Expression of Reg and cytokeratin 20 during ductal cell differentiation and proliferation in a mouse model of autoimmune diabetes.
OBJECTIVE
To evaluate the existence of beta-cell differentiation and proliferation in the low-dose streptozotocin (ld-STZ) mouse model of autoimmune diabetes.


METHODS
We studied the expression of Reg protein and cytokeratin 20 (CK20), the presence of proliferative phenomena (judged by the incorporation of bromodeoxyuridine (BrdU)), and the co-expression of Reg, CK20 or BrdU with insulin.


METHODS
Diabetes was induced in male C57Bl6/J mice by administration of ld-STZ. The animals were killed at days 10 and 23 from the beginning of the induction of disease. Five animals were used at each time point and each group was evaluated for blood glucose concentrations, insulitis, expression of Reg and CK20 pancreatic proteins and BrdU incorporation, together with staining for insulin by immunohistochemistry and laser confocal microscopy.


RESULTS
All mice treated with ld-STZ were hyperglycemic and histological investigation showed a mild or severe insulitis both at day 10 and at day 23. At day 10, immunochemistry revealed an intense expression of Reg and CK20 in pancreatic ducts in ld-STZ mice, but not in control mice. Reg and CK20 immunoreactive cells were also positive for insulin. In contrast, at day 23, pancreatic sections reacted weakly with anti-Reg and anti-CK20 antibody; co-localization with insulin was observed for both Reg and CK20. The incorporation of BrdU was observed only in insulin-positive cells in pancreatic sections from mice killed at day 10.


CONCLUSIONS
These observations show an islet regeneration mechanism in response to an autoimmune attack, and that the ld-STZ mouse is a suitable model in which to evaluate intervention strategies.
Anastasi
E
E
Endocrinology, Clinica Medica 2, University of Rome 'La Sapienza', Italy. eanastasi@uniroma1.it
Ponte
E
E
Gradini
R
R
Bulotta
A
A
Sale
P
P
Tiberti
C
C
Okamoto
H
H
Dotta
F
F
Di Mario
U
U
D000818:Animals; D001327:Autoimmune Diseases / Q000378:metabolism / Q000473:pathology; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002454:Cell Differentiation; D002455:Cell Division; D003921:Diabetes Mellitus, Experimental / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D005455:Fluorescent Antibody Technique; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D007515:Islets of Langerhans / Q000276:immunology / Q000473:pathology; D053554:Keratin-20; D051847:Lithostathine; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins; D010179:Pancreas / Q000473:pathology; D010183:Pancreatic Ducts / Q000473:pathology*
D001786:Blood Glucose; D002135:Calcium-Binding Proteins; D007381:Intermediate Filament Proteins; D053554:Keratin-20; C489975:Krt20 protein, mouse; D051847:Lithostathine; D009419:Nerve Tissue Proteins; C497615:Reg1 protein, mouse