The Extracellular Matrix Glycoprotein Tenascin-C and Matrix Metalloproteinases Modify Cerebellar Structural Plasticity by Exposure to an Enriched Environment
TnC MMP-9 Shape Cerebellar Structural Plasticity
Vera Stamenkovic1, Stefan Stamenkovic1, Tomasz Jaworski2, Maciej Gawlak3, Milos Jovanovic1, Igor Jakovcevski4,5, Grzegorz M. Wilczynski6, Leszek Kaczmarek2, Melitta Schachner7,8, Lidija Radenovic1, Pavle R. Andjus1
1Center for Laser Microscopy, Department of Physiology and Biochemistry, Faculty of Biology, University of Belgrade, 11000 Belgrade, Serbia, 2Laboratory of Neurobiology, Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland, 3Laboratory of Physiology and Pathophysiology, Center for Preclinical Research and Technology, The Medical University of Warsaw, 02-097 Warsaw, Poland, 4Experimental Neurophysiology, University Hospital Cologne, 50931 Köln, Germany, 5Experimental Neurophysiology, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany,6Laboratory of Neuromorphology, Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland,7W. M. Keck Center for Collaborative Neuroscience and Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA, 8Center for Neuroscience, Shantou University Medical College, Shantou, Guangdong 515041, People’s Republic of China.
Corresponding author: Pavle R. Andjus. E-mail: , tel: + 38111-3032356, fax: + 38111-3032356 or -2638500
SUPPLEMENTARY INFORMATION
SUPPLEMENTARY MATERIALS AND METHODS
Quantification of compartmental ISZ signal intensity: Toseparate extracellular from nuclear ISZ signal further analysis of the images was performed using the ImageJ software. The nuclear ISZ signal intensity was analyzed in the regions of interest (ROIs) which included all nuclei in the image frame. To quantify extracellular ISZ signal intensity we first created a mask image containing all previously marked ROIs. After exclusion of the nuclear signal from the analysis and copying the selection boundaries from the mask image to the original confocal image, we measured mean fluorescence in the extracellular space. Tresholding was done by subtracting the background fluorescence signal obtained from the non-stained region of the image (area between LDCN neurons). Quantification of the cytosolic ISZ signal showed that fluorescence in this compartment was matched with the background fluorescence, thus it was excluded from the analysis by tresholding.
SUPPLEMENTARYFIGURE
Supplementary Fig 1 Compartmental analysis of ISZ signal intensity. Quantitative representations of mean pixel intensity of nuclear (a) and extracellular (b) ISZ signal in the LDCN. All values arenormalized to the average intensity of TnC +/+ SC for the corresponding compartment. Two-way ANOVA;*p< 0.05, ***p< 0.001; n = 4 miceper experimental group, 2-3 sections per animal. For abbreviations see the list in the main paper