Pancreatic Islet RNA Extraction v1

Author(s):  
Islet and Pancreas Analysis Core

This SOP defines the methods used by the Vanderbilt Diabetes Center Islet and Pancreas Analysis (IPA) Core for RNA extraction of pancreatic islets isolated from mouse or human tissue.

2021 ◽  
Author(s):  
Islet and Pancreas Analysis Core

This SOP defines the methods used by the Vanderbilt Diabetes Center Islet and Pancreas Analysis (IPA) Core for static incubation of pancreatic islets isolated from mouse or human tissue. See also our islet isolation protocol.


2021 ◽  
Author(s):  
Islet and Pancreas Analysis Core

This SOP defines the methods used by the Vanderbilt Diabetes Center Islet and Pancreas Analysis (IPA) Core for isolation of pancreatic islets from wild type and transgenic mice.


1997 ◽  
Vol 34 (5) ◽  
pp. 387-393 ◽  
Author(s):  
G. A. Andrews ◽  
N. C. Myers ◽  
C. Chard-Bergstrom

Twenty-two pancreatic islet cell tumors and normal pancreatic islets from ferrets were evaluated by immunohistochemistry for expression of the peptide hormones insulin, somatostatin, glucagon, and pancreatic polypeptide (PP) and the neuroendocrine markers chromogranin A (CgA) and neuron-specific enolase (NSE). In normal pancreatic islets, the majority of cells stained strongly with CgA and NSE. A cells, B cells, D cells, and PP cells stained strongly with glucagon, insulin, somatostatin, and PP, respectively. All 22 tumors stained with CgA and NSE. The proportion of cells within tumors staining for CgA was variable, but more than half of the cells stained positively in 18 of the tumors. The intensity of staining for CgA was strong (reactivity equivalent to or greater than normal islet cells in adjacent tissue) in 11 moderate in six, and weak in five of the tumors. All tumors stained for NSE, with ≥50% of the cells staining in 21 of the tumors, and the intensity of staining was strong in 18 of the tumors. Twenty of 22 tumors stained positively for insulin, with ≥50% of the cells staining in 19 of them. The intensity of staining for insulin was strong in 12, moderate in seven, and weak in one of the tumors. Approximately ≤1% of the cells in 15 of 22 tumors stained for somatostatin, five tumors stained for pancreatic polypeptide, and three tumors stained for glucagon. These data indicate that the majority of islet cell tumors of ferrets express immunohistochemically detectable insulin. CgA and NSE are both useful general markers for such tumors, including those that are insulin negative. Commercially available antisera to CgA, NSE, insulin, glucagon, somatostatin, and PP work well in formalin-fixed, paraffin-embedded tissue for immunophenotyping islet cell tumors in the ferret.


1988 ◽  
Vol 255 (2) ◽  
pp. E189-E200
Author(s):  
M. J. Achs ◽  
D. Garfinkel

Pancreatic islets detect glucose level by phosphorylating it and converting the glycolytic rate to a signal to secrete insulin. Insulin secretion is greater from the alpha- than from the beta-anomer when the D-glucose level is below 22 mM. D-mannose behaves similarly but at nearly twofold higher concentrations. Two explanations have been proposed: 1) glucokinase, which has the same anomeric preference, is the principal hexose phosphorylating enzyme and limits glycolytic rate. 2) Phosphofructokinase limits glycolysis and hexokinase is the principal enzyme phosphorylating hexose; hexosediphosphate activators of phosphofructokinase are more readily synthesized from alpha-anomers of hexose phosphates. We have simulated both alternatives with a detailed anomerically specific model of the hexose-metabolizing glycolytic enzymes. The pathway preference for alpha-anomer of both hexoses was adequately reproduced with anomerically active limiting glucokinase. The other mechanism did not reproduce the observed pathway preference.


2004 ◽  
Vol 10 (7) ◽  
pp. 965-977
Author(s):  
Kwang-Woong Lee ◽  
Suk Koo Lee ◽  
Jae-Won Joh ◽  
Sung-Joo Kim ◽  
Byung-Boong Lee ◽  
...  

1989 ◽  
Vol 9 (1) ◽  
pp. 55-61 ◽  
Author(s):  
Ramon Gomis ◽  
Angeles Casanovas ◽  
Willy J. Malaisse

Rat pancreatic islet homogenates catalyze the incorporation of [2,5−3-H]histamine into endogenous proteins recovered in both the stacking gel and a Mr 84000 protein separated by polyacrylamide electrophoresis. The labelling of these proteins represents a Ca2+-dependent process inhibited by glycine methylester, but not sarcosine methylester, and enhanced after preincubation of the islets at a high concentration of D-glucose. Although transglutaminase activity is found in both soluble and particlate subcelluler fractions, the endogenous transglutaminase substrates were located mainly in paarticulate, possibly membrane-associated, material.


2021 ◽  
Author(s):  
Joanne Boldison ◽  
Terri C. Thayer ◽  
Joanne Davies ◽  
F. Susan Wong

The non-obese diabetic (NOD) mouse develops spontaneous type 1 diabetes, with some features of disease that are very similar to the human disease. However, a proportion of NOD mice are naturally-protected from developing diabetes, and currently studies characterising this cohort are very limited. Here, using both immunofluorescence and multi-parameter flow cytometry we focus on the pancreatic islet morphology and immune infiltrate observed in naturally-protected NOD mice. We show that naturally-protected NOD mice are characterised by an increased frequency of insulin-containing, smaller sized, pancreatic islets. Although mice remain diabetes free, florid immune infiltrate remains. However, this immune infiltrate is skewed towards a regulatory phenotype in both T and B-cell compartments. Pancreatic islets have an increased frequency of IL-10 producing B cells and associated cell surface markers. Resident memory CD69<sup>+</sup>CD8<sup>+</sup> T cells show a significant shift towards reduced CD103 expression, while CD4<sup>+</sup> T cells have increased FoxP3<sup>+</sup>CTLA4<sup>+</sup> expression. These data indicate that naturally-protected NOD mice have a unique islet signature and provide new insight into regulatory mechanisms within pancreatic islets.<br>


Sains Medika ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 48
Author(s):  
Muhammad Naufal Widyatmaka ◽  
Akhmad Ismail

Introduction: Some studies indicated that antlions (Myrmeleon sp.) is traditionally used by many people in Indonesia as a diabetes treatmenter. However. reseach has not been done to find the effect of antlion extract in pancreas, an important organ in diabetes mellitus pathophysiology. Objective: To prove the effect of antlion extract towards histopathological feature of pancreas in diabetic mice.Methods: In the post test only control group design, 25 Swiss mice were divided into five groups. Normal group (Nor-G), Negative control group (Neg-G), and the other 3 groups were intraperitoneally injected with Streptozotocine 200 mg/kg to induce diabetic state. In Neg-G had no antlion treatment, whereas in At-14, At-28, and At-56 were treated with antlion extract at the dose of 14, 28, and 56 mg/kg BW respectively for 24 days. Pancreas then collected and prepared to be examined under microscope with 400x magnification. The score of pancreatic islet injury was determined by assessing structural damage and inflammatory cells infiltration. Kruskal-Wallis and Mann-Whitney were adopted as statistical analysis.Results: Mann-Whitney analysis indicated that the score of pancreatic islets injury in At-56 was significant lower compared to that of Neg-G, p<0.05. The score of pancreatic islets injury in At-56 also significant lower compared to that of At-14 and At-28, p<0.05. In contrary, when compared to that of Nor-G, the pancreatic islets injury in At-56 was not signicantly lower, p>0.05.Conclusion: Antlion extract treatment at the dose of 56 mg/kg BW was capable of reducing pancreatic islet injury in diabetic mice induced by streptozotocine.


2011 ◽  
Vol 300 (2) ◽  
pp. E410-E421 ◽  
Author(s):  
A. D. Dobrian ◽  
Q. Ma ◽  
J. W. Lindsay ◽  
K. A. Leone ◽  
K. Ma ◽  
...  

Adipose tissue inflammation and reduced pancreatic β-cell function are key issues in the development of cardiovascular disease and progressive metabolic dysfunction in type 2 diabetes mellitus. The aim of this study was to determine the effect of the DPP IV inhibitor sitagliptin on adipose tissue and pancreatic islet inflammation in a diet-induced obesity model. C57Bl/6J mice were placed on a high-fat (60% kcal fat) diet for 12 wk, with or without sitagliptin (4 g/kg) as a food admix. Sitagliptin significantly reduced fasting blood glucose by 21% as well as insulin by ∼25%. Sitagliptin treatment reduced body weight without changes in overall body mass index or in the epididymal and retroperitoneal fat mass. However, sitagliptin treatment led to triple the number of small adipocytes despite reducing the number of the very large adipocytes. Sitagliptin significantly reduced inflammation in the adipose tissue and pancreatic islet. Macrophage infiltration in adipose tissue evaluated by immunostaining for Mac2 was reduced by sitagliptin ( P < 0.01), as was the percentage of CD11b+/F4/80+ cells in the stromal vascular fraction ( P < 0.02). Sitagliptin also reduced adipocyte mRNA expression of inflammatory genes, including IL-6, TNFα, IL-12(p35), and IL-12(p40), 2.5- to fivefold as well as 12-lipoxygenase protein expression. Pancreatic islets were isolated from animals after treatments. Sitagliptin significantly reduced mRNA expression of the following inflammatory cytokines: MCP-1 (3.3-fold), IL-6 (2-fold), IL-12(p40) (2.2-fold), IL-12(p35) (5-fold, P < 0.01), and IP-10 (2-fold). Collectively, the results indicate that sitagliptin has anti-inflammatory effects in adipose tissue and in pancreatic islets that accompany the insulinotropic effect.


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