Abstract
Before being released, nucleotides are stored in secretory vesicles through the vesicular nucleotide transporter (VNUT). Once released, extracellular ATP participates in neuronal differentiation processes. Thus, the expression of a functional VNUT could be an additional component of the purinergic system which regulates neuronal differentiation and axonal elongation. In vitro expression of VNUT decreases neuritogenesis in N2a cells differentiated by retinoic acid treatment, whereas silencing of VNUT expression increases the number and length of neurites in these cells. These results highlight the role of VNUT in the neuritogenic process because this transporter regulates the ATP content in neurosecretory vesicles.Electronic supplementary materialThe online version of this article (doi:10.1007/s11302-015-9449-4) contains supplementary material, which is available to authorized users.
Highlights
Neurotransmission and neuroendocrine signaling depends on the regulated release of a large variety of vesicular-stored signaling molecules
In order to validate the molecular tools used in this study, N2a cells were transfected with the designed constructs VNUTmyc (Fig. 1a) and small hairpin RNAs (shRNA) either control or designed against vesicular nucleotide transporter (VNUT)
After 24 h, the transfection efficiency was about 85 % and mRNA and protein levels of VNUT were analyzed by quantitative real-time PCR (qPCR) (Fig. 1b) and Western blotting (Fig. 1c), respectively
Summary
Neurotransmission and neuroendocrine signaling depends on the regulated release of a large variety of vesicular-stored signaling molecules. Non-peptidergic compounds require specific vesicular transporters, all of them are members of the solute carrier family (SLC) [1, 2]. It is widely accepted that a single secretory vesicle contains more than one neurotransmitter and requires the presence of the corresponding specific transporters in the membrane vesicles. ATP and a large variety of nucleotidic compounds are among the most frequent substances co-stored with the classical neurotransmitters earlier discovered, such as acetylcoline, catecholamines, serotonine, and even glutamate, or other aminoacidic compounds [3, 4]. To explain the complexity of signaling events upon vesicular release, G. Burnstock coined the term of co-transmission [5]
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