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Proceedings of the National Academy of... Aug 2023Endothelial cells (ECs) line the lumen of all blood vessels and regulate functions, including contractility. Physiological stimuli, such as acetylcholine (ACh) and...
Endothelial cells (ECs) line the lumen of all blood vessels and regulate functions, including contractility. Physiological stimuli, such as acetylcholine (ACh) and intravascular flow, activate transient receptor potential vanilloid 4 (TRPV4) channels, which stimulate small (SK3)- and intermediate (IK)-conductance Ca-activated potassium channels in ECs to produce vasodilation. Whether physiological vasodilators also modulate the surface abundance of these ion channels in ECs to elicit functional responses is unclear. Here, we show that ACh and intravascular flow stimulate rapid anterograde trafficking of an intracellular pool of SK3 channels in ECs of resistance-size arteries, which increases surface SK3 protein more than two-fold. In contrast, ACh and flow do not alter the surface abundance of IK or TRPV4 channels. ACh triggers SK3 channel trafficking by activating TRPV4-mediated Ca influx, which stimulates Rab11A, a Rab GTPase associated with recycling endosomes. Superresolution microscopy data demonstrate that SK3 trafficking specifically increases the size of surface SK3 clusters which overlap with TRPV4 clusters. We also show that Rab11A-dependent trafficking of SK3 channels is an essential contributor to vasodilator-induced SK current activation in ECs and vasorelaxation. In summary, our data demonstrate that vasodilators activate Rab11A, which rapidly delivers an intracellular pool of SK3 channels to the vicinity of surface TRPV4 channels in ECs. This trafficking mechanism increases surface SK3 cluster size, elevates SK3 current density, and produces vasodilation. These data also demonstrate that SK3 and IK channels are differentially regulated by trafficking-dependent and -independent signaling mechanisms in endothelial cells.
Topics: Vasodilator Agents; TRPV Cation Channels; Endothelial Cells; Small-Conductance Calcium-Activated Potassium Channels; Arteries; Vasodilation; Acetylcholine; Endothelium, Vascular
PubMed: 37494394
DOI: 10.1073/pnas.2303238120 -
The Journal of Physiology Feb 2023The present study aimed to determine the isoform-specific role of the NADPH oxidases (NOX) in the endothelium-mediated vascular dysfunction associated with ageing....
The present study aimed to determine the isoform-specific role of the NADPH oxidases (NOX) in the endothelium-mediated vascular dysfunction associated with ageing. Endothelium-dependent [intraluminal flow- and acetylcholine (ACh)-induced] vasodilatation in human skeletal muscle feed arteries (SMFAs) of young (24 ± 1 years, n = 16), middle aged (45 ± 1 years, n = 18) and old (76 ± 2 years, n = 21) subjects was assessed in vitro with and without the inhibition of NOX1 (ML090), NOX2 (gp91) and NOX4 (plumbagin). To identify the role of nitric oxide (NO) bioavailability in these responses, NO synthase blockade (l- -monomethyl arginine citrate) was utilized. SMFA NOX1, NOX2 and NOX4 protein expression was determined by western blotting. Age related endothelium-dependent vasodilatory dysfunction was evident in response to flow (young: 69 ± 3; middle aged: 51 ± 3; old: 27 ± 3%, P < 0.05) and ACh (young: 89 ± 2; middle aged: 72 ± 3; old: 45 ± 4%, P < 0.05). NOX1 inhibition had no effect on SMFA vasodilatation, whereas NOX2 inhibition restored flow- and ACh-induced vasodilatation in the middle aged and the old SMFAs (middle aged + gp91: 69 ± 3; 86 ± 3, old + gp91: 65 ± 5; 83 ± 2%, P < 0.05) and NOX4 inhibition tended to restore these vasodilatory responses in these two groups, but neither achieved statistical significance (P ≈ 0.06). l- -monomethyl arginine citrate negated the restorative effects of NOX2 and NOX4 blockade. Only NOX2 and NOX4 protein expression was significantly greater in the two older groups and inversely related to vascular function (r = 0.48 to 0.93, P < 0.05). NOX2 and, to a lesser extent, NOX4 appear to play an important, probably NO-mediated, role in age-related endothelial dysfunction. KEY POINTS: The present study aimed to determine the isoform-specific role of the NADPH oxidases (NOX) in the endothelium-mediated vascular dysfunction associated with ageing. Age related endothelium-dependent vasodilatory dysfunction was evident in skeletal muscle feed arteries in response to both flow and acetylcholine. NOX2 inhibition (gp91) restored endothelium-dependent vasodilatation in the middle aged and the old skeletal muscle feed arteries, and NOX4 inhibition (plumbagin) tended to restore these vasodilatory responses in these two groups. Nitric oxide synthase inhibition negated the restorative effects of NOX2 and NOX4 blockade. NOX2 and NOX4 protein expression was significantly greater in the two older groups and inversely related to vascular function. NOX2 and, to a lesser extent, NOX4 appear to play an important, probably nitric oxide-mediated, role in age-related endothelial dysfunction and could be important therapeutic targets to maintain vascular health with ageing.
Topics: Middle Aged; Humans; NADPH Oxidases; NADPH Oxidase 4; Nitric Oxide; Acetylcholine; Aging; Vascular Diseases; Endothelium, Vascular; Nitric Oxide Synthase; Arginine
PubMed: 36416565
DOI: 10.1113/JP283208 -
Journal of Pharmacological and... 2023The central nervous system of hard ticks (Ixodidae) consists of a concentrated merged nerve mass known as the synganglion. Although knowledge of tick neurobiology has...
The central nervous system of hard ticks (Ixodidae) consists of a concentrated merged nerve mass known as the synganglion. Although knowledge of tick neurobiology has dramatically improved over the last two decades, this is the first time that isolation and electrophysiological recordings have been carried out on tick neurons from the synganglion. Method: We developed a simple protocol for synganglion neuron isolation and used a whole-cell patch clamp to measure ionic currents induced by acetylcholine, nicotine and muscarine. Relatively large neurons (∼ 25 μm and ∼ 35 μm) were isolated and 1 mM acetylcholine was used to induce strong inward currents of -0.38 ± 0.1 nA and - 1.04 ± 0.1 nA, respectively, with the corresponding cell capacitances being at around 142 pF and 188 pF. In addition, successive application of 1 mM acetylcholine through ∼25 μm and ∼ 35 μm cells for increasing amounts of time resulted in a rapid reduction in current amplitudes. We also found that acetylcholine-evoked currents were associated with a reversible increase in intracellular calcium levels for each neuronal type. In contrast, 1 mM muscarine and nicotine induced a strong and non-reversible increase in intracellular calcium levels. This study serves as a proof of concept for the mechanical isolation of tick synganglion neurons followed by their electrophysiological recording. This approach will aid investigations into the pharmacological properties of tick neurons and provides the tools needed for the identification of drug-targeted sites and effective tick control measures.
Topics: Animals; Ixodes; Nicotine; Acetylcholine; Calcium; Muscarine; Neurons
PubMed: 37866797
DOI: 10.1016/j.vascn.2023.107473 -
BMC Oral Health May 2022Sex hormones influence circulation, periodontitis, and wound healing. The aim of the study was to compare the endothelium-dependent and independent vasodilation in human...
BACKGROUND
Sex hormones influence circulation, periodontitis, and wound healing. The aim of the study was to compare the endothelium-dependent and independent vasodilation in human gingiva in men and women.
METHODS
Gingival blood flow was evaluated in twelve male and twelve female subjects with healthy gingiva and no systemic conditions after acetylcholine or nitric oxide donor (NitroPOHL). Agonists were administered into the gingival sulcus at the right secondary incisor (test site). Regional gingival blood flow (GBF) was imaged by Laser Speckle Contrast Imager from the marginal gingiva to the mucogingival junction in four consecutive regions (coronal, midway1, midway2 and apical). Blood flow was expressed in Laser Speckle Perfusion Unit (LSPU). The absolute maximal blood flow change (Dmax), the area under the blood flow curve (AUC), and the time to peak (TTP) were calculated.
RESULTS
Males had higher baseline GBF than females (257 ± 18.2 vs. 225 ± 18.8 LSPU, p < 0.001). Acetylcholine and NitroPOHL significantly increased the GBF in all test regions. The Dmax after the acetylcholine was reduced apically compared to the coronal (90 ± 13 LSPU vs. 117 ± 7 LSPU, p < 0.01), but it was similar after NitroPOHL (78 ± 9 LSPU vs. 86 ± 6 LSPU, p = 0.398) in both sexes. The Dmax and AUC were higher, and the TTP was smaller in men in most regions after acetylcholine but not after NitroPOHL.
CONCLUSION
In the human gingiva, the endothelium-independent vasodilation propagates without attenuation in the line of the vascular supply in both sexes. At the same time, the endothelium-dependent ascending vasodilation attenuates similarly in men and women. However, men had more pronounced endothelium-dependent vasodilation than women. Therefore, it might contribute to the increased severity of periodontal disease in men.
TRIAL REGISTRATION
The study was registered with ClinicalTrials.gov on 09.06.2021 (NCT04918563).
Topics: Acetylcholine; Endothelium; Female; Gingiva; Humans; Male; Regional Blood Flow; Vasodilation
PubMed: 35562729
DOI: 10.1186/s12903-022-02186-2 -
Journal of Neurochemistry Sep 2021Cholinergic signaling is crucial in cognitive processes, and degenerating cholinergic projections are a pathological hallmark in dementia. Use of cholinesterase... (Review)
Review
Cholinergic signaling is crucial in cognitive processes, and degenerating cholinergic projections are a pathological hallmark in dementia. Use of cholinesterase inhibitors is currently the main treatment option to alleviate symptoms of Alzheimer's disease and has been postulated as a therapeutic strategy in acute brain damage (stroke and traumatic brain injury). However, the benefits of this treatment are still not clear. Importantly, cholinergic receptors are expressed both by neurons and by astrocytes and microglia, and binding of acetylcholine to the α7 nicotinic receptor in glial cells results in anti-inflammatory response. Similarly, the brain fine-tunes the peripheral immune response over the cholinergic anti-inflammatory axis. All of these processes are of importance for the outcome of acute and chronic neurological disease. Here, we summarize the main findings about the role of cholinergic signaling in brain disorders and provide insights into the complexity of molecular regulators of cholinergic responses, such as microRNAs and transfer RNA fragments, both of which may fine-tune the orchestra of cholinergic mRNAs. The available data suggest that these small noncoding RNA regulators may include promising biomarkers for predicting disease course and assessing treatment responses and might also serve as drug targets to attenuate signaling cascades during overwhelming inflammation and to ameliorate regenerative capacities of neuroinflammation.
Topics: Acetylcholine; Animals; Central Nervous System Diseases; Cholinergic Agents; Cholinergic Neurons; Cholinesterase Inhibitors; Humans; RNA; Signal Transduction
PubMed: 33638173
DOI: 10.1111/jnc.15332 -
Molecules (Basel, Switzerland) Jan 2023As an indispensable component in human beings, the acetylcholine system regulates multiple physiological processes not only in neuronal tissues but also in nonneuronal... (Review)
Review
As an indispensable component in human beings, the acetylcholine system regulates multiple physiological processes not only in neuronal tissues but also in nonneuronal tissues. However, since the concept of the "Nonneuronal cholinergic system (NNCS)" has been proposed, the role of the acetylcholine system in nonneuronal tissues has received increasing attention. A growing body of research shows that the acetylcholine system also participates in modulating inflammatory responses, regulating contraction and mucus secretion of respiratory tracts, and influencing the metastasis and invasion of lung cancer. In addition, the susceptibility and severity of respiratory tract infections caused by pathogens such as Mycobacterium Tuberculosis and the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) can also correlate with the regulation of the acetylcholine system. In this review, we summarized the major roles of the acetylcholine system in respiratory diseases. Despite existing achievements in the field of the acetylcholine system, we hope that more in-depth investigations on this topic will be conducted to unearth more possible pharmaceutical applications for the treatment of diverse respiratory diseases.
Topics: Humans; COVID-19; Acetylcholine; SARS-CoV-2; Respiratory System; Respiratory Tract Infections
PubMed: 36770805
DOI: 10.3390/molecules28031139 -
Neuroscience May 2022Dendrites represent the "reception hub" of the neuron as they collect thousands of different inputs and send a coherent response to the cell body. A considerable portion... (Review)
Review
Dendrites represent the "reception hub" of the neuron as they collect thousands of different inputs and send a coherent response to the cell body. A considerable portion of these signals, especially in vivo, arises from neuromodulatory sources, which affect dendritic computations and cellular activity. In this context, acetylcholine (ACh) exerts a coordinating role of different brain structures, contributing to goal-driven behaviors and sleep-wake cycles. Specifically, cholinergic neurons from the medial septum-diagonal band of Broca complex send numerous projections to glutamatergic principal cells and GABAergic inhibitory neurons in the hippocampus, differentially entraining them during network oscillations. Interneurons display abundant expression of cholinergic receptors and marked responses to stimulation by ACh. Nonetheless, the precise localization of ACh inputs is largely unknown, and evidence for cholinergic modulation of interneuronal dendritic signaling remains elusive. In this article, we review evidence that suggests modulatory effects of ACh on dendritic computations in three hippocampal interneuron subtypes: fast-spiking parvalbumin-positive (PV) cells, somatostatin-expressing (SOM) oriens lacunosum moleculare cells and vasoactive intestinal polypeptide-expressing (VIP) interneuron-selective interneurons. We consider the distribution of cholinergic receptors on these interneurons, including information about their specific somatodendritic location, and discuss how the action of these receptors can modulate dendritic Ca signaling and activity of interneurons. The implications of ACh-dependent Ca signaling for dendritic plasticity are also discussed. We propose that cholinergic modulation can shape the dendritic integration and plasticity in interneurons in a cell type-specific manner, and the elucidation of these mechanisms will be required to understand the contribution of each cell type to large-scale network activity.
Topics: Acetylcholine; Cholinergic Agents; Hippocampus; Interneurons; Parvalbumins; Receptors, Cholinergic; Vasoactive Intestinal Peptide
PubMed: 34129910
DOI: 10.1016/j.neuroscience.2021.06.011 -
Environmental Research Jun 2023Neurotoxic pesticides are a group of chemicals that pose a severe threat to both human health and the environment. These molecules are also known to accumulate in the... (Review)
Review
Neurotoxic pesticides are a group of chemicals that pose a severe threat to both human health and the environment. These molecules are also known to accumulate in the food chain and persist in the environment, which can lead to long-term exposure and adverse effects on non-target organisms. The detrimental effects of these pesticides on neurotransmitter levels and function can lead to a range of neurological and behavioral symptoms, which are closely associated with neurodegenerative diseases. Hence, the accurate and reliable detection of these neurotoxic pesticides and associated neurotransmitters is essential for clinical applications, such as diagnosis and treatment. Over the past few decades, acetylcholinesterase (AchE) biosensors have emerged as a sensitive and reliable tool for the electrochemical detection of neurotoxic pesticides and acetylcholine. These biosensors can be tailored to utilize the high specificity and sensitivity of AchE, enabling the detection of these chemicals. Additionally, enzyme immobilization and the incorporation of nanoparticles have further improved the detection capabilities of these biosensors. AchE biosensors have shown tremendous potential in various fields, including environmental monitoring, clinical diagnosis, and pesticide residue analysis. This review summarizes the advancements in AchE biosensors for electrochemical detection of neurotoxic pesticides and acetylcholine over the past two decades.
Topics: Humans; Pesticides; Acetylcholinesterase; Acetylcholine; Pesticide Residues; Biosensing Techniques
PubMed: 36948285
DOI: 10.1016/j.envres.2023.115724 -
Biochemical Society Transactions Apr 2023Barbeau's seesaw hypothesis of dopamine-acetylcholine balance has predominated movement disorders literature for years. Both the simplicity of the explanation and the... (Review)
Review
Barbeau's seesaw hypothesis of dopamine-acetylcholine balance has predominated movement disorders literature for years. Both the simplicity of the explanation and the matching efficacy of anticholinergic treatment in movement disorders seem to support this hypothesis. However, evidence from translational and clinical studies in movement disorders indicates that many features of this simple balance are lost, broken, or absent from movement disorders models or in imaging studies of patients with these disorders. This review reappraises the dopamine-acetylcholine balance hypothesis in light of recent evidence and describes how the Gαi/o coupled muscarinic M4 receptor acts in opposition to dopamine signaling in the basal ganglia. We highlight how M4 signaling can ameliorate or exacerbate movement disorders symptoms and physiological correlates of these symptoms in specific disease states. Furthermore, we propose future directions for investigation of this mechanisms to fully understand the potential efficacy of M4 targeting therapeutics in movement disorders. Overall, initial evidence suggest that M4 is a promising pharmaceutical target to ameliorate motor symptoms of hypo- and hyper-dopaminergic disorders.
Topics: Humans; Acetylcholine; Receptor, Muscarinic M4; Dopamine; Movement Disorders; Cholinergic Agents
PubMed: 37013974
DOI: 10.1042/BST20220525 -
The Journal of Neuroscience : the... Oct 2023Alzheimer's disease (AD) is associated with amyloidosis and dysfunction of the cholinergic system, which is crucial for learning and memory. However, the nature of...
Alzheimer's disease (AD) is associated with amyloidosis and dysfunction of the cholinergic system, which is crucial for learning and memory. However, the nature of acetylcholine signaling within regions of cholinergic-dependent plasticity and how it changes with experience is poorly understood, much less the impact of amyloidosis on this signaling. Therefore, we optically measure the release profile of acetylcholine to unexpected, predicted, and predictive events in visual cortex (VC)-a site of known cholinergic-dependent plasticity-in a preclinical mouse model of AD that develops amyloidosis. We find that acetylcholine exhibits reinforcement signaling qualities, reporting behaviorally relevant outcomes and displaying release profiles to predictive and predicted events that change as a consequence of experience. We identify three stages of amyloidosis occurring before the degeneration of cholinergic synapses within VC and observe that cholinergic responses in amyloid-bearing mice become impaired over these stages, diverging progressively from age- and sex-matched littermate controls. In particular, amyloidosis degrades the signaling of unexpected rewards and punishments, and attenuates the experience-dependent (1) increase of cholinergic responses to outcome predictive visual cues, and (2) decrease of cholinergic responses to predicted outcomes. Hyperactive spontaneous acetylcholine release occurring transiently at the onset of impaired cholinergic signaling is also observed, further implicating disrupted cholinergic activity as an early functional biomarker in AD. Our findings suggest that acetylcholine acts as a reinforcement signal that is impaired by amyloidosis before pathologic degeneration of the cholinergic system, providing a deeper understanding of the effects of amyloidosis on acetylcholine signaling and informing future interventions for AD. The cholinergic system is especially vulnerable to the neurotoxic effects of amyloidosis, a hallmark of Alzheimer's disease (AD). Though amyloid-induced cholinergic synaptic loss is thought in part to account for learning and memory impairments in AD, little is known regarding how amyloid impacts signaling of the cholinergic system before its anatomic degeneration. Optical measurement of acetylcholine (ACh) release in a mouse model of AD that develops amyloidosis reveals that ACh signals reinforcement and outcome prediction that is disrupted by amyloidosis before cholinergic degeneration. These observations have important scientific and clinical implications: they implicate ACh signaling as an early functional biomarker, provide a deeper understanding of the action of acetylcholine, and inform on when and how intervention may best ameliorate cognitive decline in AD.
Topics: Mice; Animals; Alzheimer Disease; Acetylcholine; Amyloidosis; Amyloid; Cholinergic Agents; Biomarkers; Amyloid beta-Peptides
PubMed: 37648452
DOI: 10.1523/JNEUROSCI.0967-23.2023