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Methodenartikel

Generation of Spontaneous and On-Demand Ictal Events in Mouse Brain Cortical Slices

698 weergaven

8 juli 2025

In dit artikel

Samenvatting

Source: Chang, M. et al. Generation and On-Demand Initiation of Acute Ictal Activity in Rodent and Human Tissue. J. Vis. Exp. (2019)

This video demonstrates the induction of ictal events in coronal cortical slices from a transgenic mouse expressing light-activated cation channels in cortical excitatory pyramidal neurons. The slice is placed in a recording chamber, and a recording electrode is inserted into the superficial cortical layer. Upon perfusion with a seizure-inducing drug that blocks potassium channels, the neurons become hyperexcitable and generate spontaneous ictal events. To generate on-demand ictal events, a brief light pulse is used to activate the cation channels in pyramidal neurons, triggering the release of excitatory neurotransmitters, which induce the postsynaptic neurons to generate ictal events for a short duration.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Preparation of dissection solutions and artificial cerebrospinal fluid

  1. Carbogenate ultrapure water (which is water filtered with a resistivity of 18.2 MΩ·cm) with carbogen (95% O2/5% CO2) for 5 min using air stone bubble diffusers (aerators for aquariums).
    Note: Air stones can be connected to the carbogen tank's regulator via standard silicone tubing. If air stones are not available, seal shut the end of the silic....

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Openbaarmakingen

No conflicts of interest declared.

Materialen

Lijst van materialen gebruikt in dit artikel
NaamBedrijfCatalogusnummerOpmerkingen
Sodium pentobarbitalN/AN/APurchased through the Toronto Western Hospital's Suppliers
1 mm syringeN/AN/APurchased through UT Med Store
25G 5/8" sterile needleN/AN/APurchased through UT Med Store
Single edge razor blade (2x)N/AN/APurchased through UT Med Store
Instant adhesive glueN/AN/APurchased through UT Med Store
Lens paperN/AN/APurchased through UT Med Store
Glass petri dish (2x)N/AN/APurchased through UT Med Store
Splinter forceps (2x)N/AN/APurchased through UT Med Store
PVC handle micro spatulaN/AN/APurchased through UT Med Store
Micro spoon with flat endN/AN/APurchased through UT Med Store
Detailing brush 5/0N/AN/APurcahsed from a boutique art store
Wide bore transfer pipetteN/AN/APurchased through UT Med Store
Dental TweezerN/AN/APurchased through UT Med Store
Thermometer (digital)N/AN/APurchased on Amazon.ca
Check carbogen tank (95%O2/5%CO2N/AN/APurchased through the Toronto Western Hospital's Suppliers
VibratomeLeicaN/APurchased through the Toronto Western Hospital's Suppliers
brain slice incubation chamber (a.k.a. brain slice keeper) Scientific Systems Design IncN/A
Sodium Chloride (NaCl)N/AN/APurchased through UT Med Store
Sodium BicarbonateN/AN/APurchased through UT Med Store
DextroseN/AN/APurchased through UT Med Store
Potassium Chloride (KCl)N/AN/APurchased through UT Med Store
Magnesium Sulfate (MgSO4.H2O)N/AN/APurchased through UT Med Store
Sodium phosphate monobasic monohydrate (HNaPO4·H2O)N/AN/APurchased through UT Med Store
Calcium Chloride (CaCl2·2H2O)N/AN/APurchased through UT Med Store
SucroseN/AN/APurchased through UT Med Store

Trefwoorden

Preparatie van corticale coupeselektrofysiologische registratieoptogenetische stimulatieinductie van insultengeneratie van ictale eventstransgeen muismodellichtgeactiveerde kationenkanalenblokkade van spanningsafhankelijke kaliumkanalenafgifte van exciterende neurotransmittersactivatie van postsynaptische neuronen