Dendritic Action Potentials Research Articles

Cellular Mechanisms for Direction Selectivity in the Retina

Direction selectivity represents a fundamental computation found across multiple sensory systems. In the mammalian visual system, direction selectivity appears first in the retina, where excitatory and inhibitory interneurons release neurotransmitter most rapidly during movement in a preferred direction. Two parallel sets of interneuron signals are integrated by a direction-selective ganglion cell, which creates a direction preference for both bright and dark moving objects. Direction selectivity of synaptic input becomes amplified by action potentials in the ganglion cell dendrites. Recent work has elucidated direction-selective mechanisms in inhibitory circuitry, but mechanisms in excitatory circuitry remain unexplained.

Intracellular long-wavelength voltage-sensitive dyes for studying the dynamics of action potentials in axons and thin dendrites

In CNS neurons most of synaptic integration takes place in thin dendritic branches that are difficult to study with conventional physiological recording techniques (electrodes). When cellular compartments are too small, or too many, for electrode recordings, optical methods bring considerable advantages. Here we focused our experimental effort on the development and utilization of new kinds of voltage-sensitive dyes (VSD). The new VSDs have bluish appearance in organic solvents, and hence are dubbed “blue dyes”. They have preferred excitation windows for voltage recording that are shifted to longer wavelengths (∼660nm). Excitation in deep red light and emission in the near-infrared render “blue VSDs” potentially useful in measurements from fluorescent structures below the tissue surface because light scattering is minimized at longer wavelengths. Seven new molecules were systematically tested using intracellular injection. In comparison to the previously used red dye (JPW-3028) the blue dyes have better sensitivity (ΔF/F) by approximately 40%. Blue dyes take little time to fill the dendritic tree, and in this aspect they are comparable with the fastest red dye JPW-3028. Based on our results, blue VSDs are well suited for experimental exploration of thin neuronal processes in semi intact preparations (brain slice). In some cases only six sweeps of temporal averaging were needed to acquire excellent records of individual action potentials in basal and oblique dendritic branches, or in axons and axon collaterals up to 200μm away from the cell body. Signal-to-noise ratio of these recordings was ∼10. The combination of blue dyes and laser illumination approach imposed little photodynamic damage and allowed the total number of recording sweeps per cell to exceed 100. Using these dyes and a spot laser illumination technique, we demonstrate the first recording of action potentials in the oblique dendrite and distal axonal segment of the same pyramidal cell.

Dendritic signals from rat hippocampal CA1 pyramidal neurons during coincident pre‐ and post‐synaptic activity: a combined voltage‐ and calcium‐imaging study

The non-linear and spatially inhomogeneous interactions of dendritic membrane potential signals that represent the first step in the induction of activity-dependent long-term synaptic plasticity are not fully understood, particularly in dendritic regions which are beyond the reach of electrode measurements. We combined voltage-sensitive-dye recordings and Ca(2+) imaging of hippocampal CA1 pyramidal neurons to study large regions of the dendritic arbor, including branches of small diameter (distal apical and oblique dendrites). Dendritic membrane potential transients were monitored at high spatial resolution and correlated with supra-linear [Ca(2+)](i) changes during one cycle of a repetitive patterned stimulation protocol that typically results in the induction of long-term potentiation (LTP). While the increase in the peak membrane depolarization during coincident pre- and post-synaptic activity was required for the induction of supra-linear [Ca(2+)](i) signals shown to be necessary for LTP, the change in the baseline-to-peak amplitude of the backpropagating dendritic action potential (bAP) was not critical in this process. At different dendritic locations, the baseline-to-peak amplitude of the bAP could be either increased, decreased or unaltered at sites where EPSP-AP pairing evoked supra-linear summation of [Ca(2+)](i) transients. We suggest that modulations in the bAP baseline-to-peak amplitude by local EPSPs act as a mechanism that brings the membrane potential into the optimal range for Ca(2+) influx through NMDA receptors (0 to -15 mV); this may require either boosting or the reduction of the bAP, depending on the initial size of both signals.

Etomidate Reduces Initiation of Backpropagating Dendritic Action Potentials: Implications for Sensory Processing and Synaptic Plasticity During Anesthesia

Anesthetics may induce specific changes that alter the balance of activity within neural networks. Here we describe the effects of the GABA(A) receptor potentiating anesthetic etomidate on sensory processing, studied in a cerebellum-like structure, the electrosensory lateral line lobe (ELL) of mormyrid fish, in vitro. Previous studies have shown that the ELL integrates sensory input and removes predictable features by comparing reafferent sensory signals with a descending electromotor command-driven corollary signal that arrives in part through parallel fiber synapses with the apical dendrites of GABAergic interneurons. These synapses show spike timing-dependent depression when presynaptic activation is associated with postsynaptic backpropagating dendritic action potentials. Under etomidate, almost all neurons become tonically hyperpolarized. The threshold for action potential initiation increased for both synaptic activation and direct intracellular depolarization. Synaptically evoked inhibitory postsynaptic potentials (IPSPs) were also strongly potentiated and prolonged. Current source density analysis showed that backpropagation of action potentials through the apical dendritic arborization in the molecular layer was reduced but could be restored by increasing stimulus strength. These effects of etomidate were blocked by bicuculline or picrotoxin. It is concluded that etomidate affects both tonic and phasic inhibitory conductances at GABA(A) receptors and that increased shunting inhibition at the level of the proximal dendrites also contributes to increasing the threshold for action potential backpropagation. When stimulus strength is sufficient to evoke backpropagation, repetitive association of synaptic excitation with postsynaptic action potential initiation still results in synaptic depression, showing that etomidate does not interfere with the molecular mechanism underlying plastic modulation.

The Synaptic Mechanism of Direction Selectivity in Distal Processes of Starburst Amacrine Cells

Patch-clamp recordings revealed that distal processes of starburst amacrine cells (SACs) received largely excitatory synaptic input from the receptive field center and nearly purely inhibitory inputs from the surround during both stationary and moving light stimulations. The direct surround inhibition was mediated mainly by reciprocal GABA(A) synapses between opposing SACs, which provided leading and prolonged inhibition during centripetal stimulus motion. Simultaneous Ca(2+) imaging and current-clamp recording during apparent-motion stimulation further demonstrated the contributions of both centrifugal excitation and GABA(A/C)-receptor-mediated centripetal inhibition to the direction-selective Ca(2+) responses in SAC distal processes. Thus, by placing GABA release sites in electrotonically semi-isolated distal processes and endowing these sites with reciprocal GABA(A) synapses, SACs use a radial-symmetric center-surround receptive field structure to build a polar-asymmetric circuitry. This circuitry may integrate at least three levels of interactions--center excitation, surround inhibition, and reciprocal inhibitions that amplify the center--surround antagonism-to generate robust direction selectivity in the distal processes.

Calcium Spikes in Basal Dendrites of Layer 5 Pyramidal Neurons during Action Potential Bursts

Patch-clamp recording from dendrites has lead to a significant increase in our understanding of the mechanisms underlying signal integration and propagation in neurons. The majority of synaptic input to neurons, however, is made onto small-diameter dendrites, currently beyond the scope of patch-clamp recording techniques. Here we use both calcium and voltage imaging to investigate propagation of action potentials (APs) in fine basal dendrites of cortical layer 5 pyramidal neurons. High-frequency (200 Hz) AP bursts caused supralinear increases in dendritic calcium at distal, but not proximal, basal locations. Supralinear increases in dendritic calcium were also observed at distal basal locations during AP trains above a critical frequency (approximately 100 Hz). Using voltage imaging, we show that single APs undergo significant attenuation as they propagate into basal dendrites, whereas AP bursts lead to generation of dendritic calcium spikes. Focal and bath application of 4-AP increased the amplitude of calcium transients evoked by APs at distal, but not proximal, locations, suggesting that A-type potassium channels regulate AP backpropagation into basal dendrites. Finally, we show that pairing EPSPs with AP bursts is an effective means of activating synaptic NMDA receptors in basal dendrites. The experimental observations on the role of A-type potassium channels in regulation of AP backpropagation in basal dendrites, as well as the generation of dendritic calcium spikes during AP bursts, were reproduced in a morphologically realistic neuronal model with uniform distributions of dendritic sodium, calcium, and potassium channels. Together, these findings have important implications for understanding dendritic integration and synaptic plasticity in cortical basal dendrites.

BDNF enhances dendritic Ca 2+ signals evoked by coincident EPSPs and back-propagating action potentials in CA1 pyramidal neurons

BDNF, a member of the neurotrophin family, is emerging as a key modulator of synaptic structure and function in the CNS. Due to the critical role of postsynaptic Ca 2+ signals in dendritic development and synaptic plasticity, we tested whether long-term exposure to BDNF affects Ca 2+ elevations evoked by coincident excitatory postsynaptic potentials (EPSPs) and back-propagating action potentials (bAPs) in spiny dendrites of CA1 pyramidal neurons within hippocampal slice cultures. In control neurons, a train of 5 coincident EPSPs and bAPs evoked Ca 2+ elevations in oblique radial branches of the main apical dendrite that were of similar amplitude than those evoked by a train of 5 bAPs alone. On the other hand, dendritic Ca 2+ signals evoked by coincident EPSPs and bAPs were always larger than those triggered by bAPs in CA1 neurons exposed to BDNF for 48 h. This difference was not observed after blockade of NMDA receptors (NMDARs) with D,L-APV, but only in BDNF-treated neurons, suggesting that Ca 2+ signals in oblique radial dendrites include a synaptic NMDAR-dependent component. Co-treatment with the receptor tyrosine kinase inhibitor k-252a prevented the effect of BDNF on coincident dendritic Ca 2+ signals, suggesting the involvement of neurotrophin Trk receptors. These results indicate that long-term exposure to BDNF enhances Ca 2+ signaling during coincident pre- and postsynaptic activity in small spiny dendrites of CA1 pyramidal neurons, representing a potential functional consequence of neurotrophin-mediated dendritic remodeling in developing neurons.

Voltage-gated ion channels in dendrites of hippocampal pyramidal neurons

The properties and distribution of voltage-gated ion channels contribute to electrical signaling in neuronal dendrites. The apical dendrites of CA1 pyramidal neurons in hippocampus express a wide variety of sodium, calcium, potassium, and other voltage-gated channels. In this report, we provide some new evidence for the role of the delayed-rectifier K(+) channel in shaping the dendritic action potential at different membrane potentials.

The GABAB1b Isoform Mediates Long-Lasting Inhibition of Dendritic Ca2+ Spikes in Layer 5 Somatosensory Pyramidal Neurons

The apical tuft of layer 5 pyramidal neurons is innervated by a large number of inhibitory inputs with unknown functions. Here, we studied the functional consequences and underlying molecular mechanisms of apical inhibition on dendritic spike activity. Extracellular stimulation of layer 1, during blockade of glutamatergic transmission, inhibited the dendritic Ca2+ spike for up to 400 ms. Activation of metabotropic GABAB receptors was responsible for a gradual and long-lasting inhibitory effect, whereas GABAA receptors mediated a short-lasting (approximately 150 ms) inhibition. Our results suggest that the mechanism underlying the GABAB inhibition of Ca2+ spikes involves direct blockade of dendritic Ca2+ channels. By using knockout mice for the two predominant GABAB1 isoforms, GABAB1a and GABAB1b, we showed that postsynaptic inhibition of Ca2+ spikes is mediated by GABAB1b, whereas presynaptic inhibition of GABA release is mediated by GABAB1a. We conclude that the molecular subtypes of GABAB receptors play strategically different physiological roles in neocortical neurons.

Direction-Selective Dendritic Action Potentials in Rabbit Retina

Dendritic spikes that propagate toward the soma are well documented, but their physiological role remains uncertain. Our in vitro patch-clamp recordings and two-photon calcium imaging show that direction-selective retinal ganglion cells (DSGCs) utilize orthograde dendritic spikes during physiological activity. DSGCs signal the direction of image motion. Excitatory subthreshold postsynaptic potentials are observed in DSGCs for motion in all directions and provide a weakly tuned directional signal. However, spikes are generated over only a narrow range of motion angles, indicating that spike generation greatly enhances directional tuning. Our results indicate that spikes are initiated at multiple sites within the dendritic arbors of DSGCs and that each dendritic spike initiates a somatic spike. We propose that dendritic spike failure, produced by local inhibitory inputs, might be a critical factor that enhances directional tuning of somatic spikes.

Inhibitory glutamate receptors in spider peripheral mechanosensory neurons

Most mechanosensory neurons are inhibited by GABAergic efferent neurons. This inhibition is often presynaptic and mediated by ionotropic GABA receptors at the axon terminals. GABA receptor activation opens Cl- channels, leading to membrane depolarization and an increase in membrane conductance. In many invertebrate preparations, efferent neurons that innervate mechanosensory afferents contain glutamate in addition to GABA, suggesting that the sensory neurons are also modulated by glutamate. However, the effects of glutamate on these neurons are not well understood. Peripheral parts of the spider (Cupiennius salei) mechanosensory neurons are surrounded by efferent fibers immunoreactive to antibodies against GABA and glutamate. GABA and its analogue muscimol were shown to effectively inhibit spider mechanosensory neurons innervating lyriform slit sensilla VS-3 that detects cuticular strains in the leg. Here, we show that glutamate also inhibits the VS-3 neurons, but its effects are different from those of GABA or muscimol, suggesting that it acts on a different group of receptors. GABA and muscimol always depolarized these neurons and the inhibitory effect was strongly correlated with the amount of depolarization. In contrast, glutamate inhibited the VS-3 neurons even when it did not depolarize them. In addition, while glutamate inhibited both the axonal action potentials elicited with electrical stimulation and dendritic action potentials produced by mechanical stimulation, muscimol only inhibited the axonal action potentials. Therefore, the inhibitory glutamate receptors in the VS-3 neurons are distinct from and differently distributed than the GABA receptors, providing a subtle control of the neurons' sensitivity in varying behavioural situations.

Effect of dendritic backpropagating action potential on neural interaction

We elucidate the effect of dendritic backpropagating action potentials on neural interactions. We propose a neural oscillator model that mimics the backpropagating potentials upon the dendrite. We show that backpropagating potentials change the stability structure of the system from monostable to bistable. Such bistability causes the multiple phase clustering of the neural population. The phase clustering is a cogent hypothesis to explain the information representation binding distributed codes in the brain.

Action Potential Backpropagation and Multiglomerular Signaling in the Rat Vomeronasal System

In the accessory olfactory bulb (AOB), sensory neurons expressing a given vomeronasal receptor (VR) gene send divergent projections to many glomeruli, and second-order neurons (mitral cells) link to multiple glomeruli via branched primary dendrites. We used calcium imaging and paired somadendritic patch-clamp recording to track backpropagated action potentials (APs) in rat AOB primary dendrites. In cells loaded with 150 microm Calcium Orange, somatic spikes elicited fluorescence transients over the entire primary dendritic tree, and the relative fluorescence increment DeltaF/F(0) increased along all branches from soma to glomeruli. Backpropagation was reliant on Na+ channels: in 1 microm TTX, somatic AP commands evoked dendritic Ca2+ transients that declined steeply with distance. In paired soma- dendritic whole-cell recordings, backpropagated APs were unattenuated up to approximately 200 microm from the soma, whereas subthreshold voltage transients decayed markedly. Computational modeling indicated that the large distal Ca2+ transients are consistent with active, not passive, backpropagation. Genetic tracing in the AOB has suggested homotypic connectivity with individual mitral cell dendritic arbors projecting only to glomeruli targeted by sensory neurons expressing the same VR gene. Non-decremental, non-dichotomous backpropagation in AOB primary dendrites ensures fast, reliable communication between mitral cells and their homotypic glomeruli, binding them into functional modules in accordance with their VR-coded inputs.

Multiple spatiotemporal patterns of dendritic Ca 2+ signals in goldfish retinal amacrine cells

Although it has been reported that dendritic neurotransmitter releases from amacrine cells are regulated by the intracellular Ca 2+ concentration ([Ca 2+] i), their spatiotemporal patterns are not well explained. Fast Ca 2+ imagings of amacrine cells in the horizontal slice preparation of goldfish retinas under whole-cell patch-clamp recordings were undertaken to better investigate the spatiotemporal patterns of dendritic [Ca 2+] i. We found that amacrine cell dendrites showed inhomogeneous [Ca 2+] i increases in both Na + spiking cells and cells without Na + spikes. The spatiotemporal properties of inhomogeneous [Ca 2+] i increases were classified into three patterns: local, regional and global. Local [Ca 2+] i increases were observed in very discrete regions and appeared as discontinuous patches, presumably evoked by local excitatory postsynaptic potentials. Regional [Ca 2+] i increases were observed in either a single or a small number of dendrites, presumably reflecting the result of dendritic action potentials. Global [Ca 2+] i increases were observed in the entire dendrites of a cell and were mediated by Na + action potentials or multiple Na + action potentials riding on slow depolarization. Ca 2+-mediated potentials also evoked global [Ca 2+] i increase in cells without Na + spikes. These spatiotemporal dynamics of dendritic Ca 2+ signals may reflect multiple modes of synaptic integration on the dendrites of amacrine cells.

Epilepsy and Dendritic Ion Channels

Little is known about the changes in ion channel function in the neuronal membrane after a region of the brain has become epileptic. Bernard et al . (see the Perspective by Staley) show that the electrical properties of hippocampal pyramidal cell dendrites are altered in a rat model of temporal lobe epilepsy. Back-propagation of dendritic action potentials is enhanced in these animals because of a loss of A-type K + channel function. This change appears ultimately to alter the input-output characteristics of the dendrites that enhance excitability and susceptibility to seizures. C. Bernard, A. Anderson, A. Becker, N. P. Poolos, H. Beck, D. Johnston, Acquired dendritic channelopathy in temporal lobe epilepsy. Science 305 , 532-535 (2004). [Abstract] [Full Text] K. Staley, Epileptic neurons go wireless. Science 305 , 482-483 (2004). [Summary] [Full Text]

Targeting mechanisms of high voltage-activated Ca2+ channels.

Functional voltage-dependent Ca2+ channel complexes are assembled by three to four subunits: alpha1, beta, alpha2delta subunits (C. Leveque et al., 1994, J. Biol Chem. 269, 6306-6312; M. W. McEnery et al., 1991, Proc. Natl. Acad. Sci. U.S.A. 88, 11095-11099) and at least in muscle cells also y subunits (B. M. Curtis and W. A. Catterall, 1984, Biochemistry 23, 2113-2118). Ca2+ channels mediate the voltage-dependent Ca2+ influx in subcellular compartments, triggering such diverse processes as neurotransmitter release, dendritic action potentials, excitation-contraction, and excitation-transcription coupling. The targeting of biophysically defined Ca2+ channel complexes to the correct subcellular structures is, thus, critical to proper cell and physiological functioning. Despite their importance, surprisingly little is known about the targeting mechanisms by which Ca2+ channel complexes are transported to their site of function. Here we summarize what we know about the targeting of Ca2+ channel complexes through the cell to the plasma membrane and subcellular structures.

Ca2+ imaging of mouse neocortical interneurone dendrites: Ia-type K+ channels control action potential backpropagation

GABAergic interneurones are essential in cortical processing, yet the functional properties of their dendrites are still poorly understood. In this first study, we combined two-photon calcium imaging with whole-cell recording and anatomical reconstructions to examine the calcium dynamics during action potential (AP) backpropagation in three types of V1 supragranular interneurones: parvalbumin-positive fast spikers (FS), calretinin-positive irregular spikers (IS), and adapting cells (AD). Somatically generated APs actively backpropagated into the dendritic tree and evoked instantaneous calcium accumulations. Although voltage-gated calcium channels were expressed throughout the dendritic arbor, calcium signals during backpropagation of both single APs and AP trains were restricted to proximal dendrites. This spatial control of AP backpropagation was mediated by Ia-type potassium currents and could be mitigated by by previous synaptic activity. Further, we observed supralinear summation of calcium signals in synaptically activated dendritic compartments. Together, these findings indicate that in interneurons, dendritic AP propagation is synaptically regulated. We propose that interneurones have a perisomatic and a distal dendritic functional compartment, with different integrative functions.

Ca2+ imaging of mouse neocortical interneurone dendrites: Ia-type K+ channels control action potential backpropagation.

GABAergic interneurones are essential in cortical processing, yet the functional properties of their dendrites are still poorly understood. In this first study, we combined two-photon calcium imaging with whole-cell recording and anatomical reconstructions to examine the calcium dynamics during action potential (AP) backpropagation in three types of V1 supragranular interneurones: parvalbumin-positive fast spikers (FS), calretinin-positive irregular spikers (IS), and adapting cells (AD). Somatically generated APs actively backpropagated into the dendritic tree and evoked instantaneous calcium accumulations. Although voltage-gated calcium channels were expressed throughout the dendritic arbor, calcium signals during backpropagation of both single APs and AP trains were restricted to proximal dendrites. This spatial control of AP backpropagation was mediated by Ia-type potassium currents and could be mitigated by by previous synaptic activity. Further, we observed supralinear summation of calcium signals in synaptically activated dendritic compartments. Together, these findings indicate that in interneurons, dendritic AP propagation is synaptically regulated. We propose that interneurones have a perisomatic and a distal dendritic functional compartment, with different integrative functions.

Regulation of backpropagating action potentials in mitral cell lateral dendrites by A-type potassium currents.

Dendrodendritic synapses, distributed along mitral cell lateral dendrites, provide powerful and extensive inhibition in the olfactory bulb. Activation of inhibition depends on effective penetration of action potentials into dendrites. Although action potentials backpropagate with remarkable fidelity in apical dendrites, this issue is controversial for lateral dendrites. We used paired somatic and dendritic recordings to measure action potentials in proximal dendritic segments (0-200 microm from soma) and action potential-generated calcium transients to monitor activity in distal dendritic segments (200-600 microm from soma). Somatically elicited action potentials were attenuated in proximal lateral dendrites. The attenuation was not due to impaired access resistance in dendrites or to basal synaptic activity. However, a single somatically elicited action potential was sufficient to evoke a calcium transient throughout the lateral dendrite, suggesting that action potentials reach distal dendritic compartments. Block of A-type potassium channels (I(A)) with 4-aminopyridine (10 mM) prevented action potential attenuation in direct recordings and significantly increased dendritic calcium transients, particularly in distal dendritic compartments. Our results suggest that I(A) may regulate inhibition in the olfactory bulb by controlling action potential amplitudes in lateral dendrites.

Solution Structure and Function of the “Tandem Inactivation Domain” of the Neuronal A-type Potassium Channel Kv1.4

Cumulative inactivation of voltage-gated (Kv) K(+) channels shapes the presynaptic action potential and determines timing and strength of synaptic transmission. Kv1.4 channels exhibit rapid "ball-and-chain"-type inactivation gating. Different from all other Kvalpha subunits, Kv1.4 harbors two inactivation domains at its N terminus. Here we report the solution structure and function of this "tandem inactivation domain" using NMR spectroscopy and patch clamp recordings. Inactivation domain 1 (ID1, residues 1-38) consists of a flexible N terminus anchored at a 5-turn helix, whereas ID2 (residues 40-50) is a 2.5-turn helix made up of small hydrophobic amino acids. Functional analysis suggests that only ID1 may work as a pore-occluding ball domain, whereas ID2 most likely acts as a "docking domain" that attaches ID1 to the cytoplasmic face of the channel. Deletion of ID2 slows inactivation considerably and largely impairs cumulative inactivation. Together, the concerted action of ID1 and ID2 may promote rapid inactivation of Kv1.4 that is crucial for the channel function in short term plasticity.