Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

Summary

There is accumulating evidence, that ischemic preconditioning (PC) – a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

Abstract

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

Introduction

Ischemic stroke is a disease with a high mortality and an enormous socio-economic burden ¹. Despite intensive experimental and clinical scientific efforts throughout recent decades, treatment options for acute ischemic stroke patients remain very limited ². In contrast to that, increase in the percentage of elderly people in developed countries will dramatically increase incidences and prevalences of patients with ischemic stroke in the next decades ³. Therefore, there is a pressing need for new treatment strategies in patients with ischemic stroke.

One approach is to gain further understanding in mechanism....

Protocol

All procedures described in this article were performed in accordance with the guidelines and regulations of the Landesamt für Gesundheit und Soziales, Berlin, Germany.

Pre-surgical procedures

1. Preparation of Monofilaments

1. For preparation of intraluminal filaments, 8-0 nylon monofilament sutures were used and were cut to a length of 13 mm each.
2. For coating of the monofilaments, Xantopren M mucosa, (Heraeus Kulzer GmbH, Hanau, Germany) was used activated by adding a drop of activator solution (Activator, Universal plus, Heraeus Kulzer GmbH, Hanau, Germany).
3. The monofila....

Results

Our BCCAO protocol resulted in immediate and profound (>90%) CBF reductions when both CCA's were occluded (Figure 1). Reopening of the CCA's led to complete reperfusion as monitored by LDF (Figure 1). After PC, animals showed no signs of functional disability. Furthermore, there was no mortality. TUNEL staining performed 72 hr after BCCAO in a separate group of animals showed no signs of delayed cell death in the brain after BCCAO (data not shown).

When an.......

Discussion

In our study, we have shown that three sequences of BCCAO, each lasting for one minute and followed by reperfusion for five minutes, are an adequate ischemic PC stimulus to induce early IT.

Furthermore, we have demonstrated that the preconditioning protocol used is a safe and minor invasive procedure with no noteworthy postoperative morbidity, no signs of apoptosis in brain and no mortality. Several different preconditioning protocols have been described previously ^6-7,12. Others .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Binocular surgical microscope	Zeiss	Stemi 2000 C
Light source for microscope	Zeiss	SteREO CL 1500 ECO
Heating pad with rectal probe	FST	21061-10
Stereotactic frame	David Kopf	Model 930
Scissors	FST	91460-11
Dumont forceps #5	FST	11251-10
Dumont forceps #7	FST	11271-30
Mircovascular clamp	FST	00398-02
Clamp applicator	FST	00072-14
Springscissors	FST	15372-62
Needle holder	FST	12010-14
Needles	Feuerstein, Suprama	BER 562-20
5-0 silk suture	Feuerstein, Suprama
7-0 silk suture	Feuerstein,Suprama
8-0 silk suture	Feuerstein, Suprama