Source Paper
Targeting CRHR1 Signaling in Experimental Infantile Epileptic Spasms Syndrome: Evidence for Route-Dependent Efficacy
Chachua T, Yum MS, Chern CR, Vieira K, Velíšková J et al.
Children (Basel) • 2026
NMDA-Induced Spasm Model
Objective: To induce spasms using NMDA and evaluate the role of CRHR1 antagonists in modulating spasm frequency and onset latency
This is a NMDA-Induced Spasm Model protocol using Rat as the model organism. The procedure involves 7 procedural steps, 4 equipment items, 5 materials. Extracted from a 2026 paper published in Children (Basel).
Model and subjects
Rat • Sprague-Dawley • Both males and females • P15 (postnatal day 15) • Not specified
Study window
~5 hours hands-on
Core workflow
Surgery for cannula implantation • Drug administration preparation • Intracranial drug microinfusion
Primary readouts
- Number of spasms within 90 minutes (primary outcome)
- Latency to onset of spasms from time of NMDA injection (secondary outcome)
- Presence of preceding symptoms (tail twisting, arching) as NMDA administration confirmation
Key equipment and reagents
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Protocol Steps
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Surgery for cannula implantation
On P13, anesthetize pups with isoflurane and implant guide cannula using stereotaxic coordinates
View evidence from paper
“On P13, rat pups (both males and females) were anesthetized by isoflurane inhalation”
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”
Intracranial drug microinfusion
Gently immobilize animals and slowly microinfuse 0.5 µL of drug solution over 1 minute through implanted 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”
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”
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”
Behavioral observation
Place animals in separate cages on heating pad and observe for 90 minutes, recording spasm symptoms
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”
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”