Direct evidence of abca1-mediated efflux of cholesterol at the mouse blood–brain barrier

• Published in: Molecular and cellular biochemistry Vol. 357; no. 1-2; pp. 397 - 404
• Main Authors: Do, Tuan Minh, Ouellet, Mélissa, Calon, Frédéric, Chimini, Giovanna, Chacun, Hélène, Farinotti, Robert, Bourasset, Fanchon
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01-11-2011; Springer; Springer Nature B.V
• Subjects: Analysis; Animals; ATP Binding Cassette Transporter 1; ATP Binding Cassette Transporter, Sub-Family G; ATP-Binding Cassette Transporters - drug effects; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - metabolism; Biochemistry; Biomedical and Life Sciences; Blood; Blood-Brain Barrier - metabolism; Brain; Brain - metabolism; Brain - surgery; Cardiology; Cholesterol; Cholesterol - metabolism; Life Sciences; Medical Biochemistry; Mice; Mice, Inbred C57BL; Mice, Transgenic; Oncology; Probucol - pharmacology; Rodents; Abca1; Abcb1; Mouse; In situ brain perfusion; Abcg4; Cholesterol; Blood–brain barrier; Abcg2
• ISSN: 0300-8177; 1573-4919
• DOI: 10.1007/s11010-011-0910-6

We investigated the expression and function of Abca1 in wild-type C57BL/6,
abca1
(+/+), and
abca1
(−/−) mice brain capillaries forming the blood–brain barrier (BBB). We first demonstrated by quantitative RT-PCR and Western immunoblot that Abca1 was expressed and enriched in the wild-type mouse brain capillaries. In
abca1
(−/−) mice, we reported that the lack of Abca1 resulted in an 1.6-fold increase of the Abcg4 expression level compared to
abca1
(+/+) mice. Next, using the in situ brain perfusion technique, we showed that the [
3
H]cholesterol brain uptake clearance (Cl
up
, μl/s/g brain), was significantly increased (107%) in
abca1
(−/−) mice compared to
abca1
(+/+) mice, meaning that the deficiency of Abca1 conducted to a significant decrease of the cholesterol efflux at the BBB level. In addition, the co-perfusion of probucol (Abca1 inhibitor) with [
3
H]cholesterol resulted in an increase of [
3
H]cholesterol Cl
up
(115%) in
abca1
(+/+) but not in
abca1
(−/−) mice, meaning that probucol inhibited selectively the efflux function of Abca1. In conclusion, our results demonstrated that Abca1 was expressed in the mouse brain capillaries and that Abca1 functions as an efflux transporter through the mouse BBB.