Fingolimod--a sphingosine-like molecule inhibits vesicle mobility and secretion in astrocytes.
In the brain, astrocytes signal to the neighboring cells by the release of chemical messengers (gliotransmitters) via regulated exocytosis. Recent studies uncovered a potential role of signaling lipids in modulation of exocytosis. Hence, we investigated whether sphingosine and the structural analog fingolimod/FTY720, a recently introduced therapeutic for multiple sclerosis, affect (i) intracellular vesicle mobility and (ii) vesicle cargo discharge from cultured rat astrocytes. Distinct types of vesicles, peptidergic, glutamatergic, and endosomes/lysosomes, were fluorescently prelabeled by cell transfection with plasmids encoding atrial natriuretic peptide tagged with mutant green fluorescent protein and vesicular glutamate transporter tagged with enhanced green fluorescent protein or by LysoTracker staining, respectively. The confocal and total internal reflection fluorescence microscopies were used to monitor vesicle mobility in the cytoplasm and near the basal plasma membrane, respectively. Sphingosine and FTY720, but not the membrane impermeable lipid analogs, dose-dependently attenuated vesicle mobility in the subcellular regions studied, and significantly inhibited stimulated exocytotic peptide and glutamate release. We conclude that in astrocytes, cell permeable sphingosine-like lipids affect regulated exocytosis by attenuating vesicle mobility, thereby preventing effective vesicle access/interaction with the plasma membrane docking/release sites.
Celica d.o.o., Biomedical Center, Technology Park 24, Ljubljana, Slovenia.
Saša
S
Trkov
Matjaž
M
Stenovec
Marko
M
Kreft
Maja
M
Potokar
Vladimir
V
Parpura
Bazbek
B
Davletov
Robert
R
Zorec
D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005089:Exocytosis / Q000187:drug effects* / Q000502:physiology; D000068876:Fingolimod Hydrochloride; D018698:Glutamic Acid / Q000378:metabolism; D011409:Propylene Glycols / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D022161:Transport Vesicles / Q000187:drug effects* / Q000378:metabolism
D011409:Propylene Glycols; D018698:Glutamic Acid; D000068876:Fingolimod Hydrochloride; D013110:Sphingosine; D002118:Calcium