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Adenovirus-mediated gene expression in isolated rat pancreatic acini and individual pancreatic acinar cells.
- Source :
-
Pflugers Archiv : European journal of physiology [Pflugers Arch] 1998 Oct; Vol. 436 (5), pp. 782-7. - Publication Year :
- 1998
-
Abstract
- In this study we have examined the feasibility of using replication-deficient recombinant adenoviral vectors to transfer and express genes in pancreatic acinar cells in vitro. We infected primary cultures of both isolated pancreatic acini and individual acinar cells with a recombinant adenovirus containing the coding sequence for beta-galactosidase. Our data demonstrate that recombinant adenoviruses readily infect pancreatic acinar cells in vitro. Close to 100% infection and maximal beta-galactosidase expression were obtained, when acini or acinar cells were infected with 5x10(6) or 10(6) plaque-forming units (pfu) of virus per millitre of acini or acinar cell suspension, respectively. Examination of the time-course of beta-galactosidase expression showed that there was a lag of approximately 6 h before beta-galactosidase levels increased. Thereafter beta-galactosidase expression increased rapidly. By 20 h post-infection beta-galactosidase activity had increased from undetectable levels to 2.5-3.0 units/mg of cellular protein. Acini/acinar cells maintained a robust secretory response after adenoviral infection. The cholecystokinin-octapeptide (CCK8) dose/response curves for amylase secretion for acini and acinar cells infected with 5x10(5) and 1x10(5) pfu/ml of virus, respectively, were biphasic, with maximal amylase secretion being stimulated by 1 nM CCK8. In addition, the dose/response curves were identical to those obtained from control, sham-infected, acini/acinar cells. Our findings indicate that replication-deficient recombinant adenoviral vectors will be excellent tools to transfer and express genes in isolated pancreatic acini or acinar cells.
- Subjects :
- Amylases analysis
Amylases drug effects
Amylases metabolism
Animals
Cells, Cultured
Cholecystokinin pharmacology
Dose-Response Relationship, Drug
Exocytosis
Gene Transfer Techniques
Genetic Vectors
Models, Biological
Pancreas metabolism
Rats
Recombinant Proteins biosynthesis
Recombinant Proteins genetics
Transfection methods
Transgenes genetics
beta-Galactosidase analysis
beta-Galactosidase biosynthesis
beta-Galactosidase genetics
Adenoviridae genetics
Gene Expression genetics
Pancreas cytology
Subjects
Details
- Language :
- English
- ISSN :
- 0031-6768
- Volume :
- 436
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Pflugers Archiv : European journal of physiology
- Publication Type :
- Academic Journal
- Accession number :
- 9716713
- Full Text :
- https://doi.org/10.1007/s004240050702