Recording Neuronal Field Excitatory Postsynaptic Potentials in Acute Hippocampal Slices

Abstract

Source: Weng, W., et al. Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System. J. Vis. Exp. (2018).

This video demonstrates the procedure for setting up a brain slice in a submersion chamber to record synaptic activity. The process involves securing the slice on lens paper, maintaining a continuous flow of artificial cerebrospinal fluid (aCSF) to ensure neuronal viability, and accurately positioning the stimulation and recording electrodes relative to a specific brain region of interest. This setup allows for precise monitoring of baseline synaptic activity and the effects of electrical stimulation.

Protocol

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

1. Recording Synaptic Responses in a Submersion Slice Chamber

NOTE: See the Table of Materials.

1. Pre-warm the aCSF solution in a water bath to about 28-30 °C (this will prevent bubble formation in the recording chamber).
2. Start the recycling system (4-5 mL/min) and activate the inline heater to equilibrate the system for at least 1 h.
3. Presoak a small piece of lens cleaning paper and a nylon mesh fixed on a....

Results

Figure 1: Components of outflow-carbogenation for experiments in a submersion slice chamber. (A) Presentation of the submersion slice chamber. The U-shaped platinum wire with nylon fibers holding a hippocampal slice is depicted. The sil.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Electrophysiology equipment and materials
Vertical Pipette Puller	Narishige, Japan	PC-10
Vibration isolation table	Meirits, Japan	ADZ-A0806
Submerged type recording chamber	Warner Instruments	RC-26GLP
Thermostatic water bath	Zhongcheng Yiqi, China	HH-1
4 Axis Micromanipulator	Sutter, USA	MP-285, MP-225
Platinum Wire	World Precision Instruments	PTP406
Amplifier	Molecular Devices, USA	Multiclamp 700B
Data Acquisition System	Molecular Devices, USA	Digidata 1440A
Anaysis software	Molecular Devices, USA	Clampex 10.2
Fluorescence Microscope	Nikon, Japan	FN1
LED light source	Lumen Dynamics Group, Canada	X-cite 120LED
Micropipettes	Harvard apparatus	GC150TF	extracelluar recording
Borosilicate micropipettes	Sutter, USA	BF150-86	patch clamp
Tungsten electrode	A-M Systems, USA	575500
Peristaltic pump	Longer, China	BT00-300T
Tubes for peristaltic pump	ISMATEC, Wertheim, Germany	SC0309	1x inflow, ID: 1.02mm
Tubes for peristaltic pump	ISMATEC, Wertheim, Germany	SC0319	2x tubes for outflow, ID: 2.79 mm
CCD camera	PCO, Germany	pco.edge sCMOS
Lens cleaning paper	Kodak
50 ml conical centrifuge tube	Thermo scientific	339652
Prechamber	Warner Instruments	BSC-PC
Inline heater	Warner Instruments	SF-28
Temperature Controller	Warner Instruments	TC-324B
Reagents required
NaCl	Sinopharm Chemical Reagent, China	10019318
KCl	Sinopharm Chemical Reagent, China	10016318
KH2PO4	Sinopharm Chemical Reagent, China	10017618
MgCl2·6H2O	Sinopharm Chemical Reagent, China	10012818
CaCl2	Sinopharm Chemical Reagent, China	10005861
NaHCO3	Sinopharm Chemical Reagent, China	10018960
Glucose	Sinopharm Chemical Reagent, China	10010518
NaH2PO4	Sinopharm Chemical Reagent, China	20040718
HEPES	Sigma	H3375
Sodium pyruvate	Sigma	A4043
MgSO4	Sinopharm Chemical Reagent, China	20025118
NaOH	Sinopharm Chemical Reagent, China	10019718