Patch-Clamp Investigations and Compartmental Modeling of Rod Bipolar Axon Terminals in an In Vitro Thin-Slice Preparation of the Mammalian Retina

2007 ◽  
Vol 97 (2) ◽  
pp. 1171-1187 ◽  
Author(s):  
Leif Oltedal ◽  
Svein Harald Mørkve ◽  
Margaret Lin Veruki ◽  
Espen Hartveit
Keyword(s):  
Synaptic Transmission ◽  
Computer Simulations ◽  
Bipolar Cells ◽  
Slice Preparation ◽  
Whole Cell ◽  
Axon Terminals ◽  
Postsynaptic Currents ◽  
Whole Cell Recordings ◽  
Rod Bipolar Cells

To extend the usefulness of rod bipolar cells for studies of chemical synaptic transmission, we have performed electrophysiological recordings from rod bipolar axon terminals in an in vitro slice preparation of the rat retina. Whole cell recordings from axon terminals and cell bodies were used to investigate the passive membrane properties of rod bipolar cells and analyzed with a two-compartment equivalent electrical circuit model developed by Mennerick et al. For both terminal- and soma-end recordings, capacitive current decays were well fitted by biexponential functions. Computer simulations of simplified models of rod bipolar cells demonstrated that estimates of the capacitance of the axon terminal compartment can depend critically on the recording location, with terminal-end recordings giving the best estimates. Computer simulations and whole cell recordings demonstrated that terminal-end recordings can yield more accurate estimates of the peak amplitude and kinetic properties of postsynaptic currents generated at the axon terminals due to increased electrotonic filtering of these currents when recorded at the soma. Finally, we present whole cell and outside-out patch recordings from axon terminals with responses evoked by GABA and glycine, spontaneous inhibitory postsynaptic currents, voltage-gated Ca2+ currents, and depolarization-evoked reciprocal synaptic responses, verifying that the recorded axon terminals are involved in normal pre- and postsynaptic relationships. These results demonstrate that axon terminals of rod bipolar cells are directly accessible to whole cell and outside-out patch recordings, extending the usefulness of this preparation for detailed studies of pre- and postsynaptic mechanisms of synaptic transmission in the CNS.

GABA responses of rod bipolar cells in rabbit retinal slices

2000 ◽  
Vol 17 (3) ◽  
pp. 381-389 ◽  
Author(s):  
GREGORY S. McGILLEM ◽  
THOMAS C. ROTOLO ◽  
RAMON F. DACHEUX
Keyword(s):  
Bipolar Cell ◽  
Axon Terminal ◽  
Bipolar Cells ◽  
Slice Preparation ◽  
Pharmacological Agents ◽  
Axon Terminals ◽  
Receptor Input ◽  
Sr 95531 ◽  
Whole Cell Recordings ◽  
Rod Bipolar Cells

GABAergic responses of rabbit rod bipolar cells were reexamined by using whole-cell recordings in the superfused slice preparation to determine if there is GABAC receptor input to their axon terminal and to characterize the contribution that GABAA and GABAC receptors make to the total GABA current on the axon terminals of these cells. Pharmacological agents specifically blocking GABAA and GABAC receptor currents demonstrated that 37% of the GABA-activated current was blocked by either the GABAA antagonists bicuculline or SR-95531, whereas the remaining 63% of the GABA current was blocked by a mixture of bicuculline and the GABAC antagonist TPMPA. This indicated that GABAC receptors were present on the axon terminal of the rabbit rod bipolar cell and that they were responsible for mediating the bicuculline insensitive GABA current.

Properties of synaptic transmission from photoreceptors to bipolar cells in the mudpuppy retina

1993 ◽  
Vol 69 (2) ◽  
pp. 352-360 ◽  
Author(s):  
H. G. Kim ◽  
R. F. Miller
Keyword(s):  
Kynurenic Acid ◽  
Response Times ◽  
Plexiform Layer ◽  
Bipolar Cells ◽  
Slice Preparation ◽  
Peak Response ◽  
Rods And Cones ◽  
Significant Difference ◽  
Pharmacological Studies ◽  
Whole Cell Recordings

1. Simultaneous, whole-cell recordings were obtained from synaptically coupled photoreceptor/bipolar cell pairs, by the use of direct visualization in a superfused, mudpuppy retinal slice preparation. 2. OFF-bipolar cells (BPs) generated sign-conserving responses when extrinsic current was injected into rods and cones, whereas ON-BPs generated a sign-reversing response. OFF-BPs (n = 24) responded faster than ON-BPs (n = 12), in terms of response latency (27.8 vs. 80.6 ms) and peak response times (50.5 vs. 159.8 ms) when current was injected into photoreceptors. We did not detect any significant difference between rod- versus cone-mediated latency or peak response times in the ON- and OFF-BP subtypes. 3. Rod and cone inputs to OFF-BPs were blocked by kynurenic acid (Kyn), but the doses required were significantly higher for rod inputs: the IC50 (the concentration at which an antagonist blocks 50% of the responses) for Kyn was 0.3 mM for cone inputs and 1 mM for rod inputs. 4. Rod inputs to OFF-BPs showed the same Kyn sensitivity as rod inputs to horizontal cells (HCs). However, cone inputs to HCs (IC50 < 200 microM) were more sensitive to Kyn than those to OFF-BPs. 5. The pharmacological studies presented here, together with previous studies, suggest that the sign-conserving pathway in the outer plexiform layer of the mudpuppy retina involves at least three subtypes of glutamate receptors: 1) cone-activated receptors of HCs; 2) cone-activated receptors of OFF-BPs; and 3) rod-activated receptors found in HCs and BPs.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of Hyposmolar Solutions on Membrane Currents of Hippocampal Interneurons and Mossy Cells In Vitro

1998 ◽  
Vol 79 (2) ◽  
pp. 1108-1112 ◽  
Author(s):  
Scott C. Baraban ◽  
Philip A. Schwartzkroin
Keyword(s):  
Voltage Clamp ◽  
Hippocampal Slices ◽  
Cell Voltage ◽  
Potassium Currents ◽  
Membrane Currents ◽  
Excitatory Postsynaptic Currents ◽  
Whole Cell ◽  
Mossy Cells ◽  
Postsynaptic Currents

Baraban, Scott C. and Philip A. Schwartzkroin. Effects of hyposmolar solutions on membrane currents of hippocampal interneurons and mossy cells in vitro. J. Neurophysiol. 79: 1108–1112, 1998. Whole cell voltage-clamp recordings in rat hippocampal slices were used to investigate the effect of changes in extracellular osmolarity on voltage-activated potassium currents. Currents were evoked from oriens/alveus (O/A) interneurons, hilar interneurons, and mossy cells. Hyposmolar external solutions produced a significant potentiation of K+ current recorded from O/A and hilar interneurons, but not from mossy cells. Hyposmolar solutions also dramatically potentiated the spontaneous excitatory postsynaptic currents recorded from mossy cells. These results suggest that hippocampal excitability can be modulated by the complex actions exerted by changes in extracellular osmolarity.

Enhancement of Whole Cell Synaptic Currents by Low Osmolarity and by Low [NaCl] in Rat Hippocampal Slices

1997 ◽  
Vol 77 (5) ◽  
pp. 2349-2359 ◽  
Author(s):  
Rong Huang ◽  
Daniel F. Bossut ◽  
George G. Somjen
Keyword(s):  
Synaptic Transmission ◽  
Time Constant ◽  
Hippocampal Slices ◽  
Input Resistance ◽  
Tissue Slices ◽  
Synaptic Currents ◽  
Whole Cell ◽  
Input Capacitance ◽  
Postsynaptic Currents ◽  
Inward Currents

Huang, Rong, Daniel F. Bossut, and George G. Somjen. Enhancement of whole cell synaptic currents by low osmolarity and by low [NaCl] in rat hippocampal slices. J. Neurophysiol. 77: 2349–2359, 1997. We recorded whole cell currents of patch-clamped neurons in stratum pyramidale of CA1 region of rat hippocampal tissue slices. Synaptic currents were evoked by orthodromic stimulation while holding potential of the neuron was varied from hyperpolarized to depolarized levels. Extracellular osmolarity (πo) was lowered by superfusion with artificial cerebrospinal fluid in which NaCl concentration ([NaCl]) was reduced. The effect of low extracellular NaCl was tested in additional trials in which NaCl was substituted by isosmolar fructose. Both lowering of πo and isosmotic lowering of extracellular [NaCl] ([NaCl]o) caused reversible increase of excitatory postsynaptic currents. The effect of lowering πo was concentration dependent, and it was significantly stronger than the effect of equivalent isosmotic lowering of [NaCl]o. Inhibitory postsynaptic currents also increased in many but not in all cases. Lowering of πo caused a prolongation of the time constant of relaxation of the capacitive charging current induced by small hyperpolarizing voltage steps. A virtual input capacitance, calculated by dividing this time constant by the input resistance, increased during hypotonic exposure. Isosmotic lowering of [NaCl]o had no effect on time constant or input capacitance. Depolarizing voltage commands evoked spikelike inward currents presumably representing Na+-dependent action potentials generated outside the voltage-clamped region of the cell. These current spikes became smaller in low πo and in low [NaCl]o. Broader, voltage-dependent, presumably Ca2+-mediated inward currents became more prominent during hypotonic exposure. We conclude that lowering of [NaCl]o causes enhancement of excitatory synaptic transmission. Transmission may be facilitated by the uptake of Ca2+ into presynaptic terminals as well as into postsynaptic target neurons, induced by the low [NaCl]o. Lowering of πo enhances synaptic transmission more than does a corresponding isosmotic lowering of [NaCl]. The excess increase recorded from the cell soma in low πo may in part be due to changing electrotonic length caused by the swelling of dendrites.

Synaptically Evoked GABA Transporter Currents in Neocortical Glia

2002 ◽  
Vol 88 (6) ◽  
pp. 2899-2908 ◽  
Author(s):  
Gregory A. Kinney ◽  
William J. Spain
Keyword(s):  
Synaptic Transmission ◽  
Time Course ◽  
Glutamate Transporter ◽  
Repetitive Stimulation ◽  
Slice Preparation ◽  
Gaba Transporter ◽  
Gaba Transporters ◽  
Decay Times ◽  
Slow Rise

The presence, magnitude, and time course of GABA transporter currents were investigated in electrophysiologically characterized neocortical astrocytes in an in vitro slice preparation. On stimulation with a bipolar-tungsten stimulating electrode placed nearby, the majority of cells tested displayed long-lasting GABA transporter currents using both single and repetitive stimulation protocols. Using subtype-specific GABA transporter antagonists, long-lasting GABA transporter currents were identified in neocortical astrocytes that originated from at least two subtypes of GABA transporters: GAT-1 and GAT-2/3. These transporter currents displayed slow rise times and long decay times, contrasting the time course observed for glutamate transporter currents, and are indicative of a long extracellular time course of GABA as well as a role for glial GABA transporters during synaptic transmission.

scholarly journals Transducin translocation contributes to rod survival and enhances synaptic transmission from rods to rod bipolar cells

2013 ◽  
Vol 110 (30) ◽  
pp. 12468-12473 ◽  
Author(s):  
A. Majumder ◽  
J. Pahlberg ◽  
K. K. Boyd ◽  
V. Kerov ◽  
S. Kolandaivelu ◽  
...  
Keyword(s):  
Synaptic Transmission ◽  
Bipolar Cells ◽  
Rod Bipolar Cells

scholarly journals Different Contributions of GABAA and GABAC Receptors to Rod and Cone Bipolar Cells in a Rat Retinal Slice Preparation

1998 ◽  
Vol 79 (3) ◽  
pp. 1384-1395 ◽  
Author(s):  
Thomas Euler ◽  
Heinz Wässle
Keyword(s):  
Receptor Antagonist ◽  
Glutamate Receptors ◽  
Metabotropic Glutamate Receptors ◽  
Phosphinic Acid ◽  
Bipolar Cells ◽  
Resting Potential ◽  
Slice Preparation ◽  
Metabotropic Glutamate ◽  
Rod Bipolar Cells ◽  
Cone Bipolar Cells

Euler, Thomas and Heinz Wässle. Different contributions ofGABAA and GABAC receptors to rod and cone bipolar cells in a rat retinal slice preparation. J. Neurophysiol. 79: 1384–1395, 1998. Whole cell currents were recorded from rod and cone bipolar cells in a slice preparation of the rat retina. Use of the gramicidin D perforated-patch technique prevented loss of intracellular compounds. The recorded cells were identified morphologically by injection with Lucifer yellow. During the recordings, the cells were isolated synaptically by extracellular cobalt. To distinguish the γ-aminobutyric acid (GABA) receptors pharmacologically, theGABAA receptor antagonist, bicuculline, and the GABAC receptor antagonist, 3-aminopropyl(methyl)phosphinic acid, were used. In all bipolar cells tested, application of GABA induced postsynaptic chloride currents that hyperpolarized the cells from their resting potential of about −40 mV. GABA was applied at different concentrations to allow for the different affinity of GABA at GABAA and GABAC receptors. At a GABA concentration of 25 μM, in the case of rod bipolar cells, ∼70% of the current was found to be mediated by GABAC receptors. In the case of cone bipolar cells, only ∼20% of the current was mediated by GABAC receptors. Furthermore, this GABAC-mediated fraction varied among the different morphological types of cone bipolar cells, supporting the hypothesis of distinct functional roles for the different types of cone bipolar cells. There is evidence that the efficacy of GABAC receptors is modulated by glutamate through metabotropic glutamate receptors. We tested this hypothesis by applying agonists of metabotropic glutamate receptors (mGluR)1/5 to rod bipolar cells. The specific agonist (±)-trans-azetidine-2,4-dicarboxylic acid and the potent mGluR agonist quisqualic acid reduced the amplitude of the GABAC responses by 10–30%. This suggests a functional role for the modulation of GABAC receptors by the metabotropic glutamate receptors mGluR1/5.

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