A standardized and automated method of perineuronal net analysis using Wisteria floribunda agglutinin staining intensity
Open accessAbstract
Perineuronal nets (PNNs) are aggregations of extracellular matrix molecules that are critical for plasticity. Their altered development or changes during adulthood appear to contribute to a wide range of diseases/disorders of the brain. An increasing number of studies examining the contribution of PNN to various behaviors and types of plasticity have analyzed the fluorescence intensity of Wisteria floribunda agglutinin (WFA) as an indirect measure of the maturity of PNNs, with brighter WFA staining corresponding to a more mature PNN and dim WFA staining corresponding to an immature PNN. However, a clearly-defined and unified method for assessing the intensity of PNNs is critical to allow us to make comparisons across studies and to advance our understanding of how PNN plasticity contributes to normal brain function and brain disease states. Here we examined methods of PNN intensity quantification and demonstrate that creating a region of interest around each PNN and subtracting appropriate background is a viable method for PNN intensity quantification that can be automated. This method produces less variability and bias across experiments compared to other published analyses, and this method increases reproducibility and reliability of PNN intensity measures, which is critical for comparisons across studies in this emerging field.
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Mean fluorescence intensity of WFA, ACAN, GFAP, and Cat-315 was performed based on an established method for PNN quantification and applied using a stereological approach.
“Mean fluorescence intensity (MFI) of WFA, ACAN, GFAP, and Cat-315 were performed based on an established method for PNN quantification (Slaker et al. 2016) and applied using a stereological approach.”
From Perineuronal net deglycosylation associates with tauopathy‐induced gliosis and neurodegeneration (2024) · cites this paper as Slaker et al. 2016
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Self-citationPerineuronal net deglycosylation associates with tauopathy‐induced gliosis and neurodegeneration
Logsdon, A. F., Foresi, B., Hu, S. J., Quah, E., Meuret, C. J., Le, J. P., Hendrickson, A. S., Redford, I. K., Kumar, A., Phan, B. A., et al. (2024) · Journal of Neurochemistry · doi:10.1111/jnc.16067 · Cited by 16 · see citing works
- Authors
- Slaker, M. L., Harkness, J. H., & Sorg, B. A.
- Affiliations
- Washington State University Vancouver (US)
- Corresponding author
- Sorg, Barbara A.
- Venue
- IBRO Reports
- Publisher
- Elsevier BV
- Published
- 2016-12
- Volume
- 1
- Pages
- 54-60
- ISSN
- 2451-8301 (ISSN-L)
- PMC
- PMC5507617
- OpenAlex
- W2529600742
- Cited by
- 86
- References
- 10
- Citations by year
- 2026: 9 · 2025: 10 · 2024: 11 · 2023: 10 · 2022: 8 · 2021: 10 · 2020: 7 · 2019: 12 · 2018: 7 · 2017: 1 · 2016: 1
- FWCI
- 3.94
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- IBRO Reports (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Washington State University
- University of Washington
- Universitat Jaume I
- National Institutes of Health (040965)
- National Institute on Drug Abuse (R01DA040965, R01DA033404)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Keywords
- perineuronal nets, Wisteria floribunda agglutinin, extracellular matrix, automated image analysis, region of interest, background subtraction, neuroplasticity, reproducibility
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