Evidence of pituitary adenylate cyclase activating polypeptide (PACAP) in pancreatic islet cells by confocal microscopy.
Several studies have shown that pituitary adenylate cyclase activating polypeptide (PACAP) stimulates at very low concentration insulin release from pancreatic beta cells. In addition, PACAP has been evidenced in pancreatic nervous fibers surrounding the islets, the core of the islet, and the capillaries. The aim of the present study was to demonstrate internalization of PACAP in pancreatic islet cells. Pancreatic islets were obtained from Wistar rat pancreata by modified Lacy's isolation method. The isolated islets were incubated in the presence of Fluo-PACAP 27, a fluorescent ligand specific for PACAP receptors. At the end of incubation the islets were fixed in paraformaldehyde and then observed by confocal microscope. Fluo-PACAP 27 was internalized into pancreatic islet cells, and this process was time- and temperature-dependent (37 degrees C). The fluorescent molecules converged toward the nucleus where an intense fluorescence was evidenced after 60 minutes. Incubation with phenyl arsine oxide as well as with PACAP 6-38, a receptor antagonist, prevented the internalization process. Further studies are required to explain the internalization process of PACAP 27 into the nucleus of pancreatic islet cells.
Petruzzo
P
P
Department of Surgical Sciences, University of Cagliari, Italy. brotzu@vaxca1.unica.it
Cappai
A
A
Spiga
S
S
Picciau
S
S
Serra
G
G
Fattore
L
L
Onali
P
P
Brotzu
G
G
D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure; D002469:Cell Separation; D004705:Endocytosis; D007091:Image Processing, Computer-Assisted; D007515:Islets of Langerhans / Q000737:chemistry* / Q000648:ultrastructure; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009479:Neuropeptides / Q000032:analysis*; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D051381:Rats; D017208:Rats, Wistar; D013048:Specimen Handling
C495403:Adcyap1 protein, rat; D001152:Arsenicals; D009479:Neuropeptides; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; C029341:oxophenylarsine