Kimberly E. Miller

Postsynaptic neural activity regulates neuronal addition in the adult avian song control system.

Significance Neural activity in the adult brain plays a key role in mediating experience-dependent neural plasticity. We show that inhibiting electrical activity in the song nucleus, robust nucleus of the arcopallium, in adult bird brain decreases the number of new projection neurons added to the afferent nucleus HVC. Our results are consistent with the general principle of activity-based target selection of newborn neurons during nervous system development and support the idea that developmental and adult plasticity exploit similar mechanisms. Understanding mechanisms influencing the incorporation of new neurons into established neural circuits in adult brains is critical for our basic understanding of neural plasticity and for exploiting the clinical potential of neuronal replacement to repair damage associated with injury and neurodegenerative diseases. A striking feature of the nervous system is that it shows extensive plasticity of structure and function that allows animals to adjust to changes in their environment. Neural activity plays a key role in mediating experience-dependent neural plasticity and, thus, creates a link between the external environment, the nervous system, and behavior. One dramatic example of neural plasticity is ongoing neurogenesis in the adult brain. The role of neural activity in modulating neuronal addition, however, has not been well studied at the level of neural circuits. The avian song control system allows us to investigate how activity influences neuronal addition to a neural circuit that regulates song, a learned sensorimotor social behavior. In adult white-crowned sparrows, new neurons are added continually to the song nucleus HVC (proper name) and project their axons to its target nucleus, the robust nucleus of the arcopallium (RA). We report here that electrical activity in RA regulates neuronal addition to HVC. Decreasing neural activity in RA by intracerebral infusion of the GABAA receptor agonist muscimol decreased the number of new HVC neurons by 56%. Our results suggest that postsynaptic electrical activity influences the addition of new neurons into a functional neural circuit in adult birds.

Stimulus-specific adaptation in specialized neurons in the inferior colliculus of the big brown bat, Eptesicus fuscus

The inferior colliculus (IC) of the big brown bat (Eptesicus fuscus) contains specialized neurons that respond exclusively to highly specific spectrotemporal patterns such as sinusoidally frequency modulated (SFM) signals or directional frequency modulated sweeps (FM). Other specialized cells with I-shaped frequency response areas (FRAs) are tuned to very narrow frequency bands (1-2 kHz) in an amplitude-tolerant manner. In contrast, non-specialized neurons respond to any stimulus with energy in their frequency response area. IC neurons in several mammalian species, including bats, demonstrate stimulus-specific adaptation (SSA), a reduction in response to a high-probability stimulus. To evaluate the relation between stimulus selectivity and SSA, we presented sounds using an oddball stimulus paradigm and recorded extracellular responses of IC neurons. SFM-selective cells (n = 10), FM-selective cells (n = 7), and cells with I-shaped FRAs (n = 13) did not show SSA under any of the conditions tested (NSSI = 0.009, 0.033, 0.020 respectively). However, non-specialized neurons (n = 52) exhibited various levels of SSA (NSSI = 0.163), with a subset of these cells displaying strong adaptation. These findings suggest that SSA is not a ubiquitous characteristic of all neurons in the bat IC, but is present only in a subset of non-specialized neurons.

Extreme Postnatal Scaling in Bat Feeding Performance: A View of Ecomorphology from Ontogenetic and Macroevolutionary Perspectives

Ecomorphology studies focus on understanding how anatomical and behavioral diversity result in differences in performance, ecology, and fitness. In mammals, the determinate growth of the skeleton entails that bite performance should change throughout ontogeny until the feeding apparatus attains its adult size and morphology. Then, interspecific differences in adult phenotypes are expected to drive food resource partitioning and patterns of lineage diversification. However, Formal tests of these predictions are lacking for the majority of mammal groups, and thus our understanding of mammalian ecomorphology remains incomplete. By focusing on a fundamental measure of feeding performance, bite force, and capitalizing on the extraordinary morphological and dietary diversity of bats, we discuss how the intersection of ontogenetic and macroevolutionary changes in feeding performance may impact ecological diversity in these mammals. We integrate data on cranial morphology and bite force gathered through longitudinal studies of captive animals and comparative studies of free-ranging individuals. We demonstrate that ontogenetic trajectories and evolutionary changes in bite force are highly dependent on changes in body and head size, and that bats exhibit dramatic, allometric increases in bite force during ontogeny. Interspecific variation in bite force is highly dependent on differences in cranial morphology and function, highlighting selection for ecological specialization. While more research is needed to determine how ontogenetic changes in size and bite force specifically impact food resource use and fitness in bats, interspecific diversity in cranial morphology and bite performance seem to closely match functional differences in diet. Altogether, these results suggest direct ecomorphological relationships at ontogenetic and macroevolutionary scales in bats.

Comparison of Auditory Responses in the Medial Geniculate and Pontine Gray of the Big Brown Bat, Eptesicus fuscus

The inferior colliculus has been well studied for its role of transmitting information from the brainstem to the thalamocortical system. However, it is also the source of a major pathway to the cerebellum, via the pontine gray (PG). We compared auditory responses from single neurons in the medial geniculate body (MGB) and PG of the awake big brown bat. MGB neurons were selective for a variety of stimulus types whereas PG neurons only responded to pure tones or simple FM sweeps. Best frequencies (BF) in MGB ranged from 8 kHz to > 80 kHz. BFs of PG neurons were all above 20 kHz with a high proportion above 60 kHz. The mean response latency was 19 ms for MGB neurons and 11 ms for PG neurons. MGB and PG contained neurons with a variety of discharge patterns but the most striking difference was the proportion of neurons with responses that lasted longer than the stimulus duration (MGB 13%, PG 58%). Both nuclei contained duration-sensitive neurons; the majority of those in MGB were band pass whereas in the PG they were long pass. Over half of the neurons in both nuclei were binaural. Differences between these nuclei are consistent with the idea that the thalamocortical pathway performs integration over time for cognitive analysis, thereby increasing selectivity and lengthening latency, while the colliculo-pontine pathway, which is more concerned with sensory-motor control, provides rapid input and a lasting trace of an auditory event.

Dopaminergic modulation of basal ganglia output through coupled excitation–inhibition

Significance Trial-and-error learning requires variation in successive trials, but the source of such variability is unknown. We describe a unique striatal glutamatergic neuron in the zebra finch. This neuron exerts a potent, dopamine-regulated action on pallidal output neurons that modifies neuronal firing statistics in the circuit known to contribute to vocal variability. A simple model reveals how this microcircuit could be influenced by social context and striatal dopamine to switch between firing patterns that modify song variability essential for vocal learning. Learning and maintenance of skilled movements require exploration of motor space and selection of appropriate actions. Vocal learning and social context-dependent plasticity in songbirds depend on a basal ganglia circuit, which actively generates vocal variability. Dopamine in the basal ganglia reduces trial-to-trial neural variability when the bird engages in courtship song. Here, we present evidence for a unique, tonically active, excitatory interneuron in the songbird basal ganglia that makes strong synaptic connections onto output pallidal neurons, often linked in time with inhibitory events. Dopamine receptor activity modulates the coupling of these excitatory and inhibitory events in vitro, which results in a dynamic change in the synchrony of a modeled population of basal ganglia output neurons receiving excitatory and inhibitory inputs. The excitatory interneuron thus serves as one biophysical mechanism for the introduction or modulation of neural variability in this circuit.

Adult Neurogenesis Leads to the Functional Reconstruction of a Telencephalic Neural Circuit

Seasonally breeding songbirds exhibit pronounced annual changes in song behavior, and in the morphology and physiology of the telencephalic neural circuit underlying production of learned song. Each breeding season, new adult-born neurons are added to the pallial nucleus HVC in response to seasonal changes in steroid hormone levels, and send long axonal projections to their target nucleus, the robust nucleus of the arcopallium (RA). We investigated the role that adult neurogenesis plays in the seasonal reconstruction of this circuit. We labeled newborn HVC neurons with BrdU, and RA-projecting HVC neurons (HVCRA) with retrograde tracer injected in RA of adult male white-crowned sparrows (Zonotrichia leucophrys gambelii) in breeding or nonbreeding conditions. We found that there were many more HVCRA neurons in breeding than nonbreeding birds. Furthermore, we observed that more newborn HVC neurons were back-filled by the tracer in breeding animals. Behaviorally, song structure degraded as the HVC-RA circuit degenerated, and recovered as the circuit regenerated, in close correlation with the number of new HVCRA neurons. These results support the hypothesis that the HVC-RA circuit degenerates in nonbreeding birds, and that newborn neurons reconstruct the circuit in breeding birds, leading to functional recovery of song behavior. SIGNIFICANCE STATEMENT We investigated the role that adult neurogenesis plays in the seasonal reconstruction of a telencephalic neural circuit that controls song behavior in white-crowned sparrows. We showed that nonbreeding birds had a 36%–49% reduction in the number of projection neurons compared with breeding birds, and the regeneration of the circuit in the breeding season is due to the integration of adult-born projection neurons. Additionally, song structure degraded as the circuit degenerated and recovered as the circuit regenerated, in close correlation with new projection neuron number. This study demonstrates that steroid hormones can help reestablish functional neuronal circuits following degeneration in the adult brain and shows non–injury-induced degeneration and reconstruction of a neural circuit critical for producing a learned behavior.

Expression of the potassium-chloride co-transporter, KCC2, within the avian song system

Songbirds learn to produce vocalizations early in life by listening to, then copying the songs of conspecific males. The anterior forebrain pathway, homologous to a basal ganglia‐forebrain circuit, is essential for song learning. The projection between the striato‐pallidal structure, Area X, and the medial portion of the dorsolateral thalamic nucleus (DLM) is strongly hyperpolarizing in adults, due to a very negative chloride reversal potential (Person & Perkel, Neuron 46:129–140, 2005). The chloride reversal potential is determined, in part, by the expression level of a neuron‐specific potassium‐chloride cotransporter, KCC2, which is developmentally upregulated in mammals. To determine whether a similar upregulation in KCC2 expression occurs at the Area X to DLM synapse during development, we examined the expression level of KCC2 in adult zebra finches across the song system as well as during development in the Area X – DLM synapse. We demonstrate that KCC2 is expressed in a subset of neurons throughout the song system, including HVC (used as a proper name), robust nucleus of the arcopallium (RA), lateral magnocellular nucleus of the anterior nidopallium (LMAN), Area X, and DLM. The majority of pallidal‐like projection neurons in Area X showed KCC2 immunoreactivity. In adults, KCC2 expression was robust within DLM, and was upregulated between 14 and 24 days post hatching, before the onset of song learning. Light and electron microscopic analysis indicated that KCC2 immunoreactivity is strongly associated with the plasma membrane. Thus, in the song system as in the mammalian brain, KCC2 expression is well placed to modulate the GABAA reversal potential.

Seasonal variations in auditory processing in the inferior colliculus of Eptesicus fuscus

Eptesicus fuscus is typical of temperate zone bats in that both sexes undergo marked seasonal changes in behavior, endocrine status, and reproductive status. Acoustic communication plays a key role in many seasonal behaviors. For example, males emit specialized vocalizations during mating in the fall, and females use different specialized vocalizations to communicate with infants in late spring. Bats of both sexes use echolocation for foraging during times of activity, but engage in little sound-directed behavior during torpor and hibernation in winter. Auditory processing might be expected to reflect these marked seasonal changes. To explore the possibility that seasonal changes in hormonal status could drive functional plasticity in the central auditory system, we examined responses of single neurons in the inferior colliculus throughout the year. The average first spike latency in females varied seasonally, almost doubling in spring compared to other times of year. First spike latencies in males remained relatively stable throughout the year. Latency jitter for both sexes was higher in winter and spring than in summer or fall. Females had more burst responders than other discharge patterns throughout the year whereas males had more transient responders at all times of year except fall, when burst responses were the predominant type. The percentage of simple discharge patterns (sustained and transient) was higher in males than females in the spring and higher in females than males in the fall. In females, the percentage of shortpass duration-tuned neurons doubled in summer and remained elevated through fall and early winter. In males, the percentage of shortpass duration-tuned cells increased in spring and the percentage of bandpass duration-tuned cells doubled in the fall. These findings suggest that there are clear seasonal changes in basic response characteristics of midbrain auditory neurons in Eptesicus, especially in temporal response properties and duration sensitivity. Moreover, the pattern of changes is different in males and females, suggesting that hormone-driven plasticity adjusts central auditory processing to fit the characteristics of vocalizations specific to seasonal behavioral patterns.