Articulo
Constitutive activity of dopamine receptor type 1 (D1R) increases Ca<sub>V</sub>2.2 currents in PFC neurons
Registro en:
issn:1540-7748
issn:0022-1295
Autor
McCarthy, Clara Inés
Chou Freed, Cambria
Rodríguez, Silvia Susana
Yaneff, Agustín
Davio, Carlos
Raingo, Jesica
Institución
Resumen
Alterations in dopamine receptor type 1 (D1R) density are associated with cognitive deficits of aging and schizophrenia. In the prefrontal cortex (PFC), D1R plays a critical role in the regulation of working memory, which is impaired in these cognitive deficit states, but the cellular events triggered by changes in D1R expression remain unknown. A previous report demonstrated that interaction between voltage-gated calcium channel type 2.2 (Ca<sub>V</sub>2.2) and D1R stimulates Ca<sub>V</sub>2.2 postsynaptic surface location in medial PFC pyramidal neurons. Here, we show that in addition to the occurrence of the physical receptor-channel interaction, constitutive D1R activity mediates up-regulation of functional Ca<sub>V</sub>2.2 surface density. We performed patch-clamp experiments on transfected HEK293T cells and wild-type C57BL/6 mouse brain slices, as well as imaging experiments and cAMP measurements. We found that D1R coexpression led to ∼60% increase in Ca<sub>V</sub>2.2 currents in HEK293T cells. This effect was occluded by preincubation with a D1/D5R inverse agonist, chlorpromazine, and by replacing D1R with a D1R mutant lacking constitutive activity. Moreover, D1R-induced increase in Ca<sub>V</sub>2.2 currents required basally active Gs protein, as well as D1R-Ca<sub>V</sub>2.2 interaction. In mice, intraperitoneal administration of chlorpromazine reduced native Ca<sub>V</sub> currents' sensitivity to ω-conotoxin-GVIA and their size by ∼49% in layer V/VI pyramidal neurons from medial PFC, indicating a selective effect on Ca<sub>V</sub>2.2. Additionally, we found that reducing D1/D5R constitutive activity correlates with a decrease in the agonist-induced D1/D5R inhibitory effect on native Ca<sub>V</sub> currents. Our results could be interpreted as a stimulatory effect of D1R constitutive activity on the number of Ca<sub>V</sub>2.2 channels available for dopamine-mediated modulation. Our results contribute to the understanding of the physiological role of D1R constitutive activity and may explain the noncanonical postsynaptic distribution of functional Ca<sub>V</sub>2.2 in PFC neurons. Instituto Multidisciplinario de Biología Celular