Investigating Auditory Neuronal Responses in an Awake Mouse in the Presence of an Inhibitory Neurotransmitter Blocker

Samenvatting

Source: Ayala, Y. A., et al. Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice. J. Vis. Exp. (2016).

This video demonstrates the study of neuronal activity in a specific region of the mouse brain that processes auditory stimuli. The technique involves applying an inhibitory neurotransmission blocker and comparing neuronal responses to auditory stimuli before and after the drug is applied to the localized brain region.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Drugs for Microiontophoresis

NOTE: The drugs used for microiontophoresis must have an electrical charge when dissolved in water. Check the literature to see if the drug of interest is appropriate for this procedure. Here, the procedure for gabazine, an antagonist of the GABA_A receptor, is described.

1. Filter distilled water using a 0.2 μm syringe filter to sterilize it and ensure it contains no solutes. Using....

Resultaten

Figure 1. Accessories for Awake Mice Recordings. (A) Headpost. (B) Custom-made cushioned foam restrainer. Place the mouse in between both pieces, leaving the head outside. (C) Modifications to the stereotaxic frame. The headpost holder (top bar) assists during .......

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Tungsten wire	Harvard Apparatus LTD	33-0099	0.005 inches x 3 inches
Borosilicate glass capillary	Harvard Apparatus LTD	30-0053	Borosilicate standard wall without filament, 1.5 mm OD, 0.86 mm ID, 100 mm long
Multibarrel glass capillaries	World Precision Instruments	5B120F-4	5-barrel capillary, 4 inches long, 1.2 mm OD, with filament
Diaplus	DiaDent	2001-2101	Light-curing adhesive, used to attach the tungsten electrode to the glas multibarrel pipette
G-Bond	GC Corporation	2277	Light-curing adhesive, used to attach the headpost to the animal's skull
Charisma	Heraeus Kulzer	66000087	Light-curing composite, used to reinforce the bond of the headpost with the skull
Araldit Cristal	Ceys		2-component expoxy, used to further secure the attachment of the tungsten electrode to the glass multibarrel pipette
Heating blanket	Cibertec	RTC1
Stereotactic frame	Narishige	SR-6N	Modified for mice
Microiontophoretic device	Harvard Apparatus LTD	Neurophore BH-2	Including IP-2 iontophoresis pumps (one for each drug delivery channel) and a balance module
Multibarrel glass pipette puller	Narishige	Model PE-21
LED lamp	Technoflux	CV-215	5 W, 430-485 nm
MicroFil	World Precision Instruments	MF34G-5	Flexible plastic needle, 34 AWG
Imalgene	Merial		Ketamine, 100 mg/mL
Rompun	Bayer		Xylazine, 20 mg/mL
Gabazine / SR-95531	Sigma	S106	Prepare ~ 1000µl of 20 mM gabazine in distilled water and adjust the pH to 4