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  • 3T3-L1 cells and perivascular adipocytes are not equivalent in amine transporter expression.

3T3-L1 cells and perivascular adipocytes are not equivalent in amine transporter expression.

FEBS letters (2016-12-08)
Alex Ismail, Nadia Ayala-Lopez, Maleeha Ahmad, Stephanie W Watts
ABSTRACT

Rat perivascular adipose tissue (PVAT) stores, takes up, and releases norepinephrine (NE; Ayala-Lopez et al. (2014) Pharmacol Res Perspect 2, e00041). We hypothesized that 3T3-L1 adipocytes would exhibit similar behaviors and, thus, could serve as a model for PVAT adipocytes. However, basal levels of NE were not detected in 3T3-L1 adipocytes. While incubation of 3T3-L1 adipocytes with exogenous NE increased their cellular NE content, the mRNA expression of several NE transporters [e.g., norepinephrine transporter (NET)] were not detected in these cells. Similarly, we observed expression of the vesicular monoamine transporter 1 (VMAT1) in 3T3-L1 adipocytes by qRT-PCR and immunostaining, but stimulation of the cells with tyramine (100 μm) did not cause a significant release of NE. These studies support that 3T3-L1 adipocytes are not an adequate model of perivascular adipocytes for studying NE handling.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagenase from Clostridium histolyticum, Type IA, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid, For general use
Sigma-Aldrich
Ro 41-0960, solid
Sigma-Aldrich
Anti-Vesicular Monoamine Transporter 1 Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Bicinchoninic Acid Kit for Protein Determination, for 200-1000 μg/ml protein