Visualization of sympathetic neural innervation in human white adipose tissue.


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Authors
Cacciottolo, Tessa 
Henning, Elana 
Mendes de Oliveira, Edson  ORCID logo  https://orcid.org/0000-0002-7330-7826
Keogh, Julia M 
Abstract

Obesity, defined as an excess of adipose tissue that adversely affects health, is a major cause of morbidity and mortality. However, to date, understanding the structure and function of human adipose tissue has been limited by the inability to visualize cellular components due to the innate structure of adipocytes, which are characterized by large lipid droplets. Combining the iDISCO and the CUBIC protocols for whole tissue staining and optical clearing, we developed a protocol to enable immunostaining and clearing of human subcutaneous white adipose tissue (WAT) obtained from individuals with severe obesity. We were able to perform immunolabelling of sympathetic nerve terminals in whole WAT and subsequent optical clearing by eliminating lipids to render the opaque tissue completely transparent. We then used light sheet confocal microscopy to visualize sympathetic innervation of human WAT from obese individuals in a three-dimensional manner. We demonstrate the visualization of sympathetic nerve terminals in human WAT. This protocol can be modified to visualize other structures such as blood vessels involved in the development, maintenance and function of human adipose tissue in health and disease.

Description

Peer reviewed: True


Funder: Bernard Wolfe Health Neuroscience Endowment


Funder: Botnar Fondation


Funder: NIHR Cambridge Biomedical Research Center


Funder: NIHR Senior Investigator Award

Keywords
human adipose tissue, obesity, sympathetic innervation, three-dimensional microscopy, whole tissue immunolabelling, Adipocytes, Adipose Tissue, Adipose Tissue, White, Humans, Obesity, Sympathetic Nervous System
Journal Title
Open Biol
Conference Name
Journal ISSN
2046-2441
2046-2441
Volume Title
Publisher
The Royal Society
Sponsorship
Wellcome Trust (207462/Z/17/Z)
Wellcome Trust (208363/Z/17/Z)