Source: Bèchet, N. B., et al. Direct Cannula Implantation in the Cisterna Magna of Pigs. J. Vis. Exp. (2021).
In this video, we describe a method to access the cisterna magna in a pig model. The cisterna magna is a preferred site for studies concerning cerebrospinal fluid.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animal Preparation
2. Surgery
NOTE: All through the surgery, it is necessary to have at least one assistant to suction the light bleeding and cauterize any severed vessels.
Figure 1. Cisterna magna cannulation in pigs. (A) Pig prepped prior to the start of the surgery and marked where dermal incisions will be performed starting from the occipital crest (OC) then posterior to thoracic vertebrae (TV) and lateral to each ear base (EB). (B) Head in the relaxed position with the trapezius, semispinalis capitus biventer and semispinalis capitus complexus muscles retracted, thus exposing cisterna magna (CM). (C) Head flexed manually to increase access to CM for cannulation and injection. (D) Close-up image of a cannula inserted into CM after injection and fixed in place with the dental cement (DC). (E) Dorsal, ventral, and lateral brain surfaces, respectively, after fluorescent imaging with accompanying structural white light images. Areas of interest that are visible at these surfaces include the interhemispheric fissure (IHS), temporal lobe (TL), and lateral fissure (LF). (F) Structural white light image of the artery and veins on the brain surface. (G) Fluorescent image of (F) showing the tracer distribution along the surface artery. (H) Macroscopic slices from the anterior and posterior cerebral regions show two-dimensional tracer dispersion and distribution in fissures (LF, IHS) and subcortical structures like the striatum (STR) and hippocampus (HPC). (I-J). Confocal images showing the tracer in the PVS, bounded by lectin-stained endothelial cells internally and AQP4 on astrocyte foot processes externally. (K-L). Confocal images showing the tracer in the PVS, bounded by endothelial cells internally with astrocyte foot processes stained for glial fibrillary acidic protein (GFAP) visible forming an outer boundary. (M) Confocal image showing the tracer in the PVS around an arteriole stained for smooth muscle actin (SMA) with tracer also visible in and around, surrounding the brain parenchyma. CM, cisterna magna; DC, dental cement; EB, ear base; GFAP, glial fibrillary acidic protein; HPC, hippocampus; IHS, interhemispheric fissure; LF, lateral fissure; OLB, olfactory bulb; OC, occipital crest; STR, striatum; TL, temporal lobe; TV, thoracic vertebrae.
The authors have nothing to disclose.
Anatomical forceps | NA | NA | |
Marker pen | NA | NA | |
No. 21 Scalpel blade | Agnthos | BB521 | |
No. 4 Scalpel handle | Agnthos | 10004-13 | |
Self-retaining retractors | NA | NA | |
Surgical forceps | NA | NA |