Pathological and immunological consequences of basic experimental animal
Contact: Dorte Bratbo Sørensen
Annually, millions of mice are used for experimental purposes and these experiments often include sampling of blood throughout the trial period. However, there is a lack of comparative studies investigating the impact of blood sampling on the mice with regards to both resulting tissue damages and immunological changes and stress. The overall aim of the project is thus to describe the local and systemic pathological and immunological changes and possible stress introduced in mice after blood sampling by six different basic procedures, namely retrobulbar venous plexus puncture (periorbital puncture), vena submandibularis puncture (cheek blood), vena sublingualis puncture (tongue blood), vena saphena puncture, lateral tail incision and amputation of tail tip.
All six blood sampling techniques are performed by skilled technicians and the mice are euthanasized after either 6, 10 or 24 hours or 2, 4, 6, 8, 10 and 12 days after the procedure. After euthanasia, a complete necropsy is performed and blood, selected internal organs and tissue samples from the local area of blood sampling are collected. The pathological damages induced as a consequence of the procedure and how fast these changes are normalized are described. The expression of major mediators of local acute inflammatory events and systemic inflammatory responses are analyzed using qPCR for gene expression profiling of 15 selected immune-related genes and ELISA (enzyme-linked immunosorbent assay) is used for quantifying relevant blood protein inflammation markers. Moreover, the Optical projection tomography (OPT) scanning technique is used to construct a complete three-dimensional image of the distribution of inflammatory cells surrounding the puncture site 6 and 24 hours and 10 days after sublingual vene puncture. Last, the level of stress hormone (corticosterone) is measured either 6, 12 or 24 hours after blood sampling.
This study will hence provide a comprehensive description of the effects of blood sampling in laboratory mice, forming the basis for a more detailed and useful guideline for choosing the optimal blood sampling technique for a specific study.