Recording of Mesenteric Afferent Nerve Activity in an Ex Vivo Mouse Jejunum Segment

Samenvatting

Source: Nullens, S., et al. In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments. J. Vis. Exp. (2016).

This video shows the process of recording electrical signals from mesenteric afferent nerves in a dissected mouse jejunal segment to study the effects of tissue pressure and movement changes on afferent nerve activity.

Protocol

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

1. Preparation of the jejunal afferent nerve

1. Perform rodent euthanasia of the adolescent or adult rodent that has been approved prior to the experiment by the local Ethical Committee (e.g., terminal sedation followed by cardiac puncture, cervical dislocation, etc.).
   NOTE: We used cervical dislocation to sacrifice the animals, thus resulting in experiments without the need for further anesthesia or post-surgical care as tissues a....

Resultaten

Figure 1: Schematic Overview of the Purpose-built Recording Chamber and Suction Electrode. Detailed overview of the technical setup with the suction electrode and the recording chamber in place.

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Sodium chloride (NaCl)	VWR Chemicals	2,78,10,295	compound Krebs solution
Potassium chloride (KCl)	Acros organics	196770010	compound Krebs solution
Sodium dihydrogen phosphate (NaH2PO4)	VWR Chemicals	1,06,34,61,000	compound Krebs solution
Sodium bicarbonate (NaHCO3)	Merck	1,06,32,91,000	compound Krebs solution
Magnesium sulfate (MgSO4)	Merck	1,05,88,61,000	compound Krebs solution
Calcium chloride (CaCl2)	Merck	2,38,11,000	compound Krebs solution
D-glucose	VWR Chemicals	1011175P	compound Krebs solution
Distilled water			compound Krebs solution
PVC tubing	Scientific Laboratory Supplies		The intestinal segment should be mounted over PVC tubing
Silicone tubing	Scientific Laboratory Supplies		The rest of the tubing, ideally silicone-based - more easily dislodging of debris in the tubing
Silk thread	Pearsall Limited	10B15S220	Attachment of the segment over the PVC tubing
Syringe driver	Harvard Apparatus	55-2222	Intraluminal infusion of Krebs
Binocular - including 10x magnification in oculair	Zeiss STEMI 2000		Optimal visualization for the dissection of the afferent nerve
Homeothermic Blanket Control Unit	Harvard Apparatus	507214	Heating of the organ chamber
Custom made organ bath with Sylgard covered bottom
Spike2 software			Recording and analysis of the data
Insect pins, 500 pieces, stainless steel, diameter 0.2 mm	Austerlitz insect pins minutiens		Dissection of the afferent nerve
Tweezer Dumont #5 inox 11cm	World Precision Instrument	500341	Dissection of the afferent nerve
Scissors, spring, 14 cm	World Precision Instrument	15905	Dissection of the afferent nerve
DB digitimer	NL 108T2/10		pressure transducer
Micromanipulator	Narishige	M-3333	3D manipulation of the suction electrode
Micromanipulator	X-4 rotating block		3D manipulation of the suction electrode
Micromanipulator	GJ-8 magnetic stand		3D manipulation of the suction electrode
LightSource	Euromex Microscopes Holland EK-1		Optimal visualization for the dissection of the afferent nerve
CED 1401 Recording Apparatus			Recording of afferent nerve activity
Humbug 50/60Hz Noise Eliminator	Quest Scientific Instruments		Elimination of background noise
Infusion Pump	Gibson Minipuls 2		Infusion of the organ chamber in which the segment is mounted
Microelectrode Holder Half Cells 1.5 mm	World Precision Instrument	MEH2SW	Suction electrode for isolation of the afferent fiber
Borosilicate Glass Capillaries, 300 pc; 1.5/0.84 OD/ID	World Precision Instrument	1B150-4	Capillary for the isolation of the afferent nerve