Combining pharmacology and whole-cell patch recording from CNS neurons, in vivo

Rose, Garrett S.; Alluri, Rishi K.; Vasquez-Opazo, Gustavo A.; Odom, Stephen E.; Graham, Jalina A.; Leary, Christopher J.

doi:10.1016/j.jneumeth.2012.12.003

Cited by 12 publications

(7 citation statements)

References 19 publications

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“…The time courses and magnitudes of conductances should provide a better understanding of how excitation and inhibition are integrated to generate responses selectively to particular temporal parameters of sounds. A particularly powerful and promising direction for future work is to perform whole-cell recording in conjunction with iontophoretic application of pharmacological agents (Rose et al, 2013 ). In this methodology, glutamate iontophoresis is used to position a multibarrel pipette close to the neuron from which whole-cell recordings are being made.…”

Section: Discussionmentioning

confidence: 99%

Time computations in anuran auditory systems

Rose

2014

Front. Physiol.

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Temporal computations are important in the acoustic communication of anurans. In many cases, calls between closely related species are nearly identical spectrally but differ markedly in temporal structure. Depending on the species, calls can differ in pulse duration, shape and/or rate (i.e., amplitude modulation), direction and rate of frequency modulation, and overall call duration. Also, behavioral studies have shown that anurans are able to discriminate between calls that differ in temporal structure. In the peripheral auditory system, temporal information is coded primarily in the spatiotemporal patterns of activity of auditory-nerve fibers. However, major transformations in the representation of temporal information occur in the central auditory system. In this review I summarize recent advances in understanding how temporal information is represented in the anuran midbrain, with particular emphasis on mechanisms that underlie selectivity for pulse duration and pulse rate (i.e., intervals between onsets of successive pulses). Two types of neurons have been identified that show selectivity for pulse rate: long-interval cells respond well to slow pulse rates but fail to spike or respond phasically to fast pulse rates; conversely, interval-counting neurons respond to intermediate or fast pulse rates, but only after a threshold number of pulses, presented at optimal intervals, have occurred. Duration-selectivity is manifest as short-pass, band-pass or long-pass tuning. Whole-cell patch recordings, in vivo, suggest that excitation and inhibition are integrated in diverse ways to generate temporal selectivity. In many cases, activity-related enhancement or depression of excitatory or inhibitory processes appear to contribute to selective responses.

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Section: Discussionmentioning

confidence: 99%

Time computations in anuran auditory systems

Rose

2014

Front. Physiol.

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“…Approximately 50-100 nA constant current was used to iontophoretically deliver pharmacological compounds (negative for glutamate, NBQX, and CPP and positive for other agents). As described previously (43), two electrodes were concurrently used in these experiments. We iontophoresed glutamate to activate recorded cells, thereby evaluating whether the multibarrel pipette was sufficiently close to the recording pipette for conducting pharmacological manipulations.…”

Section: Methodsmentioning

confidence: 99%

Phasic, suprathreshold excitation and sustained inhibition underlie neuronal selectivity for short-duration sounds

Alluri

Rose

Hanson

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Sound duration is important in acoustic communication, including speech recognition in humans. Although duration-selective auditory neurons have been found, the underlying mechanisms are unclear.To investigate these mechanisms we combined in vivo whole-cell patch recordings from midbrain neurons, extraction of excitatory and inhibitory conductances, and focal pharmacological manipulations. We show that selectivity for short-duration stimuli results from integration of short-latency, sustained inhibition with delayed, phasic excitation; active membrane properties appeared to amplify responses to effective stimuli. Blocking GABA A receptors attenuated stimulus-related inhibition, revealed suprathreshold excitation at all stimulus durations, and decreased short-pass selectivity without changing resting potentials. Blocking AMPA and NMDA receptors to attenuate excitation confirmed that inhibition tracks stimulus duration and revealed no evidence of postinhibitory rebound depolarization inherent to coincidence models of duration selectivity. These results strongly support an anticoincidence mechanism of short-pass selectivity, wherein inhibition and suprathreshold excitation show greatest temporal overlap for long duration stimuli.whole-cell | GABA | synaptic conductance | gabazine | inferior colliculus A principal goal in neuroscience is to understand the computational mechanisms that underlie selectivity for particular types of information. Sensory neurons have been identified that show selectivity for biologically relevant stimulus features; however, the underlying mechanisms are, in most cases, poorly understood. A notable exception involves mechanisms of selectivity for sound duration, particularly in bat and anuran auditory systems (1-3). Duration-selective neurons were first found in the midbrain torus semicircularis of anurans (4, 5), homolog of the mammalian inferior colliculus (IC) and referred to here as the IC an . In anurans and many mammalian species, midbrain neurons have been identified that show short-pass, band-pass, or long-pass duration selectivity (3, 6-11). These neurons code for temporal properties of acoustic signals that are important in communication and echolocation. Processing sound duration is also critical for human speech communication, and deficiencies in the neural processing of this and other temporal information feature prominently in disorders of speech recognition (12, 13). Hence, understanding the mechanisms of duration selectivity is of considerable importance.Several models of duration selectivity have been proposed. The first model, derived from extracellular recordings in the anuran IC (4), incorporates delayed, onset excitation and shortlatency offset excitation that coincide for the optimal stimulus duration (Fig. 1A). Subsequent extracellular recordings from IC neurons in bats showed that blocking receptors of inhibitory neurotransmitters eliminated, or greatly attenuated, duration selectivity (14-16). Further, the temporal pattern of discharges shifted from offset to onse...

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“…This technique has been successfully used to investigate glutamatergic and GABAergic synaptic transmission in different brain regions in vitro and in vivo 9,[20][21][22] . Micro-iontophoresis has been used for more than 60 years, but in early years it was mostly used to either locally apply neurotransmitters and drugs on slow or intermediate timescales 23 or for microinjection of substances into cells 24 .…”

Section: Discussionmentioning

confidence: 99%

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

Müller

Remy

2013

JoVE

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Citation: Müller, C., Remy, S. Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration. J. Vis. Exp. (77), e50701, doi:10.3791/50701 (2013). AbstractOne of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of the dendritic tree, which eventually leads to neuronal output of action potentials at the axon. The influence of diverse spatial and temporal parameters of specific synaptic input on neuronal output is currently under investigation, e.g. the distance-dependent attenuation of dendritic inputs, the location-dependent interaction of spatially segregated inputs, the influence of GABAergig inhibition on excitatory integration, linear and non-linear integration modes, and many more.With fast micro-iontophoresis of glutamate and GABA it is possible to precisely investigate the spatial and temporal integration of glutamatergic excitation and GABAergic inhibition. Critical technical requirements are either a triggered fluorescent lamp, light-emitting diode (LED), or a twophoton scanning microscope to visualize dendritic branches without introducing significant photo-damage of the tissue. Furthermore, it is very important to have a micro-iontophoresis amplifier that allows for fast capacitance compensation of high resistance pipettes. Another crucial point is that no transmitter is involuntarily released by the pipette during the experiment.Once established, this technique will give reliable and reproducible signals with a high neurotransmitter and location specificity. Compared to glutamate and GABA uncaging, fast iontophoresis allows using both transmitters at the same time but at very distant locations without limitation to the field of view. There are also advantages compared to focal electrical stimulation of axons: with micro-iontophoresis the location of the input site is definitely known and it is sure that only the neurotransmitter of interest is released. However it has to be considered that with micro-iontophoresis only the postsynapse is activated and presynaptic aspects of neurotransmitter release are not resolved. In this article we demonstrate how to set up micro-iontophoresis in brain slice experiments. Video LinkThe video component of this article can be found at

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Combining pharmacology and whole-cell patch recording from CNS neurons, in vivo

Cited by 12 publications

References 19 publications

Time computations in anuran auditory systems

Time computations in anuran auditory systems

Phasic, suprathreshold excitation and sustained inhibition underlie neuronal selectivity for short-duration sounds

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

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