Evaluation of Blood-CSF Barrier Transport by Quantitative Real Time Fluorescence Microscopy

Sun, Austin; Wang, Joanne

doi:10.1007/s11095-022-03251-9

Cited by 2 publications

(7 citation statements)

References 43 publications

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“…previously described (Sun and Wang, 2022). The CP tissue was either snap frozen for mRNA analysis or transferred to pre-gassed, aCSF for transport imaging studies.…”

Section: Animals and Choroid Plexus Tissue Collectionmentioning

confidence: 99%

“…Transport studies were conducted in freshly isolated mouse CP as described previously (Hu et al, 2022;Sun and Wang, 2022). Isolated CPs maintained vitality and transport activity for up to 2-3 hours, and all transport studies were performed within 2 hours after tissue isolation (Sun and Wang, 2022). Transport studies were initiated by adding a fluorescent substrate into the aCSF.…”

Section: Confocal Imaging and Transport Studies In Freshly Isolated C...mentioning

confidence: 99%

“…For real-time recording, fluorescent signals were recorded every minute for 20 minutes. Image acquisition were performed using a Zeiss LSM 710 confocal microscope as previously described (Sun and Wang, 2022).…”

Section: Confocal Imaging and Transport Studies In Freshly Isolated C...mentioning

confidence: 99%

“…Our laboratory recently applied confocal microscopy and live-tissue imaging techniques to study the transcellular transport mechanisms for organic cations and organic anions at the BCSFB (Hu et al, 2022;Sun and Wang, 2022). Using live CP tissues isolated from the mouse brain, we observed that the CPE cells lack an intact transcellular transport system for organic cations but possess a highly functional CSFto-blood transport system for large and amphipathic organic anions.…”

Section: Introductionmentioning

confidence: 99%

“…Using quantitative analysis and real-time imaging, we further showed that apical uptake of fluorescent organic anions (e.g. fluorescein methotrexate (FL-MTX), fluorescein-cyclic AMP (Fluo-cAMP)), is the rate-limiting step in the overall CSF-to-blood transport process and highly sensitive to the OATP inhibitor rifampin (Hu et al, 2022;Sun and Wang, 2022). However, definitive data establishing the role of OATPs in CPE cell apical uptake is lacking, and the specific OATP isoform(s) involved in this rate-limiting clearance step remain unknown.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Mechanisms of Organic Anion Transporting Polypeptide–Mediated Organic Anion Clearance at the Blood–Cerebrospinal Fluid Barrier

Sun,

Hagenbuch,

Kelly

et al. 2023

Molecular Pharmacology

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The blood-cerebrospinal fluid barrier (BCSFB), formed by the choroid plexus epithelial (CPE) cells, plays an active role in removing drugs and metabolic wastes from the brain. Recent functional studies in isolated mouse choroid plexus (CP) tissues suggested the existence of organic anion transporting polypeptides (OATPs, encoded by SLCOs) at the apical membrane of BCSFB, which may clear large organic anions from the cerebrospinal fluid (CSF). However, the specific OATP isoform involved is unclear. Using quantitative fluorescence imaging, we showed that the fluorescent anions, sulforhodamine101 (SR101), fluorescein methotrexate (FL-MTX), and 8fluorescein-cAMP (Fluo-cAMP), are actively transported from the CSF to the subepithelial space in CP tissues isolated from wild-type mice. In contrast, transepithelial transport of these compounds across the CPE cells was abolished in Oatp1a/1b -/mice due to impaired apical uptake. Using transporter-expressing cell lines, SR101, FL-MTX and fluo-cAMP were additionally shown to be transported by mouse OATP1A5 and its human counterpart OATP1A2. Kinetic analysis showed that estrone-3-sulfate and SR101 are transported by OATP1A2 and OATP1A5 with similar K m values. Immunofluorescence staining further revealed the presence of OATP1A2 protein in human CP tissues. Together, our results suggest that large organic anions in the CSF are actively transported into CPE cells by apical OATP1A2 (OATP1A5 in mice), then subsequently effluxed into the blood by basolateral multidrug resistance associated proteins (MRPs). As OATP1A2 transports a wide array of endogenous compounds and xenobiotics, the presence of this transporter at the BCSFB may imply a novel route for removing neurohormones, drugs and toxins from the CSF.

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“…previously described (Sun and Wang, 2022). The CP tissue was either snap frozen for mRNA analysis or transferred to pre-gassed, aCSF for transport imaging studies.…”

Section: Animals and Choroid Plexus Tissue Collectionmentioning

confidence: 99%

Section: Confocal Imaging and Transport Studies In Freshly Isolated C...mentioning

confidence: 99%

Section: Confocal Imaging and Transport Studies In Freshly Isolated C...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Molecular Mechanisms of Organic Anion Transporting Polypeptide–Mediated Organic Anion Clearance at the Blood–Cerebrospinal Fluid Barrier

Sun,

Hagenbuch,

Kelly

et al. 2023

Molecular Pharmacology

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Functional Evaluation of P-gp and Bcrp at the Murine Blood-Cerebrospinal Fluid Barrier

Sun,

Wang

2023

Pharm Res

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Evaluation of Blood-CSF Barrier Transport by Quantitative Real Time Fluorescence Microscopy

Cited by 2 publications

References 43 publications

Molecular Mechanisms of Organic Anion Transporting Polypeptide–Mediated Organic Anion Clearance at the Blood–Cerebrospinal Fluid Barrier

Molecular Mechanisms of Organic Anion Transporting Polypeptide–Mediated Organic Anion Clearance at the Blood–Cerebrospinal Fluid Barrier

Functional Evaluation of P-gp and Bcrp at the Murine Blood-Cerebrospinal Fluid Barrier

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