Long-Term Synaptic Changes Induced in the Cerebellar Cortex by Fear Conditioning

AbstractTo better understand learning mechanisms, one needs to study synaptic plasticity induced by behavioral training. Recently, it has been demonstrated that the cerebellum is involved in the consolidation of fear memory. Nevertheless, how the cerebellum contributes to emotional behavior is far from known. In cerebellar slices at 10 min and 24 hr following fear conditioning, we found a long-lasting potentiation of the synapse between parallel fibers and Purkinje cells in vermal lobules V-VI, but not in the climbing fiber synapses. The mechanism is postsynaptic, due to an increased AMPA response. In addition, in hotfoot mice with a primary deficiency of the parallel fiber to Purkinje cell synapse, cued (but not contextual) fear conditioning is affected. We propose that this synapse plays an important role in the learned fear and that its long-term potentiation may represent a contribution to the neural substrate of fear memory

Modulation, plasticity and pathophysiology of the parallel fiber-purkinje cell synapse

The parallel fiber-Purkinje cell (PF-PC) synapse represents the point of maximal signal divergence in the cerebellar cortex with an estimated number of about 60 billion synaptic contacts in the rat and 100,000 billions in humans. At the same time, the Purkinje cell dendritic tree is a site of remarkable convergence of more than 100,000 parallel fiber synapses. Parallel fiber activity generates fast postsynaptic currents via α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, and slower signals, mediated by mGlu1 receptors, resulting in Purkinje cell depolarization accompanied by sharp calcium elevation within dendritic regions. Long-term depression (LTD) and long-term potentiation (LTP) have been widely described for the PF-PC synapse and have been proposed as mechanisms for motor learning. The mechanisms of induction for LTP and LTD involve different signaling mechanisms within the presynaptic terminal and/or at the postsynaptic site, promoting enduring modification in the neurotransmitter release and change in responsiveness to the neurotransmitter. The PF-PC synapse is finely modulated by several neurotransmitters, including serotonin, noradrenaline and acetylcholine. The ability of these neuromodulators to gate LTP and LTD at the PF-PC synapse could, at least in part, explain their effect on cerebellar-dependent learning and memory paradigms. Overall, these findings have important implications for understanding the cerebellar involvement in a series of pathological conditions, ranging from ataxia to autism. For example, PF-PC synapse dysfunctions have been identified in several murine models of spino-cerebellar ataxia (SCA) types 1, 3, 5 and 27. In some cases, the defect is specific for the AMPA receptor signaling (SCA27), while in others the mGlu1 pathway is affected (SCA1, 3, 5). Interestingly, the PF-PC synapse has been shown to be hyper-functional in a mutant mouse model of autism spectrum disorder, with a selective deletion of Pten in Purkinje cells. However, the full range of methodological approaches, that allowed the discovery of the physiological principles of PF-PC synapse function, has not yet been completely exploited to investigate the pathophysiological mechanisms of diseases involving the cerebellum. We, therefore, propose to extend the spectrum of experimental investigations to tackle this problem

Effects of the administration of Elovl5-dependent fatty acids on a spino-cerebellar ataxia 38 mouse model

BACKGROUND: Spinocerebellar ataxia 38 (SCA38) is a rare autosomal neurological disorder characterized by ataxia and cerebellar atrophy. SCA38 is caused by mutations of ELOVL5 gene. ELOVL5 gene encodes a protein, which elongates long chain polyunsaturated fatty acids (PUFAs). Knockout mice lacking Elovl5 recapitulate SCA38 symptoms, including motor coordination impairment and disruption of cerebellar architecture. We asked whether, in Elovl5 knockout mice (Elovl5(−/−)), a diet with both ω3 and ω6 PUFAs downstream Elovl5 can prevent the development of SCA38 symptoms, and at which age such treatment is more effective. Elovl5(−/−) mice were fed either with a diet without or containing PUFAs downstream the Elovl5 enzyme, starting at different ages. Motor behavior was assessed by the balance beam test and cerebellar structure by morphometric analysis. RESULTS: The administration from birth of the diet containing PUFAs downstream Elovl5 led to a significant amelioration of the motor performance in the beam test of Elovl5(−/−) mice, with a reduction of foot slip errors at 6 months from 2.2 ± 0.3 to 1.3 ± 0.2 and at 8 months from 3.1 ± 0.5 to 1.9 ± 0.3. On the contrary, administration at 1 month of age or later had no effect on the motor impairment. The cerebellar Purkinje cell layer and the white matter area of Elovl5(−/ −)mice were not rescued even by the administration of diet from birth, suggesting that the improvement of motor performance in the beam test was due to a functional recovery of the cerebellar circuitry. CONCLUSIONS: These results suggest that the dietary intervention in SCA38, whenever possible, should be started from birth or as early as possible

Mouse brain expression patterns of Spg7, Afg3l1, and Afg3l2 transcripts, encoding for the mitochondrial m-AAA protease

<p>Abstract</p> <p>Background</p> <p>The <it>m</it>-AAA (<b>A</b>TPases <b>A</b>ssociated with a variety of cellular <b>A</b>ctivities) is an evolutionary conserved metalloprotease complex located in the internal mitochondrial membrane. In the mouse, it is a hetero-oligomer variably formed by the <it>Spg7</it>, <it>Afg3l1</it>, and <it>Afg3l2 </it>encoded proteins, or a homo-oligomer formed by either Afg3l1 or Afg3l2. In humans, <it>AFG3L2 </it>and <it>SPG7 </it>genes are conserved, whereas <it>AFG3L1 </it>became a pseudogene. Both <it>AFG3L2 </it>and <it>SPG7 </it>are involved in a neurodegenerative disease, namely the autosomal dominant spinocerebellar ataxia SCA28 and a recessive form of spastic paraplegia, respectively.</p> <p>Results</p> <p>Using quantitative RT-PCR, we measured the expression levels of <it>Spg7</it>, <it>Afg3l1</it>, and <it>Afg3l2 </it>in the mouse brain. In all regions <it>Afg3l2 </it>is the most abundant transcript, followed by <it>Spg7</it>, and <it>Afg3l1</it>, with a ratio of approximately 5:3:1 in whole-brain mRNA. Using <it>in-situ </it>hybridization, we showed that <it>Spg7</it>, <it>Afg3l1 </it>and <it>Afg3l2 </it>have a similar cellular pattern of expression, with high levels in mitral cells, Purkinje cells, deep cerebellar nuclei cells, neocortical and hippocampal pyramidal neurons, and brainstem motor neurons. However, in some neuronal types, differences in the level of expression of these genes were present, suggesting distinct degrees of contribution of their proteins.</p> <p>Conclusions</p> <p>Neurons involved in SCA28 and hereditary spastic paraplegia display high levels of expression, but similar or even higher expression is also present in other types of neurons, not involved in these diseases, suggesting that the selective cell sensitivity should be attributed to other, still unknown, mechanisms.</p

Resurgent Current and Voltage Sensor Trapping Enhanced Activation by a β-Scorpion Toxin Solely in Nav1.6 Channel SIGNIFICANCE IN MICE PURKINJE NEURONS

Abstract Resurgent currents are functionally crucial in sustaining the high frequency firing of cerebellar Purkinje neurons expressing Nav1.6 channels. β-Scorpion toxins, such as CssIV, induce a left shift in the voltage-dependent activation of Nav1.2 channels by "trapping" the IIS4 voltage sensor segment. We found that the dangerous Cn2 β-scorpion peptide induces both the left shift voltage-dependent activation and a transient resurgent current only in human Nav1.6 channels (among 1.1-1.7), whereas CssIV did not induce the resurgent current. Cn2 also produced both actions in mouse Purkinje cells. These findings suggest that only distinct β-toxins produce resurgent currents. We suggest that the novel and unique selectivity of Cn2 could make it a model drug to replace deep brain stimulation of the subthalamic nucleus in patients with Parkinson disease

Genetic deletion of fibroblast growth factor 14 recapitulates phenotypic alterations underlying cognitive impairment associated with schizophrenia

Cognitive processing is highly dependent on the functional integrity of gamma-amino-butyric acid (GABA) interneurons in the brain. These cells regulate excitability and synaptic plasticity of principal neurons balancing the excitatory/inhibitory tone of cortical networks. Reduced function of parvalbumin (PV) interneurons and disruption of GABAergic synapses in the cortical circuitry result in desynchronized network activity associated with cognitive impairment across many psychiatric disorders, including schizophrenia. However, the mechanisms underlying these complex phenotypes are still poorly understood. Here we show that in animal models, genetic deletion of fibroblast growth factor 14 (Fgf14), a regulator of neuronal excitability and synaptic transmission, leads to loss of PV interneurons in the CA1 hippocampal region, a critical area for cognitive function. Strikingly, this cellular phenotype associates with decreased expression of glutamic acid decarboxylase 67 (GAD67) and vesicular GABA transporter (VGAT) and also coincides with disrupted CA1 inhibitory circuitry, reduced in vivo gamma frequency oscillations and impaired working memory. Bioinformatics analysis of schizophrenia transcriptomics revealed functional co-clustering of FGF14 and genes enriched within the GABAergic pathway along with correlatively decreased expression of FGF14, PVALB, GAD67 and VGAT in the disease context. These results indicate that Fgf14(-/-) mice recapitulate salient molecular, cellular, functional and behavioral features associated with human cognitive impairment, and FGF14 loss of function might be associated with the biology of complex brain disorders such as schizophrenia

Algoritmos primais-duais de ponto fixo aplicados ao problema Ridge Regression

In this work we propose algorithms for solving a fixed-point general primal-dual formulation applied to the Ridge Regression problem. We study the primal formulation for regularized least squares problems, especially L2-norm, named Ridge Regression and then describe convex duality for that class of problems. Our strategy was to consider together primal and dual formulations and minimize the duality gap between them. We established the primal-dual fixed point algorithm, named SRP and a reformulation for this method, the main contribution of the thesis, which was more efficient and robust, called acc-SRP method or accelerated version of the SRP method. The theoretical study of the algorithms was done through the analysis of the spectral properties of the associated iteration matrices. We proved the linear convergence of algorithms and some numerical examples comparing two variants for each algorithm proposed were presented. We also showed that our best method, acc-SRP, has excellent numerical performance for solving very ill-conditioned problems, when compared to the conjugate gradient method, which makes it computationally more attractive.Neste trabalho propomos algoritmos para resolver uma formulação primal-dual geral de ponto fixo aplicada ao problema de Ridge Regression. Estudamos a formulação primal para problemas de quadrados mínimos regularizado, em especial na norma L2, nomeados Ridge Regression e descrevemos a dualidade convexa para essa classe de problemas. Nossa estratégia foi considerar as formulações primal e dual conjuntamente, e minimizar o gap de dualidade entre elas. Estabelecemos o algoritmo de ponto fixo primal-dual, nomeado SRP e uma reformulação para esse método, contribuição principal da tese, a qual mostrou-se mais eficaz e robusta, designada por método acc-SRP, ou versão acelerada do método SRP. O estudo teórico dos algoritmos foi feito por meio da análise de propriedades espectrais das matrizes de iteração associadas. Provamos a convergência linear dos algoritmos e apresentamos alguns exemplos numéricos comparando duas variantes para cada algoritmo proposto. Mostramos também que o nosso melhor método, acc-SRP, possui excelente desempenho numérico na resolução de problemas muito mal-condicionados quando comparado ao Método de Gradientes Conjugados, o que o torna computacionalmente mais atraente