Visualization and analysis of whole depot adipose tissue neural innervation

Jake W. Willows; Magdalena Blaszkiewicz; Amy Lamore; Samuel Borer; Amanda L. Dubois; Emma Garner; William P. Breeding; Karissa B. Tilbury; Andre Khalil; Kristy L. Townsend

iScience (Oct 2021)

Visualization and analysis of whole depot adipose tissue neural innervation

Jake W. Willows,
Magdalena Blaszkiewicz,
Amy Lamore,
Samuel Borer,
Amanda L. Dubois,
Emma Garner,
William P. Breeding,
Karissa B. Tilbury,
Andre Khalil,
Kristy L. Townsend

Affiliations

Jake W. Willows: School of Biology and Ecology, University of Maine, Orono, ME, USA; Department of Neurological Surgery, The Ohio State University, 1014 Biomedical Research Tower, 460 W. 12th Avenue, Columbus, OH, USA
Magdalena Blaszkiewicz: School of Biology and Ecology, University of Maine, Orono, ME, USA; Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA; Department of Neurological Surgery, The Ohio State University, 1014 Biomedical Research Tower, 460 W. 12th Avenue, Columbus, OH, USA
Amy Lamore: School of Molecular and Biomedical Sciences, University of Maine, Orono, ME, USA
Samuel Borer: School of Biology and Ecology, University of Maine, Orono, ME, USA
Amanda L. Dubois: School of Molecular and Biomedical Sciences, University of Maine, Orono, ME, USA
Emma Garner: School of Biology and Ecology, University of Maine, Orono, ME, USA
William P. Breeding: Department of Chemical and Biomedical Engineering, University of Maine, Orono, ME, USA
Karissa B. Tilbury: Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA; Department of Chemical and Biomedical Engineering, University of Maine, Orono, ME, USA
Andre Khalil: Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA; Department of Chemical and Biomedical Engineering, University of Maine, Orono, ME, USA; CompuMAINE Laboratory, University of Maine, Orono, ME, USA
Kristy L. Townsend: School of Biology and Ecology, University of Maine, Orono, ME, USA; Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA; School of Molecular and Biomedical Sciences, University of Maine, Orono, ME, USA; Department of Chemical and Biomedical Engineering, University of Maine, Orono, ME, USA; Department of Neurological Surgery, The Ohio State University, 1014 Biomedical Research Tower, 460 W. 12th Avenue, Columbus, OH, USA; Corresponding author

Journal volume & issue: Vol. 24, no. 10
p. 103127

Abstract

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Summary: Little is known about the diversity and function of adipose tissue nerves, due in part to the inability to effectively visualize the tissue’s diverse nerve subtypes and the patterns of innervation across an intact depot. The tools to image and quantify adipose tissue innervation are currently limited. Here, we present a method of tissue processing that decreases tissue thickness in the z-axis while leaving cells intact for subsequent immunostaining. This was combined with autofluorescence quenching techniques to permit intact whole tissues to be mounted on slides and imaged by confocal microscopy, with a complementary means to perform whole tissue neurite density quantification after capture of tiled z-stack images. Additionally, we demonstrate how to visualize nerve terminals (the neuro-adipose nexus) in intact blocks of adipose tissue without z-depth reduction. We have included examples of data demonstrating nerve subtypes, neurovascular interactions, label-free imaging of collagen, and nerve bundle digital cross-sections.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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