PrismPlus: a mouse line expressing distinct fluorophores in four different brain cell types

Janak Gaire; Heui Chang Lee; Ray Ward; Seth Currlin; Andrew J Woolley; Jason E Coleman; Justin C Williams; Kevin J Otto

doi:10.1038/s41598-018-25208-y

PrismPlus: a mouse line expressing distinct fluorophores in four different brain cell types

Sci Rep. 2018 May 8;8(1):7182. doi: 10.1038/s41598-018-25208-y.

Authors

Janak Gaire¹, Heui Chang Lee^{2

3}, Ray Ward², Seth Currlin¹, Andrew J Woolley³, Jason E Coleman⁴, Justin C Williams^{5

6}, Kevin J Otto^{7

8

9

10

11

12}

Affiliations

¹ Department of Neuroscience, University of Florida, Gainesville, FL, USA.
² J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, USA.
³ Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA.
⁴ Department of Pediatrics and Child Health Research Institute, University of Florida, Gainesville, FL, USA.
⁵ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
⁶ Department of Neurological Surgery, University of Wisconsin-Madison, Madison, WI, USA.
⁷ Department of Neuroscience, University of Florida, Gainesville, FL, USA. [email protected].
⁸ J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, USA. [email protected].
⁹ Department of Materials Science and Engineering, University of Florida, Gainesville, FL, USA. [email protected].
¹⁰ Department of Neurology, University of Florida, Gainesville, FL, USA. [email protected].
¹¹ Nanoscience Institute for Medical and Engineering Technology, University of Florida, Gainesville, FL, USA. [email protected].
¹² Department of Electrical and Computer Engineering, University of Florida, Gainesville, FL, USA. [email protected].

Abstract

To screen the complex central nervous system (CNS) injury responses, we created a quadruple-labelled 'PrismPlus' mouse line with a genetically encoded distinct fluorescent tag in oligodendrocytes, microglia, neurons, and astrocytes. Cx3cr1-gfp and Prism mice originally developed by Jung et al., 2000 and Dougherty et al., 2012, respectively, were cross-bred. First, we confirmed the presence of fluorophores in appropriate cell types in PrismPlus mice. PrismPlus mice were then used to examine the cellular responses to brain implanted micro-devices. We observed an increase in microglial response at earlier time points as compared to 4 weeks, a progressive astrocytic response, and fewer neurons at the vicinity of an implanted device. These results are similar to what has been described in literature using other rodent strains, previously attainable only through time-consuming and variable immunohistochemistry methods. Finally, we demonstrate the compatibility of PrismPlus brain tissue with CLARITY, an advanced tissue clearing technique, opening the door to future thick tissue imaging studies. This report confirms PrismPlus transgenic fluorescence and highlights the utility of these mice to study CNS injuries. The work herein seeks to establish a novel transgenic mouse line to improve experimental scope, consistency, and efficiency for CNS researchers.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Astrocytes / metabolism*
Astrocytes / ultrastructure
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Brain / metabolism
Brain / pathology
Brain Injuries, Traumatic / diagnosis
Brain Injuries, Traumatic / genetics*
Brain Injuries, Traumatic / metabolism
Brain Injuries, Traumatic / pathology
CX3C Chemokine Receptor 1 / genetics
CX3C Chemokine Receptor 1 / metabolism
Crosses, Genetic
Disease Models, Animal
Electrodes, Implanted
Female
Founder Effect*
Gene Expression
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Male
Mice
Mice, Transgenic
Microglia / metabolism*
Microglia / ultrastructure
Neurons / metabolism*
Neurons / ultrastructure
Oligodendroglia / metabolism*
Oligodendroglia / ultrastructure
Recombinant Fusion Proteins / genetics*
Recombinant Fusion Proteins / metabolism
Repressor Proteins / genetics
Repressor Proteins / metabolism
Transgenes

Substances

Bacterial Proteins
CX3C Chemokine Receptor 1
Cx3cr1 protein, mouse
Cyan Fluorescent Protein
Luminescent Proteins
PRDM6, protein, mouse
Recombinant Fusion Proteins
Repressor Proteins
fluorescent protein 583
yellow fluorescent protein, Bacteria
Green Fluorescent Proteins