Visualization and Analysis of Whole Depot Adipose Tissue Neural Innervation

Overview

Journal iScience

Publisher Cell Press

Date 2021 Oct 8

PMID 34622172

Citations 17

Authors

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

Soon will be listed here.

Abstract

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.

Citing Articles

Clearing the path for whole-mount labeling and quantification of neuron and vessel density in adipose tissue.

Rauchenwald T, Benedikt-Kuhnast P, Eder S, Grabner G, Forstreiter S, Lang M J Cell Sci. 2025; 138(3).

PMID: 39878039 PMC: 11832183. DOI: 10.1242/jcs.263438.

Perivascular adipose tissue: a central player in the triad of diabetes, obesity, and cardiovascular health.

Queiroz M, Sena C Cardiovasc Diabetol. 2024; 23(1):455.

PMID: 39732729 PMC: 11682657. DOI: 10.1186/s12933-024-02549-9.

The early transition to cold-induced browning in mouse subcutaneous white adipose tissue (scWAT) involves proteins related to nerve remodeling, cytoskeleton, mitochondria, and immune cells.

Blaszkiewicz M, Johnson C, Willows J, Gardner M, Taplin D, Freitas M Adipocyte. 2024; 13(1):2428938.

PMID: 39641403 PMC: 11633174. DOI: 10.1080/21623945.2024.2428938.

Phenotypic and functional disparities in perivascular adipose tissue.

Guo E, Liu D, Zhu Z Front Physiol. 2024; 15:1499340.

PMID: 39588268 PMC: 11586278. DOI: 10.3389/fphys.2024.1499340.

Interplay between the brain and adipose tissue: a metabolic conversation.

Diaz-Castro F, Morselli E, Claret M EMBO Rep. 2024; 25(12):5277-5293.

PMID: 39558137 PMC: 11624209. DOI: 10.1038/s44319-024-00321-4.

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