Serotonin Storage Pools

We studied binding of serotonin to protein(s) derived from rat basophil leukemia (RBL) cells and mast cells. We found two types of serotonin binding protein in RBL cells. These proteins differed from one another in molecular weight and eluted in separate peaks from sephadex G-200 columns. Peak I protein (Kr) = 1 .9 X 10-6 M) was a glycoprotein that bound to concanavalin A (Con A) ; Peak II protein (KD1 = 4.5 X 10-I'M ; Ko2 = 3.9 X 10-6 M) did not bind to Con A. Moreover, binding of [3H]serotonin to protein of peak I was sensitive to inhibition by reserpine, while binding of [3H]serotonin to protein of peak II resisted inhibition by that drug . Other differences between the two types of binding protein were found, the most significant of which was the far more vigorous conditions of homogenization required to extract peak I than peak II protein . Neither peak I nor peak II protein resembled the serotonin binding protein (SBP) that is found in serotonergic neurons of the brain and gut. Electron microscope radioautographic analysis of the intracellular distribution of [3H]serotonin taken up in vitro by RBL cells or in vivo by murine mast cells indicated that essentially all of the labeled amine was located in cytoplasmic granules . No evidence for a pool in the cytosol was found and all granules were capable of becoming labeled. The presence of two types of intracellular serotonin binding proteins in these cells may indicate that there are two intracellular storage compartments for the amine. Both may be intragranular, but peak I protein may be associated with the granular membrane while peak II protein may be more free within the granular core . Different storage proteins may help to explain the differential release of amines from mast cell granules . The cellular biology ofserotonergic neurons is difficult to study in the central nervous system (CNS) because the tissue is complex and because glial cells take up and bind serotonin (10, 18). As a result, the mechanisms involved in the storage and release of serotonin by serotonergic neurons are still rather poorly understood (11) . To circumvent some of the problems inherent in studying the CNS, serotonin uptake and release have been examined in several simpler model systems . The myenteric plexus of the gut, for instance, contains serotonergic neurons that share many of the characteristics of their CNS counterparts (12): they both take up serotonin by a similar mechanism and both contain a highly specific serotonin binding protein (SBP) . Since SBP has also been found to be moved proximodistally in serotonergic axons by fast transport and to be concentrated in synaptic vesicles (29), it seems likely that SBP is a component of serotonergic synaptic vesicles and is, at least in part, released together with the transmitter by exocytosis (21) . Non-neuronal cells have also been studied in the hope that they will manifest serotonergic mechanisms identical to those of neurons and thus serve as proxies for neurons . Such cells include platelets, thyroid parafolfcular cells, and mast cells . THE JOURNAL OF CELL BIOLOGY " VOLUME 93 JUNE 1982 638-647 ©The Rockefeller University Press " 0021-9525/82/06/0638/10 $1 .00 on M ay 4, 2017 D ow nladed fom Published June 1, 1982