Liisa A. Tremere

Estradiol Shapes Auditory Processing in the Adult Brain by Regulating Inhibitory Transmission and Plasticity-Associated Gene Expression

Estradiol impacts a wide variety of brain processes, including sex differentiation, mood, and learning. Here we show that estradiol regulates auditory processing of acoustic signals in the vertebrate brain, more specifically in the caudomedial nidopallium (NCM), the songbird analog of the mammalian auditory association cortex. Multielectrode recordings coupled with local pharmacological manipulations in awake animals reveal that both exogenous and locally generated estradiol increase auditory-evoked activity in NCM. This enhancement in neuronal responses is mediated by suppression of local inhibitory transmission. Surprisingly, we also found that estradiol is both necessary and sufficient for the induction of multiple mitogen-activated protein kinase (MAPK)-dependent genes thought to be required for synaptic plasticity and memorization of birdsong. Specifically, we show that local blockade of estrogen receptors or aromatase activity in awake birds decrease song-induced MAPK-dependent gene expression. Infusions of estradiol in acoustically isolated birds induce transcriptional activation of these genes to levels comparable with song-stimulated animals. Our results reveal acute and rapid nongenomic functions for estradiol in central auditory physiology and suggest that such roles may be ubiquitously expressed across sensory systems.

Synaptic Activation of Presynaptic Glutamate Transporter Currents in Nerve Terminals

Glutamate uptake by high-affinity transporters is responsible for limiting the activation of postsynaptic receptors and maintaining low levels of ambient glutamate. The reuptake process generates membrane currents, which can be activated by synaptically released glutamate in glial cells and some postsynaptic neurons. However, less is known about presynaptic transporter currents because the small size of synaptic boutons precludes direct recordings. Here, we have recorded from two giant nerve terminals: bipolar cell synaptic terminals in goldfish retina and the calyx of Held in rat auditory brainstem. Exocytosis was evoked by brief depolarizations and measured as an increase in membrane capacitance. In isolated bipolar cell terminals, exocytosis was associated with an anion (NO3- or Cl-) current. The current peaked 2.8 msec after the start of the depolarization and decayed with a mean time constant of 8.5 msec. It was inhibited by the nontransportable glutamate transporter antagonist sc-threo-β-benzyloxyaspartate (TBOA) but was insensitive to the GLT1/EAAT2 subtype-selective antagonist dihydrokainate and was affected by extracellular pH buffering. A TBOA-sensitive anion current was also evoked by application of exogenous glutamate to bipolar cell terminals. The large single-channel conductance, derived from noise analysis, and previous immunolocalization studies suggest that synaptically released glutamate activates EAAT5-type transporters in bipolar cell terminals. In contrast, neither exocytosis nor exogenous glutamate evoked a transporter current in the calyx of Held. Glutamate transporter currents with rapid kinetics are therefore identified and characterized in bipolar cell terminals, providing a valuable system for investigating the function and modulation of presynaptic glutamate transporters.

Brain-Generated Estradiol Drives Long-Term Optimization of Auditory Coding to Enhance the Discrimination of Communication Signals

Auditory processing and hearing-related pathologies are heavily influenced by steroid hormones in a variety of vertebrate species, including humans. The hormone estradiol has been recently shown to directly modulate the gain of central auditory neurons, in real time, by controlling the strength of inhibitory transmission via a nongenomic mechanism. The functional relevance of this modulation, however, remains unknown. Here we show that estradiol generated in the songbird homolog of the mammalian auditory association cortex, rapidly enhances the effectiveness of the neural coding of complex, learned acoustic signals in awake zebra finches. Specifically, estradiol increases mutual information rates, coding efficiency, and the neural discrimination of songs. These effects are mediated by estradiols modulation of both the rate and temporal coding of auditory signals. Interference with the local action or production of estradiol in the auditory forebrain of freely behaving animals disrupts behavioral responses to songs, but not to other behaviorally relevant communication signals. Our findings directly show that estradiol is a key regulator of auditory function in the adult vertebrate brain.

GABAergic neurons participate in the brain's response to birdsong auditory stimulation

Birdsong is a learned vocal behaviour that requires intact hearing for its development in juveniles and for its maintenance during adulthood. However, the functional organization of the brain circuits involved in the perceptual processing of song has remained obscure. Here we provide evidence that GABAergic mechanisms are an important component of these circuits and participate in the auditory processing of birdsong. We first cloned a zebra finch homologue of the gene encoding the 65‐kDa isoform of glutamic acid decarboxylase (zGAD‐65), a specific GABAergic marker, and conducted an expression analysis by in situ hybridization to identify GABAergic cells and to map their distribution throughout auditory telencephalic areas. The results showed that field L2, the caudomedial nidopallium (NCM) and the caudomedial mesopallium (CMM) contain a high number of GABAergic cells. Using patch‐clamp brain slice recordings, we found abundant GABAergic mIPSCs in NCM. Pharmacological antagonism of mIPSCs induced large EPSC bursts, suggesting that tonic inhibition helps to stabilize NCM against runaway excitation via activation of GABA‐A receptors. Next, using double fluorescence in situ hybridization and double immunocytochemical labelling, we demonstrated that large numbers of GABAergic cells in NCM and CMM show inducible expression of the transcriptional regulator ZENK in response to song auditory stimulation. These data provide direct evidence that GABAergic neurons in auditory brain regions are activated by song stimulation. Altogether, our results suggest that GABAergic mechanisms participate in auditory processing and perception, and might contribute to the memorization of birdsong.

Inhibitory network interactions shape the auditory processing of natural communication signals in the songbird auditory forebrain.

The role of GABA in the central processing of complex auditory signals is not fully understood. We have studied the involvement of GABA A-mediated inhibition in the processing of birdsong, a learned vocal communication signal requiring intact hearing for its development and maintenance. We focused on caudomedial nidopallium (NCM), an area analogous to parts of the mammalian auditory cortex with selective responses to birdsong. We present evidence that GABA A-mediated inhibition plays a pronounced role in NCMs auditory processing of birdsong. Using immunocytochemistry, we show that approximately half of NCMs neurons are GABAergic. Whole cell patch-clamp recordings in a slice preparation demonstrate that, at rest, spontaneously active GABAergic synapses inhibit excitatory inputs onto NCM neurons via GABA A receptors. Multi-electrode electrophysiological recordings in awake birds show that local blockade of GABA A-mediated inhibition in NCM markedly affects the temporal pattern of song-evoked responses in NCM without modifications in frequency tuning. Surprisingly, this blockade increases the phasic and largely suppresses the tonic response component, reflecting dynamic relationships of inhibitory networks that could include disinhibition. Thus processing of learned natural communication sounds in songbirds, and possibly other vocal learners, may depend on complex interactions of inhibitory networks.

Immediate Early Genes in Sensory Processing, Cognitive Performance and Neurological Disorders

Preface.- Contributors.- The Use of Immediate Early Genes as Mapping Tools for Neuronal Activation: Concepts and Methods.- Part I Immediate Early Gene Expression as Part of Sensory Processing: Regulation of Immediate Early Genes in the Visual Cortex.- Immediate Early Gene Regulation in the Auditory System.- Immediate Early Genes and Sensory Maps of Olfactory and Gustatory Function.- Immediate Early Gene Expression in the Primary Somatosensory Cortex: Focus on the Barrel Cortex.- Immediate Early Genes Induced in Models of Acute and Chronic Pain.- Part II Immediate Early Gene Expression in Complex Systems and Higher Order Cognitive Function: Mapping Sleep-Wake Control With the Transcription Factor c-Fos.- c-Fos and Zif268 in Learning and Memory - Studies on Expression and Function.- Immediate Early Genes and the Mapping of Environmental Representations in Hippocampal Neural Networks.- Neural Dysfunction and Cognitive Impairment Resulting From Inactivation of the Egr-Family Transcription Factor Zif268.- Part III Immediate Early Genes in Neurological Disorder: Clinical Implications: The Contribution of Immediate Early Genes to the Understanding of Brain Processing of Stressors.- Transcriptional Control of Nerve Cell Death, Survival and Repair.- Immediate Early Genes, Inducible Transcription Factors and Stress Kinases in Alzheimers Disease.- Parkinsons Disease, the Dopamine Pathway and Immediate Early Genes.

Control of central auditory processing by a brain-generated oestrogen

Recent discoveries show that behaviourally relevant sensory experience drives the production of oestradiol — the classic sex steroid oestrogen — in auditory neurons in the adult brain of both males and females. This brain-generated oestrogen markedly enhances the efficiency of the neural coding of acoustic cues and shapes auditory-based behaviours on a timescale that is relevant for sensory processing and congruent with the action of rapid neuromodulators. These findings are re-shaping our current understanding of the mechanistic framework that supports sensory processing and the functional roles of hormones in the brain, and have implications for multiple health issues.

Antibody for human p75 LNTR identifies cholinergic basal forebrain of non-primate species.

192-IgG is an antibody directed against the p75 low affinity nerve growth factor receptor in rats, whereas ME 20.4 was raised against the analogous protein in humans. Coupled to saporin, 192-IgG and ME 20.4 have been used to lesion basal forebrain neurons in rats and primates, respectively. We compared the cross-reactivity of 192-IgG and ME 20.4 in the basal forebrain of rat, human, dog, cat, raccoon, pig, and rabbit. We found excellent species cross-reactivity of ME 20.4 in dog, raccoon, cat, pig and rabbit. In contrast, 192-IgG did not label neurons in any species other than rat. Our findings suggest that ME 20.4-saporin could be used to produce cholinergic basal forebrain lesions in several non-primate species.

Light-induced Egr-1 expression in the striate cortex of the opossum.

In the present study, immunocytochemistry was used to assess the expression of Egr-1 nuclear protein across selected regions of the opossum visual system. In light-deprived (LD) animals, only a few scattered cell nuclei were found throughout the striate cortex (V1). Exposure to light promoted a significant increase in the density of Egr-1 labeled nuclei in V1. Laminar distribution of immunoreactive nuclei in light-stimulated animals (LS) tended to vary with topography: the lateral region, which corresponds to the central representation of the visual field, appeared to have higher density of cells expressing protein in the supragranular layers, as compared to the medial region, which corresponds to the representation of the peripheral field of vision. Finally, LS animals displayed a narrow band of labeled cell nuclei in the intergeniculate leaflet (IGL) and throughout the anteroposterior extent of the superior colliculus (SC). In contrast, almost no Egr-1 immunolabeling was found in the IGL and SC of LD animals. Our report is the first demonstration of light-regulated expression of the Egr-1 gene in the opossum visual system and provides evidence that the expression of an activity-dependent gene related to neural plasticity is evolutionarily conserved in the visual cortex of the mammalian lineage.

NGFI-A IMMUNOREACTIVITY IN THE PRIMATE RETINA: IMPLICATIONS FOR GENETIC REGULATION OF PLASTICITY

In rodents, enriched environments drive the expression of the immediate early gene NGFI-A, a regulator of the plasticity marker Synapsin I. Both proteins have been implicated as mediators of plasticity in the rat mammalian retina. In the present work immunocytochemistry directed against these proteins was used to explore their basal activity in the retina of a more visual species, the New World monkey Cebus apella. In contrast to rat, monkey retina displayed high basal expression of both NGFI-A and Synapsin I. The greatest number of NGFI-A-expressing cells was observed within the inner nuclear layer, although NGFI-A positive nuclei were also found in the ganglion cell layer. High levels of Synapsin I were found in the inner plexiform layer and outer plexiform layer. Our findings are consistent with the postulate that the retinas of highly visual animals may experience ongoing reorganization as part of normal visual processing, and that NGFI-A and Synapsin I may be well positioned to regulate some of these changes.