TY - JOUR
T1 - Ex vivo nanoscale abluminal mapping of putative cargo receptors at the blood-brain barrier of expanded brain capillaries
AU - Holst, Mikkel Roland
AU - Richner, Mette
AU - Arenshøj, Pernille Olsgaard
AU - Alam, Parvez
AU - Hyldig, Kathrine
AU - Nielsen, Morten Schallburg
N1 - Publisher Copyright:
© The Author(s) 2024.
PY - 2024/12
Y1 - 2024/12
N2 - Receptor mediated transport of therapeutic antibodies through the blood-brain barrier (BBB) give promise for drug delivery to alleviate brain diseases. We developed a low-cost method to obtain nanoscale localization data of putative cargo receptors. We combine existing ex vivo isolation methods with expansion microscopy (ExM) to analyze receptor localizations in brain microcapillaries. Using this approach, we show how to analyze receptor localizations in endothelial cells of brain microcapillaries in relation to the abluminal marker collagen IV. By choosing the thinnest capillaries, microcapillaries for analysis, we ensure the validity of collagen IV as an abluminal marker. With this tool, we confirm transferrin receptors as well as sortilin to be both luminally and abluminally localized. Furthermore, we identify basigin to be an abluminal receptor. Our methodology can be adapted to analyze different types of isolated brain capillaries and we anticipate that this approach will be very useful for the research community to gain new insight into cargo receptor trafficking in the slim brain endothelial cells to elucidate novel paths for future drug design.
AB - Receptor mediated transport of therapeutic antibodies through the blood-brain barrier (BBB) give promise for drug delivery to alleviate brain diseases. We developed a low-cost method to obtain nanoscale localization data of putative cargo receptors. We combine existing ex vivo isolation methods with expansion microscopy (ExM) to analyze receptor localizations in brain microcapillaries. Using this approach, we show how to analyze receptor localizations in endothelial cells of brain microcapillaries in relation to the abluminal marker collagen IV. By choosing the thinnest capillaries, microcapillaries for analysis, we ensure the validity of collagen IV as an abluminal marker. With this tool, we confirm transferrin receptors as well as sortilin to be both luminally and abluminally localized. Furthermore, we identify basigin to be an abluminal receptor. Our methodology can be adapted to analyze different types of isolated brain capillaries and we anticipate that this approach will be very useful for the research community to gain new insight into cargo receptor trafficking in the slim brain endothelial cells to elucidate novel paths for future drug design.
KW - Basigin
KW - Blood‒brain barrier
KW - Cargo receptor
KW - Machine leaning and expansion microscopy
KW - Porcine brain endothelial cells
KW - Sortilin
KW - Transferrin receptor
UR - http://www.scopus.com/inward/record.url?scp=85206279037&partnerID=8YFLogxK
U2 - 10.1186/s12987-024-00585-x
DO - 10.1186/s12987-024-00585-x
M3 - Journal article
C2 - 39402596
AN - SCOPUS:85206279037
SN - 2045-8118
VL - 21
JO - Fluids and Barriers of the CNS
JF - Fluids and Barriers of the CNS
IS - 1
M1 - 80
ER -