behavioralDog (narcoleptic) and MouseNot specified for dogs; DAT knock-out mice and wild-type controls implied
Objective: Monitor sleep-wake cycle through polygraphic recordings to measure non-rapid eye movement sleep and wakefulness in narcoleptic dogs and genetically modified mice
Materials & Equipment Checklist
8 items
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Not specified • Not mentioned • Not mentioned • Not mentioned
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Protocol Steps
View Abstract
The role of dopamine in sleep regulation and in mediating the effects of wake-promoting therapeutics is controversial. In this study, polygraphic recordings and caudate microdialysate dopamine measurements in narcoleptic dogs revealed that the wake-promoting antinarcoleptic compounds modafinil and amphetamine increase extracellular dopamine in a hypocretin receptor 2-independent manner. In mice, deletion of the dopamine transporter (DAT) gene reduced non-rapid eye movement sleep time and increased wakefulness consolidation independently from locomotor effects. DAT knock-out mice were also unresponsive to the normally robust wake-promoting action of modafinil, methamphetamine, and the selective DAT blocker GBR12909 but were hypersensitive to the wake-promoting effects of caffeine. Thus, dopamine transporters play an important role in sleep regulation and are necessary for the specific wake-promoting action of amphetamines and modafinil.
1
Polygraphic Recording Setup
Establish polygraphic recording system in narcoleptic dogs to monitor sleep-wake cycle parameters
Not specifiedNot specified
Note: Recording captures non-rapid eye movement sleep and wakefulness states
View evidence from paper
“polygraphic recordings and caudate microdialysate dopamine measurements in narcoleptic dogs”
2
Baseline Sleep-Wake Monitoring
Record baseline sleep-wake cycle data prior to drug administration
Not specifiedNot specified
Note: Establishes control measurements for comparison
View evidence from paper
“polygraphic recordings...revealed that the wake-promoting antinarcoleptic compounds modafinil and amphetamine”
3
Drug Administration and Concurrent Recording
Administer wake-promoting compounds (modafinil, amphetamine, methamphetamine, GBR12909, or caffeine) while simultaneously recording polygraphic data and measuring dopamine levels
Not specifiedNot specified
Note: Compounds tested include modafinil, amphetamine, methamphetamine, selective DAT blocker GBR12909, and caffeine
View evidence from paper
“wake-promoting antinarcoleptic compounds modafinil and amphetamine increase extracellular dopamine in a hypocretin receptor 2-independent manner”
4
Dopamine Measurement
Measure caudate microdialysate dopamine levels during drug administration in narcoleptic dogs
Not specifiedNot specified
Note: Dopamine measurements conducted concurrently with polygraphic recordings
View evidence from paper
“caudate microdialysate dopamine measurements in narcoleptic dogs revealed that the wake-promoting antinarcoleptic compounds modafinil and amphetamine increase extracellular dopamine”
5
Mouse Sleep-Wake Analysis
In mice, measure non-rapid eye movement sleep time and wakefulness consolidation in DAT knock-out versus wild-type controls
Not specifiedNot specified
Note: DAT deletion reduced NREM sleep time and increased wakefulness consolidation independently from locomotor effects
View evidence from paper
“deletion of the dopamine transporter (DAT) gene reduced non-rapid eye movement sleep time and increased wakefulness consolidation independently from locomotor effects”
6
Drug Response Testing in Mice
Test responsiveness of DAT knock-out mice to wake-promoting compounds (modafinil, methamphetamine, GBR12909, and caffeine)
Not specifiedNot specified
Note: DAT knock-out mice showed no response to modafinil, methamphetamine, and GBR12909 but were hypersensitive to caffeine
View evidence from paper
“DAT knock-out mice were also unresponsive to the normally robust wake-promoting action of modafinil, methamphetamine, and the selective DAT blocker GBR12909 but were hypersensitive to the wake-promoting effects of caffeine”
Subjects / Specimens
Species
Dog (narcoleptic) and Mouse
Strain
Not specified for dogs; DAT knock-out mice and wild-type controls implied
Age
Not specified
Sex
unknown
Weight
Not specified
Study included narcoleptic dogs and mice with dopamine transporter gene deletion