Brain Uptake of GLP-1 Analogs in Rats

Rats received a systemic infusion of either saline or exendin-(9-39) (GLP-1 receptor antagonist, 30 nmol/kg/min, Abcam, Cambridge, United Kingdom) via the jugular vein catheter for 20 min (Figure 1A). In the last 10 min, rats also received 30 pmol/kg/min of either GLP-1-FAM (AnaSpec, Fremont, CA, United States) or exendin-4-FAM (AnaSpec, Fremont, CA, United States) together with Dextran-TRITC (Sigma-Aldrich, St. Louis, MO, United States) (70 kD, vascular space marker, 30 pmol/kg/min). The infusion doses were based on our prior studies in rats that GLP-1 at 30 pmol/kg/min potently increased muscle microvascular perfusion and glucose use (Chai et al., 2012 (link), 2014 (link); Dong et al., 2013 (link)). After collecting plasma samples, rats were sacrificed via anesthetic overdose and various brain regions (cerebellum, cortex, hippocampus, hypothalamus, and brain stem) collected, homogenized and lysed in lysis buffer. Fluorescence from TRITC and FAM were measured in both plasma and brain lysates. Protein concentrations were determined in the brain lysates. Brain uptake of GLP-1-FAM and exendin-4-FAM were calculated using the following formula:

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Fu Z., Gong L., Liu J., Wu J., Barrett E.J., Aylor K.W, & Liu Z. (2020). Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process. Frontiers in Physiology, 11, 555.

Publication 2020

exendin-(9-39) exendin-4-FAM Dextran-TRITC

Anesthetic Brain Brain stem Buffer Catheter Cerebellum Cortex Dextran Exendin 4 Fluorescence Glp 1 Glp 1 receptor Glucose Hippocampus Hypothalamus Jugular vein Muscle Overdose Perfusion Plasma Protein Rats Saline Tritc Vascular

Corresponding Organization : University of Virginia Health System

Other organizations : Third Xiangya Hospital, Central South University, Xiangya Hospital Central South University

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Variable analysis

independent variables

Systemic infusion of either saline or exendin-(9-39) (GLP-1 receptor antagonist, 30 nmol/kg/min)
Infusion of either GLP-1-FAM (30 pmol/kg/min) or exendin-4-FAM (30 pmol/kg/min)

dependent variables

Fluorescence from TRITC and FAM measured in plasma and brain lysates
Brain uptake of GLP-1-FAM and exendin-4-FAM

control variables

Infusion via jugular vein catheter for 20 min
Dextran-TRITC (70 kD, vascular space marker, 30 pmol/kg/min) infusion

positive controls

Prior studies showing that GLP-1 at 30 pmol/kg/min potently increased muscle microvascular perfusion and glucose use

negative controls

Saline infusion

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