Labeling of Blood Vessels in the Teleost Brain and Pituitary Using Cardiac Perfusion with a DiI-fixative

• Published in: Journal of visualized experiments no. 148
• Main Authors: Fontaine, Romain, Weltzien, Finn-Arne
• Format: Journal Article
• Language: English
• Published: United States 13.06.2019
• Subjects: adults; Animals; Animals, Genetically Modified; blood; blood vessels; brain; Brain - blood supply; Cardiac Surgical Procedures - methods; Danio rerio; endocrinology; Fixatives - chemistry; fluorescent proteins; genetically modified organisms; heart; neurophysiology; nutrients; Oryzias; Oryzias latipes; oxygen; Pituitary Gland - blood supply; tissues; vertebrates; Zebrafish
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/59768

Blood vessels innervate all tissues in vertebrates, enabling their survival by providing the necessary nutrients, oxygen, and hormonal signals. It is one of the first organs to start functioning during development. Mechanisms of blood vessel formation have become a subject of high scientific and clinical interest. In adults however, it is difficult to visualize the vasculature in most living animals due to their localization deep within other tissues. Nevertheless, visualization of blood vessels remains important for several studies such as endocrinology and neurobiology. While several transgenic lines have been developed in zebrafish, with blood vessels directly visualized through expression of fluorescent proteins, no such tools exist for other teleost species. Using medaka (Oryzias latipes) as a model, the current protocol presents a quick and direct technique to label blood vessels in brain and pituitary by perfusing through the heart with fixative containing DiI. This protocol allows improvement of our understanding on how brain and pituitary cells interact with blood vasculature in whole tissue or thick tissue slices.