1
Surgery for cannula implantation
On P13, anesthetize pups with isoflurane and implant guide cannula using stereotaxic coordinates
Double Cannula System Pack
Matches: Microinfusion guide cannula
View evidence from paper
“On P13, rat pups (both males and females) were anesthetized by isoflurane inhalation”
2
Drug administration preparation
On P15, prepare drug solutions and administer either intracranially or systemically 1 hour prior to NMDA
View evidence from paper
“Both CP376395 and SN003 microinfusions were completed 1 h prior to NMDA induction of spasms”
3
Intracranial drug microinfusion
Gently immobilize animals and slowly microinfuse 0.5 µL of drug solution over 1 minute through implanted cannula
Double Cannula System Pack
Matches: Microinfusion guide cannula
View evidence from paper
“Drug solutions at varying concentrations were prepared in a fixed total volume of 0.5 µL and were slowly microinfused over 1 min”
4
Systemic drug administration
Inject CP376395 at 3 mg/kg i.p. or SN003 at 0.7 mg/kg i.p. 1 hour prior to NMDA
View evidence from paper
“CP376395 was diluted in normal saline and injected at 3 mg/kg, whereas SN003 was first diluted in 100% ethanol, then diluted with distilled water to a 1% ethanol solution and injected at 0.7 mg/kg”
5
NMDA spasm induction
Administer NMDA at 15 mg/kg i.p. dissolved in normal saline on P15
View evidence from paper
“NMDA was administered i.p. on P15 at a dose of 15 mg/kg dissolved in normal saline”
6
Behavioral observation
Place animals in separate cages on heating pad and observe for 90 minutes, recording spasm symptoms
Rodent Heating Pad
Matches: Heating pad
View evidence from paper
“Immediately after the i.p. injection, the animals were placed in separate cages on a heating pad and observed for symptomatology of NMDA syndrome”
7
Symptom monitoring
Record latency to onset of tail twisting, arching, and flexion spasms, and count total spasms within 90 minutes
View evidence from paper
“Besides marking latency to onset of tail twisting (snake-like tail movements starting at the tip), arching, and flexion spasms (emprosthotonus, hyperflexion position on the side lasting < 10 s), we also counted the total number of spasms within the 90 min observation period”
