ABSTRACT
BACKGROUND: Coronavirus disease 2019 (COVID-19) is spreading fast all around the world with more than fourteen millions of detected infected cases and more than 600.000 deaths by 20th July 2020. While scientist are working to find a vaccine, current epidemiological data shows that the most common comorbidities for patients with the worst prognosis, hypertension and diabetes, are often treated with angiotensin converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs). BODY: Both ACE inhibitors and ARBs induce overexpression of the angiotensin converting enzyme 2 (ACE-2) receptor, which has been identified as the main receptor used by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to enter into the alveolar cells of the lungs. While cannabinoids are known to reduce hypertension, the studies testing the hypotensive effects of cannabinoids never addressed their effects on ACE-2 receptors. However, some studies have linked the endocannabinoid system (ECS) with the renin angiotensin system (RAS), including a cross-modulation between the cannabinoid receptor 1 (CB1) and angiotensin II levels. CONCLUSION: Since there are around 192 million people using cannabis worldwide, we believe that the mechanism underlying the hypotensive properties of cannabinoids should be urgently studied to understand if they can also lead to ACE-2 overexpression as other antihypertensive drugs do.
ABSTRACT
Gene therapy in human disease has expanded rapidly in recent years with the development of safer and more effective viral vectors, and presents a novel approach to the treatment of epilepsy. Studies in animals models have demonstrated that overexpression of inhibitory peptides can modify seizure threshold, prevent the development of epilepsy, and modify established epilepsy. More recently there has been a flurry of studies using optogenetics in which light-activated channels expressed in neurons can transiently change neuronal excitability on exposure to light, thereby enabling the development of closed loop systems to detect and stop seizure activity. The treatment of status epilepticus presents its own challenges. Because of both the delay in gene expression following transfection and also the necessity of using focal transfection, there are a limited number of situations in which gene therapy can be used in status epilepticus. One such condition is epilepsia partialis continua (EPC). We have used gene therapy in a model of EPC and have shown that we can "cure" the condition. Recent evidence suggesting that gene therapy targeting subcortical regions can modify generalized or more diffuse epilepsies, indicates that the range of situations in status epilepticus in which gene therapy could be used will expand.
Subject(s)
Genetic Therapy , Status Epilepticus/therapy , Animals , Brain/physiopathology , Disease Models, Animal , Genetic Therapy/methods , Humans , Neurons/physiology , Status Epilepticus/geneticsABSTRACT
Neocortical epilepsy is frequently drug-resistant. Surgery to remove the epileptogenic zone is only feasible in a minority of cases, leaving many patients without an effective treatment. We report the potential efficacy of gene therapy in focal neocortical epilepsy using a rodent model in which epilepsy is induced by tetanus toxin injection in the motor cortex. By applying several complementary methods that use continuous wireless electroencephalographic monitoring to quantify epileptic activity, we observed increases in high frequency activity and in the occurrence of epileptiform events. Pyramidal neurons in the epileptic focus showed enhanced intrinsic excitability consistent with seizure generation. Optogenetic inhibition of a subset of principal neurons transduced with halorhodopsin targeted to the epileptic focus by lentiviral delivery was sufficient to attenuate electroencephalographic seizures. Local lentiviral overexpression of the potassium channel Kv1.1 reduced the intrinsic excitability of transduced pyramidal neurons. Coinjection of this Kv1.1 lentivirus with tetanus toxin fully prevented the occurrence of electroencephalographic seizures. Finally, administration of the Kv1.1 lentivirus to an established epileptic focus progressively suppressed epileptic activity over several weeks without detectable behavioral side effects. Thus, gene therapy in a rodent model can be used to suppress seizures acutely, prevent their occurrence after an epileptogenic stimulus, and successfully treat established focal epilepsy.
Subject(s)
Epilepsies, Partial/genetics , Epilepsies, Partial/therapy , Genetic Therapy , Kv1.1 Potassium Channel/genetics , Kv1.1 Potassium Channel/therapeutic use , Neocortex/pathology , Optogenetics , Animals , Disease Models, Animal , Electroencephalography , Epilepsies, Partial/pathology , Epilepsies, Partial/physiopathology , Lentivirus/genetics , Male , Neocortex/metabolism , Neocortex/physiopathology , Neurons/pathology , Pyramidal Cells/pathology , Rats , Rats, Sprague-Dawley , Tetanus Toxin/administration & dosageABSTRACT
Heterozygous mutations of KCNA1, the gene encoding potassium channel Kv1.1 subunits, cause episodic ataxia type 1 (EA1), which is characterized by paroxysmal cerebellar incoordination and interictal myokymia. Some mutations are also associated with epilepsy. Although Kv1.1-containing potassium channels play important roles in neuronal excitability and neurotransmitter release, it is not known how mutations associated with different clinical features affect the input-output relationships of individual neurons. We transduced rat hippocampal neurons, which were cultured on glial micro-islands, with lentiviruses expressing wild-type or mutant human KCNA1, and injected either depolarizing currents to evoke action potentials or depolarizing voltage commands to evoke autaptic currents. alpha-Dendrotoxin and tetraethylammonium allowed a pharmacological dissection of potassium currents underlying excitability and neurotransmission. Overexpression of wild-type Kv1.1 decreased both neuronal excitability and neurotransmitter release. By contrast, the C-terminus-truncated R417stop mutant, which is associated with severe drug-resistant EA1, had the opposite effect: increased excitability and release probability. Another mutant, T226R, which is associated with EA1 that is complicated by contractures and epilepsy, had no detectable effect on neuronal excitability; however, in common with R417stop, it markedly enhanced neurotransmitter release. The results provide direct evidence that EA1 mutations increase neurotransmitter release, and provide an insight into mechanisms underlying the phenotypic differences that are associated with different mutations.
Subject(s)
Ataxia/genetics , Mutation , Neurons/metabolism , Animals , Animals, Newborn , Ataxia/physiopathology , Disease Models, Animal , Heterozygote , Hippocampus/metabolism , Humans , Kv1.1 Potassium Channel/genetics , Lentivirus/genetics , Neurotransmitter Agents/metabolism , Promoter Regions, Genetic , Protein Structure, Tertiary , RatsABSTRACT
Motivated by its success as a therapeutic treatment in other neurological disorders, most notably Parkinson's disease, Deep Brain Stimulation (DBS) is currently being trialled in a number of patients with drug unresponsive epilepsies. However, the mechanisms by which DBS interferes with neuronal activity linked to the disorder are not well understood. Furthermore, there is a need to identify optimized values of parameters (for example in amplitude/frequency space) of the stimulation protocol with which one aims to achieve the desired outcome. In this paper we characterise the system response to stimulation, to gain an understanding of the role different brain regions play in generating the output observed in EEG. We perform a number of experiments in healthy rats, where the ventral-lateral thalamic nucleus is stimulated using a train of square-waves with different frequency and amplitudes. The response to stimulation in the motor cortex is recorded and the drive-response relationship over frequency/amplitude space is considered. Subsequently, we compare the experimental data with simulations of a mean-field model, finding good agreement between the output of the model and the experimental data--both in the time and frequency domains--when considering a transition to oscillatory activity in the cortex as the frequency of stimulation is increased. Overall, our study suggests that mean-field models can appropriately characterise the stimulus-response relationship of DBS in healthy animals. In this way, it constitutes a first step towards the goal of developing a closed-loop feedback control protocol for suppressing epileptic activity, by adaptively adjusting the stimulation protocol in response to EEG activity.
Subject(s)
Cerebral Cortex/physiology , Deep Brain Stimulation/methods , Models, Neurological , Ventral Thalamic Nuclei/physiology , Afferent Pathways/physiology , Animals , Biophysics , Electric Stimulation/methods , Electroencephalography , Evoked Potentials/physiology , Male , Mathematical Computing , Models, Animal , Rats , Rats, Sprague-Dawley , Spectrum AnalysisABSTRACT
Long-term potentiation (LTP), which approximates Hebb's postulate of associative learning, typically requires depolarization-dependent glutamate receptors of the NMDA (N-methyl-D-aspartate) subtype. However, in some neurons, LTP depends instead on calcium-permeable AMPA-type receptors. This is paradoxical because intracellular polyamines block such receptors during depolarization. We report that LTP at synapses on hippocampal interneurons mediating feedback inhibition is "anti-Hebbian":Itis induced by presynaptic activity but prevented by postsynaptic depolarization. Anti-Hebbian LTP may occur in interneurons that are silent during periods of intense pyramidal cell firing, such as sharp waves, and lead to their altered activation during theta activity.
Subject(s)
Hippocampus/physiology , Interneurons/physiology , Long-Term Potentiation , Neural Inhibition/physiology , Pyramidal Cells/physiology , Synapses/physiology , Animals , Electric Stimulation , Excitatory Postsynaptic Potentials , Hippocampus/cytology , Male , Membrane Potentials , Patch-Clamp Techniques , Rats , Rats, Sprague-Dawley , Receptors, AMPA/physiology , Receptors, N-Methyl-D-Aspartate/physiology , Spermine/analogs & derivatives , Spermine/pharmacologyABSTRACT
GABA(A) receptors can mediate both phasic (synaptic) and tonic (extrasynaptic) forms of inhibition. It has been proposed that tonic inhibition plays a critical part in controlling neuronal and network excitability. Although tonic GABA(A) receptor-mediated currents have been well characterized in rodents, their existence in human tissue has yet to be demonstrated. Here we show that tonic currents can be recorded from human tissue obtained from patients undergoing temporal lobectomies. Tonic GABA(A) receptor-mediated currents were present in pyramidal cells and interneurons in layer V-VI of temporal neocortex and granule cells in the dentate gyrus. These tonic currents have cell type-specific pharmacologies, opening up the possibility of targeted therapeutics.
Subject(s)
Cerebral Cortex/metabolism , Dentate Gyrus/metabolism , Membrane Potentials , Receptors, GABA-A/metabolism , Animals , Anti-Anxiety Agents/pharmacology , Cerebral Cortex/cytology , Dentate Gyrus/cytology , Desoxycorticosterone/analogs & derivatives , Desoxycorticosterone/pharmacology , Electrophysiology , GABA Antagonists/pharmacology , Humans , Interneurons/cytology , Interneurons/metabolism , Neurons/cytology , Neurons/drug effects , Neurons/metabolism , Picrotoxin/pharmacology , Pyridazines/pharmacologyABSTRACT
Cortical information processing requires a delicate balance of excitatory and inhibitory signaling. How is this balance preserved during hippocampal memory encoding, which involves NMDA receptor-dependent long term potentiation (LTP)? This form of LTP occurs at synapses between pyramidal neurons but has not been detected in feed-forward inhibitory interneurons. We show that paired pre- and postsynaptic activity evokes pathway-specific LTP in half of rat stratum radiatum interneurons if cytoplasmic integrity is preserved. LTP occurs in aspiny feed-forward interneurons and propagates to pyramidal neurons as an enhancement of disynaptic inhibition. We also show that when LTP is restricted to synapses on pyramidal neurons, the temporal fidelity of synaptic integration and action potential generation in pyramidal cells is compromised. However, when LTP also occurs at synapses on feed-forward interneurons, temporal fidelity is preserved. We propose that Hebbian LTP at synapses driving disynaptic inhibition is necessary to maintain information processing without degradation during memory encoding.
Subject(s)
Discrimination, Psychological/physiology , Hippocampus/physiology , Interneurons/physiology , Long-Term Potentiation/physiology , Memory/physiology , Neural Inhibition/physiology , Animals , Cytoplasm/physiology , Electrophysiology , In Vitro Techniques , Male , Presynaptic Terminals , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/physiology , Synapses , Time FactorsABSTRACT
The stability of neuronal networks is thought to depend on synaptic transmission which provides activity-dependent maintenance signals for both synapses and neurons. Here, we tested the relationship between presynaptic secretion and neuronal maintenance using munc18-1-null mutant mice as a model. These mutants have a specific defect in secretion from synaptic and large dense-cored vesicles [Verhage et al. (2000), Science, 287, 864-869; Voets et al. (2001), Neuron, 31, 581-591]. Neuronal networks in these mutants develop normally up to synapse formation but eventually degenerate. The proposed relationship between secretion and neuronal maintenance was tested in low-density and organotypic cultures and, in vivo, by conditional cell-specific inactivation of the munc18-1 gene. Dissociated munc18-1-deficient neurons died within 4 days in vitro (DIV). Application of trophic factors, insulin or BDNF delayed degeneration up to 7 DIV. In organotypic cultures, munc18-1-deficient neurons survived until 9 DIV. On glial feeders, these neurons survived up to 10 DIV and 14 DIV when insulin was applied. Co-culturing dissociated mutant neurons with wild-type neurons did not prolong survival beyond 4 DIV, but coculturing mutant slices with wild-type slices prolonged survival up to 19 DIV. Cell-specific deletion of munc18-1 expression in cerebellar Purkinje cells in vivo resulted in the specific loss of these neurons without affecting connected or surrounding neurons. Together, these data allow three conclusions. First, the lack of synaptic activity cannot explain the degeneration in munc18-1-null mutants. Second, trophic support delays but cannot prevent degeneration. Third, a cell-intrinsic yet unknown function of munc18-1 is essential for prolonged survival.