Characterization of Voltage-Gated Ionic Channels in Cholinergic Amacrine Cells in the Mouse Retina

2007 ◽  
Vol 97 (6) ◽  
pp. 4225-4234 ◽  
Author(s):  
Makoto Kaneda ◽  
Koichi Ito ◽  
Yosuke Morishima ◽  
Yasuhide Shigematsu ◽  
Yukio Shimoda
Keyword(s):  
Amacrine Cells ◽  
Ionic Channels ◽  
Signal Propagation ◽  
Membrane Properties ◽  
Mouse Retina ◽  
Delayed Rectifier ◽  
Ca Channel ◽  
Voltage Gated ◽  
A Current ◽  
Cholinergic Amacrine Cells

Recent studies have shown that cholinergic amacrine cells possess unique membrane properties. However, voltage-gated ionic channels in cholinergic amacrine cells have not been characterized systematically. In this study, using electrophysiological and immunohistochemical techniques, we examined voltage-gated ionic channels in a transgenic mouse line the cholinergic amacrine cells of which were selectively labeled with green fluorescent protein (GFP). Voltage-gated K+ currents contained a 4-aminopyridine-sensitive current (A current) and a tetraethylammonium-sensitive current (delayed rectifier K+ current). Voltage-gated Ca2+ currents contained a ω-conotoxin GVIA-sensitive component (N-type) and a ω-Aga IVA-sensitive component (P/Q-type). Tetrodotoxin-sensitive Na+ currents and dihydropyridine-sensitive Ca2+ currents (L-type) were not observed. Immunoreactivity for the Na channel subunit (Pan Nav), the K channel subunits (the A-current subunits [Kv. 3.3 and Kv 3.4]) and the Ca channel subunits (α1A [P/Q-type], α1B [N-type] and α1C [L-type]) was detected in the membrane fraction of the mouse retina by Western blot analysis. Immunoreactivity for the Kv. 3.3, Kv 3.4, α1A [P/Q-type], and α1B [N-type] was colocalized with the GFP signals. Immunoreactivity for α1C [L-type] was not colocalized with the GFP signals. Immunoreactivity for Pan Nav did not exist on the membrane surface of the GFP-positive cells. Our findings indicate that signal propagation in cholinergic amacrine cells is mediated by a combination of two types of voltage-gated K+ currents (the A current and the delayed rectifier K+ current) and two types of voltage-gated Ca2+ currents (the P/Q-type and the N-type) in the mouse retina.

I A in Kenyon Cells of the Mushroom Body of Honeybees Resembles Shaker Currents: Kinetics, Modulation by K+, and Simulation

1999 ◽  
Vol 81 (4) ◽  
pp. 1749-1759 ◽  
Author(s):  
Corinna Pelz ◽  
Johannes Jander ◽  
Hendrik Rosenboom ◽  
Martin Hammer ◽  
Randolf Menzel
Keyword(s):  
Action Potential ◽  
Time Constant ◽  
Mushroom Body ◽  
Delayed Rectifier ◽  
Patch Clamp Technique ◽  
Time Constants ◽  
Voltage Gated ◽  
Kenyon Cells ◽  
Recovery From Inactivation ◽  
A Current

I A in Kenyon cells of the mushroom body of honeybees resembles shaker currents: kinetics, modulation by K+, and simulation. Cultured Kenyon cells from the mushroom body of the honeybee, Apis mellifera, show a voltage-gated, fast transient K+ current that is sensitive to 4-aminopyridine, an A current. The kinetic properties of this A current and its modulation by extracellular K+ ions were investigated in vitro with the whole cell patch-clamp technique. The A current was isolated from other voltage-gated currents either pharmacologically or with suitable voltage-clamp protocols. Hodgkin- and Huxley-style mathematical equations were used for the description of this current and for the simulation of action potentials in a Kenyon cell model. Activation and inactivation of the A current are fast and voltage dependent with time constants of 0.4 ± 0.1 ms (means ± SE) at +45 mV and 3.0 ± 1.6 ms at +45 mV, respectively. The pronounced voltage dependence of the inactivation kinetics indicates that at least a part of this current of the honeybee Kenyon cells is a shaker-like current. Deactivation and recovery from inactivation also show voltage dependency. The time constant of deactivation has a value of 0.4 ± 0.1 ms at −75 mV. Recovery from inactivation needs a double-exponential function to be fitted adequately; the resulting time constants are 18 ± 3.1 ms for the fast and 745 ± 107 ms for the slow process at −75 mV. Half-maximal activation of the A current occurs at −0.7 ± 2.9 mV, and half-maximal inactivation occurs at −54.7 ± 2.4 mV. An increase in the extracellular K+concentration increases the conductance and accelerates the recovery from inactivation of the A current, affecting the slow but not the fast time constant. With respect to these modulations the current under investigation resembles some of the shaker-like currents. The data of the A current were incorporated into a reduced computational model of the voltage-gated currents of Kenyon cells. In addition, the model contained a delayed rectifier K+ current, a Na+current, and a leakage current. The model is able to generate an action potential on current injection. The model predicts that the A current causes repolarization of the action potential but not a delay in the initiation of the action potential. It further predicts that the activation of the delayed rectifier K+ current is too slow to contribute markedly to repolarization during a single action potential. Because of its fast activation, the A current reduces the amplitude of the net depolarizing current and thus reduces the peak amplitude and the duration of the action potential.

scholarly journals Inhibitory inputs tune the light response properties of dopaminergic amacrine cells in mouse retina

2013 ◽  
Vol 110 (2) ◽  
pp. 536-552 ◽  
Author(s):  
G. S. Newkirk ◽  
M. Hoon ◽  
R. O. Wong ◽  
P. B. Detwiler
Keyword(s):  
Visual Processing ◽  
Fluorescent Protein ◽  
Bright Light ◽  
Light Response ◽  
Amacrine Cells ◽  
Membrane Properties ◽  
Mouse Retina ◽  
Light Onset ◽  
Threshold Response ◽  
Green Fluorescent

Dopamine (DA) is a neuromodulator that in the retina adjusts the circuitry for visual processing in dim and bright light conditions. It is synthesized and released from retinal interneurons called dopaminergic amacrine cells (DACs), whose basic physiology is not yet been fully characterized. To investigate their cellular and input properties as well as light responses, DACs were targeted for whole cell recording in isolated retina using two-photon fluorescence microscopy in a mouse line where the dopamine receptor 2 promoter drives green fluorescent protein (GFP) expression. Differences in membrane properties gave rise to cell-to-cell variation in the pattern of resting spontaneous spike activity ranging from silent to rhythmic to periodic burst discharge. All recorded DACs were light sensitive and generated responses that varied with intensity. The threshold response to light onset was a hyperpolarizing potential change initiated by rod photoreceptors that was blocked by strychnine, indicating a glycinergic amacrine input onto DACs at light onset. With increasing light intensity, the ON response acquired an excitatory component that grew to dominate the response to the strongest stimuli. Responses to bright light (photopic) stimuli also included an inhibitory OFF response mediated by GABAergic amacrine cells driven by the cone OFF pathway. DACs expressed GABA (GABAAα1 and GABAAα3) and glycine (α2) receptor clusters on soma, axon, and dendrites consistent with the light response being shaped by dual inhibitory inputs that may serve to tune spike discharge for optimal DA release.

scholarly journals Quantitative Assessment of the Distributions of Membrane Conductances Involved in Action Potential Backpropagation Along Basal Dendrites

2009 ◽  
Vol 101 (3) ◽  
pp. 1524-1541 ◽  
Author(s):  
Corey D. Acker ◽  
Srdjan D. Antic
Keyword(s):  
Cortical Neurons ◽  
Action Potentials ◽  
Synaptic Integration ◽  
Membrane Properties ◽  
Delayed Rectifier ◽  
Published Data ◽  
Membrane Excitability ◽  
Voltage Gated ◽  
Prefrontal Cortical ◽  
Basal Dendrites

Basal dendrites of prefrontal cortical neurons receive strong synaptic drive from recurrent excitatory synaptic inputs. Synaptic integration within basal dendrites is therefore likely to play an important role in cortical information processing. Both synaptic integration and synaptic plasticity depend crucially on dendritic membrane excitability and the backpropagation of action potentials. We carried out multisite voltage-sensitive dye imaging of membrane potential transients from thin basal branches of prefrontal cortical pyramidal neurons before and after application of channel blockers. We found that backpropagating action potentials (bAPs) are predominantly controlled by voltage-gated sodium and A-type potassium channels. In contrast, pharmacologically blocking the delayed rectifier potassium, voltage-gated calcium, or Ih conductance had little effect on dendritic AP propagation. Optically recorded bAP waveforms were quantified and multicompartmental modeling was used to link the observed behavior with the underlying biophysical properties. The best-fit model included a nonuniform sodium channel distribution with decreasing conductance with distance from the soma, together with a nonuniform (increasing) A-type potassium conductance. AP amplitudes decline with distance in this model, but to a lesser extent than previously thought. We used this model to explore the mechanisms underlying two sets of published data involving high-frequency trains of APs and the local generation of sodium spikelets. We also explored the conditions under which IA down-regulation would produce branch strength potentiation in the proposed model. Finally, we discuss the hypothesis that a fraction of basal branches may have different membrane properties compared with sister branches in the same dendritic tree.

scholarly journals Spatial patterning of cholinergic amacrine cells in the mouse retina

2008 ◽  
Vol 508 (2) ◽  
pp. SPC1-SPC1
Author(s):  
Irene E. Whitney ◽  
Patrick W. Keeley ◽  
Mary A. Raven ◽  
Benjamin E. Reese
Keyword(s):  
Amacrine Cells ◽  
Mouse Retina ◽  
Spatial Patterning ◽  
Cholinergic Amacrine Cells

scholarly journals Spatial patterning of cholinergic amacrine cells in the mouse retina

2008 ◽  
Vol 508 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Irene E. Whitney ◽  
Patrick W. Keeley ◽  
Mary A. Raven ◽  
Benjamin E. Reese
Keyword(s):  
Amacrine Cells ◽  
Mouse Retina ◽  
Spatial Patterning ◽  
Cholinergic Amacrine Cells

scholarly journals Novel channel-mediated choline transport in cholinergic neurons of the mouse retina

2017 ◽  
Vol 118 (4) ◽  
pp. 1952-1961 ◽  
Author(s):  
Toshiyuki Ishii ◽  
Kohei Homma ◽  
Asuka Mano ◽  
Takumi Akagi ◽  
Yasuhide Shigematsu ◽  
...  
Keyword(s):  
Purinergic Receptors ◽  
Expression System ◽  
Cholinergic Neurons ◽  
Amacrine Cells ◽  
Mouse Retina ◽  
Choline Uptake ◽  
Transport Pathway ◽  
Choline Transport ◽  
High Affinity ◽  
Cholinergic Amacrine Cells

Choline uptake into the presynaptic terminal of cholinergic neurons is mediated by the high-affinity choline transporter and is essential for acetylcholine synthesis. In a previous study, we reported that P2X2 purinoceptors are selectively expressed in OFF-cholinergic amacrine cells of the mouse retina. Under specific conditions, P2X2 purinoceptors acquire permeability to large cations, such as N-methyl-d-glucamine, and therefore potentially could act as a noncanonical pathway for choline entry into neurons. We tested this hypothesis in OFF-cholinergic amacrine cells of the mouse retina. ATP-induced choline currents were observed in OFF-cholinergic amacrine cells, but not in ON-cholinergic amacrine cells, in mouse retinal slice preparations. High-affinity choline transporters are expressed at higher levels in ON-cholinergic amacrine cells than in OFF-cholinergic amacrine cells. In dissociated preparations of cholinergic amacrine cells, ATP-activated cation currents arose from permeation of extracellular choline. We also examined the pharmacological properties of choline currents. Pharmacologically, α,β-methylene ATP did not produce a cation current, whereas ATPγS and benzoyl-benzoyl-ATP (BzATP) activated choline currents. However, the amplitude of the choline current activated by BzATP was very small. The choline current activated by ATP was strongly inhibited by pyridoxalphosphate-6-azophenyl-2′,4′-sulfonic acid. Accordingly, P2X2 purinoceptors expressed in HEK-293T cells were permeable to choline and similarly functioned as a choline uptake pathway. Our physiological and pharmacological findings support the hypothesis that P2 purinoceptors, including P2X2 purinoceptors, function as a novel choline transport pathway and may provide a new regulatory mechanism for cholinergic signaling transmission at synapses in OFF-cholinergic amacrine cells of the mouse retina. NEW & NOTEWORTHY Choline transport across the membrane is exerted by both the high-affinity and low-affinity choline transporters. We found that choline can permeate P2 purinergic receptors, including P2X2 purinoceptors, in cholinergic neurons of the retina. Our findings show the presence of a novel choline transport pathway in cholinergic neurons. Our findings also indicate that the permeability of P2X2 purinergic receptors to choline observed in the heterologous expression system may have a physiological relevance in vivo.

scholarly journals Cellular positioning and dendritic field size of cholinergic amacrine cells are impervious to early ablation of neighboring cells in the mouse retina

2004 ◽  
Vol 21 (1) ◽  
pp. 13-22 ◽  
Author(s):  
REZA FARAJIAN ◽  
MARY A. RAVEN ◽  
KAREN CUSATO ◽  
BENJAMIN E. REESE
Keyword(s):  
Lucifer Yellow ◽  
Single Cells ◽  
Amacrine Cells ◽  
Field Size ◽  
Mouse Retina ◽  
Dendritic Field ◽  
Normal Retina ◽  
Field Area ◽  
Cholinergic Amacrine Cells ◽  
Neighbor Relationships

We have examined the role of neighbor relationships between cholinergic amacrine cells upon their positioning and dendritic field size by producing partial ablations of this population of cells during early development. We first determined the effectiveness ofl-glutamate as an excitotoxin for ablating cholinergic amacrine cells in the developing mouse retina. Subcutaneous injections (4 mg/g) made on P-3 and thereafter were found to produce a near-complete elimination, while injections at P-2 were ineffective. Lower doses on P-3 produced only partial reductions, and were subsequently used to examine the effect of partial ablation upon mosaic organization and dendritic growth of the remaining cells. Four different Voronoi-based measures of mosaic geometry were examined inl-glutamate-treated and normal (saline-treated) retinas. Partial depletions of around 40% produced cholinergic mosaics that, when scaled for density, approximated the mosaic geometry of the normal retina. Separate comparisons simulating a 40% random deletion of the normal retina produced mosaics that were no different from those experimentally depleted retinas. Consequently, no evidence was found for positional regulation in the absence of normal neighbor relationships. Single cells in the ganglion cell layer were intracellularly filled with Lucifer Yellow to examine the morphology and dendritic field extent following partial ablation of the cholinergic amacrine cells. No discernable effect was found on their starburst morphology, and total dendritic field area, number of primary dendrites, and branch frequency were not significantly different. Cholinergic amacrine cells normally increase their dendritic field area after P-3 in excess of retinal expansion; despite this, the present results show that this growth is not controlled by the density of neighboring processes.

scholarly journals Random spatial patterning of cone bipolar cell mosaics in the mouse retina

2017 ◽  
Vol 34 ◽  
Author(s):  
PATRICK W. KEELEY ◽  
JASON J. KIM ◽  
SAMMY C.S. LEE ◽  
SILKE HAVERKAMP ◽  
BENJAMIN E. REESE
Keyword(s):  
Bipolar Cell ◽  
Amacrine Cells ◽  
Cell Types ◽  
Bipolar Cells ◽  
Statistical Properties ◽  
Horizontal Cells ◽  
Mouse Retina ◽  
Dendritic Arbors ◽  
Cone Bipolar Cells ◽  
Cholinergic Amacrine Cells

AbstractRetinal bipolar cells spread their dendritic arbors to tile the retinal surface, extending them to the tips of the dendritic fields of their homotypic neighbors, minimizing dendritic overlap. Such uniform nonredundant dendritic coverage of these populations would suggest a degree of spatial order in the properties of their somal distributions, yet few studies have examined the patterning in retinal bipolar cell mosaics. The present study examined the organization of two types of cone bipolar cells in the mouse retina, the Type 2 cells and the Type 4 cells, and compared their spatial statistical properties with those of the horizontal cells and the cholinergic amacrine cells, as well as to random simulations of cells matched in density and constrained by soma size. The Delauney tessellation of each field was computed, from which nearest neighbor distances and Voronoi domain areas were extracted, permitting a calculation of their respective regularity indexes (RIs). The spatial autocorrelation of the field was also computed, from which the effective radius and packing factor (PF) were determined. Both cone bipolar cell types were found to be less regular and less efficiently packed than either the horizontal cells or cholinergic amacrine cells. Furthermore, while the latter two cell types had RIs and PFs in excess of those for their matched random simulations, the two types of cone bipolar cells had spatial statistical properties comparable to random distributions. An analysis of single labeled cone bipolar cells revealed dendritic arbors frequently skewed to one side of the soma, as would be expected from a randomly distributed population of cells with dendrites that tile. Taken together, these results suggest that, unlike the horizontal cells or cholinergic amacrine cells which minimize proximity to one another, cone bipolar cell types are constrained only by their physical size.

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