A Vector-Based Method to Analyze the Topography of Glial Networks
Abstract
:1. Introduction
2. Results
2.1. Astrocyte Networks
2.2. Analysis of Network Topography
2.3. Meta-Analysis of “Vector Means”
2.4. Performance of Approaches
3. Discussion
3.1. Intensity-Based Cell Detection Method
3.2. Comparison of Approaches
3.3. Astrocyte Maturation and Network Topography
3.4. Tracer Coupling versus Electrical Coupling
3.5. Conclusions
4. Materials and Methods
4.1. Preparation of Acute Brainstem Slices
4.2. Tracer Loading
4.3. Visualization of Coupled Cells
4.4. Confocal Microscopy
4.5. Reconstruction of Gap Junction Networks
4.6. Analysis of Network Topography
4.6.1. Manual “YX Ratio”
4.6.2. Automated “YX Ratio”
4.6.3. “Intensity + Coordinates”
4.6.4. “Intensity Profiles”
4.6.5. “Vector Sum”
4.6.6. “Vector Means”
4.6.7. Meta-Analysis
4.6.8. Generation of Artificial Networks In Silico
4.7. Statistics
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ACSF | Artificial cerebrospinal fluid |
BSA | Bovine serum albumin |
Ctx | Cortex |
Cx | Connexin |
FWHM | Full-width at half-maximum |
GJ | Gap junction |
HC | Hippocampus |
IC | Inferior colliculus |
LSO | Lateral superior olive |
Nb+ | Neurobiotin-positive |
NGS | Normal goat serum |
nPA | Nonpassive astrocyte |
PA | Passive astrocyte |
PBS | Phosphate buffered solution |
PFA | Paraformaldehyde |
R | Ratio |
R2 | Regression coefficient |
RT | Room temperature |
ROI | Region of interest |
SR101 | Sulforhodamine 101 |
Th | Thalamus |
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Study | Approach | nPA (classes) | PA (classes) | p | ||||
---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 1 | 2 | 3 | |||
This study (LSO) | YX ratio (manual) | 79% | 14% | 7% | 40% | 50% | 10% | 1.6096 × 10−14 |
YX ratio (automatic) | 64% | 36% | 0% | 50% | 30% | 20% | 3.4229 × 10−6 | |
Intensity + coordinates | 93% | 0% | 7% | 60% | 20% | 20% | 9.0177 × 10−11 | |
Intensity profiles | 57% | 43% | 0% | 30% | 70% | 0% | 3.1601 × 10−9 | |
Vector means | 79% | 21% | 0% | 30% | 60% | 10% | 2.3325 × 10−25 | |
Previous studies | ||||||||
[4] (LSO) | YX ratio (manual) | 58% | 33% | 8% | 39% | 39% | 22% | 6.7850 × 10−5 |
[5] (IC) | YX ratio (manual) | 71% | 21% | 7% | 43% | 43% | 13% | 1.0375 × 10−7 |
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Eitelmann, S.; Hirtz, J.J.; Stephan, J. A Vector-Based Method to Analyze the Topography of Glial Networks. Int. J. Mol. Sci. 2019, 20, 2821. https://doi.org/10.3390/ijms20112821
Eitelmann S, Hirtz JJ, Stephan J. A Vector-Based Method to Analyze the Topography of Glial Networks. International Journal of Molecular Sciences. 2019; 20(11):2821. https://doi.org/10.3390/ijms20112821
Chicago/Turabian StyleEitelmann, Sara, Jan J. Hirtz, and Jonathan Stephan. 2019. "A Vector-Based Method to Analyze the Topography of Glial Networks" International Journal of Molecular Sciences 20, no. 11: 2821. https://doi.org/10.3390/ijms20112821