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. 2015 Jul 22;1(6):678–694. doi: 10.1016/j.jcmgh.2015.07.006

Figure 1.

Figure 1

Pancreatic levels of lysosome-associated membrane protein 1 (LAMP-1) and LAMP-2 decrease in rodent models and human pancreatitis. Pancreatitis was induced in rats by administration of cerulein (CR) or l-arginine (Arg), or by a combination of ethanol diet (EtOH) and low-dose CR; and in mice, by CR or feeding choline-deficient ethionine supplemented (CDE) diet (see Materials and Methods). (A, B). Immunoblot analysis of pancreatic LAMP levels in fully developed pancreatitis and at the onset (after 30 minutes) of CR pancreatitis, using Abs against the C terminus of LAMP-1 (Abcam) and LAMP-2 (Sigma). In this and other figures, extracellular signal-activated kinases 1/2 (ERK1/2), β-actin, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are loading controls; each lane represents an individual animal; the data are representative of several immunoblots from at least three animals for each condition; the narrow white space on the immunoblot indicates that the lanes are on the same blot but not contiguous. (B) The densitometric intensity of LAMP band was normalized to that of ERK in the same sample, and the mean LAMP/ERK ratio in pancreatitis group was further normalized to that in control group. Values are mean ± SEM (n = 4 per group). (C). Immunogold electron microscopy labeling shows LAMP-1 in lysosomes (L) and autolysosomes (AL) in pancreas of rats with CR pancreatitis and control (saline). The asterisks point to remnants of sequestered organelles. Scale bar: 500 nm. (D, E). Immunofluorescence analysis of LAMP-2 (using the C-terminal Ab) and cathepsin D (CatD) in human pancreas. Nuclei staining with 4′,6-diamidino-2-phenylindole (DAPI) (blue; not shown in “merge” panels). DIC, differential interference contrast, which prominently displays zymogen granules area in acinar cells. Scale bar: 10 μm (all images). (E) LAMP-2 punctate staining and colocalization of CatD with LAMP-2 in human normal pancreas and pancreatitis were quantified with ImageJ. Values are mean ± SEM from four normal and seven pancreatitis tissue samples from different patients. *P < .05 versus normal pancreas.