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Dopamine Promotes Ascorbate Release from Retinal Neurons: Role of D1 Receptors and the Exchange Protein Directly Activated by cAMP type 2 (EPAC2)

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Abstract

Ascorbate, the reduced form of vitamin C, is highly concentrated in the central nervous system (CNS), including the retina, where it plays important physiological functions. In the CNS, the plasma membrane transporter sodium vitamin C co-transporter 2 (SVCT2) is responsible for ascorbate transport in neurons. The neurotransmitter dopamine (DA), acting through D1- and D2-like receptor subfamilies and classically coupled to adenylyl cyclase, is known to modulate synaptic transmission in the retina. Here, we reveal that DA controls the release of ascorbate from retinal neurons. Using primary retinal cultures, we show that this DA effect is dose-dependent, occurring by the reversal of the SVCT2, and could be elicited by brief and repetitive pulses of DA. The DA effect in inducing ascorbate release occurs by the activation of D1R and is independent of PKA. Moreover, the exchange protein directly activated by cAMP type 2 (EPAC2) is present in retinal neurons and its specific knockdown using shRNAs abrogates the D1R-induced ascorbate release. Confirming the physiological relevance of this pathway, activation of D1R or EPAC2 also triggered ascorbate release ex vivo in acute preparations of the intact retina. Overall, DA plays pivotal roles in regulating ascorbate homeostasis through an unanticipated signaling pathway involving D1R/adenylyl cyclase/cAMP/EPAC2, thereby suggesting that vitamin C might fine-tune dopaminergic neurotransmission in the retina.

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Acknowledgements

This work is dedicated to Fernando Garcia de Mello for his advisory and pioneering contributions to the study of dopamine system in the retina. We greatly acknowledge Luzeli R. de Assis and Sarah de A. Rodrigues for the technical assistance and Dr. Karin C. Calaza for critically reviewing the manuscript.

Funding

This work was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES, Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro—FAPERJ, Pró-Reitoria de Pesquisa, Pós-Graduação e Inovação da Universidade Federal Fluminense—PROPPI/UFF, and the Instituto Nacional de Neurociência Translacional—INNT. Thaísa Godinho da Encarnação and Felipe Nascimento Santiago were recipients of graduate student fellowships from CAPES and Caio Eduardo Nogueira was recipient of an undergraduate fellowship from CNPq. Camila Cabral Portugal and Renato Socodato hold postdoctoral fellowships from Fundação para a Ciência e Tecnologia—FCT Portugal (Refs SFRH/BPD/91962/2012 and SFRH/BPD/91833/2012, respectively). Roberto Paes-de-Carvalho is a research fellow from CNPq and FAPERJ.

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  1. Thaísa Godinho da Encarnação and Camila Cabral Portugal contributed equally to this work.

Authors and Affiliations

  1. Department of Neurobiology and Program of Neurosciences, Institute of Biology, Fluminense Federal University, Outeiro de São João Batista s/n, 24020-141 Niterói, Rio de Janeiro, Brazil

    Thaísa Godinho da Encarnação, Caio Eduardo Nogueira, Felipe Nascimento Santiago & Roberto Paes-de-Carvalho

  2. Instituto de Investigação e Inovação em Saúde and Instituto de Biologia Molecular e Celular (IBMC), Universidade do Porto, Rua Alfredo Allen, 208, 4200-135, Porto, Portugal

    Camila Cabral Portugal & Renato Socodato

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  1. Thaísa Godinho da Encarnação
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Correspondence to Roberto Paes-de-Carvalho.

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da Encarnação, T.G., Portugal, C.C., Nogueira, C.E. et al. Dopamine Promotes Ascorbate Release from Retinal Neurons: Role of D1 Receptors and the Exchange Protein Directly Activated by cAMP type 2 (EPAC2). Mol Neurobiol 55, 7858–7871 (2018). https://doi.org/10.1007/s12035-018-0962-7

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