Controlled Infusion of Aβ Peptide-Specific Antibodies and Aβ Oligomers into the Rat Hippocampus

Abstract

Source: Sajadi, A., et al. Neurodegeneration in an Animal Model of Chronic Amyloid-beta Oligomer Infusion Is Counteracted by Antibody Treatment Infused with Osmotic Pumps. J. Vis. Exp. (2016).

This video demonstrates a technique for the continuous infusion of amyloid-beta (Aβ) peptide-specific antibodies into the hippocampus of a rat. The study employs implanting a bilateral cannula system for a controlled infusion of Aβ peptide-specific antibodies coupled with the infusion of soluble Aβ oligomers (Aβo) into the rat hippocampus to study the interactions between the oligomers and the antibodies.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Catheter Preparation before Stereotaxic Surgery Cut PE50 catheters (6 cm in length) that will be used to connect the cannula with osmotic pumps (see Figure 1). Fill PE50 catheters with artificial cerebrospinal fluid (aCSF) and seal both ends of the catheters. Keep the catheter at 4 °C until used.</…

Representative Results

Figure 1. Photo of the Animal with Bilateral Cannula During Aβo Infusion. A solution of Aβo (0.2 µg/µl; 1 µl) was injected in awake and freely moving rats (once a day for 6 days) using PE50 catheters connected to an internal cannula inserted into a guide cannula. Treatment with control (Ctl) IgG1 or 6E10 antibody was infused directly at the site of A&#946…

Materials

Artificial Cerebrospinal Fluid (aCSF)	Harvard Apparatus	59-7316
PE50 Catheter thin wall	Plastics one	C232CT
Ketamine Hydrochloride (100 mg/mL)	Bioniche	1989529
Xylaxine Hydrochloride (100 mg/mL)	Bimeda	8XYL004C
Meloxicam (5 mg/mL)	Norbrook	215670I01
Solution of chlorhexidine gluconate 2% and isopropyl alcohol 2%	Carefusion	260100C
Lidocaine Hydrochloride	Alveda Pharma	0122AG01
Bupivacaine Hydrochloride	Hospira	1559
Ophthalmic ointment	Baussh and Lomb inc.	2125706
Stereotaxic frame	Stoelting	51600
Stereotaxic cannula holder arm	Harvard Apparatus	72-4837
Drill	Dremel	8050-N/18
Guide Cannula	Plastics one	326OPG/spc
Injection Cannula	Plastics one	C315I/spc
Dummy Cannula	Plastics one	C315DC/spc
Suture thread coated vicryl rapide 4-0	Ethicon	VR2297
Dental Acrylic Cement	Harvard Apparatus	72-6906
Screws	JI Morris Company	P0090CE125
6E10 antibody (mouse IgG1 isotype)	BioLegend	803003
Mouse IgG1 isotype control antibody	Abcam	AB18447
Alzet osmotic pumps model 1007D	Durect corporation	290
Isoflurane	Baxter	CA2L9100
Amyloid-beta 1-42	rPeptide	A-1163-1
Hamilton syringe (10 µL)	Fisher Scientique	14815279
Infusion Syringe Pump CMA 402	Harvard Apparatus	CMA8003110
Syringe 1 mL	BD	309659
Needle 21G	Terumo	NN-2125R
Snuggle	Lomir Biomedical	RTS04