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In vitro secretion of zymogens by bovine pancreatic acini and ultra-structural analysis of exocytosis.

ABSTRACT: The aim of this study is to establish a bovine pancreatic acinar cell culture model with longer viability and functionality. The cells could be maintained in a functional state for upto 20 days with normal morphology. Cells were positive for amylase as observed by immunofluorescence staining. Acinar cells are spherical and range about 2-3 µm in diameter. The porosome formed by exocytosis and heterogenous enzyme granules of size ranging 100-300 nm were seen on the surface of cells by electron microscopy. The activity of the enzymes was high on day 15 and the activity profile of the enzymes is in the order: protease>lipase>amylase and the enzymes were identified by SDS-PAGE. Long-term culture of bovine pancreatic acini could be useful in studying the pathogenesis of pancreatitis. Since the bovine genome shares about 80% identity with the human genome, the cells derived from bovine pancreas can be engineered and used as a potential xenotransplant to treat conditions like pancreatitis as the tissue source is abundantly available.

SUBMITTER: Jayaveni S

PROVIDER: S-EPMC5600341 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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<i>In vitro</i> secretion of zymogens by bovine pancreatic acini and ultra-structural analysis of exocytosis.

Jayaveni Sivalingam S Nithyanandham Kamaraj K Rose Chellan C

Biochemistry and biophysics reports 20151223

The aim of this study is to establish a bovine pancreatic acinar cell culture model with longer viability and functionality. The cells could be maintained in a functional state for upto 20 days with normal morphology. Cells were positive for amylase as observed by immunofluorescence staining. Acinar cells are spherical and range about 2-3 µm in diameter. The porosome formed by exocytosis and heterogenous enzyme granules of size ranging 100-300 nm were seen on the surface of cells by electron mic ...[more]

PMID: 28955829

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Project description:BackgroundHuman African trypanosomiasis is a lethal disease caused by the extracellular parasite Trypanosoma brucei. The proteins secreted by T. brucei inhibit the maturation of dendritic cells and their ability to induce lymphocytic allogenic responses. To better understand the pathogenic process, we combined different approaches to characterize these secreted proteins.ResultsOverall, 444 proteins were identified using mass spectrometry, the largest parasite secretome described to date. Functional analysis of these proteins revealed a strong bias toward folding and degradation processes and to a lesser extent toward nucleotide metabolism. These features were shared by different strains of T. brucei, but distinguished the secretome from published T. brucei whole proteome or glycosome. In addition, several proteins had not been previously described in Trypanosoma and some constitute novel potential therapeutic targets or diagnostic markers. Interestingly, a high proportion of these secreted proteins are known to have alternative roles once secreted. Furthermore, bioinformatic analysis showed that a significant proportion of proteins in the secretome lack transit peptide and are probably not secreted through the classical sorting pathway. Membrane vesicles from secretion buffer and infested rat serum were purified on sucrose gradient and electron microscopy pictures have shown 50- to 100-nm vesicles budding from the coated plasma membrane. Mass spectrometry confirmed the presence of Trypanosoma proteins in these microvesicles, showing that an active exocytosis might occur beyond the flagellar pocket.ConclusionsThis study brings out several unexpected features of the secreted proteins and opens novel perspectives concerning the survival strategy of Trypanosoma as well as possible ways to control the disease. In addition, concordant lines of evidence support the original hypothesis of the involvement of microvesicle-like bodies in the survival strategy allowing Trypanosoma to exchange proteins at least between parasites and/or to manipulate the host immune system.

Dataset Information

In vitro secretion of zymogens by bovine pancreatic acini and ultra-structural analysis of exocytosis.

Publications

<i>In vitro</i> secretion of zymogens by bovine pancreatic acini and ultra-structural analysis of exocytosis.

Similar Datasets

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