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1.
Nat Genet ; 53(4): 455-466, 2021 04.
Article in English | MEDLINE | ID: mdl-33795864

ABSTRACT

Single-nucleus assay for transposase-accessible chromatin using sequencing (snATAC-seq) creates new opportunities to dissect cell type-specific mechanisms of complex diseases. Since pancreatic islets are central to type 2 diabetes (T2D), we profiled 15,298 islet cells by using combinatorial barcoding snATAC-seq and identified 12 clusters, including multiple alpha, beta and delta cell states. We cataloged 228,873 accessible chromatin sites and identified transcription factors underlying lineage- and state-specific regulation. We observed state-specific enrichment of fasting glucose and T2D genome-wide association studies for beta cells and enrichment for other endocrine cell types. At T2D signals localized to islet-accessible chromatin, we prioritized variants with predicted regulatory function and co-accessibility with target genes. A causal T2D variant rs231361 at the KCNQ1 locus had predicted effects on a beta cell enhancer co-accessible with INS and genome editing in embryonic stem cell-derived beta cells affected INS levels. Together our findings demonstrate the power of single-cell epigenomics for interpreting complex disease genetics.


Subject(s)
Chromatin/chemistry , Diabetes Mellitus, Type 2/genetics , Glucagon-Secreting Cells/metabolism , Insulin-Secreting Cells/metabolism , KCNQ1 Potassium Channel/genetics , Pancreatic Polypeptide-Secreting Cells/metabolism , Somatostatin-Secreting Cells/metabolism , Blood Glucose/metabolism , Cell Differentiation , Chromatin/metabolism , Diabetes Mellitus, Type 2/metabolism , Diabetes Mellitus, Type 2/pathology , Epigenomics , Fasting , Gene Expression Profiling , Genome-Wide Association Study , Glucagon-Secreting Cells/pathology , High-Throughput Nucleotide Sequencing , Human Embryonic Stem Cells/cytology , Humans , Insulin-Secreting Cells/pathology , KCNQ1 Potassium Channel/metabolism , Multigene Family , Pancreatic Polypeptide-Secreting Cells/pathology , Polymorphism, Genetic , Single-Cell Analysis , Somatostatin-Secreting Cells/pathology , Transcription Factors/classification , Transcription Factors/genetics , Transcription Factors/metabolism
2.
Diabetologia ; 63(10): 1966-1973, 2020 10.
Article in English | MEDLINE | ID: mdl-32894306

ABSTRACT

For much of the last century, our knowledge regarding the pancreas in type 1 and type 2 diabetes was largely derived from autopsy studies of individuals with these disorders or investigations utilising rodent models of either disease. While many important insights emanated from these efforts, the mode for investigation has increasingly seen change due to the availability of transplant-quality organ-donor tissues, improvements in pancreatic imaging, advances in metabolic assessments of living patients, genetic analyses, technological advances for laboratory investigation and more. As a result, many long-standing notions regarding the role for and the changes that occur in the pancreas in individuals with these disorders have come under question, while, at the same time, new issues (e.g., beta cell persistence, disease heterogeneity, exocrine contributions) have arisen. In this article, we will consider the vital role of the pancreas in human health and physiology, including discussion of its anatomical features and dual (exocrine and endocrine) functions. Specifically, we convey changes that occur in the pancreas of those with either type 1 or type 2 diabetes, with careful attention to the facets that may contribute to the pathogenesis of either disorder. Finally, we discuss the emerging unknowns with the belief that understanding the role of the pancreas in type 1 and type 2 diabetes will lead to improvements in disease diagnosis, understanding of disease heterogeneity and optimisation of treatments at a personalised level. Graphical abstract.


Subject(s)
Diabetes Mellitus, Type 1/metabolism , Diabetes Mellitus, Type 2/metabolism , Pancreas/metabolism , Adipose Tissue/pathology , Amyloidosis/metabolism , Amyloidosis/pathology , Autoimmunity/immunology , Diabetes Mellitus, Type 1/immunology , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Humans , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Islet Amyloid Polypeptide/metabolism , Islets of Langerhans/blood supply , Islets of Langerhans/metabolism , Islets of Langerhans/pathology , Islets of Langerhans/physiopathology , Pancreas/pathology , Pancreas/physiopathology , Pancreas, Exocrine/metabolism , Pancreas, Exocrine/pathology , Pancreas, Exocrine/physiopathology , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology
3.
Front Immunol ; 10: 1442, 2019.
Article in English | MEDLINE | ID: mdl-31293592

ABSTRACT

The CD83 is a type I membrane protein and part of the immunoglobulin superfamily of receptors. CD83 is involved in the regulation of antigen presentation and dendritic cell dependent allogeneic T cell proliferation. A soluble form of CD83 inhibits dendritic cell maturation and function. Furthermore, CD83 is expressed on activated B cells, T cells, and in particular on regulatory T cells. Previous studies on murine CD83 demonstrated this molecule to be involved in several immune-regulatory processes, comprising that CD83 plays a key role in the development und function of different immune cells. In order to get further insights into the function of the human CD83 and to provide preclinical tools to guide the function of CD83/sCD83 for therapeutic purposes we generated Bacterial Artificial Chromosomes (BAC) transgenic mice. BACs are excellent tools for manipulating large DNA fragments and are utilized to engineer transgenic mice by pronuclear injection. Two different founders of BAC transgenic mice expressing human CD83 (BAC-hCD83tg mice) were generated and were examined for the hCD83 expression on different immune cells as well as both the in vitro and in vivo role of human CD83 (hCD83) in health and disease. Here, we found the hCD83 molecule to be present on activated DCs, B cells and subtypes of CD4+ T cells. CD8+ T cells, on the other hand, showed almost no hCD83 expression. To address the function of hCD83, we performed in vitro mixed lymphocyte reactions (MLR) as well as suppression assays and we used the in vivo model of experimental autoimmune encephalomyelitis (EAE) comparing wild-type and hCD83-BAC mice. Results herein showed a clearly diminished capacity of hCD83-BAC-derived T cells to proliferate accompanied by an enhanced activation and suppressive activity of hCD83-BAC-derived Tregs. Furthermore, hCD83-BAC mice were found to recover faster from EAE-associated symptoms than wild-type mice, encouraging the relevance also of the hCD83 as a key molecule for the regulatory phenotype of Tregs in vitro and in vivo.


Subject(s)
Antigens, CD/immunology , Encephalomyelitis, Autoimmune, Experimental/immunology , Immunoglobulins/immunology , Membrane Glycoproteins/immunology , T-Lymphocytes, Regulatory/immunology , Animals , Antigens, CD/genetics , B-Lymphocytes/immunology , B-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/pathology , Encephalomyelitis, Autoimmune, Experimental/genetics , Encephalomyelitis, Autoimmune, Experimental/pathology , Humans , Immunoglobulins/genetics , Membrane Glycoproteins/genetics , Mice , Mice, Transgenic , Somatostatin-Secreting Cells/immunology , Somatostatin-Secreting Cells/pathology , T-Lymphocytes, Regulatory/pathology , CD83 Antigen
4.
Diabetes Obes Metab ; 19 Suppl 1: 124-136, 2017 09.
Article in English | MEDLINE | ID: mdl-28880471

ABSTRACT

The progressive loss of pancreatic ß-cell mass that occurs in both type 1 and type 2 diabetes is a primary factor driving efforts to identify strategies for effectively increasing, enhancing or restoring ß-cell mass. While factors that seem to influence ß-cell proliferation in specific contexts have been described, reliable stimulation of human ß-cell proliferation has remained a challenge. Importantly, ß-cells exist in the context of a complex, integrated pancreatic islet microenvironment where they interact with other endocrine cells, vascular endothelial cells, extracellular matrix, neuronal projections and islet macrophages. This review highlights different components of the pancreatic microenvironment, and reviews what is known about how signaling that occurs between ß-cells and these other components influences ß-cell proliferation. Future efforts to further define the role of the pancreatic islet microenvironment on ß-cell proliferation may lead to the development of successful approaches to increase or restore ß-cell mass in diabetes.


Subject(s)
Cell Communication , Cell Proliferation , Cellular Microenvironment , Insulin-Secreting Cells/metabolism , Islets of Langerhans/cytology , Models, Biological , Animals , Apoptosis , Diabetes Mellitus, Type 1/metabolism , Diabetes Mellitus, Type 1/pathology , Diabetes Mellitus, Type 2/metabolism , Diabetes Mellitus, Type 2/pathology , Extracellular Matrix/immunology , Extracellular Matrix/metabolism , Extracellular Matrix/pathology , Glucagon-Secreting Cells/cytology , Glucagon-Secreting Cells/immunology , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Humans , Insulin-Secreting Cells/cytology , Insulin-Secreting Cells/immunology , Insulin-Secreting Cells/pathology , Islets of Langerhans/blood supply , Islets of Langerhans/innervation , Islets of Langerhans/pathology , Macrophages/cytology , Macrophages/immunology , Macrophages/metabolism , Macrophages/pathology , Pancreatic Polypeptide-Secreting Cells/cytology , Pancreatic Polypeptide-Secreting Cells/immunology , Pancreatic Polypeptide-Secreting Cells/metabolism , Pancreatic Polypeptide-Secreting Cells/pathology , Somatostatin-Secreting Cells/cytology , Somatostatin-Secreting Cells/immunology , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology , Species Specificity
5.
J Diabetes Res ; 2015: 385395, 2015.
Article in English | MEDLINE | ID: mdl-26236746

ABSTRACT

Reduced beta cell mass in pancreatic islets (PI) of Goto-Kakizaki (GK) rats is frequently observed in this diabetic model, but knowledge on delta cells is scarce. Aiming to compare delta cell physiology/pathology of GK to Wistar rats, we found that delta cell number increased over time as did somatostatin mRNA and delta cells distribution in PI is different in GK rats. Subtle changes in 6-week-old GK rats were found. With maturation and aging of GK rats, disturbed cytoarchitecture occurred with irregular beta cells accompanied by delta cell hyperplasia and loss of pancreatic polypeptide (PPY) positivity. Unlike the constant glucose-stimulation index for insulin PI release in Wistar rats, this index declined with GK age, whereas for somatostatin it increased with age. A decrease of GK rat PPY serum levels was found. GK rat body weight decreased with increasing hyperglycemia. Somatostatin analog octreotide completely blocked insulin secretion, impaired proliferation at low autocrine insulin, and decreased PPY secretion and mitochondrial DNA in INS-1E cells. In conclusion, in GK rats PI, significant local delta cell hyperplasia and suspected paracrine effect of somatostatin diminish beta cell viability and contribute to the deterioration of beta cell mass. Altered PPY-secreting cells distribution amends another component of GK PI's pathophysiology.


Subject(s)
Aging , Diabetes Mellitus, Type 2/pathology , Insulin Resistance , Somatostatin-Secreting Cells/pathology , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/metabolism , Gene Expression Regulation, Developmental , Hyperplasia , Immunohistochemistry , Insulin/metabolism , Insulin Antagonists/pharmacology , Insulin Secretion , Insulin-Secreting Cells/drug effects , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Octreotide/pharmacology , Pancreatic Polypeptide/antagonists & inhibitors , Pancreatic Polypeptide/genetics , Pancreatic Polypeptide/metabolism , RNA, Messenger/metabolism , Rats, Inbred Strains , Rats, Wistar , Somatostatin/antagonists & inhibitors , Somatostatin/genetics , Somatostatin/metabolism , Somatostatin-Secreting Cells/drug effects , Somatostatin-Secreting Cells/metabolism
6.
Diabetologia ; 58(8): 1814-26, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26049399

ABSTRACT

AIMS/HYPOTHESIS: The cellular composition of the islet of Langerhans is essential to ensure its physiological function. Morphophysiological islet abnormalities are present in type 2 diabetes but the relationship between fasting plasma glucose (FPG) and islet cell composition, particularly the role of delta cells, is unknown. We explored these questions in pancreases from baboons (Papio hamadryas) with FPG ranging from normal to type 2 diabetic values. METHODS: We measured the volumes of alpha, beta and delta cells and amyloid in pancreatic islets of 40 baboons (Group 1 [G1]: FPG < 4.44 mmol/l [n = 10]; G2: FPG = 4.44-5.26 mmol/l [n = 9]; G3: FPG = 5.27-6.94 mmol/l [n = 9]; G4: FPG > 6.94 mmol/l [n = 12]) and correlated islet composition with metabolic and hormonal variables. We also performed confocal microscopy including TUNEL, caspase-3, and anti-caspase cleavage product of cytokeratin 18 (M30) immunostaining, electron microscopy, and immuno-electron microscopy with anti-somatostatin antibodies in baboon pancreases. RESULTS: Amyloidosis preceded the decrease in beta cell volume. Alpha cell volume increased ∼ 50% in G3 and G4 (p < 0.05), while delta cell volume decreased in these groups by 31% and 39%, respectively (p < 0.05). In G4, glucagon levels were higher, while insulin and HOMA index of beta cell function were lower than in the other groups. Immunostaining of G4 pancreatic sections with TUNEL, caspase-3 and M30 showed apoptosis of beta and delta cells, which was also confirmed by immuno-electron microscopy with anti-somatostatin antibodies. CONCLUSIONS/INTERPRETATION: In diabetic baboons, changes in islet composition correlate with amyloid deposition, with increased alpha cell and decreased beta and delta cell volume and number due to apoptosis. These data argue for an important role of delta cells in type 2 diabetes.


Subject(s)
Cell Death , Diabetes Mellitus, Type 2/pathology , Insulin Resistance/physiology , Islets of Langerhans/pathology , Somatostatin-Secreting Cells/pathology , Animals , Blood Glucose/metabolism , Caspase 3/metabolism , Diabetes Mellitus, Type 2/metabolism , Disease Progression , Female , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Islets of Langerhans/metabolism , Male , Papio hamadryas , Somatostatin-Secreting Cells/metabolism
7.
Endocrine ; 49(3): 693-702, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25605478

ABSTRACT

Previous studies describing the symptomatic onset of type 1 diabetes (T1D) and rate of beta-cell loss (C-peptide) support the notion that childhood onset T1D exhibits more severe beta-cell depletion compared to adult onset T1D. To test this notion, we performed whole pancreas analyses in two T1D cases, one of childhood onset (7-year old, onset at 1.5-year) along with an adult onset case (43-year old with onset at 27-year). Both cases were matched for age and gender with control subjects. Striking regional differences in beta-cell loss were observed in both T1D cases, with severity of loss in the order of tail > body > head regions. In contrast, pancreatic alpha- and delta-cell mass was similar in controls and T1D patients. In the childhood onset T1D case, no intra-islet beta-cells were detected while in the adult onset case, beta-cell containing islets were found, exclusively in the head region. In the latter case, considerable numbers of small cellular clusters negative for three major endocrine hormones were observed, in islets with or without beta-cells. Ultrastructural analysis suggests these cells correspond to degenerating beta-cells, with empty granular membranes and abnormal morphology of nuclei with intranuclear pseudo-inclusions, adjacent to healthy alpha- and delta-cells. These results support a hypothesis that during T1D development in childhood, beta-cells are more susceptible to autoimmune destruction or immune attack is more severe, while beta-cell death in the adult onset T1D may be more protracted and incomplete. In addition, T1D may be associated with the formation of "empty" beta-cells, an interesting population of cells that may represent a key facet to the disorder's pathogenesis.


Subject(s)
Diabetes Mellitus, Type 1/pathology , Insulin-Secreting Cells/pathology , Adult , Age of Onset , Child , Female , Glucagon-Secreting Cells/pathology , Glucagon-Secreting Cells/ultrastructure , Humans , Immunohistochemistry , Infant , Insulin-Secreting Cells/ultrastructure , Male , Pancreas/pathology , Pancreatic Function Tests , Somatostatin-Secreting Cells/pathology , Somatostatin-Secreting Cells/ultrastructure
8.
World J Gastroenterol ; 20(9): 2383-91, 2014 Mar 07.
Article in English | MEDLINE | ID: mdl-24605036

ABSTRACT

AIM: To study the ileal endocrine cell types in irritable bowel syndrome (IBS) patients. METHODS: Ninety-eight patients with IBS (77 females and 21 males; mean age 35 years, range 18-66 years) were included, of which 35 patients had diarrhea (IBS-D), 31 patients had a mixture of both diarrhea and constipation (IBS-M), and 32 patients had constipation (IBS-C) as the predominant symptoms. The controls were 38 subjects (26 females and 12 males; mean age 40 years, range 18-65 years) who had submitted to colonoscopy for the following reasons: gastrointestinal bleeding, where the source of bleeding was identified as hemorrhoids (n = 24) or angiodysplasia (n = 3), and health worries resulting from a relative being diagnosed with colon carcinoma (n = 11). The patients were asked to complete the: Birmingham IBS symptom questionnaire. Ileal biopsy specimens from all subjects were immunostained using the avidin-biotin-complex method for serotonin, peptide YY (PYY), pancreatic polypeptide (PP), enteroglucagon, and somatostatin cells. The cell densities were quantified by computerized image analysis, using Olympus cellSens imaging software. RESULTS: The gender and age distributions did not differ significantly between the patients and the controls (P = 0.27 and P = 0.18, respectively). The total score of Birmingham IBS symptom questionnaire was 21 ± 0.8, and the three underlying dimensions: pain, diarrhea, and constipation were 7.2 ± 0.4, 6.6 ± 0.4, and 7.2 ± 0.4, respectively. The density of serotonin cells in the ileum was 40.6 ± 3.6 cells/mm² in the controls, and 11.5 ± 1.2, 10.7 ± 5.6, 10.0 ± 1.9, and 13.9 ± 1.4 cells/mm² in the all IBS patients (IBS-total), IBS-D, IBS-M, and IBS-C patients, respectively. The density in the controls differed significantly from those in the IBS-total, IBS-D, IBS-M, and IBS-C groups (P < 0.0001, P = 0.0001, P = 0.0001, and P < 0.0001, respectively). There was a significant inverse correlation between the serotonin cell density and the pain dimension of Birmingham IBS symptom questionnaire (r = -0.6, P = 0.0002). The density of PYY cells was 26.7 ± 1.6 cells/mm(2) in the controls, and 33.1 ± 1.4, 27.5 ± 1.4, 34.1 ± 2.5, and 41.7 ± 3.1 cells/mm² in the IBS-total, IBS-D, IBS-M, and IBS-C patients, respectively. This density differed significantly between patients with IBS-total and IBS-C and the controls (P = 0.03 and < 0.0001, respectively), but not between controls and, IBS-D, and IBS-M patients (P = 0.8, and P = 0.1, respectively). The density of PYY cells correlated significantly with the degree of constipation as recorded by the Birmingham IBS symptom questionnaire (r = 0.6, P = 0.0002). There were few PP-, enteroglucagon-, and somatostatin-immunoreactive cells in the biopsy material examined, which made it impossible to reliably quantify these cells. CONCLUSION: The decrease of ileal serotonin cells is associated with the visceral hypersensitivity seen in all IBS subtypes. The increased density of PYY cells in IBS-C might contribute to the constipation experienced by these patients.


Subject(s)
Endocrine Cells/pathology , Ileum/pathology , Irritable Bowel Syndrome/pathology , Adolescent , Adult , Aged , Biomarkers/analysis , Biopsy , Case-Control Studies , Colonoscopy , Constipation/etiology , Diarrhea/etiology , Endocrine Cells/chemistry , Female , Glucagon-Like Peptides/analysis , Humans , Hyperalgesia/etiology , Ileum/chemistry , Image Interpretation, Computer-Assisted , Immunohistochemistry , Irritable Bowel Syndrome/complications , Irritable Bowel Syndrome/metabolism , Male , Middle Aged , Pain Measurement , Pancreatic Polypeptide/analysis , Peptide YY/analysis , Serotonin/analysis , Somatostatin/analysis , Somatostatin-Secreting Cells/chemistry , Somatostatin-Secreting Cells/pathology , Surveys and Questionnaires , Visceral Pain/etiology , Young Adult
9.
PLoS One ; 8(8): e72213, 2013.
Article in English | MEDLINE | ID: mdl-23977255

ABSTRACT

Mutations in the human homolog of the Vhlh gene [encoding the von-Hippel Lindau (VHL) protein] lead to tumor development. In mice, depletion of Vhlh in pancreatic ß-cells causes perturbed glucose homeostasis, but the role of this gene in other pancreatic cells is poorly understood. To investigate the function of VHL/HIF pathway in pancreatic cells, we inactivated Vhlh in the pancreatic epithelium as well as in the endocrine and exocrine lineages. Our results show that embryonic depletion of Vhlh within the pancreatic epithelium causes postnatal lethality due to severe hypoglycemia. The hypoglycemia is recapitulated in mice with endocrine-specific removal of Vhlh, while animals with loss of Vhlh predominantly in the exocrine compartment survive to adulthood with no overt defects in glucose metabolism. Mice with hypoglycemia display diminished insulin release in response to elevated glucose. Significantly, the glucagon response is impaired both in vivo (circulating glucagon levels) as well as in an in vitro secretion assay in isolated islets. Hypoxia also impairs glucagon secretion in a glucagon-expressing cell line in culture. Our results reveal a novel role for the hypoxia/HIF pathway in islet hormone secretion and maintenance of the fine balance that allows for the establishment of normoglycemia.


Subject(s)
Endocrine System/metabolism , Glucagon-Secreting Cells/metabolism , Hypoxia-Inducible Factor 1/genetics , Hypoxia/genetics , Somatostatin-Secreting Cells/metabolism , Von Hippel-Lindau Tumor Suppressor Protein/genetics , Animals , Cells, Cultured , Embryo, Mammalian , Endocrine System/pathology , Gene Deletion , Gene Expression Regulation, Developmental , Glucagon/metabolism , Glucagon-Secreting Cells/pathology , Glucose/metabolism , Homeostasis/genetics , Hypoxia/metabolism , Hypoxia/pathology , Hypoxia-Inducible Factor 1/metabolism , Insulin/metabolism , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Mice , Mice, Knockout , Pancreatic Polypeptide-Secreting Cells/metabolism , Pancreatic Polypeptide-Secreting Cells/pathology , Signal Transduction , Somatostatin-Secreting Cells/pathology , Von Hippel-Lindau Tumor Suppressor Protein/metabolism
10.
Endocrinology ; 154(10): 3796-806, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23913443

ABSTRACT

Selenoproteins are involved in the regulation of redox status, which affects several cellular processes, including cell survival and homeostasis. Considerable interest has arisen recently concerning the role of selenoproteins in the regulation of glucose metabolism. Here, we found that selenoprotein T (SelT), a new thioredoxin-like protein of the endoplasmic reticulum, is present at high levels in human and mouse pancreas as revealed by immunofluorescence and quantitative PCR. Confocal immunohistochemistry studies revealed that SelT is mostly confined to insulin- and somatostatin-producing cells in mouse and human islets. To elucidate the role of SelT in ß-cells, we generated, using a Cre-Lox strategy, a conditional pancreatic ß-cell SelT-knockout C57BL/6J mice (SelT-insKO) in which SelT gene disruption is under the control of the rat insulin promoter Cre gene. Glucose administration revealed that male SelT-insKO mice display impaired glucose tolerance. Although insulin sensitivity was not modified in the mutant mice, the ratio of glucose to insulin was significantly higher in the SelT-insKO mice compared with wild-type littermates, pointing to a deficit in insulin production/secretion in mutant mice. In addition, morphometric analysis showed that islets from SelT-insKO mice were smaller and that their number was significantly increased compared with islets from their wild-type littermates. Finally, we found that SelT is up-regulated by pituitary adenylate cyclase-activating polypeptide (PACAP) in ß-pancreatic cells and that SelT could act by facilitating a feed-forward mechanism to potentiate insulin secretion induced by the neuropeptide. Our findings are the first to show that the PACAP-regulated SelT is localized in pancreatic ß- and δ-cells and is involved in the control of glucose homeostasis.


Subject(s)
Gene Expression Regulation , Glucose Intolerance/metabolism , Insulin-Secreting Cells/metabolism , Pituitary Adenylate Cyclase-Activating Polypeptide/metabolism , Selenoproteins/metabolism , Animals , Blood Glucose , Cell Line , Crosses, Genetic , Gene Silencing , Glucose Intolerance/pathology , Humans , Insulin/metabolism , Insulin Secretion , Insulin-Secreting Cells/pathology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , Selenoproteins/antagonists & inhibitors , Selenoproteins/genetics , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology , Tissue Culture Techniques
11.
J Endocrinol ; 216(1): 13-20, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23092878

ABSTRACT

Type 2 diabetes is characterized by impaired insulin secretion from pancreatic ß-cells. Quantification of the islet area in addition to the insulin-positive area is important for detailed understanding of pancreatic islet histopathology. Here we show computerized automatic recognition of the islets of Langerhans as a novel high-throughput method to quantify islet histopathology. We utilized state-of-the-art tissue pattern recognition software to enable automatic recognition of islets, eliminating the need to laboriously trace islet borders by hand. After training by a histologist, the software successfully recognized even irregularly shaped islets with depleted insulin immunostaining, which were quite difficult to automatically recognize. The results from automated image analysis were highly correlated with those from manual image analysis. To establish whether this automated, rapid, and objective determination of islet area will facilitate studies of islet histopathology, we showed the beneficial effect of chronic exendin-4, a glucagon-like peptide-1 analog, treatment on islet histopathology in Zucker diabetic fatty (ZDF) rats. Automated image analysis provided qualitative and quantitative evidence that exendin-4 treatment ameliorated the loss of pancreatic insulin content and gave rise to islet hypertrophy. We also showed that glucagon-positive α-cell area was decreased significantly in ZDF rat islets with disorganized structure. This study is the first to demonstrate the utility of automatic quantification of digital images to study pancreatic islet histopathology. The proposed method will facilitate evaluations in preclinical drug efficacy studies as well as elucidation of the pathophysiology of diabetes.


Subject(s)
Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/pathology , Hypoglycemic Agents/therapeutic use , Islets of Langerhans/drug effects , Islets of Langerhans/pathology , Pattern Recognition, Automated , Peptides/therapeutic use , Venoms/therapeutic use , Animals , Artificial Intelligence , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/metabolism , Exenatide , Expert Systems , Glucagon/metabolism , Glucagon-Like Peptide 1/analogs & derivatives , Glucagon-Secreting Cells/drug effects , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , High-Throughput Screening Assays , Insulin/blood , Insulin/metabolism , Insulin-Secreting Cells/drug effects , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Islets of Langerhans/metabolism , Male , Obesity/complications , Rats , Rats, Zucker , Somatostatin/metabolism , Somatostatin-Secreting Cells/drug effects , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology
12.
Endocr Pract ; 19(2): 301-12, 2013.
Article in English | MEDLINE | ID: mdl-23186955

ABSTRACT

OBJECTIVE: To elucidate why diabetes is so difficult to treat despite the present tools and pharmacologic armamentarium and to provide insights into emerging therapies by describing human and rodent data that demonstrates the ability to transform progenitor cells within the adult pancreas into new islets. METHODS: A literature review focused on the distinctions between human and rodent islets. RESULTS: We are beginning to elucidate important differences between the architecture and composition of the islets of Langerhans in humans and rodents. In contrast to rodent islets, human islets are more heterogeneous in cellular composition and have more prominent intra-islet vascularity, with smooth muscle-containing blood vessels that are not present in rodent islets. Some studies report that more than 70% of human beta cells have direct physical contact with other cell types, whereas others describe that smaller human islets possess features more typical of rodents, while larger islets exhibit greater vascularity and a cellular distribution distinct from centrally clustered beta cells surrounded by a mantle of alpha and delta cells found in rodents. CONCLUSIONS: The differences between the islets of mice and men may influence why treatments hailed as reversing diabetes among rodents have not been successfully translated into humans. Increased understanding of the complexities within the human islet may yield unique insights into reversing diabetes in humans.


Subject(s)
Diabetes Mellitus, Type 1/pathology , Diabetes Mellitus, Type 1/therapy , Diabetes Mellitus, Type 2/pathology , Diabetes Mellitus, Type 2/therapy , Disease Models, Animal , Islets of Langerhans/pathology , Animals , Diabetes Mellitus, Type 1/drug therapy , Diabetes Mellitus, Type 1/metabolism , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , Glucagon-Secreting Cells/drug effects , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Humans , Hypoglycemic Agents/therapeutic use , Insulin-Secreting Cells/drug effects , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Islets of Langerhans/blood supply , Islets of Langerhans/metabolism , Mice , Somatostatin-Secreting Cells/drug effects , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology , Species Specificity , Stem Cells/pathology
13.
Chirurg ; 82(7): 574-82, 2011 Jul.
Article in German | MEDLINE | ID: mdl-21691871

ABSTRACT

Neuroendocrine tumors (NETs) of the stomach are the most frequent among all neuroendocrine neoplasms in the digestive tract. The diagnosis and classification are complicated by the fact that these tumors have to be categorized not only by common staging and grading but also according to their pathophysiological background (types). The types differ in their biological behaviour (aggressiveness) which influences therapeutic concepts. This article explains and summarizes the etiology and classification of gastric NETs and offers a precise concept for diagnosis and treatment to improve clinical outcome.


Subject(s)
Gastroscopy , Neuroendocrine Tumors/surgery , Stomach Neoplasms/surgery , Enterochromaffin Cells/pathology , Enterochromaffin-like Cells/pathology , Gastric Mucosa/pathology , Gastrin-Secreting Cells/pathology , Humans , Neoplasm Grading , Neoplasm Invasiveness , Neoplasm Staging , Neuroendocrine Tumors/classification , Neuroendocrine Tumors/diagnosis , Neuroendocrine Tumors/pathology , Prognosis , Somatostatin-Secreting Cells/pathology , Stomach/pathology , Stomach Neoplasms/classification , Stomach Neoplasms/diagnosis , Stomach Neoplasms/pathology
14.
Islets ; 3(4): 166-74, 2011.
Article in English | MEDLINE | ID: mdl-21633185

ABSTRACT

AIMS/HYPOTHESIS: Islet amyloid polypeptide is originally isolated as the chief constituent of amyloid deposits in type 2 diabetic islets. Islet amyloid polypeptide hyposecretion was known in type 1 diabetics and this study aimed to detect possibly reduced islet amyloid polypeptide-positive cells in type 1 diabetic islets. RESULTS: Non-diabetic control islets showed about 60% of islet cells were insulin cells, and 60% of insulin cells were positive for IAPP. In type 1 diabetic islets, islets were generally smaller than control islets, consisting of weaker positive cells for insulin and islet amyloid polypeptide. Medium-sized islets still retained some insulin positive cells, whereas islet amyloid polypeptide positive cells were much less or even absent, but some insulin-negative cells were weakly islet amyloid polypeptide positive. An occasional extra-large islet, representing regenerating islets, consisting of more than 100 islet cells revealed less than 35% insulin and 20% islet amyloid polypeptide positive cells with relatively increased glucagon and somatostatin cells. Both normal and type 1 diabetic islets revealed scattered, densely insulin and islet amyloid polypeptide positive sickle-shaped cytoplasm without granular appearance, consistent with degenerating insulin cells. METHODS: Using commercially available rabbit anti-islet amyloid polypeptide antibody, immunostaning was performed on ten cases of type 1 diabetic pancreata and eight non-diabetic controls. Both control and type 1 diabetic pancreata were systematically immunostained for insulin, glucagon, somatostatin and islet amyloid polypeptide. CONCLUSION/INTERPRETATION: Control islets consisted of about 60% insulin cells, and about 34% of islet cells were amyloid polypeptide positive with scattered and densely positive for insulin and islet amyloid polypeptide without granular appearance, consistent with degenerating ß cells. All islets, including occasional extra-large islets from type 1 diabetics, showed less insulin cells and less islet amyloid polypeptide positive cells with twice increased glucagon and somatostatin cells of the control islets, but some insulin-negative cells were positive for islet amyloid polypeptide, suggesting the presence of islet amyloid polypeptide in degenerating and extra-large regenerating islets. Thus, this immunocytochemical staining revealed generally less islet amyloid positive cells in type 1 diabetic islets, corresponding to severe hyposecretion of islet amyloid polypeptide in type 1 diabetics.


Subject(s)
Diabetes Mellitus, Type 1/metabolism , Diabetes Mellitus, Type 1/pathology , Islet Amyloid Polypeptide/metabolism , Islets of Langerhans/metabolism , Islets of Langerhans/pathology , Adolescent , Adult , Age Factors , Aged , Cell Count , Cytoplasm/metabolism , Cytoplasm/pathology , Female , Glucagon/metabolism , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Humans , Insulin/metabolism , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Islets of Langerhans/physiology , Male , Middle Aged , Organ Size , Regeneration , Somatostatin/metabolism , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology
15.
Coll Antropol ; 35(1): 5-8, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21661347

ABSTRACT

The aim of our paper was to investigate the influence of the different morphological changes on gastric mucosa on somatostatin D-cell number in antral mucosa and serum Somatostatin. We analyzed according to Sydney classification to what extent the severity of gastritis affect the observed hormonal values. somatostatin D-cell number in antral mucosa and serum Somatostatin values were compared between three groups of patients; mild, moderate and severe chronic gastritis. The average number of somatostatin cell in biopsy sample of antrum mucosa was 30.41 +/- 35.38 (N = 17) in the case of middle form, 18.69 +/- 26.65 (N = 56) in moderate and in severe case of chronic gastritis 5.23 +/- 5.93 (N = 7) cells in mm2 of mucosa. The level of somatostatin in the serum of middle form gastritis were 26.43 +/- 28.76, moderate 19.95 +/- 35.93 and severe 17.88 +/- 17.66 pg/mL. In order to determine the number of somatostatin cells in antrum mucosa and serum somatostatin with present morphological changes of mucosa, it might helpful to exclude the patients with non-ulcer dyspepsia, but with the higher risk of premalignant and malignant changes.


Subject(s)
Gastric Mucosa/cytology , Gastritis/pathology , Somatostatin-Secreting Cells/cytology , Somatostatin/blood , Cohort Studies , Gastric Mucosa/pathology , Gastritis/blood , Humans , Somatostatin-Secreting Cells/pathology
16.
Gastroenterology ; 137(6): 2052-62, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19737569

ABSTRACT

BACKGROUND & AIMS: The winged helix transcription factors Foxa1 and Foxa2 are expressed in all epithelia of the gastrointestinal tract from its embryonic origin into adulthood. In vitro studies have shown that Foxa1/a2 can transactivate the promoters of Mucin 2 (Muc2), which is expressed in goblet cells, and of preproglucagon, which is expressed in enteroendocrine cells. These findings suggest Foxa1/a2 as critical factors in the differentiation of gut epithelial cells. METHODS: Mice with intestine-specific simultaneous deletion of Foxa1 and Foxa2 were derived using the Cre-loxP system and analyzed using histologic and molecular means. RESULTS: Both Foxa1 and Foxa2 were deleted successfully in the epithelia of the small intestine and colon using Villin-Cre mice. Immunohistochemical staining showed that Foxa1/a2 mutants lack glucagon-like peptide-1- and peptide-2-expressing cells (L-cells), and have reduced numbers of somatostatin (D-cells) and peptide YY-expressing cells (L-cells). Preproglucagon, somatostatin, and peptide YY messenger RNA (mRNA) levels also were reduced significantly in Foxa1/a2 mutants. Thus, Foxa1 and Foxa2 are essential regulators of these enteroendocrine lineages in vivo. The mRNA levels of transcription factors Islet-1 and Pax6 were reduced significantly in the small intestine, showing that Foxa1 and Foxa2 impact on a transcription factor network in the enteroendocrine lineage. In addition, deletion of Foxa1/a2 caused a reduction in goblet cell number with altered expression of the secretory mucins Muc2, Mucin5b, Mucin5ac, and Mucin 6. CONCLUSIONS: The winged helix factors Foxa1 and Foxa2 are essential members of the transcription factor network that govern secretory cell differentiation in the mammalian gastrointestinal tract.


Subject(s)
Cell Differentiation , Enteroendocrine Cells/metabolism , Goblet Cells/metabolism , Hepatocyte Nuclear Factor 3-alpha/metabolism , Hepatocyte Nuclear Factor 3-beta/metabolism , Intestine, Small/metabolism , Somatostatin-Secreting Cells/metabolism , Animals , Enteroendocrine Cells/pathology , Eye Proteins/metabolism , Glucagon-Like Peptide 1/metabolism , Glucagon-Like Peptide 2/metabolism , Goblet Cells/pathology , Hepatocyte Nuclear Factor 3-alpha/deficiency , Hepatocyte Nuclear Factor 3-alpha/genetics , Hepatocyte Nuclear Factor 3-beta/deficiency , Hepatocyte Nuclear Factor 3-beta/genetics , Homeodomain Proteins/metabolism , Immunohistochemistry , Intestine, Small/pathology , LIM-Homeodomain Proteins , Male , Mice , Mice, Knockout , Mucin 5AC/metabolism , Mucin-2/metabolism , Mucin-5B/metabolism , PAX6 Transcription Factor , Paired Box Transcription Factors/metabolism , Peptide YY/metabolism , Proglucagon/metabolism , RNA, Messenger/metabolism , Repressor Proteins/metabolism , Somatostatin/metabolism , Somatostatin-Secreting Cells/pathology , Transcription Factors
17.
Endocrinology ; 150(6): 2611-7, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19213832

ABSTRACT

Recent evidence supports the idea that insulin signaling through the insulin receptor substrate/phosphatidyl-inositol 3-kinase/Akt pathway is involved in the maintenance of beta-cell mass and function. We previously identified the insulin-response element binding protein-1 (IRE-BP1) as an effector of insulin-induced Akt signaling in the liver, and showed that the 50-kDa carboxyl fragment confers the transcriptional activity of this factor. In this investigation we found that IRE-BP1 is expressed in the alpha, beta, and delta-cells of the islets of Langerhans, and is localized to the cytoplasm in beta-cells in normal rats, but is reduced and redistributed to the islet cell nuclei in obese Zucker rats. To test whether IRE-BP1 modulates beta-cell function and insulin secretion, we used the rat insulin II promoter to drive expression of the carboxyl fragment in beta-cells. Transgenic expression of IRE-BP1 in FVB mice increases nuclear IRE-BP1 expression, and produces a phenotype similar to that of type 2 diabetes, with hyperinsulinemia, hyperglycemia, and increased body weight. IRE-BP1 increased islet type I IGF receptor expression, potentially contributing to the development of islet hypertrophy. Our findings suggest that increased gene transcription mediated through IRE-BP1 may contribute to beta-cell dysfunction in insulin resistance, and allow for the hypothesis that IRE-BP1 plays a role in the pathophysiology of type 2 diabetes.


Subject(s)
Diabetes Mellitus, Type 2/metabolism , Diabetes Mellitus, Type 2/physiopathology , Insulin-Secreting Cells/metabolism , Iron Regulatory Protein 1/genetics , Iron Regulatory Protein 1/metabolism , Animals , Cells, Cultured , Diabetes Mellitus, Type 2/pathology , Disease Models, Animal , Glucagon/metabolism , Glucagon-Secreting Cells/metabolism , Glucagon-Secreting Cells/pathology , Glucose/metabolism , Hyperinsulinism/metabolism , Hyperinsulinism/pathology , Hyperinsulinism/physiopathology , Insulin/metabolism , Insulin Resistance/physiology , Insulin-Secreting Cells/pathology , Mice , Mice, Transgenic , Obesity/metabolism , Obesity/pathology , Obesity/physiopathology , Phenotype , Rats , Rats, Zucker , Somatostatin-Secreting Cells/metabolism , Somatostatin-Secreting Cells/pathology
18.
Pancreas ; 37(1): e8-e19, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18580436

ABSTRACT

OBJECTIVES: We determined if newly weaned female nonobese diabetic (NOD) mice show greater diabetes sensitivity to dose-adjusted regimens of multiple low doses of streptozotocin (Stz) than nondiabetes-prone CD-1 mice. METHODS: Female NOD mice received 5 daily doses of Stz from day 21 (0, 5, 10, 15, 20, 30, and 40 mg/kg body weight) and CD-1 mice 20, 30, and 40 mg. RESULTS: : Streptozotocin, at the 15-, 20-, 30-, and 40-mg dose, induced rapid diabetes in NOD mice. By day 100, 90% to 95% of NOD mice became diabetic after the 40- and 30-mg dose and 33% to 40% with the 15- and 20-mg dose. In comparison, only about 50% and 33% of CD-1 mice developed diabetes with the 40- and 30-mg dose, respectively, and 5.5% with the 20-mg dose. In NOD mice, the 20-mg dose also partially suppressed spontaneous diabetes. All diabetic mice displayed insulitis, variable immunostaining for insulin, and redistribution of glucagon and somatostatin cells. Glucose transporter-2 was markedly attenuated in selective beta cells. CONCLUSIONS: Newly weaned female NOD mice show heightened early sensitivity to low doses of Stz than CD-1 mice. At diabetes, several beta cells remain and show variable immunostaining for insulin and an attenuated expression for glucose transporter-2. Specific low doses of Stz may also suppress spontaneous diabetes.


Subject(s)
Diabetes Mellitus, Experimental/pathology , Insulin-Secreting Cells/pathology , Islets of Langerhans/pathology , Age Factors , Animals , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/immunology , Diabetes Mellitus, Experimental/metabolism , Dose-Response Relationship, Drug , Female , Glucagon/metabolism , Glucagon-Secreting Cells/pathology , Glucose Transporter Type 2/metabolism , Immunohistochemistry , Insulin/metabolism , Insulin-Secreting Cells/metabolism , Islets of Langerhans/immunology , Islets of Langerhans/metabolism , Macrophages/immunology , Mice , Mice, Inbred NOD , Severity of Illness Index , Somatostatin/metabolism , Somatostatin-Secreting Cells/pathology , Species Specificity , Streptozocin/administration & dosage , Weaning
19.
Chin J Integr Med ; 14(2): 111-6, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18219454

ABSTRACT

OBJECTIVE: To study the regulative action of mica monomer powder preparation on the chief and parietal cells as well as G and D cells in the gastric mucosa of the experimental atrophic gastritis (CAG) rats. METHODS: Intervention therapy was given to the experimental CAG rats at three different doses of mica monomer powder preparation to evaluate the changes of chief and parietal cells as well as G and D cells in the gastric mucosa and the histopathological changes of gastric mucosa. RESULTS: Mica monomer powder preparation at three different doses could increase the amount of chief and parietal cells as well as G and D cells in gastric mucosa of the experimental CAG rats and alleviate and control the inflammation of gastric mucosa and the atrophy of gastric mucosa glands. Especially, better effects were shown in the mid and high dose groups. CONCLUSION: Mica has the pharmacological action of protecting the gastric mucosa, enhancing blood flow of the gastric mucosa, and consequently improving the inflammatory responses of the gastric mucosa. One of the mechanisms is associated with promoting the secretion of gastric acid and gastric pepsin and regulating the neuroendocrine mechanism including gut hormone secretion (gastrin and somatostatin) by increasing the number of chief and parietal cells as well as G and D cells.


Subject(s)
Aluminum Silicates/pharmacology , Gastric Mucosa/drug effects , Gastric Mucosa/pathology , Gastritis, Atrophic/pathology , Animals , Cell Count , Chief Cells, Gastric/drug effects , Chief Cells, Gastric/pathology , Chronic Disease , Gastrin-Secreting Cells/drug effects , Gastrin-Secreting Cells/pathology , Inflammation , Parietal Cells, Gastric/drug effects , Parietal Cells, Gastric/pathology , Powders , Rats , Rats, Sprague-Dawley , Somatostatin-Secreting Cells/drug effects , Somatostatin-Secreting Cells/pathology
20.
Int J Clin Pract ; 62(7): 1044-9, 2008 Jul.
Article in English | MEDLINE | ID: mdl-17573842

ABSTRACT

BACKGROUND: Elevated gastrin concentration leading to gastritis is explained as the effect of change in the density of D and G cells. The aim of the study was to determine and compare fasting serum gastrin concentrations, G and D cell densities in gastric antrum mucosa in children with chronic gastritis and in children with no gastritis or Helicobacter pylori infection. MATERIAL AND METHODS: A total of 184 patients aged 6-18 years, with chronic abdominal pain underwent endoscopic examination. We created three groups: I--patients with chronic gastritis and H. pylori infection; II--patients with chronic gastritis but no H. pylori infection; III--patients with neither gastric mucosal abnormalities nor H. pylori infection. G and D cell densities were determined in the biopsy specimens (using Rbalpha H Gastrin & Somatostatin antibodies). Fasting serum gastrin concentrations were measured using a Beckmann gamma-counter and a GASK-PR kit. RESULTS: The mean serum gastrin concentration in group I was higher when compared with group II (p = 0.04) and group III (p = 0.019). No statistically significant differences were found between groups II and III (p = 0.91). There were no statistically significant differences in G and D cell densities between groups. CONCLUSION: The mean G/D cell ratios in groups I and III were almost identical. The mean fasting serum gastrin concentration was higher in children with both chronic gastritis and H. pylori infection compared with patients without infection or without antral inflammation. No difference in the G cell density or D cell density in children was found, regardless of the presence or absence of gastritis or H. pylori infection.


Subject(s)
Gastrins/blood , Gastritis/pathology , Pyloric Antrum/pathology , Adolescent , Cell Count , Child , Child, Preschool , Chronic Disease , Female , Gastrin-Secreting Cells/pathology , Gastritis/blood , Gastritis/microbiology , Helicobacter Infections/blood , Helicobacter Infections/pathology , Helicobacter pylori , Humans , Male , Somatostatin-Secreting Cells/pathology
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