Induction of Sca-1 via activation of STAT3 system in the duct cells of the mouse submandibular gland by ligation of the main excretory duct

To examine the very initial step that takes place immediately after tissue injury and is linked to tissue regeneration, we employed the submandibular gland (SMG), which was injured by ligation of its main excretory duct (MED). Ligation of the MED of the SMG in mice induced the expression of Sca-1, a protein marker of hematopoietic stem cells. In the normal gland, a low level of Sca-1 was expressed, which was localized predominantly in the excretory duct cells. At 1 day after ligation, Sca-1 expression increased prominently in almost all of cells in the duct system, but not in the acinar cells. The level of Sca-1 mRNA had begun to increase at 6 h after ligation and continuously rose thereafter until it reached a plateau, which occurred ∼12 h after ligation. STAT3 phosphorylated at its tyrosine-705 (p-STAT3) in the ligated gland increased immediately after ligation, and it was localized in the nuclei of all duct cells. The results of an EMSA revealed the specific binding of a nuclear extract to the sequence of the γ-interferon activation site (GAS) present in the Sca-1 promoter and confirmed that such binding increased after ligation. Thus the present study suggests that STAT3, having been phosphorylated following MED ligation, was transferred to the nucleus, where it bound to the GAS element in the promoter of Sca-1 gene, resulting in promotion of Sca-1 gene expression. Actual prevention of STAT3 phosphorylation reduced the ligation-induced Sca-1 elevation.

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Biogenesis of Catalase-Positive Rods in Excretory Duct Cells of Salivary Glands

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100090294 ◽

1976 ◽

Vol 34 ◽

pp. 62-63

Author(s):

Dwight K. Romanovicz ◽

Jacob S. Hanker

Keyword(s):

Light Microscopy ◽

Salivary Glands ◽

Submandibular Gland ◽

Excretory Duct ◽

Exocrine Glands ◽

Duct Cells ◽

Peroxidatic Activity ◽

Microscopic Studies ◽

Different Dimensions

The presence of catalase-positive rods (Fig. 1) of different dimensions, which frequently have a crystalline appearance by light microscopy, has been reported. They seem to be related to peroxisomes which were characterized morphologically and cytochemically in parotid and other exocrine glands of the rat by Hand in 1973. Our light microscopic studies of these spherical microbodies and rods of different sizes, stained by virtue of the peroxidatic activity of their catalase, indicate that they are almost entirely confined to the cells of the striated and execretory ducts of the submandibular gland in the mouse. The rods were usually noted only in the proximity of the ductal microbodies. The latter frequently showed a tendency to appear in linear close array, or even to be contiguous (Fig. 2). This suggested that the rods could be formed by the fusion of microbodies.

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Immunolocalization of Vacuolar-type H+-ATPase in Rat Submandibular Gland and Adaptive Changes Induced by Acid–Base Disturbances

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215549804600112 ◽

1998 ◽

Vol 46 (1) ◽

pp. 91-100 ◽

Cited By ~ 31

Author(s):

Eleni Roussa ◽

Frank Thévenod ◽

Ivan Sabolic ◽

Carol M. Herak–Kramberger ◽

Wolfgang Nastainczyk ◽

...

Keyword(s):

Metabolic Acidosis ◽

Submandibular Gland ◽

Excretory Duct ◽

Acid Base ◽

Chronic Metabolic Acidosis ◽

Adaptive Changes ◽

Duct Cells ◽

Rat Submandibular Gland ◽

Diffuse Distribution ◽

Apical Localization

Using antibodies against the 31-kD and 70-kD subunits of vacuolar type H+-ATPase (V-ATPase) and light microscopic immunocytochemistry, we have demonstrated the presence of this V-ATPase in rat submandibular gland. We have also investigated the adaptive changes of this transporter during acid-base disturbances such as acute and chronic metabolic acidosis or alkalosis. Our results show intracellularly distributed V-ATPase in striated, granular, and main excretory duct cells in controls, but no V-ATPase immunoreaction in acinar cells. Both acute and chronic metabolic acidosis caused a shift in V-ATPase away from diffuse distribution towards apical localization in striated and granular duct cells, suggesting that a V-ATPase could be involved in the regulation of acid–base homeostasis. In contrast, during acidosis the main excretory duct cells showed no changes in the V-ATPase distribution compared to controls. With acute and chronic metabolic alkalosis, no changes in the V-ATPase distribution occurred.

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Induction of Sca-1 in the duct cells of the mouse submandibular gland by obstruction of the main excretory duct

Journal of Oral Pathology and Medicine ◽

10.1111/j.1600-0714.2011.01011.x ◽

2011 ◽

Vol 40 (8) ◽

pp. 651-658 ◽

Cited By ~ 6

Author(s):

Nunuk Purwanti ◽

Daisuke Tsuji ◽

Ahmad Azlina ◽

Mileva Ratko Karabasil ◽

Purevjav Javkhlan ◽

...

Keyword(s):

Submandibular Gland ◽

Excretory Duct ◽

Duct Cells

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Autoradiographic localization of dihydrotestosterone binding in the major salivary glands and other androgen-responsive organs of the mouse.

Journal of Histochemistry & Cytochemistry ◽

10.1177/35.10.3624850 ◽

1987 ◽

Vol 35 (10) ◽

pp. 1053-1058 ◽

Cited By ~ 28

Author(s):

J I Morrell ◽

E W Gresik ◽

T Barka

Keyword(s):

Growth Factor ◽

Salivary Glands ◽

Submandibular Gland ◽

Cellular Localization ◽

Incidental Finding ◽

Acinar Cells ◽

Unexpected Finding ◽

Cervical Lymph Nodes ◽

Major Salivary Glands ◽

Duct Cells

Mouse submandibular glands show an androgen-dependent sexual dimorphism, reflected in higher concentrations in males than in females of bioactive peptides, such as epidermal growth factor (EGF), nerve growth factor, and renin in the cells of the granular convoluted tubules (GCT). Biochemical studies have demonstrated androgen receptors in submandibular gland and other androgen-responsive organs in mouse. We have determined the cellular localization of these receptors using steroid autoradiography. Fifteen adult gonadectomized male mice were injected intravenously with 0.13 microgram or 0.26 microgram [3H]-dihydrotestosterone (SA 135 Ci/mM); some animals were pre-treated with cyclocytidine to stimulate secretion by GCT cells. Animals were killed 15 min, 1, 2, or 3 hr after isotope injection. Steroid autoradiographs were prepared, and some were stained immunocytochemically for EGF. Of the different cell types of submandibular gland, the acinar cells most frequently and intensely concentrated [3H]-DHT; GCT cells also concentrated the hormone, as did a small number of striated duct cells. In the other major salivary glands, the only cells that concentrated the androgen were interlobular striated duct cells in sublingual gland. In prostate, anterior pituitary, and brain a large number of cells concentrated androgen, as has been previously reported. Androgen binding by the GCT cells was a predictable finding, since androgen-induced alterations in composition and form of these cells are well documented. The intense androgen concentration by the acinar cells was an unexpected finding and suggests a hitherto unknown androgen regulation of these cells. An incidental finding was intense concentration of [3H]-DHT in the nuclei of the endothelial cells of the post-capillary venules of the cervical lymph nodes.

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Protective effect of Saccharomyces cerevisiae in Rattus norvegicus Ischemic Stroke Model

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.01006 ◽

2021 ◽

pp. 5785-5789

Author(s):

Dedy Budi Kurniawan ◽

Mokhamad Fahmi Rizki Syaban ◽

Arinal Mufidah ◽

Muhammad Unzila Rafsi Zulfikri ◽

Wibi Riawan

Keyword(s):

Saccharomyces Cerevisiae ◽

Ischemic Stroke ◽

Brain Tissue ◽

Tissue Injury ◽

Intervention Group ◽

Positive Control ◽

Positive Control Group ◽

Control Group ◽

Hematopoietic Stem ◽

The Brain

Stroke is one of the leading causes of morbidity and mortality in all ages. Ischemic stroke activates excitotoxic glutamate cascade leading to brain tissue injury. Saccharomyces cerevisiae is a unicellular yeast widely found in nature. S. cerevisiae is neuroprotective and able to increase the differentiation of hematopoietic stem cells (HSCs) into neuronal cells. it may increase levels of neuroprotectant BDNF in the brain tissue, therefore increase the protection of neurons. BDNF may prevent glutamate-driven excitotoxicity by reducing glutamate levels. This study uses a randomized post-test only controlled group design. In this in vivo study, rodent models of ischemic stroke were divided into five groups comprising of the negative control group, positive control group, intervention group 1 (18mg/kgBW), intervention group 2 (36mg/kgBW) and intervention group 3 (72 mg/kgBW). Groups treated with Saccharomyces cerevisiae extract showed significantly increased BDNF levels in the brain tissue, and the expression of the glutamate level was significantly reduced (P <0.05) compared to the positive control group. Thus Saccharomyces cerevisiae has a promising potential to become a therapy against ischemic stroke disease. however further research is needed regarding the efficacy and toxicity of Saccharomyces cerevisiae.

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Three-Dimensional Localization of DNA Synthesis in Secretory Elements of Adult Female Mouse Submandibular Gland

Advances in Dental Research ◽

10.1177/08959374900040010601 ◽

1990 ◽

Vol 4 (1) ◽

pp. 34-44 ◽

Cited By ~ 13

Author(s):

P.C. Denny ◽

Y. Chai ◽

D.K. Klauser ◽

P.A. Denny

Keyword(s):

Dna Synthesis ◽

Submandibular Gland ◽

Adult Female ◽

Female Mouse ◽

Three Dimensional ◽

Acinar Cells ◽

Single Pulse ◽

Duct System ◽

Duct Cells ◽

Structural Relationships

A system based in part on three-dimensional structural relationships is described for precisely characterizing the location of cells within secretory complexes of the adult female mouse submandibular gland. The pattern of DNA synthesis during a 90-minute pulse with 3H-thymidine was characterized based upon the above system. Seventy-eight percent of all radiolabeled nuclei were found in the intercalated duct system. One-half of these were in second-order intercalated ducts. DNA synthesis was also observed in acinar cells, granular intercalated duct cells, striated granular duct cells, and granular duct cells. Some secretory complexes contained multiple radiolabeled nuclei, with some of these nuclei in a side-by-side configuration. Approximately one-half of all secretory complexes contained radiolabeled nuclei. A second survey of the frequency of complexes containing radiolabeled nuclei was conducted following four pulses at eight-hour intervals over a 26-hour period. Only about 30% of all complexes contained radiolabeled nuclei. This reduction in the frequency of radiolabeled nuclei when compared with the single pulse suggests the possibility of individual variation. However, a more prolonged period of daily injections for nine days with 3H-thymidine resulted in all but one of the secretory complexes containing radiolabeled nuclei. This latter observation suggests that cell addition in adult submandibular glands is widespread.

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CREB coactivators CRTC2 and CRTC3 modulate bone marrow hematopoiesis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1712616114 ◽

2017 ◽

Vol 114 (44) ◽

pp. 11739-11744 ◽

Cited By ~ 5

Author(s):

Jeong-Ho Kim ◽

Susan Hedrick ◽

Wen-Wei Tsai ◽

Ezra Wiater ◽

John Le Lay ◽

...

Keyword(s):

Bone Marrow ◽

Stromal Cells ◽

Camp Response Element Binding ◽

Cytokine Signaling ◽

Camp Response Element ◽

Hematopoietic Stem ◽

Small Molecule Inhibition ◽

Camp Pathway ◽

Stat3 Phosphorylation ◽

Element Binding Protein

Populations of circulating immune cells are maintained in equilibrium through signals that enhance the retention or egress of hematopoietic stem cells (HSCs) from bone marrow (BM). Prostaglandin E2 (PGE2) stimulates HSC renewal and engraftment through, for example, induction of the cAMP pathway. Triggering of PGE2 receptors increases HSC survival in part via the PKA-mediated induction of the cAMP response element-binding protein (CREB) signaling pathway. PKA stimulates cellular gene expression by phosphorylating CREB at Ser133 and by promoting the dephosphorylation of the cAMP- responsive transcriptional coactivators (CRTCs). We show here that disruption of both CRTC2 and CRTC3 causes embryonic lethality, and that a single allele of either CRTC2 or CRTC3 is sufficient for viability. CRTC2 knockout mice that express one CRTC3 allele (CRTC2/3m mice) develop neutrophilia and splenomegaly in adulthood due to the up-regulation of granulocyte-colony stimulating factor (G-CSF); these effects are reversed following administration of neutralizing anti–G-CSF antiserum. Adoptive transfer of CRTC2/3m BM conferred the splenomegaly/neutrophilia phenotype in WT recipients. Targeted disruption of both CRTC2 and CRTC3 in stromal cells with a mesenchymal Prx1-Cre transgene also promoted this phenotype. Depletion of CRTC2/3 was found to decrease the expression of Suppressor of Cytokine Signaling 3 (SOCS3), leading to increases in STAT3 phosphorylation and to the induction of CEBPβ, a key regulator of the G-CSF gene. As small molecule inhibition of JAK activity disrupted CEBPβ induction and reduced G-CSF expression in CRTC2/3m stromal cells, our results demonstrate how cross-coupling between the CREB/CRTC and JAK/STAT pathways contributes to BM homeostasis.

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CD45-targeted antibody-drug-conjugate successfully conditions for allogeneic hematopoietic stem cell transplantation

Blood ◽

10.1182/blood.2021012366 ◽

2022 ◽

Author(s):

Asim Saha ◽

Sharon Hyzy ◽

Tahirih Laforest Lamothe ◽

Katelyn J. Hammond ◽

Nicholas M Clark ◽

...

Keyword(s):

Stem Cell ◽

Hematopoietic Stem Cell Transplantation ◽

Stem Cell Transplantation ◽

Cell Transplantation ◽

Hematopoietic Stem Cell ◽

Tissue Injury ◽

Single Agent ◽

Curative Treatment ◽

Hematopoietic Stem ◽

Antibody Drug

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a potentially curative treatment for patients with non-malignant or malignant blood disorders. Its success has been limited by graft-versus-host disease (GVHD). Current genotoxic conditioning regimens mediate tissue injury and potentially incite and amplify GVHD, limiting use of this potentially curative treatment beyond malignant disorders. Minimizing genotoxic conditioning while achieving alloengraftment without global immune suppression is highly desirable. Antibody-drug-conjugates (ADCs) targeting hematopoietic cells can specifically deplete host stem and immune cells and enable alloengraftment. Here we report an anti-mouse CD45-targeted-ADC (CD45-ADC) that facilitates stable murine multi-lineage donor cell engraftment. Conditioning with CD45-ADC (3mg/kg) was effective as a single agent in both congenic and minor-mismatch transplant models resulting in full donor chimerism comparable to lethal total body irradiation (TBI). In an MHC-disparate allo-HSCT model, pre-transplant CD45-ADC (3mg/kg) combined with low-dose TBI (150cGy) and a short course of costimulatory blockade with anti-CD40 ligand antibody enabled 89% of recipients to achieve stable alloengraftment (mean value: 72%). When CD45-ADC was combined with pre-transplant TBI (50cGy) and post-transplant Rapamycin, Cytoxan or a JAK inhibitor, 90-100% of recipients achieved stable chimerism (mean: 77%, 59%, 78%, respectively). At a higher dose (5mg/kg), CD45-ADC as a single agent was sufficient for rapid, high level multi-lineage chimerism sustained through the 22 weeks observation period. Therefore, CD45-ADC has potential utility to confer the benefit of fully myeloablative conditioning but with substantially reduced toxicity when given as a single agent or at lower doses in conjunction with reduced intensity conditioning.

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1438MicroRNA editing is integral for interleukin-6 trans-signalling and leukocyte trafficking to ischemic tissues

European Heart Journal ◽

10.1093/eurheartj/ehz748.0073 ◽

2019 ◽

Vol 40 (Supplement_1) ◽

Author(s):

A Gatsiou ◽

S Tual-Chalot ◽

F Bonini ◽

V Cesarini ◽

A Ortega-Gomez ◽

...

Keyword(s):

Rna Editing ◽

Interleukin 6 ◽

Inflammatory Responses ◽

Tissue Injury ◽

Vascular Endothelial Cell ◽

Luciferase Reporter ◽

Leukocyte Trafficking ◽

Maturation Process ◽

Ischemic Disease ◽

Stat3 Phosphorylation

Abstract Background/Aim Adenosine to inosine RNA editing is an essential post-transcriptional RNA modification catalysed by adenosine deaminase acting on RNA-1 and -2 (ADAR1; ADAR2). Endothelial cells (ECs) attract and guide leukocytes to sites of ischemic tissue injury. Here we studied the role of RNA editing in ischemic disease. Methods Primary human and murine vascular endothelial cell cultures were used to assess the EC responses to interleukin-6 (IL-6) or ischemia. For the animal studies, the effect of ADAR2 in acute and chronic ischemic disease was evaluated in cremaster muscle microcirculation by intravital microscopy, in peritoneal cavity after sterile peritonitis and in gastrocnemius muscle after hind-limb ischemia by 8-colour flow cytometry and immunohistochemistry (IHC) studies of Adar2−/−/tg as well as of i(nducible)EC-ADAR2 knockout (KO) mice. For the mechanistic studies, deep RNA sequencing, qRT-PCR, western blot, confocal microscopy, target-specific microRNA (miRNA) editing studies, RNA-immunoprecipitation, miRNA/plasmid silencing/overexpression and luciferase reporter assays were used among others. For human studies, ischemic tissues derived from patients with acute or chronic ischemic heart disease were processed. Results ADAR2, but not ADAR1, expression is induced by >2-fold in hypoxic ECs and in ischemic vascular ECs in mice and humans. Unbiased gene ontology analysis of the EC transcriptome indicated that ADAR2 controls inflammatory responses and predominantly the expression of interleukin-6-signal transducer (IL6ST), the co-receptor of IL-6. Subsequently, ADAR2 controls IL-6 trans-signalling in ECs as documented by the STAT3 phosphorylation and expression of the downstream leukocyte adhesion molecules, E-selectin and VCAM-1. IL-6-inflamed cremaster muscles showed that rolling and adhesion of leukocyte subsets to vascular wall were severely impaired in Adar2−/−/tg mice. Leukocyte transmigration was also diminished by >2-fold in Adar2−/−/tg and in iEC-ADAR2 KO mice in response to IL-6 or ischemia. Similar results were obtained for leukocyte rolling, adhesion and infiltration after acute (4h) and chronic (3d; 21d) ischemia from iEC-ADAR2 KO mice and human ischemic muscle tissues. Next we studied how ADAR2 controls IL6ST expression. ADAR2-deficient vascular EC miRNAome revealed the upregulation of a conserved group of miRNAs targeting the IL6ST mRNA including miR-199a-5p and miR-335-3p. At a single-nucleotide level, ADAR2-induced RNA editing of the stem loops of the primary miR-199a1/2 and miR-335 directly disrupted Drosha recruitment to both and thus inhibited their maturation process. Accordingly, rescue experiments using miRNA-inhibitors restored IL6ST levels after ADAR2 deficiency. Conclusion Taking together, inhibition of the microRNA maturation process by ADAR2-mediated RNA editing is integral for IL-6 trans-signalling in vascular endothelium and subsequent leukocyte trafficking to ischemic tissues in mice and humans.

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