Supervisors info:
Αθανασία Σιαφάκα Καθηγήτρια, Μαίρη Μαυρή-Βαβαγιάννη Αναπληρώτρια Καθηγήτρια, Ντία Γαλανοπούλου Αναπληρώτρια Καθηγήτρια
Summary:
In the present study, the secretion of the two main degradative enzymes of the
endocannabinoid system, the fatty acid amide hydrolase (FAAH) and the
monoacylglycerol lipase (MAGL), in the protozoan Tetrahymena thermophila was
investigated. At first, the secretion of both enzymes in starvation medium was
investigated, using [3H]2-oleoylglycerol (2-OG) as substrate and in the
presence or absence of FAAH specific inhibitor AM374. Both enzymes were
secreted in the starvation medium, in a time dependent manner. The maximum
secretion was ~5 % after 4 hours of incubation and steadily declined
afterwards. Subsequently, stimulation of secretion with dibucaine leads to
exocytosis of the mycocysts of Tetrahymena. In the isolated mucus the activity
of MAGL and FAAH was determined, using [3H]2-OG as a substrate in the presence
of AM374 or MAGL specific inhibitor JZL184. In the case of MAGL, the activity
measured was equivalent with that of the cell homogenate. Immunoblot analysis
using anti-MAGL antibody revealed the presence of an immunoreactive protein at
~45kDa in the mucus fraction, the supertatant and homogenate. Finally, confocal
fluorescence microscopy showed the pericellular localization of ΜΑGL, around
the plasma membrane, suggesting its existence in the mucosysts.
Keywords:
MAGL, FAAH, Tetrahymena, lysosomal secretion, Mucocyst secretion
