Not All Lectins Are Equally Suitable for Labeling Rodent Vasculature.

Roberta Battistella,Anja Meissner,Iben Lundgaard,Anne Xiaoan Cheng,Hana Matuskova,Douglas Haswell,Marios Kritsilis,Maiken Nedergaard

doi:10.3390/ijms222111554

Abstract

The vascular system is vital for all tissues and the interest in its visualization spans many fields. A number of different plant-derived lectins are used for detection of vasculature; however, studies performing direct comparison of the labeling efficacy of different lectins and techniques are lacking. In this study, we compared the labeling efficacy of three lectins: Griffonia simplicifolia isolectin B4 (IB4); wheat germ agglutinin (WGA), and Lycopersicon esculentum agglutinin (LEA). The LEA lectin was identified as being far superior to the IB4 and WGA lectins in histological labeling of blood vessels in brain sections. A similar signal-to-noise ratio was achieved with high concentrations of the WGA lectin injected during intracardial perfusion. Lectins were also suitable for labeling vasculature in other tissues, including spinal cord, dura mater, heart, skeletal muscle, kidney, and liver tissues. In uninjured tissues, the LEA lectin was as accurate as the Tie2–eGFP reporter mice and GLUT-1 immunohistochemistry for labeling the cerebral vasculature, validating its specificity and sensitivity. However, in pathological situations, e.g., in stroke, the sensitivity of the LEA lectin decreases dramatically, limiting its applicability in such studies. This work can be used for selecting the type of lectin and labeling method for various tissues.

Highlights

The first accurate description of the cardiovascular system dates back to 1628 when the pioneering work of William Harvey revolutionized Galen’s theory by centering the movement of blood around the heart as a central pump that pushes blood through a network of conduit vessels, nourishing tissues and organs [1]
Our results showed that 98.78 ± 1.62% of the total glucose transporter-1 (GLUT-1)-labeled blood vessels were labeled with the Lycopersicon esculentum agglutinin (LEA) lectin, suggesting that sensitivity of the LEA lectin to blood vessels was unaltered by ischemic brain injury
This study provides a comprehensive analysis of the labeling efficacy of the isolectin B4 (IB4), wheat germ agglutinin (WGA), and LEA lectins in histological staining of blood vessels in brain sections and a comparison of this method with the intracardiac perfusion of the WGA lectin in labeling blood vessels in the brain and other organs

Summary

Introduction

The first accurate description of the cardiovascular system dates back to 1628 when the pioneering work of William Harvey revolutionized Galen’s theory by centering the movement of blood around the heart as a central pump that pushes blood through a network of conduit vessels, nourishing tissues and organs [1]. The early stages of cardiovascular development begin in the embryonic mesoderm when angiocysts start forming a primitive endothelial heart tube, preceding the formation of blood vessels through substantial embryonic angiogenesis, followed by pruning and remodeling of the vessels. The vasculature develops as one of the first organ systems, reorganization of the vascular system still occurs in adults during different physiological and pathological situations, such as the ovarian cycle and the formation of tumors [2]. In the brain, blood vessel angiogenesis and remodeling occur in adult life, for instance, as a response to augmented vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1 (HIF-1) signaling [3]. Angiogenesis, and neovascularization constitute a common response to tissue injury, characteristic of several neurological diseases including stroke [5], traumatic brain injury [6], and Parkinson’s disease (PD) [7,8]. It is essential for researchers to understand and investigate the integrity of the vascular system’s structure and functionality

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Conclusion