Demonstration of new sites of expression of the CCK-B/gastrin receptor in pancreatic acinar AR42J cells using immunoelectron microscopy.
The CCK-B/gastrin receptor has been characterised in both normal and tumour tissues. Endocytosis of the CCK-B/gastrin receptor has recently been demonstrated and this has similarly been described for other peptide receptors. In addition, ligand and ligand-receptor translocation to the nucleus has been demonstrated for other peptides. The aim of this study was to identify the sites of expression of the CCK-B/gastrin receptor in the known CCK-B/gastrin receptor bearing pancreatic acinar AR42J cells. The specificity of the CCK-B/gastrin receptor antibody (alpha-CCKBR-Ser antibody) was demonstrated by inhibition ELISA studies, radioligand inhibition studies and immunofluorescence binding studies on AR42J cells. Western blotting and immunogold electron microscopy techniques were used to identify the receptor in AR42J cell preparations. The affinity purified alpha-CCKBR-Ser antibody was shown to be specific for the CCK-B/gastrin receptor. The receptor was expressed on the cell membrane, in the cytoplasm and within the nucleus. Isoforms of the receptor protein identified in extra-nuclear and nuclear extracts ranged in molecular weight from 58 to 66 kDa. We conclude that the CCK-B/gastrin receptor is not only expressed on the cell membrane, but also in the cytoplasm and nucleus of AR42J pancreatic acinar cells.
Caplin
M E
ME
Department of Medicine and Histopathology, Royal Free Hospital School of Medicine, London, UK. caplin@rfhsm.ac.uk
Clarke
P
P
Grimes
S
S
Dhillon
A P
AP
Khan
K
K
Savage
K
K
Lewin
J
J
Michaeli
D
D
Pounder
R E
RE
Watson
S A
SA
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D007457:Iodine Radioisotopes; D016253:Microscopy, Immunoelectron; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D043887:Receptor, Cholecystokinin B; D011949:Receptors, Cholecystokinin / Q000276:immunology / Q000378:metabolism*
D007457:Iodine Radioisotopes; D043887:Receptor, Cholecystokinin B; D011949:Receptors, Cholecystokinin