Joshua L. Plotkin

Differential excitability and modulation of striatal medium spiny neuron dendrites.

The loss of striatal dopamine (DA) in Parkinsons disease (PD) models triggers a cell-type-specific reduction in the density of dendritic spines in D2 receptor-expressing striatopallidal medium spiny neurons (D2 MSNs). How the intrinsic properties of MSN dendrites, where the vast majority of DA receptors are found, contribute to this adaptation is not clear. To address this question, two-photon laser scanning microscopy (2PLSM) was performed in patch-clamped mouse MSNs identified in striatal slices by expression of green fluorescent protein (eGFP) controlled by DA receptor promoters. These studies revealed that single backpropagating action potentials (bAPs) produced more reliable elevations in cytosolic Ca2+ concentration at distal dendritic locations in D2 MSNs than at similar locations in D1 receptor-expressing striatonigral MSNs (D1 MSNs). In both cell types, the dendritic Ca2+ entry elicited by bAPs was enhanced by pharmacological blockade of Kv4, but not Kv1 K+ channels. Local application of DA depressed dendritic bAP-evoked Ca2+ transients, whereas application of ACh increased these Ca2+ transients in D2 MSNs, but not in D1 MSNs. After DA depletion, bAP-evoked Ca2+ transients were enhanced in distal dendrites and spines in D2 MSNs. Together, these results suggest that normally D2 MSN dendrites are more excitable than those of D1 MSNs and that DA depletion exaggerates this asymmetry, potentially contributing to adaptations in PD models.

MicroRNA-128 governs neuronal excitability and motor behavior in mice.

Not Too Much, Not Too Little The microRNA miR128 is expressed in brain neurons of the mouse. Lek Tan et al. (p. 1254) now find that miR128 is crucial to stable brain function. Mice deficient in miR128 developed hyperactivity and were susceptible to fatal seizures, whereas overexpression of miR128 correlated with reduced motor activity and reduced susceptibility to proconvulsive drugs. Experiments using ex vivo–isolated adult brain tissues suggested that miR-128 controlled motor activity by governing the signaling network that determines the intrinsic excitability and signal responsiveness of neurons. A microRNA expressed in adult neurons affects movement by modulating neuronal signaling networks and excitability. The control of motor behavior in animals and humans requires constant adaptation of neuronal networks to signals of various types and strengths. We found that microRNA-128 (miR-128), which is expressed in adult neurons, regulates motor behavior by modulating neuronal signaling networks and excitability. miR-128 governs motor activity by suppressing the expression of various ion channels and signaling components of the extracellular signal–regulated kinase ERK2 network that regulate neuronal excitability. In mice, a reduction of miR-128 expression in postnatal neurons causes increased motor activity and fatal epilepsy. Overexpression of miR-128 attenuates neuronal responsiveness, suppresses motor activity, and alleviates motor abnormalities associated with Parkinson’s–like disease and seizures in mice. These data suggest a therapeutic potential for miR-128 in the treatment of epilepsy and movement disorders.

Dopamine and synaptic plasticity in dorsal striatal circuits controlling action selection

The striatum is thought to play a central role in learning how to choose acts that lead to reward and avoid punishment. Dopamine-dependent modification of striatal synapses in the action selection circuitry has long been thought to be a key step toward this type of learning. The development of new genetic and optical tools has pushed this field forward in the last couple of years, demanding a re-evaluation of models of how experience controls dopamine-dependent synaptic plasticity and how disease states like Parkinsons disease affect the striatal circuitry.

M4 Muscarinic Receptor Signaling Ameliorates Striatal Plasticity Deficits in Models of L-DOPA-Induced Dyskinesia

A balanced interaction between dopaminergic and cholinergic signaling in the striatum is critical to goal-directed behavior. But how this interaction modulates corticostriatal synaptic plasticity underlying learned actions remains unclear--particularly in direct-pathway spiny projection neurons (dSPNs). Our studies show that in dSPNs, endogenous cholinergic signaling through M4 muscarinic receptors (M4Rs) promoted long-term depression of corticostriatal glutamatergic synapses, by suppressing regulator of G protein signaling type 4 (RGS4) activity, and blocked D1 dopamine receptor dependent long-term potentiation (LTP). Furthermore, in a mouse model of L-3,4-dihydroxyphenylalanine (L-DOPA)-induced dyskinesia (LID) in Parkinsons disease (PD), boosting M4R signaling with positive allosteric modulator (PAM) blocked aberrant LTP in dSPNs, enabled LTP reversal, and attenuated dyskinetic behaviors. An M4R PAM also was effective in a primate LID model. Taken together, these studies identify an important signaling pathway controlling striatal synaptic plasticity and point to a novel pharmacological strategy for alleviating LID in PD patients.

The role of dopamine in modulating the structure and function of striatal circuits

Dopamine (DA) is a key regulator of action selection and associative learning. The striatum has long been thought to be a major locus of DA action in this process. Although all striatal cell types express G protein-coupled receptors for DA, the effects of DA on principal medium spiny neurons (MSNs) understandably have received the most attention. In the two principal classes of MSN, DA receptor expression diverges, with striatonigral MSNs robustly expressing D(1) receptors and striatopallidal MSNs expressing D(2) receptors. In the last couple of years, our understanding of how these receptors and the intracellular signalling cascades that they couple to modulate dendritic physiology and synaptic plasticity has rapidly expanded, fuelled in large measure by the development of new optical and genetic tools. These tools also have enabled a rapid expansion of our understanding of the striatal adaptations in models of Parkinsons disease. This chapter highlights some of the major advances in these areas.

Functional and molecular development of striatal fast-spiking GABAergic interneurons and their cortical inputs

Despite their small number, fast‐spiking (FS) GABAergic interneurons play a critical role in controlling striatal output by mediating cortical feed‐forward inhibition of striatal medium‐sized spiny (MS) projection neurons. We have examined the functional development of FS interneurons and their cortical inputs, and the expression of three of their molecular markers, in the dorsolateral rat striatum between postnatal days (P)12–14 and 19–23, the time of major corticostriatal synaptogenesis. FS interneurons were visualized with infrared differential interference contrast (IR‐DIC) optics and examined with current‐clamp recording in the presence of the GABAA receptor antagonist bicuculline methiodide. FS interneurons displayed action potentials at relatively high frequencies in response to depolarizing current pulses by P12, but developmental changes occurred in action potential and afterhyperpolarization duration and amplitude and input resistance between P12–14 and P19–23, as well as an increase in maximum firing frequency in response to depolarizing current pulses. Maturation in electrophysiological properties was paralleled by increases in Kv3.1 and parvalbumin mRNA expression, while GAD‐67 mRNA levels remained constant. Furthermore, FS interneurons in the younger age group responded to stimulation of cortical afferents with excitatory postsynaptic potentials (EPSPs) of higher amplitudes and received significantly more spontaneous depolarizing inputs than did MS neurons. Thus, FS interneurons are under frequent and continuous cortical influence by the end of the 2nd postnatal week, a time when corticostriatal synapses are sparse, suggesting that they may provide a major inhibitory influence in the striatum during the period of intense developmental maturation.

Corticostriatal synaptic adaptations in Huntington's disease.

Huntington’s disease (HD) is a progressive neurodegenerative disorder that profoundly impairs corticostriatal information processing. While late stage pathology includes cell death, the appearance of motor symptoms parallels more subtle changes in neuronal function and synaptic integration. Because of the difficulty in modeling the disease and the complexity of the corticostriatal network, understanding the mechanisms driving pathology has been slow to develop. In recent years, advances in animal models and network analysis tools have begun to shed light on the circuit-specific deficits. These studies have revealed a progressive impairment of corticostriatal synaptic signaling in sub-populations of striatal neurons, turning classical excitotoxicity models of HD upside down. Disrupted brain derived neurotrophic factor signaling appears to be a key factor in this decline.

Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice

Mice carrying bacterial artificial chromosome (BAC) transgenes have become important tools for neuroscientists, providing a powerful means of dissecting complex neural circuits in the brain. Recently, it was reported that one popular line of these mice—mice possessing a BAC transgene with a D2 dopamine receptor (Drd2) promoter construct coupled to an enhanced green fluorescent protein (eGFP) reporter—had abnormal striatal gene expression, physiology, and motor behavior. Unlike most of the work using BAC mice, this interesting study relied upon mice backcrossed on the outbred Swiss Webster (SW) strain that were homozygous for the Drd2-eGFP BAC transgene. The experiments reported here were conducted to determine whether mouse strain or zygosity was a factor in the reported abnormalities. As reported, SW mice were very sensitive to transgene expression. However, in more commonly used inbred strains of mice (C57BL/6, FVB/N) that were hemizygous for the transgene, the Drd2-eGFP BAC transgene did not alter striatal gene expression, physiology, or motor behavior. Thus, the use of inbred strains of mice that are hemizygous for the Drd2 BAC transgene provides a reliable tool for studying basal ganglia function.

Regulation of dendritic calcium release in striatal spiny projection neurons

The induction of corticostriatal long-term depression (LTD) in striatal spiny projection neurons (SPNs) requires coactivation of group I metabotropic glutamate receptors (mGluRs) and L-type Ca(2+) channels. This combination leads to the postsynaptic production of endocannabinoids that act presynaptically to reduce glutamate release. Although the necessity of coactivation is agreed upon, why it is necessary in physiologically meaningful settings is not. The studies described here attempt to answer this question by using two-photon laser scanning microscopy and patch-clamp electrophysiology to interrogate the dendritic synapses of SPNs in ex vivo brain slices from transgenic mice. These experiments revealed that postsynaptic action potentials induce robust ryanodine receptor (RYR)-dependent Ca(2+)-induced-Ca(2+) release (CICR) in SPN dendritic spines. Depolarization-induced opening of voltage-gated Ca(2+) channels was necessary for CICR. CICR was more robust in indirect pathway SPNs than in direct pathway SPNs, particularly in distal dendrites. Although it did not increase intracellular Ca(2+) concentration alone, group I mGluR activation enhanced CICR and slowed Ca(2+) clearance, extending the activity-evoked intraspine transient. The mGluR modulation of CICR was sensitive to antagonism of inositol trisphosphate receptors, RYRs, src kinase, and Cav1.3 L-type Ca(2+) channels. Uncaging glutamate at individual spines effectively activated mGluRs and facilitated CICR induced by back-propagating action potentials. Disrupting CICR by antagonizing RYRs prevented the induction of corticostriatal LTD with spike-timing protocols. In contrast, mGluRs had no effect on the induction of long-term potentiation. Taken together, these results make clearer how coactivation of mGluRs and L-type Ca(2+) channels promotes the induction of activity-dependent LTD in SPNs.

Development of striatal fast-spiking GABAergic interneurons.

Fast-spiking GABAergic interneurons represent a very small portion of striatal neurons, yet they play a critical role in modulating cortical input and mediating inhibition of striatal medium-sized spiny projection neurons. Considering their pivotal role in the adult striatum, it is of importance to determine when during development these neurons acquire their characteristic properties and function. In this review we describe recent work from our laboratories indicating that fast-spiking GABAergic interneurons are under stronger cortical control than efferent neurons at postnatal day 12 but mature considerably between postnatal days 12-19 in the rat striatum. During this time period, their molecular development is under the control of GABAergic and cholinergic mechanisms. Thus, fast-spiking interneurons are poised to influence striatal function and perhaps development during the postnatal period in rats, and their properties could be influenced by commonly used pharmacological agents during a protracted developmental window. These findings point to the need for future research to better understand the functional maturation of this critical population of striatal GABAergic neurons, and the consequences of abnormal maturation of these cells.