Characterization of a Dmd EGFP reporter mouse as a tool to investigate dystrophin expression

AbstractHuLangerin-Cre-YFPf/f mice were generated to specifically mark a subset of antigen presenting immune cells, called Langerhans cells (LCs). During histological characterization of these mice, we found that, in addition to LCs an uncharacterized cell population in the central nervous system (CNS) also expressed YFP. In this study, we found that the CNS YFP+ cells were negative for microglia and astrocyte markers, but they expressed mature neuronal marker NeuN and showed neuronal localization/morphology. Thus, these mice might be used to study the ontogeny, migration and the role of a subset of CNS neurons.

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Abstract 081: Reporter Mouse Strain Provides a Novel Look at Angiotensin Type-2 Receptor Distribution in Central Nervous System Cardiovascular Control Centers

Hypertension ◽

10.1161/hyp.64.suppl_1.081 ◽

2014 ◽

Vol 64 (suppl_1) ◽

Author(s):

Annette D de Kloet ◽

Lei Wang ◽

Jacob A Ludin ◽

Helmut Hiller ◽

Justin A Smith ◽

...

Keyword(s):

Blood Pressure ◽

Fluid Balance ◽

Hypothalamic Nucleus ◽

Reporter Mouse ◽

Arterial Blood ◽

Egfp Reporter ◽

Angiotensin Type 2 Receptor ◽

The Brain ◽

Angiotensin Type

It is established that angiotensin-II acts at its type-1 receptor (AT1R) in the brain to increase sympathetic outflow and blood pressure, and modulate fluid balance. However, the role of the angiotensin type-2 receptor (AT2R) in the neural control of these processes has received far less attention, largely because of an inability to effectively localize these receptors at a cellular level in the brain. The present studies combine the use of a bacterial artificial chromosome transgenic AT2R-eGFP reporter mouse with recent advances in in situ hybridization (ISH) to circumvent this obstacle. Dual IHC/ ISH studies validated the AT2R-eGFP reporter mice by determining that eGFP and AT2R mRNA were highly co-localized within the nucleus of the solitary tract (NTS; 98.0 ± 0.18 %; 125 ± 3.6 of 127 ± 3.9 cells; n = 4). Analysis of eGFP immunoreactivity in the brain revealed localization to neurons within nuclei that regulate blood pressure and fluid balance (e.g., NTS and median preoptic nucleus [MnPO]). Additional IHC/ISH studies uncovered the phenotype of specific AT2R-eGFP cells. For example, within the NTS, AT2R-eGFP neurons primarily express glutamic acid decarboxylase-67 (GABAergic; 80 ± 2.8 %; 225 ± 12.5 of 280 ± 8.4 cells; n = 4), while only a subset express vesicular glutamate transporter-2 (glutamatergic; 18.2 ± 2.9 %; 50.8 ± 7.7 of 280 ± 8.4 cells) or AT1R (8.7 ± 1.0 %; 22 ± 2.2 of 256 ± 11.7 cells). No co-localization was observed with tyrosine hydroxylase in the NTS. Although AT2R-eGFP neurons were not observed within the paraventricular hypothalamic nucleus (PVN), eGFP was localized to efferents terminating in the PVN and to GABAergic neurons surrounding this nucleus. Retrograde neuronal tract tracing studies revealed that many eGFP-positive efferents to the PVN arise from neurons in the MnPO. Based on these neuroanatomical results, we hypothesized that activation of central AT2R would decrease blood pressure. Consistent with this hypothesis, chronic administration of the selective AT2R agonist, compound 21 (7.5 ng/h into the lateral cerebral ventricle) reduced baseline mean arterial blood pressure relative to control mice (103 ± 1.65 v. 110 ± 1.70 mmHg; n = 16; p = 0.02). These studies demonstrate that central AT2R are positioned to regulate blood pressure.

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Metabolomic characterization of experimental osteoarthritis in a non-invasive bioluminescent reporter mouse

Osteoarthritis and Cartilage ◽

10.1016/j.joca.2015.02.832 ◽

2015 ◽

Vol 23 ◽

pp. A112-A113

Author(s):

D.L. Zignego ◽

S. Mailhiot ◽

R. June

Keyword(s):

Reporter Mouse ◽

Non Invasive ◽

Experimental Osteoarthritis

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Identification and characterization of a novel gene of grouper iridovirus encoding a purine nucleoside phosphorylase

Journal of General Virology ◽

10.1099/vir.0.80249-0 ◽

2004 ◽

Vol 85 (10) ◽

pp. 2883-2892 ◽

Cited By ~ 10

Author(s):

Jing-Wen Ting ◽

Min-Feng Wu ◽

Chih-Tung Tsai ◽

Ching-Chun Lin ◽

Ing-Cherng Guo ◽

...

Keyword(s):

Purine Nucleoside Phosphorylase ◽

Enzymic Activity ◽

Host Cells ◽

Purine Nucleoside ◽

Nucleoside Phosphorylase ◽

Egfp Reporter ◽

Key Enzyme ◽

Egfp Fusion Protein ◽

Identification And Characterization

Purine nucleoside phosphorylase (PNP) is a key enzyme in the purine salvage pathway. It catalyses the reversible phosphorolysis of purine (2′-deoxy)ribonucleosides to free bases and (2′-deoxy)ribose 1-phosphates. Here, a novel piscine viral PNP gene that was identified from grouper iridovirus (GIV), a causative agent of an epizootic fish disease, is reported. This putative GIV PNP gene encodes a protein of 285 aa with a predicted molecular mass of 30 332 Da and shows high similarity to the human PNP gene. Northern and Western blot analyses of GIV-infected grouper kidney (GK) cells revealed that PNP expression increased in cells with time from 6 h post-infection. Immunocytochemistry localized GIV PNP in the cytoplasm of GIV-infected host cells. PNP–EGFP fusion protein was also observed in the cytoplasm of PNP–EGFP reporter construct-transfected GK and HeLa cells. From HPLC analysis, the recombinant GIV PNP protein was shown to catalyse the reversible phosphorolysis of purine nucleosides and could accept guanosine, inosine and adenosine as substrates. In conclusion, this is the first report of a viral PNP with enzymic activity.

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Generation and characterization of a Notch1 signaling-specific reporter mouse line

genesis ◽

10.1002/dvg.22030 ◽

2012 ◽

Vol 50 (9) ◽

pp. 700-710 ◽

Cited By ~ 5

Author(s):

Emma Smith ◽

Stéphanie Claudinot ◽

Rajwinder Lehal ◽

Luca Pellegrinet ◽

Yann Barrandon ◽

...

Keyword(s):

Mouse Line ◽

Reporter Mouse ◽

Notch1 Signaling

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Generation And Characterization Of An Nkx2-1-GFP Reporter Mouse

10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a6313 ◽

2012 ◽

Author(s):

Laertis Ikonomou ◽

Jyh-Chang Jean ◽

Letty Kwok ◽

Tyler Longmire ◽

Darrell N. Kotton

Keyword(s):

Reporter Mouse ◽

Gfp Reporter

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Impact of Diet-Induced Obesity on Intestinal Stem Cells: Hyperproliferation but Impaired Intrinsic Function That Requires Insulin/IGF1

Endocrinology ◽

10.1210/en.2014-1112 ◽

2014 ◽

Vol 155 (9) ◽

pp. 3302-3314 ◽

Cited By ~ 48

Author(s):

Amanda T. Mah ◽

Laurianne Van Landeghem ◽

Hannah E. Gavin ◽

Scott T. Magness ◽

P. Kay Lund

Keyword(s):

Stem Cells ◽

Nutrient Intake ◽

Intestinal Stem Cells ◽

Exogenous Insulin ◽

Reporter Mouse ◽

Diet Induced Obesity ◽

Egfp Reporter ◽

The Impact

Abstract Nutrient intake regulates intestinal epithelial mass and crypt proliferation. Recent findings in model organisms and rodents indicate nutrient restriction impacts intestinal stem cells (ISC). Little is known about the impact of diet-induced obesity (DIO), a model of excess nutrient intake on ISC. We used a Sox9-EGFP reporter mouse to test the hypothesis that an adaptive response to DIO or associated hyperinsulinemia involves expansion and hyperproliferation of ISC. The Sox9-EGFP reporter mouse allows study and isolation of ISC, progenitors, and differentiated lineages based on different Sox9-EGFP expression levels. Sox9-EGFP mice were fed a high-fat diet for 20 weeks to induce DIO and compared with littermates fed low-fat rodent chow. Histology, fluorescence activated cell sorting, and mRNA analyses measured impact of DIO on jejunal crypt-villus morphometry, numbers, and proliferation of different Sox9-EGFP cell populations and gene expression. An in vitro culture assay directly assessed functional capacity of isolated ISC. DIO mice exhibited significant increases in body weight, plasma glucose, insulin, and insulin-like growth factor 1 (IGF1) levels and intestinal Igf1 mRNA. DIO mice had increased villus height and crypt density but decreased intestinal length and decreased numbers of Paneth and goblet cells. In vivo, DIO resulted in a selective expansion of Sox9-EGFPLow ISC and percentage of ISC in S-phase. ISC expansion significantly correlated with plasma insulin levels. In vitro, isolated ISC from DIO mice formed fewer enteroids in standard 3D Matrigel culture compared to controls, indicating impaired ISC function. This decreased enteroid formation in isolated ISC from DIO mice was rescued by exogenous insulin, IGF1, or both. We conclude that DIO induces specific increases in ISC and ISC hyperproliferation in vivo. However, isolated ISC from DIO mice have impaired intrinsic survival and growth in vitro that can be rescued by exogenous insulin or IGF1.

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