Quantification of adipocyte numbers in transgenic mice via the Cre-LoxP recombination sites

Caroline Moser; Leon G Straub; Yael Rachamin; Christian Wolfrum

doi:10.1016/j.xpro.2021.100761

Quantification of adipocyte numbers in transgenic mice via the Cre-LoxP recombination sites

STAR Protoc. 2021 Aug 18;2(3):100761. doi: 10.1016/j.xpro.2021.100761. eCollection 2021 Sep 17.

Authors

Caroline Moser¹, Leon G Straub¹, Yael Rachamin¹, Christian Wolfrum¹

Affiliation

¹ Institute of Food Nutrition and Health, Eidgenössische Technische Hochschule Zürich (ETH), Schwerzenbach 8603, Switzerland.

Abstract

This protocol describes a method to assess adipocyte numbers within a specific depot based on their inducible genomic label. By extracting DNA from a complete adipose tissue depot stemming from two transgenic mouse lines (Adipoq-CreERT2 x ROSA26-tdRFP and Ucp1-CreERT2 x ROSA26-tdRFP), the number of adipocytes can be determined based on the quantification of the recombined LoxPRed sites. This highly sensitive system allows for the quantification of white, brown, and brite/beige adipocytes in a spatially unbiased and size-independent manner. For complete details on the use and execution of this protocol, please refer to Moser et al. (2021).

Keywords: Cell culture; Cell isolation; Cell separation/fractionation; Cell-based Assays; Metabolism; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adipocytes / cytology*
Adipocytes / physiology
Animals
Cell Count
Integrases / genetics*
Mice, Transgenic
Molecular Biology / instrumentation
Molecular Biology / methods*
Polymerase Chain Reaction
Recombination, Genetic*

Substances

Cre recombinase
Integrases