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Gait Analysis — Suellen M. Walker | ReplicateScience

Experiments/Gait Analysis

Source Paper

Effects of Intrathecal Ketamine in the Neonatal Rat

Suellen M. Walker, B. David Westin, Ronald Deumens, Marjorie Grafe, Tony L. Yaksh

Anesthesiology • 2010

Gait Analysis

behavioralratnot specified

Objective: Assessment of gait parameters in rat pups at P35 following early postnatal intrathecal ketamine or saline administration

Materials & Equipment Checklist

9 items1 from ConductScience

Gather these items before starting the experiment. Check off items as you prepare.

Equipment2

Gait analysis apparatus

not specified • not specified • not specified • not mentioned

Mechanical withdrawal threshold testing apparatus

not specified • not specified • not specified • not mentioned

Materials7

Ketamine

not specified • not specified • not specified • not mentioned

Saline

not specified • not specified • not specified • not mentioned

Fluoro-Jade C

not specified • not specified • not specified • not mentioned

Activated caspase-3 antibody

not specified • not specified • not specified • not mentioned

Ionized calcium binding adapter molecule 1 (Iba1)

not specified • not specified • not specified • not mentioned

Glial fibrillary acid protein (GFAP)

not specified • not specified • not specified • not mentioned

Carrageenan

not specified • not specified • not specified • not mentioned

1 item available from ConductScience

Estimated: $2,610.00

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View Abstract

Background Systemic ketamine can trigger apoptosis in the brain of rodents and primates during susceptible developmental periods. Clinically, spinally administered ketamine may improve the duration or quality of analgesia in children. Ketamine-induced spinal cord toxicity has been reported in adult animals but has not been systematically studied in early development. Methods In anesthetized rat pups, intrathecal ketamine was administered by lumbar percutaneous injection. Changes in mechanical withdrawal threshold evaluated dose-dependent antinociceptive and carrageenan-induced antihyperalgesic effects in rat pups at postnatal day (P) 3 and 21. After intrathecal injection of ketamine at P3, 7, or 21, spinal cords were examined for apoptosis (Fluoro-Jade C and activated caspase-3), histopathologic change, and glial responses (ionized calcium-binding adapter molecule 1 and glial fibrillary acid protein). After maximal doses of ketamine or saline at P3 or P21, sensory thresholds and gait analysis were evaluated at P35. Results Intrathecal injection of 3 mg/kg ketamine at P3 and 15 mg/kg at P21 reverses carrageenan-induced hyperalgesia. Baseline neuronal apoptosis in the spinal cord was greater at P3 than P7, predominantly in the dorsal horn. Intrathecal injection of 3-10 mg/kg ketamine in P3 pups (but not 15 mg/kg at P21) acutely increased apoptosis and microglial activation in the spinal cord and altered spinal function (reduced mechanical withdrawal threshold and altered static gait parameters) at P35. Conclusions Because acute pathology and long-term behavioral change occurred in the same dose range as antihyperalgesic effects, the therapeutic ratio of intrathecal ketamine is less than one in the neonatal rat. This measure facilitates comparison of the relative safety of spinally administered analgesic agents.

Protocol Steps

1

Anesthetize rat pups

Anesthetize rat pups prior to intrathecal injection

not specifiednot specified

Note: Anesthetic type and dose not specified

View evidence from paper

“In anesthetized rat pups, intrathecal ketamine was administered by lumbar percutaneous injection”

2

Administer intrathecal ketamine or saline

Perform lumbar percutaneous injection of ketamine or saline at maximal doses

not specifiednot specified

Note: Administration timing: P3 or P21; maximal doses used

View evidence from paper

“intrathecal ketamine was administered by lumbar percutaneous injection. Following maximal doses of ketamine or saline at P3 or P21”

3

Evaluate mechanical withdrawal threshold

Assess dose-dependent antinociceptive and carrageenan-induced anti-hyperalgesic effects

not specifiednot specified

Note: Testing performed at P3 and P21

View evidence from paper

“Changes in mechanical withdrawal threshold evaluated dose-dependent antinociceptive and carrageenan-induced anti-hyperalgesic effects in postnatal day (P)3 and 21 rat pups”

4

Examine spinal cord tissue

Collect and examine spinal cords for apoptosis, histopathological changes, and glial responses

not specifiednot specified

Note: Performed at P3, P7, or P21 following ketamine administration

View evidence from paper

“Following intrathecal ketamine at P3, 7 or 21, spinal cords were examined for apoptosis (Fluoro-Jade C and activated caspase-3), histopathological change, and glial responses”

5

Perform gait analysis

Evaluate gait parameters in rat pups at P35

not specifiednot specified

Automated gait analysis

Automated gait analysis

Matches: Gait analysis apparatus

$2,610View

Note: Performed at P35, following treatment at P3 or P21

View evidence from paper

“Following maximal doses of ketamine or saline at P3 or P21, sensory thresholds and gait analysis were evaluated at P35”

Subjects / Specimens

Species: rat
Strain: not specified
Age: P35 (postnatal day 35) at time of gait analysis; treated at P3, P7, or P21
Sex: unknown
Weight: not specified

Rat pups treated with intrathecal ketamine or saline at P3 or P21

Protocol Summary

Procedure Steps: 5
Materials: 7
Equipment: 2
Outcome Measures: 8

Equipment Available

Automated gait analysis

Estimated Total$2,610.00

Evidence-Based

14 quotes from paper

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