Insights into in vivo adipocyte differentiation through cell-specific labeling in zebrafish

Author:

Lepanto Paola1ORCID,Levin-Ferreyra Florencia1ORCID,Koziol Uriel2ORCID,Malacrida Leonel34ORCID,Badano José L.1ORCID

Affiliation:

1. Human Molecular Genetics Lab, Institut Pasteur de Montevideo, Montevideo, Mataojo 2020, CP11400, Uruguay

2. Sección Biología Celular, Facultad de Ciencias, Universidad de la República, Montevideo, Igua 4225, CP11400, Uruguay

3. Advanced Bioimaging Unit, Institut Pasteur de Montevideo and Universidad de la República, Montevideo, Mataojo 2020, CP11400, Uruguay

4. Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Av. Italia s/n, CP11600, Uruguay

Abstract

ABSTRACT White adipose tissue hyperplasia has been shown to be crucial for handling excess energy in healthy ways. Though adipogenesis mechanisms have been underscored in vitro, we lack information on how tissue and systemic factors influence the differentiation of new adipocytes. While this could be studied in zebrafish, adipocyte identification currently relies on neutral lipid labeling, thus precluding access to cells in early stages of differentiation. Here we report the generation and analysis of a zebrafish line with the transgene fabp4a(-2.7):EGFPcaax. In vivo confocal microscopy of the pancreatic and abdominal visceral depots of transgenic larvae, revealed the presence of labeled mature adipocytes as well as immature cells in earlier stages of differentiation. Through co-labeling for blood vessels, we observed a close interaction of differentiating adipocytes with endothelial cells through cell protrusions. Finally, we implemented hyperspectral imaging and spectral phasor analysis in Nile Red-labeled transgenic larvae and revealed the lipid metabolic transition towards neutral lipid accumulation of differentiating adipocytes. Altogether our work presents the characterization of a novel adipocyte-specific label in zebrafish and uncovers previously unknown aspects of in vivo adipogenesis. This article has an associated First Person interview with the first author of the paper.

Funder

Programa para el Desarrollo de las Ciencias Básicas

Agencia Nacional de Investigación e Innovación

Fondo para la Convergencia Estructural del Mercosur

Chan Zuckerberg Initiative

Publisher

The Company of Biologists

Subject

General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology

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