DAT Knockout Mouse Sleep Analysis
Objective: Assessment of sleep-wake cycle parameters and wakefulness consolidation in dopamine transporter gene-deleted mice, and evaluation of responsiveness to wake-promoting compounds
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials5
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Protocol Steps
Polygraphic Recording Setup
Establish polygraphic recordings to measure sleep-wake cycle parameters in DAT knockout mice and control animals
Note: Recordings used to assess non-rapid eye movement sleep time and wakefulness consolidation
View evidence from paper
“polygraphic recordings and caudate microdialysate dopamine measurements in narcoleptic dogs revealed”
Baseline Sleep-Wake Measurement
Record baseline sleep-wake cycle parameters in DAT knockout mice prior to drug administration
Note: Establish baseline non-rapid eye movement sleep time and wakefulness consolidation measures
View evidence from paper
“deletion of the dopamine transporter (DAT) gene reduced non-rapid eye movement sleep time and increased wakefulness consolidation”
Modafinil Administration and Response Testing
Administer modafinil to DAT knockout mice and measure wake-promoting response
Note: DAT knockout mice showed no response to modafinil despite normal responsiveness in wild-type controls
View evidence from paper
“DAT knock-out mice were also unresponsive to the normally robust wake-promoting action of modafinil”
Methamphetamine Administration and Response Testing
Administer methamphetamine to DAT knockout mice and measure wake-promoting response
Note: DAT knockout mice showed no response to methamphetamine
View evidence from paper
“DAT knock-out mice were also unresponsive to the normally robust wake-promoting action of methamphetamine”
GBR12909 Administration and Response Testing
Administer selective DAT blocker GBR12909 to DAT knockout mice and measure wake-promoting response
Note: DAT knockout mice showed no response to GBR12909
View evidence from paper
“selective DAT blocker GBR12909 but were hypersensitive to the wake-promoting effects of caffeine”
Caffeine Administration and Response Testing
Administer caffeine to DAT knockout mice and measure wake-promoting response
Note: DAT knockout mice showed hypersensitivity to caffeine wake-promoting effects
View evidence from paper
“were hypersensitive to the wake-promoting effects of caffeine”
Dopamine Measurement
Measure extracellular dopamine levels in caudate region during drug administration
Note: Microdialysis measurements to assess dopamine changes in response to wake-promoting compounds
View evidence from paper
“caudate microdialysate dopamine measurements in narcoleptic dogs revealed that the wake-promoting antinarcoleptic compounds modafinil and amphetamine increase extracellular dopamine”
Locomotor Activity Assessment
Measure locomotor activity to distinguish sleep-wake effects from general motor effects
Note: Wakefulness consolidation effects were independent 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”