Extracellular matrix molecules exhibit unique expression pattern in the climbing fiber-generating precerebellar nucleus, the inferior olive

Kecskes, Szilvia; Gaál, Botond; Rácz, Éva; Birinyi, András; Hunyadi, Andrea; Matesz, Clara

doi:10.1016/j.neuroscience.2014.09.080

Cited by 11 publications

(6 citation statements)

References 73 publications

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“…While WFA intensity is important for formulating experiments and interpreting the findings, the method by which WFA intensity is measured and analyzed has been inconsistent across studies. Some studies have used semi-quantitative methods to examine relative intensity changes, but these studies lack measures of individual PNNs (Wang et al, 2011, Deak et al, 2012, Yamada and Jinno, 2013, Racz et al, 2014, Kecskes et al, 2015). Other studies examine changes in individual PNNs by averaging the intensity of 15 pixels within a given PNN around the soma (Foscarin et al, 2011, Carulli et al, 2013, Vazquez-Sanroman et al, 2015a), and others have not included details in the methodology (Chen et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

A standardized and automated method of perineuronal net analysis using Wisteria floribunda agglutinin staining intensity

2016

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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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Section: Introductionmentioning

confidence: 99%

A standardized and automated method of perineuronal net analysis using Wisteria floribunda agglutinin staining intensity

2016

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“…Existing histological quantifications of the normal neural ECM are commonly semi-quantitative in that they assign grades to qualitative judgments (e.g., rating how intense staining appears) or in that they introduce manual error checks (where users flip through processed images and manually add/delete cells from identified populations). 8,11,19,22,23,32 Although the manual error checks in some semiautomated analyses minimize error to nearly 0% (if error is being measured relative to manual counts), this loss in time efficiency is not always desirable. For the analysis of percentages, such as that presented here, while accuracy is still important, consistency is equally important and minor errors will not be detrimental.…”

Section: Discussionmentioning

confidence: 99%

“…5,6 The ECM plays an integral role in cellular events and signaling, yet a comprehensive quantification of the spatial and regional distribution of the ECM in the brain has not yet been done, although some characterizations have been reported. 7–13 These analyses, however, are often restricted to a small area of interest or are semi-quantitative. A thorough characterization of the ECM will define both its regional and cellular distribution, including the area covered by these molecules and the percentage of major neural phenotypes that associate with these molecules.…”

Section: Introductionmentioning

confidence: 99%

Quantification of the Extracellular Matrix Molecule Thrombospondin 1 and Its Pericellular Association in the Brain Using a Semiautomated Computerized Approach

Liu

Modo

2018

J Histochem Cytochem.

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The structure and functions of the extracellular matrix (ECM), its spatial distribution and pericellular association of ECM molecules remain poorly understood. Colocalization of ECM molecules with cell phenotypes through immunohistochemistry can provide crucial insights into their juxtacrine signaling role as well as their structural relevance to tissue architecture. As manual quantification of images introduces intra- and inter-user bias and is cumbersome for high-throughput approaches, we implemented an automated high-throughput method to quantify the spatial distribution and cellular association of one ECM molecule, thrombospondin 1 (TSP1) with two major cell phenotypes, neurons, and astrocytes. The distribution of TSP1 was homogeneous throughout the striatum and cortex along the anterior-posterior axis. TSP1 occupied 8.85% of the striatum and 7.40% in the cortex. TSP1 also associated with 94.58% and 88.45% of neurons in the striatum and cortex. The association with astrocytes was significantly lower at 47.55% and 28.09%. These findings highlight the key role that TSP1 plays in neuron physiology in a healthy brain, but also highlights key regional difference in astrocytes secreting ECM molecules. The semiautomated approach implemented here will improve the throughput and reliability of measuring the distribution and cellular colocalization of ECM molecules.

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“…The immunogen is against the mouse myeloma cell line NS0‐derived recombinant human tenascin‐R isoform 1 (Glu34–Phe1358). This antibody has been tested and referenced in several publications (Xiao et al, ; Weber et al, ; Kecskes et al, ). Tenascin‐R was detected in immersion‐fixed paraffin‐embedded sections of human brainstem (https://www.rndsystems.com/products/human-tenascin-r-antibody_af3865).…”

Section: Methodsmentioning

confidence: 99%

Extracellular matrix protein expression is brain region dependent

Dauth

Grevesse

Pantazopoulos

et al. 2016

J of Comparative Neurology

105

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In the brain, extracellular matrix (ECM) components form networks that contribute to structural and functional diversity. Maladaptive remodeling of ECM networks has been reported in neurodegenerative and psychiatric disorders, suggesting that the brain microenvironment is a dynamic structure. A lack of quantitative information about ECM distribution in the brain hinders an understanding of region-specific ECM functions and the role of ECM in health and disease. We hypothesized that each ECM protein as well as specific ECM structures, such as perineuronal nets (PNNs) and interstitial matrix, are differentially distributed throughout the brain, contributing to the unique structure and function in the various regions of the brain. To test our hypothesis, we quantitatively analyzed the distribution, colocalization, and protein expression of aggrecan, brevican, and tenascin-R throughout the rat brain utilizing immunohistochemistry and mass spectrometry analysis and assessed the effect of aggrecan, brevican, and/or tenascin-R on neurite outgrowth in vitro. We focused on aggrecan, brevican, and tenascin-R as they are especially expressed in the mature brain, and have established roles in brain development, plasticity, and neurite outgrowth. The results revealed a differentiated distribution of all three proteins throughout the brain and indicated that their presence significantly reduces neurite outgrowth in a 3D in vitro environment. These results underline the importance of a unique and complex ECM distribution for brain physiology and suggest that encoding the distribution of distinct ECM proteins throughout the brain will aid in understanding their function in physiology and in turn assist in identifying their role in disease. J. Comp. Neurol. 524:1309-1336, 2016. © 2016 Wiley Periodicals, Inc.

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Extracellular matrix molecules exhibit unique expression pattern in the climbing fiber-generating precerebellar nucleus, the inferior olive

Cited by 11 publications

References 73 publications

A standardized and automated method of perineuronal net analysis using Wisteria floribunda agglutinin staining intensity

A standardized and automated method of perineuronal net analysis using Wisteria floribunda agglutinin staining intensity

Quantification of the Extracellular Matrix Molecule Thrombospondin 1 and Its Pericellular Association in the Brain Using a Semiautomated Computerized Approach

Extracellular matrix protein expression is brain region dependent

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