A Novel Technique for Visualizing and Analyzing the Cerebral Vasculature in Rodents

Arjang Salehi; Amandine Jullienne; Kara M Wendel; Mary Hamer; Jiping Tang; John H Zhang; William J Pearce; Richard A DeFazio; Zinaida S Vexler; Andre Obenaus

doi:10.1007/s12975-018-0632-0

A Novel Technique for Visualizing and Analyzing the Cerebral Vasculature in Rodents

Transl Stroke Res. 2018 May 15. doi: 10.1007/s12975-018-0632-0. Online ahead of print.

Authors

Arjang Salehi^{1

2}, Amandine Jullienne², Kara M Wendel³, Mary Hamer^{2

4}, Jiping Tang⁵, John H Zhang^{5

6

7}, William J Pearce^{5

8}, Richard A DeFazio⁹, Zinaida S Vexler¹⁰, Andre Obenaus^{11

12

13

14}

Affiliations

¹ Cell, Molecular and Developmental Biology Program, University of California, Riverside, 1140 Bachelor Hall, Riverside, CA, 92521, USA.
² Department of Basic Sciences, Loma Linda University, Loma Linda, CA, 92354, USA.
³ Department of Anatomy and Neurobiology, University of California, Irvine, Irvine, CA, 92697-4475, USA.
⁴ Department of Pediatrics, University of California, Irvine, Irvine, CA, 92697-4475, USA.
⁵ Department of Physiology and Pharmacology, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA.
⁶ Department of Anesthesiology, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA.
⁷ Department of Neurosurgery, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA.
⁸ Center for Perinatal Biology, Loma Linda University, Loma Linda, CA, 92350, USA.
⁹ Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, 48101, USA.
¹⁰ Department of Neurology, University of California, San Francisco, San Francisco, CA, 94158, USA.
¹¹ Cell, Molecular and Developmental Biology Program, University of California, Riverside, 1140 Bachelor Hall, Riverside, CA, 92521, USA. obenausa@uci.edu.
¹² Department of Basic Sciences, Loma Linda University, Loma Linda, CA, 92354, USA. obenausa@uci.edu.
¹³ Department of Anatomy and Neurobiology, University of California, Irvine, Irvine, CA, 92697-4475, USA. obenausa@uci.edu.
¹⁴ Department of Pediatrics, University of California, Irvine, Irvine, CA, 92697-4475, USA. obenausa@uci.edu.

PMID: 29766452
DOI: 10.1007/s12975-018-0632-0

Abstract

We introduce a novel protocol to stain, visualize, and analyze blood vessels from the rat and mouse cerebrum. This technique utilizes the fluorescent dye, DiI, to label the lumen of the vasculature followed by perfusion fixation. Following brain extraction, the labeled vasculature is then imaged using wide-field fluorescence microscopy for axial and coronal images and can be followed by regional confocal microscopy. Axial and coronal images can be analyzed using classical angiographic methods for vessel density, length, and other features. We also have developed a novel fractal analysis to assess vascular complexity. Our protocol has been optimized for adult rat, adult mouse, and neonatal mouse studies. The protocol is efficient, can be rapidly completed, stains cerebral vessels with a bright fluorescence, and provides valuable quantitative data. This method has a broad range of applications, and we demonstrate its use to study the vasculature in assorted models of acquired brain injury.

Keywords: Analysis; Rodents; Stroke; Traumatic brain injury; Vasculature.

Abstract

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