Cocaine-regulated trafficking of dopamine transporters in cultured neurons revealed by a pH sensitive reporter

Saenz, Jacqueline; Yao, Oscar; Khezerlou, Elnaz; Aggarwal, Meha; Zhou, Xiaofeng; Barker, David J.; DiCicco‐Bloom, Emanuel; Pan, Ping-Yue

doi:10.1016/j.isci.2022.105782

Cited by 2 publications

(8 citation statements)

References 88 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While we did not observe these clusters in the brain slices of Synj1+/mice, large varicosities are not uncommon in cultured ventral MB neurons. In our previous study, we showed that large varicosities exhibit 25% of sDAT compared to adjacent axons that exhibits 75% in wildtype neurons [42]. We next sought to determine if the reduced axonal sDAT could be due to a higher density of large varicosities in the Synj1+/-axons.…”

Section: The Number Of Large Varicosity Is Not Altered In Cultured Sy...mentioning

confidence: 96%

“…Second, we expressed a pH sensitive DAT sensor, CAGp-hDAT-pHluorin (Fig. 3E) [42] in cultured ventral MB neurons. Sequential perfusion of a membrane impermeable MES solution (pH 5.5) and a pH neutral NH 4Cl solution (pH 7.4) (Fig.…”

Section: Synj1+/-neurons Exhibit Increased Total Dat and Decreased Su...mentioning

confidence: 99%

“…Sequential perfusion of a membrane impermeable MES solution (pH 5.5) and a pH neutral NH 4Cl solution (pH 7.4) (Fig. 3E) allowed us to calculate the surface fraction and vesicular pH of the intracellular DAT-pHluorin using the Henderson-Hasselbalch equation [42,[50][51][52]. The TH immunoreactivity of the neuron was determined by post hoc analysis (raw data in Supplemental Fig.…”

Section: Synj1+/-neurons Exhibit Increased Total Dat and Decreased Su...mentioning

confidence: 99%

“…Both clathrin-dependent and independent endocytosis of DAT have been reported [32][33][34][35][36][37][38][39][40][41]. Our previous work suggested that Synj1 may participate in the cocaine-induced DAT trafficking [42], however, whether Synj1 regulates DAT recycling in physiological conditions has not been explored. Neither do we understand the interactions between DA signaling and DAT surface availability in Synj1 deficient conditions.…”

Section: Introduction (534 Words)mentioning

confidence: 99%

“…Neither do we understand the interactions between DA signaling and DAT surface availability in Synj1 deficient conditions. We now use a combination of sophisticated imaging tools including dLight [43,44] and DAT-pHluorin [42] to dissect the Synj1's impact on DA homeostasis and the surface maintenance of DAT. Our results revealed an unexpected role of Synj1's 5'-phosphatase domain in maintaining the surface maintenance of DAT by suppressing the PI(4,5)P 2-PKCβ pathway, and we provided essential insights for dysregulation of DA signaling via the dynamic availability of DAT.…”

Section: Introduction (534 Words)mentioning

confidence: 99%

See 4 more Smart Citations

Parkinson’s disease gene, Synaptojanin1, dysregulates the basal maintenance of the dopamine transporter

Pan,

Saenz,

Khezerlou

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

Missense mutations of PARK20/SYNJ1 (synaptojanin1/Synj1) have been linked to complex forms of familial parkinsonism, however, the molecular and cellular changes associated with dopaminergic dysfunction remains unknown. We now report fast depletion of evoked dopamine (DA) and altered maintenance of the axonal dopamine transporter (DAT) in the Synj1+/- neurons. While Synj1 has been traditionally known to facilitate the endocytosis of synaptic vesicles, we demonstrated that axons of cultured Synj1+/- neurons exhibit an increase of total DAT but a reduction of the surface DAT, which could be exacerbated by neuronal activity. We revealed that the loss of surface DAT is specifically associated with the impaired 5’-phosphatase activity of Synj1 and the hyperactive downstream PI(4,5)P2-PKCbeta pathway. Thus, our findings provided important mechanistic insight for Synj1-regulated DAT trafficking integral to dysfunctional DA signaling in early parkinsonism.

show abstract

Section: The Number Of Large Varicosity Is Not Altered In Cultured Sy...mentioning

confidence: 96%

Section: Synj1+/-neurons Exhibit Increased Total Dat and Decreased Su...mentioning

confidence: 99%

Section: Synj1+/-neurons Exhibit Increased Total Dat and Decreased Su...mentioning

confidence: 99%

Section: Introduction (534 Words)mentioning

confidence: 99%

Section: Introduction (534 Words)mentioning

confidence: 99%

See 3 more Smart Citations

Parkinson’s disease gene, Synaptojanin1, dysregulates the basal maintenance of the dopamine transporter

Pan,

Saenz,

Khezerlou

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Neuronal DAMPs exacerbate neurodegeneration via astrocytic RIPK3 signaling

Chang,

DaPrano,

Lindman

et al. 2024

JCI Insight

View full text Add to dashboard Cite

Astrocyte activation is a common feature of neurodegenerative diseases. However, the ways in which dying neurons influence the activity of astrocytes is poorly understood. Receptor interacting protein kinase-3 (RIPK3) signaling has recently been described as a key regulator of neuroinflammation, but whether this kinase mediates astrocytic responsiveness to neuronal death has not yet been studied.Here, we used the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of Parkinson's disease to show that activation of astrocytic RIPK3 drives dopaminergic cell death and axon damage.Transcriptomic profiling revealed that astrocytic RIPK3 promoted gene expression associated with neuroinflammation and movement disorders, and this coincided with significant engagement of damage associated molecular pattern (DAMP) signaling. In mechanistic experiments, we show that factors released from dying neurons signal through receptor for advanced glycation endproducts (RAGE) to induce astrocytic RIPK3 signaling, which conferred inflammatory and neurotoxic functional activity.These findings highlight a mechanism of neuron-glia crosstalk in which neuronal death perpetuates further neurodegeneration by engaging inflammatory astrocyte activation via RIPK3.

show abstract

Cocaine-regulated trafficking of dopamine transporters in cultured neurons revealed by a pH sensitive reporter

Cited by 2 publications

References 88 publications

Parkinson’s disease gene, Synaptojanin1, dysregulates the basal maintenance of the dopamine transporter

Parkinson’s disease gene, Synaptojanin1, dysregulates the basal maintenance of the dopamine transporter

Neuronal DAMPs exacerbate neurodegeneration via astrocytic RIPK3 signaling

Contact Info

Product

Resources

About