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

doi:10.1016/j.isci.2021.103127

Visualization and analysis of whole depot adipose tissue neural innervation

iScience. 2021 Sep 11;24(10):103127. doi: 10.1016/j.isci.2021.103127. eCollection 2021 Oct 22.

Authors

Jake W Willows^{1

2}, Magdalena Blaszkiewicz^{1

3

2}, Amy Lamore⁴, Samuel Borer¹, Amanda L Dubois⁴, Emma Garner¹, William P Breeding⁵, Karissa B Tilbury^{3

5}, Andre Khalil^{3

5

6}, Kristy L Townsend^{1

3

4

5

2}

Affiliations

¹ School of Biology and Ecology, University of Maine, Orono, ME, USA.
² Department of Neurological Surgery, The Ohio State University, 1014 Biomedical Research Tower, 460 W. 12 Avenue, Columbus, OH, USA.
³ Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA.
⁴ School of Molecular and Biomedical Sciences, University of Maine, Orono, ME, USA.
⁵ Department of Chemical and Biomedical Engineering, University of Maine, Orono, ME, USA.
⁶ CompuMAINE Laboratory, University of Maine, Orono, ME, USA.

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.

Keywords: Cell biology; Functional aspects of cell biology; Methodology in biological sciences.

Abstract

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