Targets?

Inhibition of voltage-gated L-type calcium channels by organic calcium channel blockers is a well-established pharmacodynamic concept for the treatment of hypertension and cardiac ischemia. Since decades these antihypertensives (such as the dihydropyridines amlodipine, felodipine or nifedipine) belong to the most widely prescribed drugs world-wide. Their tolerability is excellent because at therapeutic doses their pharmacological effects in humans are limited to the cardiovascular system. During the last years substantial progress has been made to reveal the physiological role of different L-type calcium channel isoforms in many other tissues, including the brain, endocrine and sensory cells. Moreover, there is accumulating evidence about their involvement in various human diseases, such as Parkinson's disease, neuropsychiatric disorders and hyperaldosteronism. In this review we discuss the pathogenetic role of L-type calcium channels, potential new indications for existing or isoform-selective compounds and strategies to minimize potential side effects.

The effects of vasoactive peptide urocortin 2 on hemodynamics in spontaneous hypertensive rat and the role of L-type calcium channel and CRFR2

BackgroundUrocortin (UCN) is a newly identified vascular-active peptide that has been shown to reverse cardiovascular remodeling and improve left ventricular (LV) function. The effects and mechanism of urocortin 2 (UCN2) in vivo on the electrical remodeling of left ventricle and the hemodynamics of hypertensive objectives have not been investigated. MethodsUCN2 (1μg/kg/d, 3.5μg/kg/d or 7μg/kg/d) was intravenously injected for 2 weeks and its effects on hemodynamics in spontaneously hypertensive rats (SHRs) observed. The whole-cell patch clamp technique was used to explore the effects of UCN2 on the electrical remodeling of left ventricular cardiomyocytes. The flow cytometry method was used to determine the content of fluorescence calcium in myocardium. ResultsUCN2 improved the systolic and diastolic function of SHRs as demonstrated by decreased left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), increased +dp/dtmax and −dp/dtmax and decreased cAMP level. UCN2 inhibited the opening of L-type calcium channel and decreased the calcium channel current of cardiomyocytes. In addition, UCN2 also decreased the contents of fluorescence calcium in SHR myocardium. However, astressin2-B (AST-2B), the antagonist of corticotropin-releasing factor receptor 2 (CRFR2), could reverse the inhibitory effects of UCN2 on calcium channel. ConclusionUCN2 can modulate electrical remodeling of the myocardium and hemodynamics in an experimental model of SHR via inhibition of L-type calcium channel and CRFR2 in cardiomyocytes.

The role of L-type voltage-gated calcium channels Cav1.2 and Cav1.3 in normal and pathological brain function.

The use of specific activators and inhibitors that penetrate the central nervous system has suggested an essential functional role of L-type calcium channels (LTCC) in several important physiological processes of the brain, including the modulation of the mesoaccumbal dopamine signalling pathway, synaptic transmission of auditory stimuli and synaptic plasticity of neutral and aversive learning and memory processes. However, the lack of selectivity of available pharmacological agents towards the most prominent LTCC isoforms in the brain, namely Cav1.2 and Cav1.3, has hampered the elucidation of the precise contribution made by each specific channel isoform within these specific physiological processes. Modern genetic approaches, both in rodents and in human, have recently enhanced our understanding of the selective functional roles of Cav1.2 and Cav1.3 channels. In rodents, the characterisation of global and conditional isoform-specific knockouts suggests a contribution of Cav1.2 channels in spatial memory formation, whereas Cav1.3 channels seem to be involved in the consolidation of fear memories and in neurodegenerative mechanisms associated with the development of Parkinson's disease. With regard to the molecular mechanisms underlying drug addiction, Cav1.3 channels are necessary for the development and Cav1.2 channels for the expression of cocaine and amphetamine behavioural sensitisation. In humans, both the identification of naturally occurring LTCC variants ("channelopathies") and unbiased genome-wide association studies have linked LTCCs to working memory performance in healthy individuals and schizophrenic patients. Individually, CACNA1C polymorphisms and CACNA1D variants have been linked to a variety of psychiatric diseases and to congenital deafness, respectively. However, the contribution of individual LTCCs and their polymorphisms to human brain function and diseases remains unclear, necessitating the use of isoform-specific pharmacological agents.

Anticonvulsant effects of bis-1,4-dihydropyridines and the probable role of L-type calcium channels suggested by docking simulations

The influx/efflux of calcium (Ca2+) ions through channels in cellular membranes plays a pivotal role in many physiological and physiopathological processes. Among these are those involved in the physiopathology of epileptic seizures. Hence, the control of permeability of ions through these channels is considered a strategy for the development of anticonvulsant drugs. According to the previous reports, dihydropyridine derivatives have proven to be Ca2+ channel blockers and have anticonvulsive properties, presumably by means of their action on L-type Ca2+ channels (LCCs). The aim of the present study was to determine the anticonvulsant effects of four bis-1,4-dihydropyridines (bis-DHPs) in male CF1 mice (25–30 g bw) using two experimental models: maximal electroshock (MES, which induces convulsions with an alternating current of 60 Hz) and pentylenetetrazole administration (PTZ, 90 mg/kg administered i.p.). Additionally, the binding mode was explored with a docking study on a 3-D model of an LCC. The four bis-DHPs herein tested showed a protective effect in relation to the number of convulsions induced by MES, the recovery time after a convulsion, and the duration of the tonic phase. However, only bis-DHPs 01–03 reduced the duration of the clonic phase, and these compounds also produced a significant protective effect against the convulsions induced by PTZ administration. This effect may be related to the interaction of bis-DHPs with a cluster of aromatic residues (Tyr1152, Tyr1463, and Phe1159) involved in blocking calcium flow, as suggested by docking studies.

The role of L-type calcium channels in the development and expression of behavioral sensitization to ethanol

Behavioral sensitization is thought to play a significant role in drug addiction. L-type calcium channels have been implicated in sensitization to stimulant and opiate drugs but it is unclear if these channels also contribute to sensitization to ethanol. The effects of three L-type calcium channel blockers, nifedipine (1–7.5mg/kg), diltiazem (12.5–50mg/kg), and verapamil (12.5 and 25mg/kg), on sensitization to ethanol (2g/kg) were examined in DBA/2J mice. All three blockers reduced but did not prevent expression of sensitization. Only nifedipine blocked acquisition of sensitization. Nifedipine and verapamil decreased blood ethanol levels. The current findings suggest L-type calcium channels do not play a substantial role in sensitization to ethanol and that the neural mechanisms underlying sensitization to ethanol are distinct from those mediating sensitization to stimulants and opiates.