MicroRNA 142-3p Mediates Post-Transcriptional Regulation of D1 Dopamine Receptor Expression

Abstract

The D1 dopamine receptor subtype is expressed in the brain, kidney and lymphocytes. D1 receptor function has been extensively studied and the receptor has been shown to modulate a wide range of physiological functions and behaviors. The expression of D1 receptor is known to change during development, disease states and chronic treatment; however, the molecular mechanisms that mediate the changes in D1 receptor expression under these circumstances are not well understood. While previous studies have identified extracellular factors and signaling mechanisms regulating the transcription of D1 receptor gene, very little is known about other regulatory mechanisms that modulate the expression of the D1 receptor gene. Here we report that the D1 receptor is post-transcriptionally regulated during postnatal mouse brain development and in the mouse CAD catecholaminergic neuronal cell line. We demonstrate that this post-transcriptional regulation is mediated by a molecular mechanism involving noncoding RNA. We show that the 1277 bp 3′untranslated region of D1 receptor mRNA is necessary and sufficient for mediating the post-transcriptional regulation. Using deletion and site-directed mutagenesis approaches, we show that the D1 receptor post-transcriptional regulation is specifically mediated by microRNA miR-142-3p interacting with a single consensus binding site in the 1277 bp 3′untranslated region of D1 receptor mRNA. Inhibiting endogenous miR-142-3p in CAD cells increased endogenous D1 receptor protein expression levels. The increase in D1 receptor protein levels was biologically significant as it resulted in enhanced D1 receptor-mediated signaling, determined by measuring the activation of both, adenylate cyclase and, the dopamine- and cAMP-regulated phosphoprotein, DARPP-32. We also show that there is an inverse correlation between miR-142-3p levels and D1 receptor protein expression in the mouse brain during postnatal development. This is the first study to demonstrate that the post-transcriptional regulation of D1 receptor expression is mediated by microRNA-induced translational suppression.

Toll-like receptor 4 enhancement of non-NMDA synaptic currents increases dentate excitability after brain injury

“…Western blots of hippocampal tissue from the injured side were performed as described previously (Tobon et al, 2012). Briefly, mice lacking the expression of TLR4 ( tlr4 −/− , Jackson Laboratory), and rats at various time points after FPI or sham-injury (4 hours, 24 hours, 3 days, 7 days and 1 month) were quickly perfused with cold artificial CSF (aCSF, 4°C) under isoflurane anesthesia, fresh hippocampal tissue was isolated under a dissecting microscope and stored at −80°C until use.…”

MicroRNAs in Schizophrenia: Implications for Synaptic Plasticity and Dopamine–Glutamate Interaction at the Postsynaptic Density. New Avenues for Antipsychotic Treatment Under a Theranostic Perspective

“…Moreover, the inhibition of endogenous miR-142-3p in mouse CAD catecholaminergic neuronal cells increased endogenous D1 receptor protein expression levels [138]. It has also been demonstrated that there is an inverse correlation between miR-142-3p levels and D1 receptor protein expression in the mouse brain during postnatal development [136]. Thus, it is conceivable that posttranslational changes in D1 receptor expression could influence NMDA receptor functioning and downstream signaling, with possible consequences in synaptic rearrangements.…”

“…Given the abovementioned functional implications of NMDA-D1 receptors heterodimers [120,135], it is noteworthy that altered D1 receptor gene and protein expression has been related to changes in brain-expressed miRNA levels [136,137]. Studies on D1 receptor expression modulation by miRNAs have shown that miR-142-3p upregulates DRD1 expression by direct binding to the 3′UTR, which leads to differential allele-specific expression of DRD1 in HEK293 cell lines [136]. Moreover, the inhibition of endogenous miR-142-3p in mouse CAD catecholaminergic neuronal cells increased endogenous D1 receptor protein expression levels [138].…”

Postnatal development of the dopaminergic signaling involved in the modulation of intestinal motility in mice

“…Scale bars: =25 μm. (26,27). D1 and D5 receptor subtypes may be coupled to different downstream signaling.…”