Liddelow, Shane A3; Dzięgielewska, Katarzyna M3; Møllgård, Kjeld7; Whish, Sophie C3; Noor, Natassya M3; Wheaton, Benjamin J3; Gehwolf, Renate5; Wagner, Andrea5; Traweger, Andreas5; Bauer, Hannelore6; Bauer, Hans-Christian5; Saunders, Norman R3
1 Section VI. Building 18.4, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, Københavns Universitet2 Morphogenesis and Differentiation Program, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, Københavns Universitet3 Department of Pharmacology & Therapeutics, University of Melbourne, Melbourne, Australia.4 Section IX. Section of Education, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, Københavns Universitet5 Paracelsus Medical University6 Department of Organismic Biology, University of Salzburg, Salzburg, Austria.7 Section IX. Section of Education, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, Københavns Universitet
To maintain the precise internal milieu of the mammalian central nervous system, well-controlled transfer of molecules from periphery into brain is required. Recently the soluble and cell-surface albumin-binding glycoprotein SPARC (secreted protein acidic and rich in cysteine) has been implicated in albumin transport into developing brain, however the exact mechanism remains unknown. We postulate that SPARC is a docking site for albumin, mediating its uptake and transfer by choroid plexus epithelial cells from blood into cerebrospinal fluid (CSF). We used in vivo physiological measurements of transfer of endogenous (mouse) and exogenous (human) albumins, in situ Proximity Ligation Assay (in situ PLA), and qRT-PCR experiments to examine the cellular mechanism mediating protein transfer across the blood-CSF interface. We report that at all developmental stages mouse albumin and SPARC gave positive signals with in situ PLAs in plasma, CSF and within individual plexus cells suggesting a possible molecular interaction. In contrast, in situ PLA experiments in brain sections from mice injected with human albumin showed positive signals for human albumin in the vascular compartment that were only rarely identifiable within choroid plexus cells and only at older ages. Concentrations of both endogenous mouse albumin and exogenous (intraperitoneally injected) human albumin were estimated in plasma and CSF and expressed as CSF/plasma concentration ratios. Human albumin was not transferred through the mouse blood-CSF barrier to the same extent as endogenous mouse albumin, confirming results from in situ PLA. During postnatal development Sparc gene expression was higher in early postnatal ages than in the adult and changed in response to altered levels of albumin in blood plasma in a differential and developmentally regulated manner. Here we propose a possible cellular route and mechanism by which albumin is transferred from blood into CSF across a sub-population of specialised choroid plexus epithelial cells.