The Na+/H+ Exchanger Nhe1 Modulates Network Excitability via GABA Release

2018 ◽  
Vol 29 (10) ◽  
pp. 4263-4276 ◽  
Author(s):  
Hartmut T Bocker ◽  
Theresa Heinrich ◽  
Lutz Liebmann ◽  
J Christopher Hennings ◽  
Eric Seemann ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Neuronal Excitability ◽  
Ph Dependence ◽  
Gaba Release ◽  
Epileptic Activity ◽  
Inhibitory Interneurons ◽  
Gabaergic Interneurons ◽  
Glutamatergic Neurons ◽  
Brain Functions ◽  
Ph Dependent

Abstract Brain functions are extremely sensitive to pH changes because of the pH-dependence of proteins involved in neuronal excitability and synaptic transmission. Here, we show that the Na+/H+ exchanger Nhe1, which uses the Na+ gradient to extrude H+, is expressed at both inhibitory and excitatory presynapses. We disrupted Nhe1 specifically in mice either in Emx1-positive glutamatergic neurons or in parvalbumin-positive cells, mainly GABAergic interneurons. While Nhe1 disruption in excitatory neurons had no effect on overall network excitability, mice with disruption of Nhe1 in parvalbumin-positive neurons displayed epileptic activity. From our electrophysiological analyses in the CA1 of the hippocampus, we conclude that the disruption in parvalbumin-positive neurons impairs the release of GABA-loaded vesicles, but increases the size of GABA quanta. The latter is most likely an indirect pH-dependent effect, as Nhe1 was not expressed in purified synaptic vesicles itself. Conclusively, our data provide first evidence that Nhe1 affects network excitability via modulation of inhibitory interneurons.

scholarly journals Defective function of GABA-containing synaptic vesicles in mice lacking the AP-3B clathrin adaptor

2004 ◽  
Vol 167 (2) ◽  
pp. 293-302 ◽  
Author(s):  
Fubito Nakatsu ◽  
Motohiro Okada ◽  
Fumiaki Mori ◽  
Noriko Kumazawa ◽  
Hiroto Iwasa ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Neuronal Excitability ◽  
Critical Role ◽  
Physiological Role ◽  
Adaptor Protein ◽  
Epileptic Seizures ◽  
Gaba Release ◽  
Long Term Potentiation ◽  
Clathrin Adaptor ◽  
And Function

AP-3 is a member of the adaptor protein (AP) complex family that regulates the vesicular transport of cargo proteins in the secretory and endocytic pathways. There are two isoforms of AP-3: the ubiquitously expressed AP-3A and the neuron-specific AP-3B. Although the physiological role of AP-3A has recently been elucidated, that of AP-3B remains unsolved. To address this question, we generated mice lacking μ3B, a subunit of AP-3B. μ3B−/− mice suffered from spontaneous epileptic seizures. Morphological abnormalities were observed at synapses in these mice. Biochemical studies demonstrated the impairment of γ-aminobutyric acid (GABA) release because of, at least in part, the reduction of vesicular GABA transporter in μ3B−/− mice. This facilitated the induction of long-term potentiation in the hippocampus and the abnormal propagation of neuronal excitability via the temporoammonic pathway. Thus, AP-3B plays a critical role in the normal formation and function of a subset of synaptic vesicles. This work adds a new aspect to the pathogenesis of epilepsy.

scholarly journals A Conditional Glutamatergic Synaptic Vesicle Marker for Drosophila

2022 ◽  
Author(s):  
Sarah J Certel ◽  
Evelyne Ruchti ◽  
Brian D McCabe ◽  
R Steven Stowers
Keyword(s):  
Nervous System ◽  
Synaptic Vesicles ◽  
Neural Circuit ◽  
Glutamate Release ◽  
Nervous System Development ◽  
Central Complex ◽  
Excitatory Neurotransmitter ◽  
Glutamatergic Neurons ◽  
Brain Functions ◽  
Conditional Expression

Abstract Glutamate is a principal neurotransmitter used extensively by the nervous systems of all vertebrate and invertebrate animals. It is primarily an excitatory neurotransmitter that has been implicated in nervous system development as well as a myriad of brain functions from the simple transmission of information between neurons to more complex aspects of nervous system function including synaptic plasticity, learning, and memory. Identification of glutamatergic neurons and their sites of glutamate release are thus essential for understanding the mechanisms of neural circuit function and how information is processed to generate behavior. Here we describe and characterize smFLAG-vGlut, a conditional marker of glutamatergic synaptic vesicles for the Drosophila model system. smFLAG-vGlut is validated for functionality, conditional expression, and specificity for glutamatergic neurons and synaptic vesicles. The utility of smFLAG-vGlut is demonstrated by glutamatergic neurotransmitter phenotyping of 26 different central complex neuron types of which nine were established to be glutamatergic. This illumination of glutamate neurotransmitter usage will enhance the modeling of central complex neural circuitry and thereby our understanding of information processing by this region of the fly brain. The use of smFLAG for glutamatergic neurotransmitter phenotyping and identification of glutamate release sites can be extended to any Drosophila neuron(s) represented by a binary transcription system driver.

scholarly journals Differential inhibition of GABA release from mouse hippocampal interneuron subtypes by the volatile anesthetic isoflurane

2020 ◽  
Author(s):  
Iris A. Speigel ◽  
Hugh C. Hemmings
Keyword(s):  
Synaptic Vesicle ◽  
Volatile Anesthetic ◽  
Gaba Release ◽  
Gabaergic Interneurons ◽  
Relative Expression ◽  
Voltage Gated ◽  
Glutamatergic Neurons ◽  
Hippocampal Interneuron ◽  
Synaptic Vesicle Exocytosis ◽  
Vesicle Exocytosis

AbstractGeneral anesthesia is critical to modern medicine and animal research, but the cellular and molecular actions of general anesthetics on the central nervous system remain poorly understood. Volatile anesthetics such as isoflurane disrupt synaptic transmission and inhibit synaptic vesicle release in a neurotransmitter-selective manner. For example, GABA release from interneurons is less sensitive to isoflurane inhibition than are glutamate or dopamine release. Hippocampal and cortical interneuron subpopulations have diverse neurophysiological and synaptic properties, and their individual subtype-specific responses to isoflurane are unknown. We used live-cell optical imaging of exocytosis using fluorescent biosensors expressed in transgenic mouse hippocampal neuron cultures to delineate interneuron subtype-specific effects of isoflurane on synaptic vesicle exocytosis. We found that a clinically relevant concentration of isoflurane (0.5 mM) differentially modulated action potential-mediated exocytosis from GABAergic interneurons: parvalbumin-expressing interneurons were inhibited to 83.1±11.7% of control, whereas somatostatin-expressing and interneurons glutamatergic neurons were inhibited to 58.6±13.3% and 64.5±8.5% of control, respectively. The role of presynaptic voltage-gated sodium channel (Nav) subtype expression in determining isoflurane sensitivity was probed by overexpression or knockdown of specific Nav subtypes, which have distinct sensitivities to isoflurane and are differentially expressed between glutamatergic and GABAergic neurons. We found that the sensitivity of exocytosis to isoflurane was determined by the relative expression of Nav1.1 (associated with lower sensitivity) and Nav1.6 (associated with higher sensitivity). Thus the selective effects of isoflurane on synaptic vesicle exocytosis from hippocampal interneuron subtypes is determined by synaptic diversity in the relative expression of Nav1.1 and Nav1.6.Significance statementThe volatile anesthetic isoflurane inhibits hippocampal GABAergic interneuron synaptic vesicle exocytosis with differences in potency between interneuron subtypes. This neuron subtype-specific pharmacology derives in part from synaptic diversity in the expression of presynaptic voltage-gated sodium channels that have different sensitivities to anesthetic modulation of channel function. GABAergic interneurons are generally more resistant to the presynaptic effects of isoflurane owing to predominant Nav1.1 expression, whereas glutamatergic neurons are more sensitive owing to predominant Nav1.6 expression, which supports heterogenous pharmacologic effects on specific neural circuits.

Rationalizing the pH-Activity Response for Escherichia Coli’s Glycinamide Ribonucleotidetransformylase Through Computational Methods

2019 ◽  
Author(s):  
Adrian Roitberg ◽  
Pancham Lal Gupta
Keyword(s):  
Transfer Reaction ◽  
Catalytic Mechanism ◽  
Ph Dependence ◽  
Ph Effects ◽  
Dependent Manner ◽  
E Coli ◽  
Gar Tfase ◽  
Activity Curve ◽  
Ph Dependent ◽  
Formyl Transfer

<div>Human Glycinamide ribonucleotide transformylase (GAR Tfase), a regulatory enzyme in the de novo purine biosynthesis pathway, has been established as an anti-cancer target. GAR Tfase catalyzes the formyl transfer reaction from the folate cofactor to the GAR ligand. In the present work, we study E. coli GAR Tfase, which has high sequence similarity with the human GAR Tfase with most functional residues conserved. E. coli GAR Tfase exhibits structural changes and the binding of ligands that varies with pH which leads to change the rate of the formyl transfer reaction in a pH-dependent manner. Thus, the inclusion of pH becomes essential for the study of its catalytic mechanism. Experimentally, the pH-dependence of the kinetic parameter kcat is measured to evaluate the pH-range of enzymatic activity. However, insufficient information about residues governing the pH-effects on the catalytic activity leads to ambiguous assignments of the general acid and base catalysts and consequently its catalytic mechanism. In the present work, we use pH-replica exchange molecular dynamics (pH-REMD) simulations to study the effects of pH on E. coli GAR Tfase enzyme. We identify the titratable residues governing the pH-dependent conformational changes in the system. Furthermore, we filter out the protonation states which are essential in maintaining the structural integrity, keeping the ligands bound and assisting the catalysis. We reproduce the experimental pH-activity curve by computing the population of key protonation states. Moreover, we provide a detailed description of residues governing the acidic and basic limbs of the pH-activity curve.</div>

Enhanced Analytic Approach for Describing pH Triggered Fast Drug Release Systems

2021 ◽  
Vol 18 ◽  
Author(s):  
Aykut Elmas ◽  
Guliz Akyuz ◽  
Ayhan Bergal ◽  
Muberra Andac ◽  
Omer Andac
Keyword(s):  
Drug Release ◽  
Ph Dependence ◽  
Slope Angle ◽  
Ph Sensitive ◽  
Analytic Model ◽  
Two Dimensional ◽  
Analytic Equation ◽  
One Dimensional ◽  
Ph Dependent ◽  
Fast Release

Background: pH sensitive dendrimers attached to nanocarriers, as one of the drug release systems, has become quite popular due to their ease of manufacture in experimental conditions and ability to generate fast drug release in the targeted area. This kind of fast release behavior cannot be represented properly most of the existing kinetic mathematical models. Besides, these models have either no pH dependence or pH dependence added separately. So, they have remained one dimensional. Objective: The aim of this study was to establish the proper analytic equation to describe the fast release of drugs from pH sensitive nanocarrier systems. Then, to combine it with the pH dependent equation for establishing a two-dimensional model for whole system. Methods: We used four common kinetic models for comparison and we fitted them to the release data. Finding that, only Higuchi and Korsmeyer-Peppas models show acceptable fit results. None of these models have pH dependence. To get a better description for pH triggered fast release, we observed the behavior of the slope angle of the release curve. Then we puroposed a new analytic equation by using relation between the slope angle and time. Result: To add a pH dependent equation, we assumed the drug release is “on” or “off” above/below specific pH value and we modified a step function to get a desired behavior. Conclusion: Our new analytic model shows good fitting, not only one-dimensional time dependent release, but also two-dimensional pH dependent release, that provides a useful analytic model to represent release profiles of pH sensitive fast drug release systems.

scholarly journals Systematic exploration of the pH dependence of a peptide hydrogel

2019 ◽  
Vol 97 (6) ◽  
pp. 430-434 ◽  
Author(s):  
Natashya Falcone ◽  
Tsuimy Shao ◽  
Xiaoyi Sun ◽  
Heinz-Bernhard Kraatz
Keyword(s):  
Mechanical Strength ◽  
Narrow Range ◽  
Ph Dependence ◽  
Stimuli Responsive ◽  
Systematic Exploration ◽  
Ph Conditions ◽  
Ph Dependent ◽  
Functional Biomaterials ◽  
Peptide Hydrogel

Stimuli-responsive peptide gels are a growing class of functional biomaterials that are involved in many applications in research. Here, we present a novel di-peptide hydrogel from the compound Boc–Phe–Trp–OH in various buffer and pH conditions. We examine the effects of different stimuli, including temperature and pH, on the mechanical strength of the gels through frequency rheology studies. We found that this hydrogelator is highly pH dependent, only forming a gel in a narrow range of pH 6–7. This hydrogelator hold promise for the development of new stimuli-responsive biomaterials for specific applications that require this type of specific stimuli.

Cholinergic modulation of synaptic inhibition in the guinea pig hippocampus in vitro: excitation of GABAergic interneurons and inhibition of GABA-release

1993 ◽  
Vol 69 (2) ◽  
pp. 626-629 ◽  
Author(s):  
J. C. Behrends ◽  
G. ten Bruggencate
Keyword(s):  
Guinea Pig ◽  
Synaptic Transmission ◽  
Pyramidal Neurons ◽  
Gaba Release ◽  
Cholinergic Receptor ◽  
Receptor Activation ◽  
Release Mechanism ◽  
Gabaergic Interneurons ◽  
Gamma Aminobutyric Acid

1. The effect of cholinergic receptor activation on gamma-aminobutyric acid (GABA)-mediated inhibitory synaptic transmission was investigated in voltage-clamped CA1 pyramidal neurons (HPNs) in the guinea pig hippocampal slice preparation. 2. The cholinergic agonist carbachol (1-10 microM) induced a prominent and sustained increase in the frequency and amplitudes of spontaneous inhibitory postsynaptic currents (IPSCs) in Cl(-)-loaded HPNs. The potentiation of spontaneous IPSCs was not dependent on excitatory synaptic transmission but was blocked by atropine (1 microM). 3. Monosynaptically evoked IPSCs were reversibly depressed by carbachol (10 microM). 4. The frequency of miniature IPSCs recorded in the presence of tetrodotoxin (0.6 or 1.2 microM) was reduced by carbachol (10 or 20 microM) in an atropine-sensitive manner. 5. We conclude that, while cholinergic receptor activation directly excites hippocampal GABAergic interneurons, it has, in addition, a suppressant effect on the synaptic release mechanism at GABAergic terminals. This dual modulatory pattern could explain the suppression of evoked IPSCs despite enhanced spontaneous transmission.

scholarly journals Control of motor coordination by astrocytic tonic GABA release through modulation of excitation/inhibition balance in cerebellum

2018 ◽  
Vol 115 (19) ◽  
pp. 5004-5009 ◽  
Author(s):  
Junsung Woo ◽  
Joo Ok Min ◽  
Dae-Si Kang ◽  
Yoo Sung Kim ◽  
Guk Hwa Jung ◽  
...  
Keyword(s):  
Synaptic Transmission ◽  
Motor Performance ◽  
Neuronal Excitability ◽  
Motor Coordination ◽  
Granule Cells ◽  
Cerebellar Granule Cells ◽  
Gaba Release ◽  
In Vivo Microdialysis ◽  
Tonic Inhibition

Tonic inhibition in the brain is mediated through an activation of extrasynaptic GABAA receptors by the tonically released GABA, resulting in a persistent GABAergic inhibitory action. It is one of the key regulators for neuronal excitability, exerting a powerful action on excitation/inhibition balance. We have previously reported that astrocytic GABA, synthesized by monoamine oxidase B (MAOB), mediates tonic inhibition via GABA-permeable bestrophin 1 (Best1) channel in the cerebellum. However, the role of astrocytic GABA in regulating neuronal excitability, synaptic transmission, and cerebellar brain function has remained elusive. Here, we report that a reduction of tonic GABA release by genetic removal or pharmacological inhibition of Best1 or MAOB caused an enhanced neuronal excitability in cerebellar granule cells (GCs), synaptic transmission at the parallel fiber-Purkinje cell (PF-PC) synapses, and motor performance on the rotarod test, whereas an augmentation of tonic GABA release by astrocyte-specific overexpression of MAOB resulted in a reduced neuronal excitability, synaptic transmission, and motor performance. The bidirectional modulation of astrocytic GABA by genetic alteration of Best1 or MAOB was confirmed by immunostaining and in vivo microdialysis. These findings indicate that astrocytes are the key player in motor coordination through tonic GABA release by modulating neuronal excitability and could be a good therapeutic target for various movement and psychiatric disorders, which show a disturbed excitation/inhibition balance.

scholarly journals pH-dependent Perforation of Macrophage Phagosomes by Listeriolysin O from Listeria monocytogenes

1997 ◽  
Vol 186 (7) ◽  
pp. 1159-1163 ◽  
Author(s):  
Kathryn E. Beauregard ◽  
Kyung-Dall Lee ◽  
R. John Collier ◽  
Joel A. Swanson
Keyword(s):  
Listeria Monocytogenes ◽  
Ph Dependence ◽  
Mouse Bone Marrow ◽  
Listeriolysin O ◽  
Bafilomycin A1 ◽  
Wild Type ◽  
Major Virulence Factor ◽  
Ph Dependent ◽  
Vacuolar Ph

The pore-forming toxin listeriolysin O (LLO) is a major virulence factor implicated in escape of Listeria monocytogenes from phagocytic vacuoles. Here we describe the pH-dependence of vacuolar perforation by LLO, using the membrane-impermeant fluorophore 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) to monitor the pH and integrity of vacuoles in mouse bone marrow–derived macrophages. Perforation was observed when acidic vacuoles containing wild-type L. monocytogenes displayed sudden increases in pH and release of HPTS into the cytosol. These changes were not seen with LLO-deficient mutants. Perforation occurred at acidic vacuolar pH (4.9–6.7) and was reduced in frequency or prevented completely when macrophages were treated with the lysosomotropic agents ammonium chloride or bafilomycin A1. We conclude that acidic pH facilitates LLO activity in vivo.

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