Controlled Infusion of Aβ Peptide-Specific Antibodies and Aβ Oligomers into the Rat Hippocampus
Published: April 30, 2024
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 Aβ…
開示
The authors have nothing to disclose.
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 |
