Our work continues to focus on the processes leading to fusion between granule and plasma membranes during exocytosis in cells, including chromaffin cells, beta cells from islet of Langerhans, nerve terminals, and mucin secreting human colon carcinoma cells. The contact and fusion event during exocytosis may be controlled by the protein synexin. We have recently cloned and sequenced the cDNA for this by the protein, and learned that it is a member of the annexin gene family. We have expressed synexin in E. coli and have found the recombinant protein to express both fusion and channel properties. Secretion mechanisms have also been studied in intact chromaffin cells, in which evidence has been found to support the concept that calcium and barium promote secretion by separate mechanisms. The acetycholine receptor in chromaffin cells has been found to be different from the classical nicotinic receptor, inasmuch as it has unexpected muscarinic character. A novel calcium channel has been discovered in human beta cells, which responds to glucose and related metabolites. This channel is named by us the """"""""G-type"""""""" calcium channel, and may play a central role in glucose activation if islets. Glucose induced insulin release is modulated by acetylcholine, and pharmacological analysis reveals that the muscarinic receptor type is M1 rather than M2. The adrenal medullas has provided a tractable system to study the interaction between endocrine cells and adherent capillaries. Endothelial cells have MAO-A, exclusively, and various steroids affect expression of this enzyme, without affecting the MAO-B which is exclusively found in chromaffin cells. A new method for measuring ascorbic acid using HPLC and coulometric electrochemical detection has been developed. The method allows measurement of the vitamin, not only in cells such as leucocytes and chromaffin cells, but also human plasma. Two transporters for ascorbic acid have been detected in human neutrophils. Millimolar concentrations of ascorbic acid have been detected in unbound form in both human neutrophils and monocytes.
