Differential contributions of voltage-gated potassium channel subunits in enhancing temporal coding in the bushy cells of the ventral cochlear nucleus.

Temporal coding precision of bushy cells in the ventral cochlear nucleus (VCN), critical for sound localization and communication, depends on the generation of rapid and temporally precise action potentials (APs). Voltage-gated potassium (Kv) channels are critically involved in this. The bushy cells in rat VCN express Kv1.1, 1.2, 1.3, 1.6, 3.1, 4.2, and 4.3 subunits. The Kv1.1 subunit contributes to the generation of a temporally precise single AP. However, the understanding of the functions of other Kv subunits expressed in the bushy cells is limited. Here, we investigated the functional diversity of Kv subunits concerning their contributions to temporal coding. We characterized the electrophysiological properties of the Kv channels with different subunits using whole cell patch-clamp recording and pharmacological methods. The neuronal firing pattern changed from single to multiple APs only when the Kv1.1 subunit was blocked. The Kv subunits, including the Kv1.1, 1.2, 1.6, or 3.1, were involved in enhancing temporal coding by lowering membrane excitability, shortening AP latencies, reducing jitter, and regulating AP kinetics. Meanwhile, all the Kv subunits contributed to rapid repolarization and sharpening peaks by narrowing half-width and accelerating fall rate, and the Kv1.1 subunit also affected the depolarization of AP. The Kv1.1, 1.2, and 1.6 subunits endowed bushy cells with a rapid time constant and a low input resistance of membrane for enhancing spike timing precision. The present results indicate that the Kv channels differentially affect intrinsic membrane properties to optimize the generation of rapid and reliable APs for temporal coding.NEW & NOTEWORTHY This study investigates the roles of Kv channels in effecting precision using electrophysiological and pharmacological methods in bushy cells. Different Kv channels have varying electrophysiological characteristics, which contribute to the interplay between changes in the membrane properties and regulation of neuronal excitability which then improve temporal coding. We conclude that the Kv channels are specialized to promote the precise and rapid coding of acoustic input by optimizing the generation of reliable APs.

Auditory Fear Conditioning Alters Sensitivity of the Medial Prefrontal Cortex but this is not based on Frequency-dependent Integration.

Although many studies have shown that the prelimbic (PL) cortex of the mPFC is involved in the formation of conditioned freezing behavior, few have considered the acoustic response characteristics of PL cortex. Importantly, the change in auditory response characteristics of the PL cortex after conditional fear learning is largely unknown. Here we used in vivo cell-attached recordings targeting the mPFC during the waking state. We confirmed that the mPFC of adult C57 mice have neurons that respond to noise and tone in the waking state, especially in the PL cortex. Interestingly, the data also confirmed that these neurons responded well to the intensity of sound but did not have frequency topological distribution characteristics. Furthermore, we found that the number of c-fos positive neurons in the PL cortex increased significantly after auditory fear conditioning. The auditory-induced local field potential recordings and in vivo cell-attached recordings demonstrated that the PL cortex was more sensitive to the auditory conditioned stimulus after the acquisition of conditioned fear. The proportion of neurons responding to noise was significantly increased, and the signal to noise ratio of the spikes were also increased. These data reveal that PL neurons themselves responded to the main information (sound intensity), while the secondary information (frequency) response was almost negligible after auditory fear conditioning. This phenomenon may be the functional basis for handling this type of emotional memory, and this response characteristic is thought to be emotional sensitization but does not change the nature of this response.

Processing of Paired Click-Tone Stimulation in the Mice Inferior Colliculus.

The inferior colliculus (IC) is known as a neuronal structure involved in the integration of acoustic information in the ascending auditory pathway. However, the processing of paired acoustic stimuli containing different sound types, especially when they are applied closely, in the IC remains poorly studied. We here firstly investigated the IC neuronal response to the paired stimuli comprising click and pure tone with different inter-stimulus (click-tone) intervals using in vivo loose-patch recordings in anesthetized BALB/c mice. It was found that the total acoustic evoked spike counts decreased under certain click-tone interval conditions on some neurons with or without click-induced supra-threshold responses. Application of click could enhance the minimum threshold of the neurons responding to the tone in a pair without changing other characteristics of the neuronal tone receptive fields. We further studied the paired acoustic stimuli evoked excitatory/inhibitory inputs, IC neurons received, by holding the membrane potential at -70/0 mV using in vivo whole-cell voltage-clamp techniques. The curvature and peak amplitude of the excitatory/inhibitory post-synaptic current (EPSC/IPSC) could be almost unchanged under different inter-stimulus interval conditions. Instead of showing the summation of synaptic inputs, most recorded neurons only had the EPSC/IPSC with the amplitude similar as the bigger one evoked by click or tone in a pair when the inter-stimulus interval was small. We speculated that the IC could inherit the paired click-tone information which had been integrated before reaching it.

Balanced Noise-Evoked Excitation and Inhibition in Awake Mice CA3.

The hippocampus is known as a neuronal structure involved in learning, memory and spatial navigation using multi-sensory cues. However, the basic features of its response to acoustic stimuli without any behavioral tasks (conditioning) remains poorly studied. Here, we investigated the CA3 response to auditory stimuli using in vivo loose-patch recordings in awake and anesthetized C57 mice. Different acoustic stimuli in addition to broadband noise such as click, FM sound and pure tone were applied to test the response of CA3 in awake animals. It was found that the wakefulness of the animal is important for the recorded neurons to respond. The CA3 neurons showed a stronger response to broadband noise rather than the other type of stimuli which suggested that auditory information arrived at CA3 via broadband pathways. Finally, we investigated the excitatory and inhibitory inputs to CA3 neurons by using in vivo whole-cell voltage-clamp techniques with the membrane potential holding at -70 and 0 mV, respectively. In awake animals, the excitatory and inhibitory inputs CA3 neurons receive induced by noise are balanced by showing stable intervals and proportional changes of their latencies and peak amplitudes as a function of the stimulation intensities.

Interdependent effects of sound duration and amplitude on neuronal onset response in mice inferior colliculus.

In this study, we adopted iso-frequency pure tone bursts to investigate the interdependent effects of sound amplitude/intensity and duration on mice inferior colliculus (IC) neuronal onset responses. On the majority of the sampled neurons (n=57, 89.1%), sound amplitude and duration had effects on the neuronal response to each other by showing complex changes of the rat-intensity function/duration selectivity types and/or best amplitudes (BAs)/durations (BDs), evaluated by spike counts. These results suggested that the balance between the excitatory and inhibitory inputs set by one acoustic parameter, amplitude or duration, affected the neuronal spike counts responses to the other. Neuronal duration selectivity types were altered easily by the low-amplitude sounds while the changes of rate-intensity function types had no obvious preferred stimulus durations. However, the first spike latencies (FSLs) of the onset response neurons were relative stable to iso-amplitude sound durations and changing systematically along with the sound levels. The superimposition of FSL and duration threshold (DT) as a function of stimulus amplitude after normalization indicated that the effects of the sound levels on FSLs are considered on DT actually.

Adenosine A1 receptors are involved in the modulation of the rhythmical respiration in neonatal rat brainstem slice in vitro.

This study was designed to investigate whether adenosine A1 receptors could modulate primary rhythmical respiration in mammals. Experiments were performed in in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of the nucleus retrofacialis (mNRF) with the hypoglossal nerve rootlets retained. The activity of the inspiration-related neurons (I neurons) in mNRF and respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve rootlets were simultaneously recorded by using microelectrodes and suction electrodes, respectively. Possible roles of adenosine A1 receptors in rhythmical respiration were investigated by administration of adenosine A1 receptor agonist R-phenylisopropyl-adenosine (R-PIA) and its specific antagonist 8-cyclopentyl-1,3- dipropylxanthine (DPCPX) into a modified Kreb's perfusion solution (MKS). DPCPX induced a significant decrease in the expiratory time and the respiratory cycles, and an increase in the discharge frequency and peak frequency of I neurons in the middle phase of inspiration. However, R-PIA significantly decreased the inspiratory time and integral amplitude as well as prolonged respiratory cycle. Moreover, the discharge frequency and the peak frequency of I neurons were decreased in the middle phase of inspiration, but not in the initial and terminal phases. The effect of R-PIA on rhythmical discharges could be partially reversed by additional application of DPCPX. These results indicate that adenosine A1-receptors are possibly involved in the modulation of rhythmical respiration through the inhibitory synaptic input from I neurons.

[Nitric oxide is involved in the modulation of central respiratory rhythm].

This experiment was expected to test whether nitric oxide (NO) exerted significant effect on the central respiratory rhythm. Experiments were performed on in vitro brainstem slice preparations from neonatal rats. These preparations include the medial region of the nucleus retrofacialis (mNRF); a part of pre-Bötinger complex, ventral respiratory group (VRG) and dorsal respiratory group (DRG). Respiratory-related burst activities were recorded from hypoglossal nerve rootlets before and during superfusion of the slice preparation with L-Arginine (L-Arg), sodium nitroprusside (SNP) or 7-nitro indazole (7-NI, an inhibitor of NO synthase). After perfusion with L-Arg and SNP, there was no significant change in respiratory rhythmical discharge activity (RRDA), but 7-NI decreased the integral amplitude of burst and inspiratory time. These results indicate that NO may take part in the inspiratory off-switching mechanism and that it also modulates the amplitude of respiratory-related bursts.

Projections between the medial region of the nucleus retrofacialis and other respiratory regions in rats brainstem.

OBJECTIVE: To study the projections between the medial region of the nucleus retrofacialis (mNRF) and other respiratory regions in rat brainstem. METHODS: Twenty-five adult SD rats were used in this study, which received anesthesia with sodium pentobarbitone. Retrograde/anterograde tracing methods were employed by unilateral microinjection into mNRF with 30% horseradish peroxidase (HRP, 0.5-1.0 microliter1) in 13 rats. Another 5 rats with injections of HRP and 2 with WGA-HRP into the regions 2 mm off mNRF served as control groups. and with 5% wheat germ agglutinin-conjugated HRP (WGA-HRP, 20-60 nl) in 5 rats (with another 2 receiving the injections as control). RESULTS: It was found that HRP-labeled neuronal somas and WGA-HRP labeled terminal fibers were found in many respiration-related regions in rat brainstem such as the nucleus tractus solitarius, nucleus ambiguous, gigantocellular reticular nucleus and so forth. CONCLUSIONS: There are comprehensive projections between mNRF and other respiratory regions in rat brainstem, and these respiratory regions may be involved in basic respiratory rhythm regulation via these connections with mNRF.