Superresolution Imaging of Cytoskeletal Networks in Fixed Brain Tissue

Hicks, Amirah‐Iman; Zhou, Suijian; Yang, Jieyi; Prager‐Khoutorsky, Masha

doi:10.1007/978-1-0716-2409-8_11

Cited by 2 publications

(3 citation statements)

References 23 publications

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“…Sections were then immediately submerged in mounting media (4% n ‐propyl gallate, 85% glycerol, and 10 mM phosphate buffer, pH 7.4) for 10 min to dehydrate the tissue without drying. This formula was refractive index‐matched to nervous tissue and Olympus silicone immersion oil (SIL300CS‐30CC) to maximize resolving power of Alexa 647 emission using a scanning confocal microscope (Akins et al., 2017; Gingrich et al., 2018; Hicks et al., 2022; Zheng et al., 2011). To ensure consistent reduction in working distance, sections were directly transferred onto coverslips (Globe Scientific Inc.; No.…”

Section: Methodsmentioning

confidence: 99%

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Characterization of Fragile X Mental Retardation Protein expression in human nociceptors and their axonal projections to the spinal dorsal horn

Mitchell

Cook

Shiers

et al. 2023

J of Comparative Neurology

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Fragile X Mental Retardation Protein (FMRP) regulates activity‐dependent RNA localization and local translation to modulate synaptic plasticity throughout the central nervous system. Mutations in the FMR1 gene that hinder or ablate FMRP function cause Fragile X Syndrome (FXS), a disorder associated with sensory processing dysfunction. FXS premutations are associated with increased FMRP expression and neurological impairments including sex dimorphic presentations of chronic pain. In mice, FMRP ablation causes dysregulated dorsal root ganglion (DRG) neuron excitability and synaptic vesicle exocytosis, spinal circuit activity, and decreased translation‐dependent nociceptive sensitization. Activity‐dependent, local translation is a key mechanism for enhancing primary nociceptor excitability that promotes pain in animals and humans. These works indicate that FMRP likely regulates nociception and pain at the level of the primary nociceptor or spinal cord. Therefore, we sought to better understand FMRP expression in the human DRG and spinal cord using immunostaining in organ donor tissues. We find that FMRP is highly expressed in DRG and spinal neuron subsets with substantia gelatinosa exhibiting the most abundant immunoreactivity in spinal synaptic fields. Here, it is expressed in nociceptor axons. FMRP puncta colocalized with Nav1.7 and TRPV1 receptor signals suggesting a pool of axoplasmic FMRP localizes to plasma membrane‐associated loci in these branches. Interestingly, FMRP puncta exhibited notable colocalization with calcitonin gene‐related peptide (CGRP) immunoreactivity selectively in female spinal cord. Our results support a regulatory role for FMRP in human nociceptor axons of the dorsal horn and implicate it in the sex dimorphic actions of CGRP signaling in nociceptive sensitization and chronic pain.

show abstract

Section: Methodsmentioning

confidence: 99%

“…to maximize resolving power of Alexa 647 emission using a scanning confocal microscope (Akins et al, 2017;Gingrich et al, 2018;Hicks et al, 2022;Zheng et al, 2011). To ensure consistent reduction in working distance, sections were directly transferred onto coverslips (Globe Scientific Inc.; No.…”

Section: Immunofluorescencementioning

confidence: 99%

Characterization of Fragile X Mental Retardation Protein expression in human nociceptors and their axonal projections to the spinal dorsal horn

Mitchell

Cook

Shiers

et al. 2023

J of Comparative Neurology

View full text Add to dashboard Cite

show abstract

“…Sections were then immediately submerged in mounting media (4% n-propyl gallate, 85% glycerol and 10 mM phosphate buffer, pH 7.4) for 10 min to dehydrate the tissue without drying. This formula was refractive index-matched to nervous tissue and Olympus silicone immersion oil (SIL300CS-30CC) to maximize resolving power of Alexa 647 emission using a scanning confocal microscope (Akins et al, 2017; Gingrich et al, 2018; Hicks et al, 2022; Zheng et al, 2011). To ensure consistent reduction in working distance, sections were directly transferred onto coverslips (Globe Scientific Inc.; No.…”

Section: Methodsmentioning

confidence: 99%

Fragile X Mental Retardation Protein (FMRP) expression in human nociceptor axons of the spinal dorsal horn— Implications for RNA targeting and localized translation

Mitchell

Cook

Shiers

et al. 2022

Preprint

View full text Add to dashboard Cite

Fragile X Mental Retardation Protein (FMRP) regulates activity-dependent RNA localization and local translation to modulate synaptic plasticity throughout the CNS. Mutations in the FMR1 gene that hinder or ablate FMRP function cause Fragile X Syndrome (FXS), a disorder associated with sensory processing dysfunction. FXS pre-mutations are associated with increased FMRP expression and neurological impairments including sex dimorphic presentations of chronic pain. In mice, FMRP ablation causes dysregulated DRG neuron excitability and synaptic vesicle exocytosis, spinal circuit activity, and decreased translation-dependent nociceptive sensitization. Activity-dependent, local translation is a key mechanism for enhancing primary nociceptor excitability which promotes pain in animals and humans. These works indicate that FMRP likely regulates nociception and pain at the level of the primary nociceptor or spinal cord. Therefore, we sought to better understand FMRP expression in the human dorsal root ganglion (DRG) and spinal cord using immunostaining in organ donor tissues. We find that FMRP is highly expressed in DRG and spinal neuron subsets with substantia gelatinosa exhibiting the most abundant immunoreactivity in spinal synaptic fields. Here, it is expressed in nociceptor axons. FMRP puncta colocalized with Nav1.7 and TRPV1 receptor signals suggesting a pool of axoplasmic FMRP localizes to plasma membrane-associated loci in these branches. Interestingly, FMRP puncta exhibited notable colocalization with calcitonin gene-related peptide (CGRP) immunoreactivity selectively in female spinal cord. Our results support a regulatory role for FMRP in human nociceptor axons of the dorsal horn and implicate it in the sex dimorphic actions of CGRP signaling in nociceptive sensitization and chronic pain.

show abstract

Superresolution Imaging of Cytoskeletal Networks in Fixed Brain Tissue

Cited by 2 publications

References 23 publications

Characterization of Fragile X Mental Retardation Protein expression in human nociceptors and their axonal projections to the spinal dorsal horn

Characterization of Fragile X Mental Retardation Protein expression in human nociceptors and their axonal projections to the spinal dorsal horn

Fragile X Mental Retardation Protein (FMRP) expression in human nociceptor axons of the spinal dorsal horn— Implications for RNA targeting and localized translation

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