scholarly journals Chromatin Immunoprecipitation (ChIP) from pancreatic acinar cells and whole pancreatic tissue

2011 ◽  
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells

scholarly journals Impaired autophagy increases susceptibility to endotoxin-induced chronic pancreatitis

2020 ◽  
Vol 11 (10) ◽  
Author(s):  
L. Xia ◽  
Z. Xu ◽  
X. Zhou ◽  
F. Bergmann ◽  
N. Grabe ◽  
...  
Keyword(s):  
Chronic Pancreatitis ◽  
Lethal Dose ◽  
Acinar Cells ◽  
Pancreatic Injury ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Chromogenic Assay ◽  
Enhanced Expression ◽  
Metabolic Endotoxemia

Abstract Chronic pancreatitis (CP) is associated with elevated plasma levels of bacterial lipopolysaccharide (LPS) and we have demonstrated reduced acinar cell autophagy in human CP tissue. Therefore, we investigated the role of autophagy in experimental endotoxin-induced pancreatic injury and aimed to identify LPS in human CP tissue. Pancreatic Atg7-deficient mice were injected with a single sub-lethal dose of LPS. Expression of autophagy, apoptosis, necroptosis, and inflammatory markers was determined 3 and 24 h later utilizing immunoblotting and immunofluorescence. The presence of LPS in pancreatic tissue from mice and from patients and healthy controls was determined using immunohistochemistry, immunoblots, and chromogenic assay. Mice lacking pancreatic autophagy exhibited local signs of inflammation and were particularly sensitive to the toxic effect of LPS injection as compared to control mice. In response to LPS, Atg7Δpan mice exhibited enhanced vacuolization of pancreatic acinar cells, increase in TLR4 expression coupled to enhanced expression of NF-κΒ, JNK, and pro-inflammatory cytokines by acinar cells and enhanced infiltration by myeloid cells (but not Atg7F/F controls). Cell death was enhanced in Atg7Δpan pancreata, but only necroptosis and trypsin activation was further amplified following LPS injection along with elevated pancreatic LPS. The presence of LPS was identified in the pancreata from all 14 CP patients examined but was absent in the pancreata from all 10 normal controls. Altogether, these results support a potential role for metabolic endotoxemia in the pathogenesis of CP. Moreover, the evidence also supports the notion that autophagy plays a major cytoprotective and anti-inflammatory role in the pancreas, and blunting metabolic endotoxemia-induced CP.

Electrogenic sodium pump in pancreatic acinar cells

1973 ◽  
Vol 184 (1074) ◽  
pp. 115-119 ◽  
Keyword(s):  
Cell Membrane ◽  
Acinar Cell ◽  
Sodium Pump ◽  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Bathing Solution ◽  
Sodium Pumps ◽  
Epithelial Tissues ◽  
Electrogenic Sodium Pump

Electrogenic sodium pumps have been described in many excitable tissues, but hitherto not with certainty in epithelial tissues. In the present work it is shown that readmission of potassium to the bathing solution of mouse pancreatic tissue previously deprived of potas­sium, causes a marked hyperpolarization of the acinar cell membrane. This hyperpolariza­tion is abolished reversibly by strophanthin-G. This indicates the presence of electrogenic sodium pumping in pancreatic acinar cells.

Critical role for NHE1 in intracellular pH regulation in pancreatic acinar cells

2003 ◽  
Vol 285 (5) ◽  
pp. G804-G812 ◽  
Author(s):  
David A. Brown ◽  
James E. Melvin ◽  
David I. Yule
Keyword(s):  
Intracellular Ph ◽  
Ph Regulation ◽  
Critical Role ◽  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Muscarinic Agonist ◽  
Targeted Disruption ◽  
General Role ◽  
Pancreatic Acini

The primary function of pancreatic acinar cells is to secrete digestive enzymes together with a NaCl-rich primary fluid which is later greatly supplemented and modified by the pancreatic duct. A Na+/H+ exchanger(s) [NHE(s)] is proposed to be integral in the process of fluid secretion both in terms of the transcellular flux of Na+ and intracellular pH (pHi) regulation. Multiple NHE isoforms have been identified in pancreatic tissue, but little is known about their individual functions in acinar cells. The Na+/H+ exchange inhibitor 5-( N-ethyl- N-isopropyl) amiloride completely blocked pHi recovery after an NH4Cl-induced acid challenge, confirming a general role for NHE in pHi regulation. The targeted disruption of the Nhe1 gene also completely abolished pHi recovery from an acid load in pancreatic acini in both [Formula: see text]-containing and [Formula: see text]-free solutions. In contrast, the disruption of either Nhe2 or Nhe3 had no effect on pHi recovery. In addition, NHE1 activity was upregulated in response to muscarinic stimulation in wild-type mice but not in NHE1-deficient mice. Fluctuations in pHi could potentially have major effects on Ca2+ signaling following secretagogue stimulation; however, the targeted disruption of Nhe1 was found to have no significant effect on intracellular Ca2+ homeostasis. These data demonstrate that NHE1 is the major regulator of pHi in both resting and muscarinic agonist-stimulated pancreatic acinar cells.

PSD-95 protects the pancreas against pathological damage through p38 MAPK signaling pathway in acute pancreatitis

2021 ◽  
pp. 153537022110032
Author(s):  
Yinan Guo ◽  
Weikai Hu ◽  
Xueyan Wang ◽  
Chunyun Li ◽  
Tianyu Cui ◽  
...  
Keyword(s):  
Acute Pancreatitis ◽  
P38 Mapk ◽  
Cell Model ◽  
Control Cell ◽  
Acinar Cells ◽  
Mapk Signaling ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Ar42j Cells ◽  
Inhibition Group

Acute pancreatitis is one of the leading causes of gastrointestinal disorder-related hospitalizations, yet its pathogenesis remains to be fully elucidated. Postsynaptic density protein-95 (PSD-95) is closely associated with tissue inflammation and injury. We aimed to investigate the expression of PSD-95 in pancreatic acinar cells, and its function in regulating the inflammatory response and pancreatic pathological damage in acute pancreatitis. A mouse model of edematous acute pancreatitis was induced with caerulein and lipopolysaccharide in C57BL/6 mice. Tat-N-dimer was injected to inhibit the PSD-95 activity separately, or simultaneously with SB203580, inhibitor of p38 MAPK phosphorylation. Rat pancreatic acinar cells AR42J were cultured with 1 μM caerulein to build a cell model of acute pancreatitis. PSD-95-knockdown and negative control cell lines were constructed by lentiviral transfection of AR42J cells. Paraffin-embedded pancreatic tissue samples were processed for routine HE staining to evaluate the pathological changes of human and mouse pancreatic tissues. Serum amylase and inflammatory cytokine levels were detected with specific ELISA kits. Immunofluorescence, immunohistochemical, Western-blot, and qRT-PCR were used to detect the expression levels of PSD-95, p38, and phosphorylated p38. Our findings showed that PSD-95 is expressed in the pancreatic tissues of humans, C57BL/6 mice, and AR42J cells, primarily in the cytoplasm. PSD-95 expression increased at 2 h, reaching the peak at 6 h in mice and 12 h in AR42J cells. IL-6, IL-8, and TNF-α increased within 2 h of disease induction. The pancreatic histopathologic score was greater in the PSD-95 inhibition group compared with the control ( P < 0.05), while it was lesser when phosphorylation of p38 MAPK was inhibited compared with the PSD-95 inhibition group ( P < 0.05). Moreover, phosphorylation of p38 MAPK increased statistically after PSD-95 knocked-down. In conclusion, PSD-95 effectively influences the pathological damage of the pancreas in acute pancreatitis by affecting the phosphorylation of p38 MAPK.

The cell-specific elastase I enhancer comprises two domains

1988 ◽  
Vol 8 (2) ◽  
pp. 893-902 ◽  
Author(s):  
F Kruse ◽  
C T Komro ◽  
C H Michnoff ◽  
R J MacDonald
Keyword(s):  
Base Pair ◽  
Regulatory Region ◽  
Acinar Cells ◽  
Chloramphenicol Acetyltransferase ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Specific Expression ◽  
Nuclear Extracts ◽  
Dna Strands ◽  
Rat Elastase

Two separate domains within the 134-base-pair rat elastase I enhancer and a third domain at the enhancer-promoter boundary are required for selective expression in pancreatic acinar cells. The domains were detected by a series of 10-base-pair substitution mutations across the elastase I gene regulatory region from positions -200 to -61. The effect of each mutant on the pancreas-specific expression of a linked chloramphenicol acetyltransferase gene was assayed by transfection into pancreatic 266-6 acinar cells and control NIH/3T3 cells. The two enhancer domains are nonredundant, because mutations in either eliminated (greater than 100-fold reduction) expression in 266-6 cells. DNase I protection studies of the elastase I enhancer-promoter region with partially purified nuclear extracts from pancreatic tissue and 266-6 cells revealed nine discrete protected regions (footprints) on both DNA strands. One of three footprints that lie within the two functional domains of the enhancer contained a sequence, conserved among several pancreas-specific genes, which when mutated decreased linked chloramphenicol acetyltransferase expression up to 170-fold in 266-6 cells. This footprint may represent a binding site for one or more pancreas-specific regulatory proteins.

scholarly journals Coxsackievirus Infection Induces Autophagy-Like Vesicles and Megaphagosomes in Pancreatic Acinar Cells In Vivo

2010 ◽  
Vol 84 (23) ◽  
pp. 12110-12124 ◽  
Author(s):  
Christopher C. Kemball ◽  
Mehrdad Alirezaei ◽  
Claudia T. Flynn ◽  
Malcolm R. Wood ◽  
Stephanie Harkins ◽  
...  
Keyword(s):  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Host Cells ◽  
Pancreatic Acinar Cells ◽  
Replication Complex ◽  
Cvb3 Infection ◽  
Infected Cells ◽  
Autophagy Pathway ◽  
The Individual

ABSTRACT Autophagy can play an important part in protecting host cells during virus infection, and several viruses have developed strategies by which to evade or even exploit this homeostatic pathway. Tissue culture studies have shown that poliovirus, an enterovirus, modulates autophagy. Herein, we report on in vivo studies that evaluate the effects on autophagy of coxsackievirus B3 (CVB3). We show that in pancreatic acinar cells, CVB3 induces the formation of abundant small autophagy-like vesicles and permits amphisome formation. However, the virus markedly, albeit incompletely, limits the fusion of autophagosomes (and/or amphisomes) with lysosomes, and, perhaps as a result, very large autophagy-related structures are formed within infected cells; we term these structures megaphagosomes. Ultrastructural analyses confirmed that double-membraned autophagy-like vesicles were present in infected pancreatic tissue and that the megaphagosomes were related to the autophagy pathway; they also revealed a highly organized lattice, the individual components of which are of a size consistent with CVB RNA polymerase; we suggest that this may represent a coxsackievirus replication complex. Thus, these in vivo studies demonstrate that CVB3 infection dramatically modifies autophagy in infected pancreatic acinar cells.

Morphological Characteristics of Infectious Pancreatic Necrosis Yirus in Trout Pancreatic Tissue

1969 ◽  
Vol 26 (8) ◽  
pp. 2247-2250 ◽  
Author(s):  
Donald Lightner ◽  
George Post
Keyword(s):  
Pancreatic Necrosis ◽  
Morphological Characteristics ◽  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Salmo Gairdneri ◽  
Thin Sections ◽  
Virus Particles ◽  
Infectious Pancreatic Necrosis ◽  
Electron Microscopical

Electron microscopical observations were made of infectious pancreatic necrosis (IPN) virus in thin sections of pancreatic acinar cells of rainbow trout (Salmo gairdneri). The virus and trout used in this study were obtained from a naturally occurring IPN epizootic in Colorado trout hatcheries and rearing units in 1968. IPN virus particles were found in large numbers in pancreatic acinar cells. Virus particles were associated most often with inclusion bodies composed of masses of virus particles and cell debris or large crystalline aggregations that were encircled by a single-layered membrane. Mature virus particles were found to average 57 nm in diameter and were hexagonal in profile.

scholarly journals The cell-specific elastase I enhancer comprises two domains.

1988 ◽  
Vol 8 (2) ◽  
pp. 893-902 ◽  
Author(s):  
F Kruse ◽  
C T Komro ◽  
C H Michnoff ◽  
R J MacDonald
Keyword(s):  
Base Pair ◽  
Regulatory Region ◽  
Acinar Cells ◽  
Chloramphenicol Acetyltransferase ◽  
Pancreatic Tissue ◽  
Pancreatic Acinar Cells ◽  
Specific Expression ◽  
Nuclear Extracts ◽  
Dna Strands ◽  
Rat Elastase

Two separate domains within the 134-base-pair rat elastase I enhancer and a third domain at the enhancer-promoter boundary are required for selective expression in pancreatic acinar cells. The domains were detected by a series of 10-base-pair substitution mutations across the elastase I gene regulatory region from positions -200 to -61. The effect of each mutant on the pancreas-specific expression of a linked chloramphenicol acetyltransferase gene was assayed by transfection into pancreatic 266-6 acinar cells and control NIH/3T3 cells. The two enhancer domains are nonredundant, because mutations in either eliminated (greater than 100-fold reduction) expression in 266-6 cells. DNase I protection studies of the elastase I enhancer-promoter region with partially purified nuclear extracts from pancreatic tissue and 266-6 cells revealed nine discrete protected regions (footprints) on both DNA strands. One of three footprints that lie within the two functional domains of the enhancer contained a sequence, conserved among several pancreas-specific genes, which when mutated decreased linked chloramphenicol acetyltransferase expression up to 170-fold in 266-6 cells. This footprint may represent a binding site for one or more pancreas-specific regulatory proteins.

scholarly journals Stable Golgi-Mitochondria Complexes and Formation of Golgi Ca2+Gradients in Pancreatic Acinar Cells

2005 ◽  
Vol 280 (16) ◽  
pp. 15794-15799 ◽  
Author(s):  
Nick J. Dolman ◽  
Julia V. Gerasimenko ◽  
Oleg V. Gerasimenko ◽  
Svetlana G. Voronina ◽  
Ole H. Petersen ◽  
...  
Keyword(s):  
Acinar Cells ◽  
Pancreatic Tissue ◽  
Outer Edge ◽  
Pancreatic Acinar Cells ◽  
Apical Region ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Strategic Position ◽  
Over Time ◽  
Close Contacts

We have determined the localization of the Golgi with respect to other organelles in living pancreatic acinar cells and the importance of this localization to the establishment of Ca2+gradients over the Golgi. Using confocal microscopy and the Golgi-specific fluorescent probe 6-((N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoyl)sphingosine, we found Golgi structures localizing to the outer edge of the secretory granular region of individual acinar cells. We also assessed Golgi positioning in acinar cells located within intact pancreatic tissue using two-photon microscopy and found a similar localization. The mitochondria segregate the Golgi from lateral regions of the plasma membrane, the nucleus, and the basal part of the cytoplasm. The Golgi is therefore placed between the principal Ca2+release sites in the apical region of the cell and the important Ca2+sink formed by the peri-granular mitochondria. During acetylcholine-induced cytosolic Ca2+signals in the apical region, large Ca2+gradients form over the Golgi (decreasing from trans- to cis-Golgi). We further describe a novel, close interaction of the peri-granular mitochondria and the Golgi apparatus. The mitochondria and the Golgi structures form very close contacts, and these contacts remain stable over time. When the cell is forced to swell, the Golgi and mitochondria remain juxtaposed up to the point of cell lysis. The strategic position of the Golgi (closer to release sites than the bulk of the mitochondrial belt) makes this organelle receptive to local apical Ca2+transients. In addition the Golgi is ideally placed to be preferentially supplied by ATP from adjacent mitochondria.

Expression of cell volume-regulated kinase h-sgk in pancreatic tissue

2000 ◽  
Vol 279 (5) ◽  
pp. G998-G1002 ◽  
Author(s):  
K. Klingel ◽  
S. Wärntges ◽  
J. Bock ◽  
C. A. Wagner ◽  
M. Sauter ◽  
...  
Keyword(s):  
Cell Volume ◽  
Phorbol Esters ◽  
Acinar Cells ◽  
Northern Blotting ◽  
Pancreatic Tissue ◽  
Normal Function ◽  
Pancreatic Acinar Cells ◽  
Mrna Levels ◽  
Serine Threonine Kinase ◽  
Intracellular Ca

Transcript levels of the human serine/threonine kinase h-sgk have been found to be highest in pancreas. In the present study, localization and regulation of h-sgk transcription in pancreatic tissue were elucidated. As was apparent from radioactive in situ hybridization, most pancreatic acinar cells expressed high levels of h-sgk mRNA. h-sgk mRNA-positive cells were also found in ductal epithelia but not in pancreatic islets. In biopsy specimens from patients with pancreatitis, h-sgk mRNA levels were decreased in acinar cells but abundant in numerous mononuclear interstitial cells within areas of pancreatic necrosis and fibrosis. As shown by Northern blotting, h-sgk transcription in DAN-G pancreatic tumor cells is upregulated by osmotic cell shrinkage, serum, phorbol esters (phorbol 12,13-didecanoate), and Ca2+ ionophore A-23187 and decreased by staurosporine and cAMP. In conclusion, h-sgk transcription is regulated not only by cell volume but also by serum, protein kinase C stimulation, cAMP, and increase of intracellular Ca2+activity. The kinase may participate not only in normal function of exocrine pancreas but also in fibrosing pancreatitis.

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