C57b6 j

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C57B6/J is a mouse strain frequently used in biomedical research. It is a widely studied inbred mouse strain that serves as a common control or background strain for generating genetically engineered mouse models.

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21 protocols using c57b6 j

Cardiac Myocyte Isolation from Genetically Modified Mice

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Several types of adult mice aged from 10 to 12 weeks were used,
including: WT C57b/6J (Jackson Lab), WT C57b/6N (Jackson Lab), Ser280Ala
KI,^{25 ,31 (link)} MMVV-KI,^{32 (link)} CaMKIIδ cardiac-specific and
global KO,^{33 (link),34 (link)} NOX2^−/− mice
(Jackson Lab) and Grx1-roGFP2 (targeted either to the cytosol or
mitochondria).^{27 (link)} All
the genetically modified mouse lines were bred on the C57b/6J background, except
for the C57b/6N line. See Online Methods for more details.
Cardiac ventricular myocytes were isolated using previously described
methods^{35 (link)} which were
approved by the UC Davis Institutional Animal Care and Use Committee (IACUC).
Freshly isolated myocytes were plated on laminin-coated glass cover slips for 30
min before dye loading. All experiments were performed at room temperature (RT)
(22–23°C) with pH 7.4.

Lu S., Liao Z., Lu X., Katschinski D.M., Mercola M., Chen J., Brown J.H., Molkentin J.D., Bossuyt J, & Bers D.M. (2020). Hyperglycemia Acutely Increases Cytosolic Reactive Oxygen Species via O-linked GlcNAcylation and CaMKII Activation in Mouse Ventricular Myocytes. Circulation research, 126(10), e80-e96.

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Vertebrate Animal Husbandry and Genetic Strains

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All animals were maintained on ad libitum rodent chow and water and housed in a climate-controlled AALAC-accredited vivarium operating under a 12-h light/dark cycle. All protocols for the use of vertebrate animals are approved by the Committee for the Human Use of Animals at Tufts University School of Medicine. Eight-week old F1 mice were bred from C57/B6J and 129S1/Sv1MJ mice in house or were purchased from Jackson Laboratories (stock 101043). Adult male Sprague-Dawley rats were purchased from Taconic Biosciences (stock SD-M). Ascl1-CreER^T2 mice (stock 012882), and the Cre reporter strain R26R(TdTomato) (stock 007909), constitutive GFP Tg(UBC-GFP) (stock 004353), and germline constitutive Tg(Sox2-Cre) (stock 004893) were purchased from Jackson Laboratories. K5-CreER^T2 mice, Neurog1-CreER^T2 mice, p63^fl/fl mice, and ΔOMP-eGFP mice have been described elsewhere and were generously provided by P. Chambon via R. Reed (Indra et al., 1999 (link)), L. Goodrich (Kim et al., 2011 (link)), A. Mills (Mills et al., 2002 (link)), and P. Mombaerts (Potter et al., 2001 (link)), respectively. Pancellular-TdT mice were generated in house by breeding R26R(TdT) mice to Tg(Sox2-Cre) mice.

Peterson J., Lin B., Barrios-Camacho C.M., Herrick D.B., Holbrook E.H., Jang W., Coleman J.H, & Schwob J.E. (2019). Activating a Reserve Neural Stem Cell Population In Vitro Enables Engraftment and Multipotency after Transplantation. Stem Cell Reports, 12(4), 680-695.

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Visualizing GABAA Receptor Subunits in RGCs

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All experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee of the University of Wisconsin-Madison and the National Institutes of Health. Animals of both sexes were used for experiments. Thy1-YFPH mice (Bleckert et al., 2014 (link); Feng et al., 2000 (link)) were used at P8, P12, P16, P21 and adult (> 1.5months) time-points. This line was crossed into the GABA_Aα3 knockout transgenic line (Yee et al., 2005 (link)) and age-matched adult (> 1.5 month) littermate control and knockout animals were used for all the GABA_Aα3 knockout analyses. To visualize GABA_Aγ2 receptor sites across the RGC arbors, the Thy1-YFPγ2 mice (Bleckert et al., 2013 (link)) were utilized and crossed into the GABA_Aα3 knockout background to assess GABA_Aγ2 puncta across RGCs in age-matched adult (> 1.5 month) littermate control and GABA_Aα3 knockout animals. For experiments involving biolistics, 1.2–1.5 month old Thy1-GABA_Aγ2YFP × GABA_Aα3 knockout and age-matched littermate control animals were used (Figure 7) and 1.2–1.5 month old age-matched GABA_Aα3 knockout-littermate control animals were used for PSD-95 transfections (Figure S7). All electrophysiology experiments were carried out on 1.5 month old wild-type animals (C57B6/J; Jackson Labs).

Sawant A., Ebbinghaus B.N., Bleckert A., Gamlin C., Yu W.Q., Berson D., Rudolph U., Sinha R, & Hoon M. (2021). Organization and emergence of a mixed GABA-glycine retinal circuit that provides inhibition to mouse ON-sustained alpha retinal ganglion cells. Cell reports, 34(11), 108858.

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Aging-Associated Changes in Microglia

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All experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of University of California San Francisco (Protocol number: 170302, Approved: July 2017–2020; Protocol number: 106982, Approved: June 2014–2017). Adult males, aged 3–6 months (young) and 20–25 months (C57B6/J) were purchased from Jackson Laboratory (Bar Harbor, ME) and the National Institute on Aging Animal Colony, respectively. Young and aged CX3CR1^GFP/+CCR2^RFP/+ (double heterozygous (Dbl-Het)), CCR2^RFP/+, and CCR2^RFP/RFP (effectively CCR2^−/−) mice were bred and aged as previously described [33 (link)] and genotyped using a commercial service (Transnetyx). Mice were group housed in environmentally-controlled conditions with a reverse light cycle (12:12 h light:dark cycle at 21 ± 1 °C) and provided food and water ad libitum. The experimental time points are presented in Supplementary Figure S5.

Chou A., Krukowski K., Morganti J.M., Riparip L.K, & Rosi S. (2018). Persistent Infiltration and Impaired Response of Peripherally-Derived Monocytes after Traumatic Brain Injury in the Aged Brain. International Journal of Molecular Sciences, 19(6), 1616.

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Immunohistochemical Analysis of DDX6 in Mouse Brain

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Eight day postnatal and 10 month old male black 6 mice (C57B/6 J, Jackson Laboratory) were anesthetized and intracardially perfused with PBS (Invitrogen) followed by 4% paraformaldehyde (PFA) and processed as published^{66 (link),67}. In brief, brains were removed, postfixed in 4% PFA overnight, washed 3x with PBS and placed in cold 30% sucrose in PBS till they sunk down (~2 days). Samples were embedded in OCT (Tissue-Tek) and cryopreserved. Sagittal cryosections (40 µm thick) were permeabilized with PBS-0.1% Triton X-100 (PBT) and then blocked with 1% BSA in PBT. Primary polyclonal goat anti-DDX6 (Abnova) antibody was incubated o/n at 4 °C. Secondary calf anti-goat Alexa488 or Alexa555 conjugated antibodies (Dianova) were incubated for 2 h at RT. Slides were mounted with Aqua-Poly/Mount (Polysciences).

Bauer K.E., Bargenda N., Schieweck R., Illig C., Segura I., Harner M, & Kiebler M.A. (2022). RNA supply drives physiological granule assembly in neurons. Nature Communications, 13, 2781.

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Age-dependent Cognitive and Neurological Changes in Mice

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Three age groups of male C57B/6J (Jackson Laboratory, Farmington, CT, USA) mice were used: 3, 17–18 and 24–25 months of age. The three month old mice are defined as young mice, comparable to humans in their early 20’s; 17–18 months old mice are defined as middle-aged, comparable to humans in their mid-50’s; and 24–25 months mice are defined as old mice, comparable to humans in their 70’s, (Flurkey et al., 2007 ). Four young, five middle-aged and six old mouse brains were collected and processed for histological examination. Three mice from each group were used for magnetic resonance imaging (MRI), and all animals were sacrificed by transcardiac perfusion under anesthesia (see below). To investigate how cognitive function changes with aging, another set of young and old mice (in total n=32) was added to participate in a Novel Object Recognition Test (NOR). Both young mice and old mice had two subgroups of eight mice each tested for either 10 min inter-trial-interval (ITI) or 2 hours ITI NOR tests.
All procedures described were approved by the Institutional Animal Care and Use Committee of Boston University (IACUC). The mice were maintained according to the NIH Guide for the Care and Use of Laboratory Animals.

Wang Y., Taylor E., Zikopoulos B., Seta F., Huang N., Hamilton J.A., Kantak K.M, & Morgan K.G. (2020). Aging-induced Microbleeds of the Mouse Thalamus Compared to Sensorimotor and Memory Defects. Neurobiology of aging, 100, 39-47.

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Bleomycin Sensitivity in HPS Mice

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Wild-type, HPS1, and HPS2 mice (C57B/6 J, 8–10 weeks old) were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed in a pathogen-free animal facility at Thomas Jefferson University. HPS1 mice have homozygous mutation of the Hps1 gene, which encodes for a protein called BLOC-3, and HPS2 mice have homozygous mutation in the adaptor protein 3b1 (Ap3b1) gene, which is a subunit of the AP-3 protein complex. In general, HPS mice are phenotypically normal, except for a light coat appearance. HPS 1 and 2 mice also have large lamellar bodies in the alveolar epithelial type II cells of their lungs. Both strains of mice are also exquisitely sensitive to bleomycin. Throughout the study period, wild-type and HPS mice were maintained on a standard chow diet (13.5% calories from fat, 58% from carbohydrates, and 28.5% from protein) and permitted to feed ad libitum. Prior to the initiation of any study, the Institutional Animal Care and Use Committee at Thomas Jefferson University approved all animal protocols.

Summer R., Krishna R., Schriner D., Cuevas-Mora K., Sales D., Para R., Roman J., Nieweld C., Gochuico B.R, & Romero F. (2019). Matrix metalloproteinase activity in the lung is increased in Hermansky-Pudlak syndrome. Orphanet Journal of Rare Diseases, 14, 162.

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Genetically Modified Mouse Models

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Mice used include WT CF-1 (Charles River Laboratories), WT C57BL/6J (The Jackson Laboratory), Ptgs1^tm1Unc (called Ptgs1 or Cox1, RRID:MGI_4366280), Ptgs2^tm1Unc (called Ptgs2 or Cox2, RRID:MGI_4366244) (C57BL/6J background) (Langenbach et al., 1995 (link); Morham et al., 1995 (link)), Sox10^tm1Weg (called Sox10+/− or Sox10 LacZ, RRID:MGI_4437131) (C3H background) (Britsch et al., 2001 (link)) (from M. Wegner (Friedrich Alexander University Erlangen Nuremberg, Erlangen, Germany) and M. Southard-Smith (Vanderbilt University, Nashville, Tennessee, USA)), and Ret^tm1Jmi (Ret (tau-EGFP-myc), Ret-TGM, RRID:MGI_3623107), called Ret+/−, C57/B6J background)(Enomoto et al., 2001 (link)). The day of the vaginal plug was called E0.5. Mice were genotyped by PCR as described in the references above.

Schill E.M., Lake J.I., Tusheva O.A., Nagy N., Bery S.K., Foster L., Avetisyan M., Johnson S.L., Stenson W.F., Goldstein A.M, & Heuckeroth R.O. (2015). Ibuprofen slows migration and inhibits bowel colonization by enteric nervous system precursors in zebrafish, chick and mouse. Developmental biology, 409(2), 473-488.

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Generation of BCR-Abl+ Leukemia

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Bone marrow cells from C57B6/J (Jackson Labs, Bar Harbor, ME, USA), Bim^−/− (Jackson Labs), Ubi-Cre-ER^T2 transgenic (Jackson Labs), and Glut1^fl/fl mice crossed to Ubi-Cre-ER^T2 mice were cultured in IL-7 (10 ng/ml) (eBioscience) in IMDM supplemented with 20% fetal bovine serum and infected with MSCV-BCR-Abl-IRES-GFP retrovirus (gift of D Fruman, UC Irvine)^{7 (link)} with polybrene (4 μg/ml) (Millipore, Billerica, MA, USA). Infected cells were cultured in methylcellulose medium containing IL-7 (Stem Cell Technologies) and individual colonies were isolated on day 7 and transferred into complete IMDM media with 20% fetal bovine serum, but no IL-7. After 7 days, viable GFP+ colonies were selected for expansion. The Institutional Animal Care and Use Committee of Duke University approved all animal protocols.

Liu T., Kishton R.J., Macintyre A.N., Gerriets V.A., Xiang H., Liu X., Abel E.D., Rizzieri D., Locasale J.W, & Rathmell J.C. (2014). Glucose transporter 1-mediated glucose uptake is limiting for B-cell acute lymphoblastic leukemia anabolic metabolism and resistance to apoptosis. Cell Death & Disease, 5(10), e1470-.

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Electrophysiological Characterization of CA1 Pyramidal Neurons

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All procedures used in this study were carried out with the approval of the University of Minnesota Institutional Animal Care and Use Committee. The mice used in this series of experiments were 3–4 months old, male wild-type C57B6/J (Jackson Labs, Bar Harbor, ME). Animals were housed in a specific-pathogen-free facility and kept on a 12 h/12 h light/ dark cycle. Animals were provided rodent chow and water ad libitum. All experiments were performed using in vitro transverse slices of hippocampus. Whole-cell current clamp recordings were performed to explore intrinsic membrane properties of CA1 pyramidal neurons exposed to SV. Extracellular dendritic field potential (fEPSP) recordings were used to probe the actions of SV on pharmacologically isolated NMDAR-mediated synaptic potentials at Schaffer collateral CA1 pyramidal cell synapses. Hippocampal slices treated with vehicle were used as controls.

Parent M.A., Hottman D.A., Cheng S., Zhang W., McMahon L.L., Yuan L.L, & Li L. (2014). Simvastatin Treatment Enhances NMDAR-Mediated Synaptic Transmission by Upregulating the Surface Distribution of the GluN2B Subunit. Cellular and molecular neurobiology, 34(5), 693-705.

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