Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function

Kearney, Patrick J.; Bolden, Nicholas C; Kahuno, Elizabeth; Conklin, Tucker L; Martin, Gilles E.; Lübec, Gert; Melikian, Haley E.

doi:10.1016/j.jbc.2023.102900

Cited by 9 publications

(23 citation statements)

References 93 publications

(133 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We (9) and others (35) previously reported that DRD2 autoreceptors significantly blunt DA transient amplitudes and accelerate DA clearance. Indeed, in control mice DA transient amplitudes were significantly smaller when evoked in ACSF as compared to those evoked in the presence of L-741,626 (25nM, Figure 2C), a DRD2-specific antagonist, as we previously reported (9). In shRit2 mice, DA transient amplitudes recorded in ACSF were not significantly different transients from control mice (Figure 2C).…”

Section: Short-term Daergic Rit2 Silencing Blunts Da Release In Malesmentioning

confidence: 70%

See 1 more Smart Citation

Rit2 silencing in dopamine neurons drives a progressive Parkinsonian phenotype

Kearney

Zhang

Tan

et al. 2023

Preprint

View full text Add to dashboard Cite

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disease and arises from dopamine (DA) neuron death selectively in the substantia nigra pars compacta (SNc) . Rit2 is a reported PD risk allele, and recent single cell transcriptomic studies identified a major RIT2 cluster in PD DA neurons, potentially linking Rit2 expression anomalies to a PD patient cohort. However, it is still unknown whether Rit2 loss itself is causative for PD or PD-like symptoms. Here we report that conditional Rit2 silencing in mouse DA neurons drove a progressive motor dysfunction that was more rapid in males than females and was rescued at early stages by either inhibiting the DA transporter (DAT) or with L-DOPA treatment. Motor dysfunction was accompanied by decreases in DA release, striatal DA content, phenotypic DAergic markers, and a loss of DA neurons, with increased pSer129-alpha synuclein expression. These results provide the first evidence that Rit2 loss is causal for SNc cell death and a PD-like phenotype, and reveal key sex-specific differences in the response to Rit2 loss.

show abstract

Section: Short-term Daergic Rit2 Silencing Blunts Da Release In Malesmentioning

confidence: 70%

“…Striata were collected by preparing 300µm coronal sections on a Vibratome as previously described (8,9). Sections were collected through the entire striatum, dorsal and ventral striata were subdissected, and slices encompassing each region were pooled for each independent mouse.…”

Section: Tissue Harvesting and Immunoblottingmentioning

confidence: 99%

Rit2 silencing in dopamine neurons drives a progressive Parkinsonian phenotype

Kearney

Zhang

Tan

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In contrast, no region-specificity was observed for KOR in this regard (Figures 3 and 4). Of note, a recent report found that DAT-surface trafficking is sensitive to D2AR stimulation with the selective D2 agonist sumanirole in both the VS and DS, albeit with different kinetics evident in the VS and DS, with short-lived effects in the VS but sustained effects on surface trafficking seen in the DS (91). Interestingly, treatment with U69,593 significantly elevated surface DAT Val559 in the DS, without promoting a further increase in phosphorylation at Thr53 (Figure 4 E and F), which contrasts with our experiments in EM4 cells (Figure 2).…”

Section: Discussionmentioning

confidence: 99%

Kappa Opioid Receptor Antagonism Restores Phosphorylation, Trafficking and Behavior induced by a Disease Associated Dopamine Transporter Variant

Mayer

Stewart

Varman

et al. 2023

Preprint

View full text Add to dashboard Cite

Aberrant dopamine (DA) signaling is implicated in schizophrenia, bipolar disorder (BPD), autism spectrum disorder (ASD), substance use disorder, and attention-deficit/hyperactivity disorder (ADHD). Treatment of these disorders remains inadequate. We established that the human DA transporter (DAT) coding variant (DAT Val559), identified in individuals with ADHD, ASD, or BPD, exhibits anomalous DA efflux (ADE) that is blocked by therapeutic amphetamines and methylphenidate. As the latter agents have high abuse liability, we exploited DAT Val559 knock-in mice to identify non-addictive agents that can normalize DAT Val559 functional and behavioral effects ex vivo and in vivo. Kappa opioid receptors (KORs) are expressed by DA neurons and modulate DA release and clearance, suggesting that targeting KORs might offset the effects of DAT Val559. We establish that enhanced DAT Thr53 phosphorylation and increased DAT surface trafficking associated with DAT Val559 expression are mimicked by KOR agonism of wildtype preparations and rescued by KOR antagonism of DAT Val559 ex vivo preparations. Importantly, KOR antagonism also corrected in vivo DA release and sex-dependent behavioral abnormalities. Given their low abuse liability, our studies with a construct valid model of human DA associated disorders reinforce considerations of KOR antagonism as a pharmacological strategy to treat DA associated brain disorders.

show abstract

“…We previously leveraged the TET-off approach to conditionally silence Rit2 in Pitx3 IRES-tTA midbrain DANs (8,9,34), and reported that DAergic Rit2 silencing in both VTA and SNc for 4-6 weeks had no significant effect on either male or female baseline locomotion in the open field (34). Given that PD is typically late onset and progressive, we aimed to more comprehensively investigate the impact of conditional Rit2 knockdown (KD) in DANs on more complex motor behaviors at either short-term (4-6 weeks; ST) or long-term (5-6 months; LT) timepoints in both male and female mice.…”

Section: Rit2 Silencing Leads To Sex-specific Progressive Motor Dysfu...mentioning

confidence: 99%

“…However, it is still completely unknown whether Rit2 loss is, itself, causative for DAN degeneration and/or PD symptomology. Here, we leveraged our previously described approach for conditional Rit2 silencing in mouse DANs (8,9,34) to directly test this possibility. Our results point to a direct role for Rit2 in DA neuron viability, and that Rit2 loss results in a progressive PD-like phenotype with sex-specific differences in males and females.…”

Section: Introductionmentioning

confidence: 99%

Rit2 silencing in dopamine neurons drives a Parkinsonian phenotype

Kearney

Zhang

Tan

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Parkinsons disease (PD) is the second most prevalent neurodegenerative disease and arises from dopamine (DA) neuron death selectively in the substantia nigra pars compacta (SNc). Rit2 is a reported PD risk allele, and recent single cell transcriptomic studies identified a major RIT2 cluster in PD DA neurons, potentially linking Rit2 expression anomalies to a PD patient cohort. However, it is still unknown whether Rit2 loss itself is causative for PD or PD-like symptoms. Here we report that conditional Rit2 silencing in mouse DA neurons drove a progressive motor dysfunction that was more rapid in males than females and was rescued at early stages by either inhibiting the DA transporter (DAT) or with L-DOPA treatment. Motor dysfunction was accompanied by decreases in DA release, striatal DA content, phenotypic DAergic markers, and a loss of DA neurons, with increased pSer129-alpha synuclein expression. These results provide the first evidence that Rit2 loss is causal for SNc cell death and a PD-like phenotype, and reveal key sex-specific differences in the response to Rit2 loss.

show abstract

Presynaptic Gq-coupled receptors drive biphasic dopamine transporter trafficking that modulates dopamine clearance and motor function

Cited by 9 publications

References 93 publications

Rit2 silencing in dopamine neurons drives a progressive Parkinsonian phenotype

Rit2 silencing in dopamine neurons drives a progressive Parkinsonian phenotype

Kappa Opioid Receptor Antagonism Restores Phosphorylation, Trafficking and Behavior induced by a Disease Associated Dopamine Transporter Variant

Rit2 silencing in dopamine neurons drives a Parkinsonian phenotype

Contact Info

Product

Resources

About