Blockade and reversal of swimming-induced paralysis in C. elegans by the antipsychotic and D2-type dopamine receptor antagonist azaperone

Refai, Osama; Blakely, Randy D.

doi:10.1016/j.neuint.2018.05.013

Cited by 18 publications

(14 citation statements)

References 62 publications

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“…It has been proposed that C. elegans movement depends not only on DA, but also on cholinergic neuron-dependent muscle contraction and GABAergic neuron-dependent relaxation. However, it has been shown that blockade of DA D2 receptors does not fully limit paralysis triggered by changes in cholinergic and GABAergic signaling (Refai & Blakely, 2019). The opposite results have also been shown, i.e.…”

Section: Discussionmentioning

confidence: 99%

Doxycycline inhibits α-synuclein-associated pathologiesin vitroandin vivo

Dominguez-Meijide

Parrales

Vasili³

et al. 2020

Preprint

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Parkinson's disease (PD) and dementia with Lewy bodies (DLB) are neurodegenerative disorders characterized by the misfolding and aggregation of alpha-synuclein (aSyn). Doxycycline, a tetracyclic antibiotic shows neuroprotective effects, initially proposed to be due to its anti-inflammatory properties. More recently, an additional mechanism by which doxycycline may exert its neuroprotective effects has been proposed as it has been shown that it inhibits amyloid aggregation. Here, we studied the effects of doxycycline on aSyn aggregation in vivo, in vitro and in a cell free system using real-time quaking induced conversion (RT-QuiC). Our results show that doxycycline decreases the number and size of aSyn aggregates in cells. In addition, doxycycline inhibits the aggregation and seeding of recombinant aSyn, and attenuates the production of mitochondrial-derived reactive oxygen species. Finally, we found doxycycline induces a cellular redistribution of the aggregates in an animal model of PD that is associated with a recovery of dopaminergic function. In summary, we provide strong evidence that doxycycline treatment may be an effective strategy against synucleinopathies.

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Section: Discussionmentioning

confidence: 99%

Doxycycline inhibits α-synuclein-associated pathologiesin vitroandin vivo

Dominguez-Meijide

Parrales

Vasili³

et al. 2020

Preprint

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“…In conclusion, with the right set of experiments, which include the use of knockout animals for key player genes of the dopaminergic system (cat-2, dat-1, dop-3) and the use of drugs depleting DA storages (reserpine), SWIP has been successfully used to elucidate the mechanism of action of drugs like AMPH 5,13 , βPEA 14, 15 and azaperone 6 . Assay set up for SWIP.…”

Section: Discussionmentioning

confidence: 99%

“…amphetamine 5 . On the other hand, pharmacological or genetic manipulations averting synthesis and release of DA and blocking DOP-3 receptor function prevent SWIP 6 . Taken together, these already published data have established SWIP as a reliable tool I) to study the behavioral effects caused by mutated proteins within dopaminergic synapses 3,4,7 and II) to be employed for forward genetic screens for the identification of novel regulatory pathways involved in DA signaling [7][8][9][10][11][12] .…”

Section: Introductionmentioning

confidence: 99%

“…Taken together, these already published data have established SWIP as a reliable tool I) to study the behavioral effects caused by mutated proteins within dopaminergic synapses 3,4,7 and II) to be employed for forward genetic screens for the identification of novel regulatory pathways involved in DA signaling [7][8][9][10][11][12] . Additionally, by providing an easily quantifiable readout of drug-induced behavior in living animals, SWIP enables the elucidation of mechanisms of action of drugs like amphetamine (AMPH) and azaperone at the dopaminergic synapses 5,6,[13][14][15] .…”

Section: Introductionmentioning

confidence: 99%

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Swimming Induced Paralysis to Assess Dopamine Signaling in <em>Caenorhabditis elegans</em>

Kudumala

Sossi

Carvelli

2019

JoVE

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The swimming assay described in this protocol is a valid tool to identify proteins regulating the dopaminergic synapses. Similarly to mammals, dopamine (DA) controls several functions in C. elegans including learning and motor activity. Conditions that stimulate DA release, e.g. amphetamine (AMPH) treatments, or that prevent DA clearance, e.g. animals lacking the DA transporter (dat-1) which are incapable of reaccumulating DA into the neurons, generate an excess of extracellular DA ultimately resulting in inhibited locomotion. This behavior is particularly evident when animals swim in water. In fact, while wild-type animals continue to swim for an extended period, dat-1 null mutants and wild-type treated with AMPH or inhibitors of the DA transporter sink to the bottom of the well and do not move. This behavior is termed "Swimming Induced Paralysis" (SWIP). Although SWIP assay is well established, a detailed description of the method is lacking. Here we describe a step-by-step guide to perform SWIP. To perform the assay, late larval stage-4 animals are placed in a glass spot plate containing control sucrose solution with or without AMPH. Animals are scored for their swimming behavior either manually by visualization under a stereoscope or automatically by recording with a camera mounted on the stereoscope. Videos are then analyzed using a tracking software, which yields a visual representation of thrashing frequency and paralysis in the form of heat maps. Both the manual and automated systems guarantee an easily quantifiable readout of the animals' swimming ability and thus facilitate screening for animals bearing mutations within the dopaminergic system or for auxiliary genes. In addition, SWIP can be used to elucidate the mechanism of action of drugs of abuse such as AMPH.

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“…Swip induced by dat-1 mutations can be prevented by growth of worms on reserpine, an inhibitor of the vesicular monoamine transporter (VMAT/CAT-1) [21], and pharmacologically reversed by treatment of dat-1 paralyzed worms by the DOP-3 antagonist azaperone [22]. To identify novel loss of function alleles in dat-1 , as well as novel determinants of DA signaling, we undertook a forward genetic screen [18] that identified lines displaying reserpine-sensitive Swip, as well as Swip sensitivity to DOP-3 genetic elimination or azaperone treatment.…”

Section: Introductionmentioning

confidence: 99%

Dopamine-dependent, swimming-induced paralysis arises as a consequence of loss of function mutations in the RUNX transcription factor RNT-1

Robinson¹,

Refai²,

Hardaway³

et al. 2019

PLoS ONE

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Dopamine (DA) is a neurotransmitter with actions across phylogeny that modulate core behaviors such as motor activity, reward, attention, and cognition. Perturbed DA signaling in humans is associated with multiple disorders, including addiction, ADHD, schizophrenia, and Parkinson’s disease. The presynaptic DA transporter exerts powerful control on DA signaling by efficient clearance of the neurotransmitter following release. As in vertebrates, Caenorhabditis elegans DAT (DAT-1) constrains DA signaling and loss of function mutations in the dat-1 gene result in slowed crawling on solid media and swimming-induced paralysis (Swip) in water. Previously, we identified a mutant line, vt34 , that exhibits robust DA-dependent Swip. vt34 exhibits biochemical and behavioral phenotypes consistent with reduced DAT-1 function though vt34; dat-1 double mutants exhibit an enhanced Swip phenotype, suggesting contributions of the vt34 -associated mutation to additional mechanisms that lead to excess DA signaling. SNP mapping and whole genome sequencing of vt34 identified the site of the molecular lesion in the gene B0412.2 that encodes the Runx transcription factor ortholog RNT-1. Unlike dat-1 animals, but similar to other loss of function rnt-1 mutants, vt34 exhibits altered male tail morphology and reduced body size. Deletion mutations in both rnt-1 and the bro-1 gene, which encodes a RNT-1 binding partner also exhibit Swip. Both vt34 and rnt-1 mutations exhibit reduced levels of dat-1 mRNA as well as the tyrosine hydroxylase ortholog cat-2 . Although reporter studies indicate that rnt-1 is expressed in DA neurons, its re-expression in DA neurons of vt34 animals fails to fully rescue Swip. Moreover, as shown for vt34 , rnt-1 mutation exhibits additivity with dat-1 in generating Swip, as do rnt-1 and bro-1 mutations, and vt34 exhibits altered capacity for acetylcholine signaling at the neuromuscular junction. Together, these findings identify a novel role for rnt-1 in limiting DA neurotransmission and suggest that loss of RNT-1 may disrupt function of both DA neurons and body wall muscle to drive Swip.

show abstract

Blockade and reversal of swimming-induced paralysis in C. elegans by the antipsychotic and D2-type dopamine receptor antagonist azaperone

Cited by 18 publications

References 62 publications

Doxycycline inhibits α-synuclein-associated pathologiesin vitroandin vivo

Doxycycline inhibits α-synuclein-associated pathologiesin vitroandin vivo

Swimming Induced Paralysis to Assess Dopamine Signaling in <em>Caenorhabditis elegans</em>

Dopamine-dependent, swimming-induced paralysis arises as a consequence of loss of function mutations in the RUNX transcription factor RNT-1

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