caa a22 4500 445313013 CHVBK 20180317142636.0 cr unu---uuuuu 170323e20111001xx s 000 0 eng 10.1007/s11095-011-0478-6 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0478-6 Assessment of P-glycoprotein Activity in the Blood-Brain Barrier (BBB) Using Near Infrared Fluorescence (NIRF) Imaging Techniques [Elektronische Daten] [Ngoc On, Fang Chen, Martha Hinton, Donald Miller] ABSTRACT: Purpose: To examine functional activity of P-glycoprotein (P-gp) in the blood-brain barrier (BBB) using near infrared fluorescence (NIRF) imaging techniques. Methods: Cellular accumulation and bi-directional permeability of the NIRF probe, rhodamine 800 (R800) was determined in MDCKMDR1 and MDCKwt monolayers under normal conditions and following P-gp inhibition with GF120918. Functional P-gp activity was also assessed in mice following administration of R800 alone and with GF230918. Quantitative analysis of R800 fluorescence in brain tissue and blood was measured ex-vivo using Odyssey Near Infrared imaging. Results: R800 accumulation was reduced in MDCKMDR1 compared to MDCKwt monolayers. Addition of GF120918, resulted in increased R800 accumulation in MDCKMDR1 monolayers. Permeability of R800 in MDCKMDR1 monolayers was significantly enhanced (4-fold) in the basolateral to apical direction under control conditions and was abolished following treatment with GF120918. With the exception of the choriod plexus, there was very little penetration of R800 into the brain under control conditions. Treatment of mice with GF120918 resulted in a nearly 4-fold increase in R800 fluorescence in the brain. In contrast, GF120918 had no effect on brain penetration of a vascular permeability marker. Conclusions: In vitro studies demonstrate the P-gp transporter properties of the NIRF probe R800. Preliminary in vivo studies confirm the P-gp transporter liabilities of R800 and suggest this probe may be useful as a molecular imaging agent for examining P-gp activity in the BBB. Springer Science+Business Media, LLC, 2011 blood-brain barrier nationallicence near infrared fluorescence nationallicence P-glycoprotein nationallicence rhodamine 800 nationallicence ATP : adenosine triphosphate nationallicence BBB : blood-brain barrier nationallicence BCRP : breast cancer resistance protein nationallicence BCSFB : blood-cerebral spinal fluid barrier nationallicence CNS : central nervous system nationallicence MDCK : Madin-Darby Canine Kidney epithelial cell line nationallicence MDR : multidrug resistance nationallicence NIRF : near infrared fluorescence nationallicence PEG : poly(ethelene) glycol nationallicence PET : positron emission topography nationallicence P-gp : P-glycoprotein nationallicence R800 : Rhodamine 800 nationallicence On Ngoc Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut Chen Fang Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut Hinton Martha Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut Miller Donald Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/10(2011-10-01), 2505-2515 0724-8741 28:10<2505 2011 28 11095 https://doi.org/10.1007/s11095-011-0478-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0478-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- On Ngoc Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut NATIONALLICENCE 700 1- Chen Fang Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut NATIONALLICENCE 700 1- Hinton Martha Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut NATIONALLICENCE 700 1- Miller Donald Department of Pharmacology & Therapeutics, University of Manitoba, 753 McDermot Ave., R3E 0T6, Winnipeg, Manitoba, Canada aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/10(2011-10-01), 2505-2515 0724-8741 28:10<2505 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer