Ruili Xie

Rethinking Tuning: In Vivo Whole-Cell Recordings of the Inferior Colliculus in Awake Bats

Tuning curves were recorded with patch electrodes from the inferior colliculus (IC) of awake bats to evaluate the tuning of the inputs to IC neurons, reflected in their synaptic tuning, compared with the tuning of their outputs, expressed in their discharge tuning. A number of unexpected features were revealed with whole-cell recordings. Among these was that most neurons responded to tones with inhibition and/or subthreshold excitation over a surprisingly broad frequency range. The synaptic tuning in many cells was at least 1.5–2.0 octaves wide and, on average, was more than twice as wide as the frequency range that evoked discharges even after inhibition was blocked. In most cells, tones evoked complex synaptic response configurations that varied with frequency, suggesting that these cells were not innervated by congruent excitatory and inhibitory projections. Synaptic tuning was not only wide but was also diverse, in which some cells were dominated by excitation (n = 20), others were dominated by excitation with sideband inhibition (n = 21), but most were dominated by inhibition with little evidence of excitation (n = 31). Another unexpected finding was that some cells responded with inhibition to the onset and offset of tones over a wide frequency range, in which the patterns of synaptic responses changed markedly with frequency. These cells never fired to tones at 50 dB sound pressure level but fired to frequency-modulated sweeps at that intensity and were directionally selective. Thus, the features revealed by whole-cell recordings show that the processing in many IC cells results from inputs spectrally broader and more complex than previously believed.

Whole cell recordings of intrinsic properties and sound-evoked responses from the inferior colliculus

Response features of inferior colliculus (IC) neurons to both current injections and tone bursts were studied with in vivo whole cell recordings in awake Mexican free-tailed bats. Of 160 cells recorded, 95% displayed one of three general types of discharge patterns in response to the injection of positive current: 1) sustained discharges; 2) adapting discharges; and 3) onset-bursting discharges. Sustained neurons were the most common type (N=78), followed by onset-bursting (N=57). The least common type was adapting (N=17). In 90 neurons the profiles of synaptic and discharge activity evoked by tones of different frequencies at 50 dB SPL were recorded. Three major tone-evoked response profiles were obtained; 1) neurons dominated by excitation (N=32) in which tones evoked excitatory post-synaptic potentials (EPSPs) or EPSPs with discharges over a range of frequencies with little or no evidence of inhibitory post-synaptic potentials (IPSPs) evoked by frequencies that flanked the excitation; 2) neurons that had an excitatory frequency region in which discharges were evoked that was flanked by frequencies that evoked predominantly IPSPs (N=26); 3) neurons in which all frequencies evoked IPSPs with little or no depolarizations (N=32). The question we asked is whether IC cells that express a particular profile of PSPs and discharges to acoustic stimulation also have the same current-evoked response profile. We show that, with one exception, the intrinsic features of an IC neuron are not correlated with the pattern of its synaptic innervation; the two features are unrelated in the majority of IC cells. The exception is a subtype of inhibitory dominated cell where most frequencies evoked IPSPs to both the onset and to the offset of the tone bursts. In those cells injected current steps always evoked an onset-bursting response.

The dominance of inhibition in the inferior colliculus.

Almost all of the processing that occurs in the various lower auditory nuclei converges upon a common target in the central nucleus of the inferior colliculus (ICc) thus making the ICc the nexus of the auditory system. A variety of new response properties are formed in the ICc through the interactions among the excitatory and inhibitory inputs that converge upon it. Here we review studies that illustrate the dominant role inhibition plays in the ICc. We begin by reviewing studies of tuning curves and show how inhibition shapes the variety of tuning curves in the ICc through sideband inhibition. We then show how inhibition shapes selective response properties for complex signals, focusing on selectivity for the sweep direction of frequency modulations (FM). In the final section we consider results from in vivo whole-cell recordings that show how parameters of the incoming excitation and inhibition interact to shape directional selectivity. We show that post-synaptic potentials (PSPs) evoked by different signals can be similar but evoke markedly different spike-counts. In these cases, spike threshold acts as a non-linear amplifier that converts small differences in PSPs into large differences in spike output. Such differences between the inputs to a cell compared to the outputs from the same cell suggest that highly selective discharge properties can be created by only minor adjustments in the synaptic strengths evoked by one or both signals. These findings also suggest that plasticity of response features may be achieved with far less modifications in circuitry than previously supposed.

Inhibitory projections from the ventral nucleus of the lateral lemniscus and superior paraolivary nucleus create directional selectivity of frequency modulations in the inferior colliculus: A comparison of bats with other mammals

This review considers four auditory brainstem nuclear groups and shows how studies of both bats and other mammals have provided insights into their response properties and the impact of their convergence in the inferior colliculus (IC). The four groups are octopus cells in the cochlear nucleus, their connections with the ventral nucleus of the lateral lemniscus (VNLL) and the superior paraolivary nucleus (SPON), and the connections of the VNLL and SPON with the IC. The theme is that the response properties of neurons in the SPON and VNLL map closely onto the synaptic response features of a unique subpopulation of cells in the IC of bats whose inputs are dominated by inhibition. We propose that the convergence of VNLL and SPON inputs generates the tuning of these IC cells, their unique temporal responses to tones, and their directional selectivities for frequency modulated (FM) sweeps. Other IC neurons form directional properties in other ways, showing that selective response properties are formed in multiple ways. In the final section we discuss why multiple formations of common response properties could amplify differences in population activity patterns evoked by signals that have similar spectrotemporal features.

Target-specific IPSC kinetics promote temporal processing in auditory parallel pathways.

The acoustic environment contains biologically relevant information on timescales from microseconds to tens of seconds. The auditory brainstem nuclei process this temporal information through parallel pathways that originate in the cochlear nucleus from different classes of cells. Although the roles of ion channels and excitatory synapses in temporal processing have been well studied, the contribution of inhibition is less well understood. Here, we show in CBA/CaJ mice that the two major projection neurons of the ventral cochlear nucleus, the bushy and T-stellate cells, receive glycinergic inhibition with different synaptic conductance time courses. Bushy cells, which provide precisely timed spike trains used in sound localization and pitch identification, receive slow inhibitory inputs. In contrast, T-stellate cells, which encode slower envelope information, receive inhibition that is eightfold faster. Both types of inhibition improved the precision of spike timing but engage different cellular mechanisms and operate on different timescales. Computer models reveal that slow IPSCs in bushy cells can improve spike timing on the scale of tens of microseconds. Although fast and slow IPSCs in T-stellate cells improve spike timing on the scale of milliseconds, only fast IPSCs can enhance the detection of narrowband acoustic signals in a complex background. Our results suggest that target-specific IPSC kinetics are critical for the segregated parallel processing of temporal information from the sensory environment.

GABAergic and glycinergic inhibitory synaptic transmission in the ventral cochlear nucleus studied in VGAT channelrhodopsin-2 mice

Both glycine and GABA mediate inhibitory synaptic transmission in the ventral cochlear nucleus (VCN). In mice, the time course of glycinergic inhibition is slow in bushy cells and fast in multipolar (stellate) cells, and is proposed to contribute to the processing of temporal cues in both cell types. Much less is known about GABAergic synaptic transmission in this circuit. Electrical stimulation of the auditory nerve or the tuberculoventral pathway evokes little GABAergic synaptic current in brain slice preparations, and spontaneous GABAergic miniature synaptic currents occur infrequently. To investigate synaptic currents carried by GABA receptors in bushy and multipolar cells, we used transgenic mice in which channelrhodopsin-2 and EYFP is driven by the vesicular GABA transporter (VGAT-ChR2-EYFP) and is expressed in both GABAergic and glycinergic neurons. Light stimulation evoked action potentials in EYFP-expressing presynaptic cells, and evoked inhibitory postsynaptic potentials (IPSPs) in non-expressing bushy and planar multipolar cells. Less than 10% of the IPSP amplitude in bushy cells arose from GABAergic synapses, whereas 40% of the IPSP in multipolar neurons was GABAergic. In voltage clamp, glycinergic IPSCs were significantly slower in bushy neurons than in multipolar neurons, whereas there was little difference in the kinetics of the GABAergic IPSCs between two cell types. During prolonged stimulation, the ratio of steady state vs. peak IPSC amplitude was significantly lower for glycinergic IPSCs. Surprisingly, the reversal potentials of GABAergic IPSCs were negative to those of glycinergic IPSCs in both bushy and multipolar neurons. In the absence of receptor blockers, repetitive light stimulation was only able to effectively evoke IPSCs up to 20 Hz in both bushy and multipolar neurons. We conclude that local GABAergic release within the VCN can differentially influence bushy and multipolar cells.

Glycinergic synaptic transmission in the cochlear nucleus of mice with normal hearing and age-related hearing loss

The principal inhibitory neurotransmitter in the mammalian cochlear nucleus (CN) is glycine. During age-related hearing loss (AHL), glycinergic inhibition becomes weaker in CN. However, it is unclear what aspects of glycinergic transmission are responsible for weaker inhibition with AHL. We examined glycinergic transmission onto bushy cells of the anteroventral CN in normal-hearing CBA/CaJ mice and in DBA/2J mice, a strain that exhibits an early onset AHL. Glycinergic synaptic transmission was examined in brain slices of mice at 10-15 postnatal days old, 20-35 days old, and at 6-7 mo old. Spontaneous inhibitory postsynaptic current (sIPSC) event frequency and amplitude were the same among all three ages in both strains of mice. However, the amplitudes of IPSCs evoked (eIPSC) from stimulating the dorsal CN were smaller, and the failure rate was higher, with increasing age due to decreased quantal content in both mouse strains, independent of hearing status. The coefficient of variation of the eIPSC amplitude also increased with age. The decay time constant (τ) of sIPSCs and eIPSCs were constant in CBA/CaJ mice at all ages, but were significantly slower in DBA/2J mice at postnatal days 20-35, following the onset of AHL, and not at earlier or later ages. Our results suggest that glycinergic inhibition at the synapses onto bushy cells becomes weaker and less reliable with age through changes in release. However, the hearing loss in DBA/2J mice is accompanied by a transiently enhanced inhibition, which could disrupt the balance of excitation and inhibition.

The Endbulbs of Held

A remarkable nerve terminal is present at the endings of the auditory nerve fibers (ANFs) in the anterior ventral cochlear nucleus (AVCN), called the “endbulb of Held.” The endbulbs are complex synaptic endings that provide a coordinated release from multiple presynaptic sites of neurotransmitter onto their target postsynaptic cells, the globular and spherical bushy cells of the cochlear nucleus. These synapses play a key role in bringing a precisely timed representation of sound into the central auditory system. Traditionally, the endbulbs, owing to their large size and the presence of multiple presynaptic release zones, were thought to provide a “secure” synapse between auditory nerve fibers and the target neurons, the globular and spherical bushy cells. However, this view has been strongly challenged by several recent observations. While the endbulbs are indeed a particularly strong synapse, they are subject to dynamic regulation of transmitter release probability and receptor sensitivity, and their ability to initiate action potentials in the postsynaptic cell is not immune to postsynaptic inhibition. Integration by convergence of endbulb synapses onto target cells is an important part of central auditory processing. In particular, cells postsynaptic to the endbulbs can fire more precisely to specific temporal features of acoustic stimuli than their individual auditory nerve fiber inputs. The endbulb synapses are found widely in mammals including humans (Adams 1986), as well as in birds (Carr and Boudreau 1991; Koppl 1994) and reptiles (Browner and Marbey 1988; Szpir et al. 1990), but their presence in amphibians is less clear (Lewis et al. 1980; Feng and Lin 1996).

HMW glutenin subunits in multiploidAegilops species: composition analysis and molecular cloning of coding sequences

TheAegilops genus contains species closely related to wheat. In common with wheat,Aegilops species accumulate high molecular weight (HMW) glutenin subunits in their endospermic tissue. In this study, we investigated the composition of HMW glutenin subunits in four multiploidAegilops species using SDS-PAGE analysis. Furthermore, by working withAe. ventricosa, we established an efficient genomic PCR condition for simultaneous amplification of DNA sequences coding for either x-or y-type HMW glutenin subunits from polyploidAegilops species. Using the genomic PCR condition, we amplified and subsequently cloned two DNA fragments that may code for HMW glutenin subunits inAe. ventricosa. Based on an analysis of the deduced amino acid sequences, we concluded that the two cloned sequences encode one x- and one y-type of HMW glutenin subunit, respectively.

Synaptic transmission at the endbulb of Held deteriorates during age‐related hearing loss

Synaptic transmission at the endbulb of Held was assessed by whole‐cell patch clamp recordings from auditory neurons in mature (2–4 months) and aged (20–26 months) mice. Synaptic transmission is degraded in aged mice, which may contribute to the decline in neural processing of the central auditory system during age‐related hearing loss. The changes in synaptic transmission in aged mice can be partially rescued by improving calcium buffering, or decreasing action potential‐evoked calcium influx. These experiments suggest potential mechanisms, such as regulating intraterminal calcium, that could be manipulated to improve the fidelity of transmission at the aged endbulb of Held.