RESUMEN
Reduced expression of the Gad1 gene-encoded 67-kDa protein isoform of glutamic acid decarboxylase (GAD67) is a hallmark of schizophrenia. GAD67 downregulation occurs in multiple interneuronal sub-populations, including the parvalbumin-positive (PVALB+) cells. To investigate the role of the PV-positive GABAergic interneurons in behavioral and molecular processes, we knocked down the Gad1 transcript using a microRNA engineered to target specifically Gad1 mRNA under the control of Pvalb bacterial artificial chromosome. Verification of construct expression was performed by immunohistochemistry. Follow-up electrophysiological studies revealed a significant reduction in γ-aminobutyric acid (GABA) release probability without alterations in postsynaptic membrane properties or changes in glutamatergic release probability in the prefrontal cortex pyramidal neurons. Behavioral characterization of our transgenic (Tg) mice uncovered that the Pvalb/Gad1 Tg mice have pronounced sensorimotor gating deficits, increased novelty-seeking and reduced fear extinction. Furthermore, NMDA (N-methyl-d-aspartate) receptor antagonism by ketamine had an opposing dose-dependent effect, suggesting that the differential dosage of ketamine might have divergent effects on behavioral processes. All behavioral studies were validated using a second cohort of animals. Our results suggest that reduction of GABAergic transmission from PVALB+ interneurons primarily impacts behavioral domains related to fear and novelty seeking and that these alterations might be related to the behavioral phenotype observed in schizophrenia.
Asunto(s)
Conducta Animal , Glutamato Descarboxilasa/genética , Interneuronas/metabolismo , Parvalbúminas/metabolismo , Esquizofrenia/genética , Animales , Encéfalo/fisiología , Modelos Animales de Enfermedad , Electrofisiología , Conducta Exploratoria , Miedo , Silenciador del Gen , Ketamina/administración & dosificación , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Transgénicos , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Filtrado Sensorial/genética , Transmisión SinápticaRESUMEN
Endocannabinoids are released 'on-demand' on the basis of physiological need, and can be pharmacologically augmented by inhibiting their catabolic degradation. The endocannabinoid anandamide is degraded by the catabolic enzyme fatty acid amide hydrolase (FAAH). Anandamide is implicated in the mediation of fear behaviors, including fear extinction, suggesting that selectively elevating brain anandamide could modulate plastic changes in fear. Here we first tested this hypothesis with preclinical experiments employing a novel, potent and selective FAAH inhibitor, AM3506 (5-(4-hydroxyphenyl)pentanesulfonyl fluoride). Systemic AM3506 administration before extinction decreased fear during a retrieval test in a mouse model of impaired extinction. AM3506 had no effects on fear in the absence of extinction training, or on various non-fear-related measures. Anandamide levels in the basolateral amygdala were increased by extinction training and augmented by systemic AM3506, whereas application of AM3506 to amygdala slices promoted long-term depression of inhibitory transmission, a form of synaptic plasticity linked to extinction. Further supporting the amygdala as effect-locus, the fear-reducing effects of systemic AM3506 were blocked by intra-amygdala infusion of a CB1 receptor antagonist and were fully recapitulated by intra-amygdala infusion of AM3506. On the basis of these preclinical findings, we hypothesized that variation in the human FAAH gene would predict individual differences in amygdala threat-processing and stress-coping traits. Consistent with this, carriers of a low-expressing FAAH variant (385A allele; rs324420) exhibited quicker habituation of amygdala reactivity to threat, and had lower scores on the personality trait of stress-reactivity. Our findings show that augmenting amygdala anandamide enables extinction-driven reductions in fear in mouse and may promote stress-coping in humans.
Asunto(s)
Amígdala del Cerebelo/metabolismo , Amígdala del Cerebelo/fisiología , Ácidos Araquidónicos/fisiología , Endocannabinoides/fisiología , Extinción Psicológica/fisiología , Miedo/fisiología , Adaptación Psicológica/fisiología , Adulto , Alcanosulfonatos/administración & dosificación , Alcanosulfonatos/farmacología , Amidohidrolasas/antagonistas & inhibidores , Amidohidrolasas/genética , Amígdala del Cerebelo/efectos de los fármacos , Animales , Ácidos Araquidónicos/metabolismo , Antagonistas de Receptores de Cannabinoides/administración & dosificación , Antagonistas de Receptores de Cannabinoides/farmacología , Condicionamiento Psicológico/efectos de los fármacos , Condicionamiento Psicológico/fisiología , Endocannabinoides/metabolismo , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/farmacología , Miedo/efectos de los fármacos , Miedo/psicología , Femenino , Neuroimagen Funcional , Estudios de Asociación Genética , Habituación Psicofisiológica/efectos de los fármacos , Habituación Psicofisiológica/fisiología , Humanos , Masculino , Ratones , Microinyecciones , Persona de Mediana Edad , Plasticidad Neuronal/efectos de los fármacos , Plasticidad Neuronal/fisiología , Personalidad/genética , Personalidad/fisiología , Fenoles/administración & dosificación , Fenoles/farmacología , Piperidinas/administración & dosificación , Piperidinas/farmacología , Polimorfismo de Nucleótido Simple , Alcamidas Poliinsaturadas/metabolismo , Pirazoles/administración & dosificación , Pirazoles/farmacología , RimonabantRESUMEN
The molecular constituents of endocannabinoid (eCB) signaling are abundantly expressed within the mammalian amygdaloid complex, consistent with the robust role of eCB signaling in the modulation of emotional behavior, learning, and stress-response physiology. Here, we detail the anatomical distribution of eCB signaling machinery in the amygdala and the role of this system in the modulation of excitatory and inhibitory neuroplasticity in this region. We also summarize recent findings demonstrating dynamic alternations in eCB signaling that occur in response to stress exposure, as well as known behavioral consequences of eCB-mediated modulation of amygdala function. Finally, we discuss how integrating anatomical and physiological data regarding eCB signaling in the amygdala could help elucidate common functional motifs of this system in relation to broader forebrain function.