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STORM Super-Resolution Imaging of CB1 Receptors in Tissue Preparations

Part of the Methods in Molecular Biology book series (MIMB,volume 2576)

Abstract

Single-molecule localization microscopy (SMLM) opened new possibilities to study the spatial arrangement of molecular distribution and disease-associated redistribution at a previously unprecedented resolution that was not achievable with optical microscopy approaches. Recent discoveries based on SMLM techniques uncovered specific nanoscale organizational principles of signaling proteins in several biological systems including the chemical synapses in the brain. Emerging data suggest that the spatial arrangement of the molecular players of the endocannabinoid system is also precisely regulated at the nanoscale level in synapses and in other neuronal and glial subcellular compartments. The precise nanoscale distribution pattern is likely to be important to subserve several specific signaling functions of this important messenger system in a cell-type- and subcellular domain-specific manner.

STochastic Optical Reconstruction Microscopy (STORM) is an especially suitable SMLM modality for cell-type-specific nanoscale molecular imaging due to its compatibility with traditional diffraction-limited microscopy approaches and classical staining methods. Here, we describe a detailed protocol for STORM imaging in mouse brain tissue samples with a focus on the CB1 cannabinoid receptor, one of the most abundant synaptic receptors in the brain. We also summarize important conceptual and methodical details that are essential for the valid interpretation of single-molecule localization microscopy data.

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Acknowledgments

This study was supported by the Frontier Program 129961 of the National Research, Development and Innovation Office, Hungary. I.K. holds the Naus Family Chair in Addiction Sciences in the Department of Psychological and Brain Sciences at Indiana University Bloomington, and his work is also supported by the National Institutes of Health (R01NS099457 and R01DA044925).

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Correspondence to István Katona .

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Zöldi, M., Katona, I. (2023). STORM Super-Resolution Imaging of CB1 Receptors in Tissue Preparations. In: Maccarrone, M. (eds) Endocannabinoid Signaling. Methods in Molecular Biology, vol 2576. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2728-0_36

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  • DOI: https://doi.org/10.1007/978-1-0716-2728-0_36

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  • Publisher Name: Humana, New York, NY

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