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11 protocols using thy1 yfp h

1

Transgenic Mouse Models for Neuroscience

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All animal procedures were performed in accordance with the regulations of the Yale University animal care committee. Experiments were performed on adult (postnatal day 73–149) Thy1-GFP-M (n = 13; #007788, The Jackson Laboratory, RRID:IMSR_JAX:007788) and Thy1-YFP-H transgenic mice (n = 3; #003782, The Jackson Laboratory, RRID:IMSR_JAX:003782). Mice of both sexes were used. Mice were housed under controlled temperature on a 12 h light/dark cycle with siblings (one to five per cage) and nesting material.
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2

Sciatic Nerve Injury Model in Thy1-YFP Mice

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Both male and female adult (8–12 weeks old) Thy1-yellow fluorescent protein mice, line H (Thy1-YFP-H; Jackson Laboratories, Bar Harbor, USA) [43 (link)], were used in these studies. All experimental procedures were approved by the IPMon Animal Care Committee and conducted according to international FELASA guidelines, national law, and ethical guidelines (Uruguayan Animal Care Committee).
The surgical procedure was carried out following sterile precautions. Mice were anesthetized with ketamine-xylazine (90–10 mg/kg) and the right sciatic nerve at the mid thigh level was exposed. Treatments were performed by direct injections at 45 mm from the tip of the third digit in 2 μl of sterile PBS, using a fine glass micropipette connected to a Hamilton syringe. Immediately after the injection and at the same position, the nerve was crushed in two different directions 30 s each time with fine forceps. The crush site was labeled with lamp black powder. The wound was closed with 5–0 mononylon Ethilon sutures (Ethicon) and disinfected. The sciatic and tibial nerves and the plantar skin were harvested at 24 h, 3, 10, and 28 days post lesion (dpl).
All nerve injections were performed in 2 μl PBS and at 10 μg/ml concentration of the following products: rCD300f-IgG2a (rat extracellular domain of CD300f fused to mouse IgG2a protein) or purified mouse myeloma IgG2a (Invitrogen, Cat. N° 026200).
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3

Generation of Transgenic Mouse Models

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The Institutional Animal Care and Use Committee of University of California Santa Cruz approved all animal care and experimental procedures. The Fmr1fl and Fmr1neo mice were obtained from Dr. David L. Nelson, Baylor College of Medicine; the global Fmr1 KO mice from Dr. Stephen T. Warren, Emory University; the mGFAP-Cre+ mice (line 73.12) from Dr. Michael V. Sofroniew, University of California Los Angeles; and the S100β-GFP mice from Dr. Wesley J. Thompson, Texas A&M University. Thy1-YFP-H and Rosa26tdTomato mice were purchased from Jackson Laboratory. All mice were backcrossed with C57BL/6 mice more than 10 generations to produce congenic strains. Male mice were used in all experiments.
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4

Transgenic Mouse Model Experiments

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Mice were group-housed under a 12 h light/dark cycle, with food and water available ad libitum. Thy1-YFPH, PV-Cre, SST-Cre, VIP-Cre, and Ai9-tdTomato mice with C57BL/6 background were purchased from the Jackson Laboratory and housed in the Laboratory Animal Unit, The University of Hong Kong. Postnatal day 30 (P30) ± 1 mice were used in this study. Animals were randomly assigned to different experimental groups. All experiments were approved and performed in accordance with University of Hong Kong Committee on the Use of Live Animals in Teaching and Research guidelines.
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5

Transgenic mouse models for neural circuit analysis

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All experiments were carried out in compliance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Our study protocol was reviewed and approved by the University of Michigan Institutional Animal Care & Use Committee. The transgenic mice used in this study were: PV-Cre (Jackson stock no. 008069), SOM-Cre (Jackson stock no. 013044), VGAT-Cre (Jackson stock no. 016962), Ai14 (Jackson stock no. 007914), and Thy1-YFP-H (Jackson stock no. 003782). P28–P56 male and female mice were used for virus injections. A total of ten mice on a C57/Bl6 genetic background were used in this study (two PV-Cre × SOM-Cre, three PV-Cre, one VGAT-Cre × Ai14, one PV-Cre × Ai14, one Thy1-YFP-H, two wild type).
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6

Visualizing Neuronal Morphology in TDP-43 Mice

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Experimental procedures utilised male and female mice and were approved by the Animal Ethics Committee of the University of Tasmania (A0016593). Experiments were performed in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes (2013) and in accordance with the Animal Research: Reporting of In Vivo Experiments (ARRIVE) Guidelines. Mice were housed in individually ventilated cages at 20 °C with littermates unless otherwise stated, on a 12-h light–dark cycle, with access to food and water ad libitum. C57/BL6 (strain 000,664), Thy1-YFPH [strain 003,782 B6.Cg-Tg(Thy1-YFP)HJrs/J] [31 (link), 32 (link)], and TDP-43 [strain 010,700 B6.Cg-Tg(Prp-TARDBPA315T)95Balo/J] mice were all purchased from Jackson Laboratories, with all lines maintained on a C57/BL6 background. To capture high-resolution fluorescence of layer V pyramidal neurons residing in layer V and extending into layer II/III the TDP-43 model was crossed with Thy1-YFPH mice. For tissue collection, animals were injected with an intraperitoneal (i.p.) terminal anaesthesia (sodium pentobarbital) and transcardially perfused with 0.01 M phosphate-buffered saline (PBS). The cortex was immediately dissected and frozen in liquid nitrogen, and stored at − 80 °C prior to experimental processing. Male and female mice were included.
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7

Thy1-YFP-H Mouse Model Study

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Adult female B6.Cg-Tg(Thy1-YFP)HJrs/J mice (referred to as Thy1-YFP-H, stock no. 003782; Jackson Laboratory, Bar Harbor, ME, USA) aged 11 to 12 months were used in this study. They were housed on a 12-hour light/dark cycle with standard rodent chow and water as desired. All animal operations were approved by the Institutional Animal Care and Use Committee of Zhongshan Ophthalmic Center and complied with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.
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8

Conditional Knockout of Kif5b in Layer V Neurons

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Mice were group housed under a 12 hr light/dark cycle, with food and water available ad libitum. C57BL/6 mice expressing CaMKIIα-Cre and yellow fluorescent protein (YFP) in layer V pyramidal neurons (Thy1-YFP-H) and CaMKIIα promoter-driven Cre transgenic mice were purchased from the Jackson Laboratory. Kif5bfl/fl mice were described previously (Cui et al., 2011 (link)). CaMKIIα promoter-driven Cre transgenic mice were used to conditionally delete exons flanked by loxP. Mice were then further crossed with Thy1-YFP-H line to allow imaging of layer V pyramidal neurons. Sample size was decided based on experiments in previous studies (Lai et al., 2012 (link); Yang et al., 2014 (link)). For animal behavioral tests and in vivo imaging experiments, results from at least two independent experiments were pooled together for analysis. Mice were group housed in The Laboratory Animal Unit, The University of Hong Kong, accredited by Association for Assessment and Accreditation of Laboratory Animal Care International. Four to five weeks old mice were used in this study unless stated otherwise. All experiments were approved and performed in accordance with University of Hong Kong Committee on the Use of Live Animals in Teaching and Research guidelines.
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9

Fluorescent Mouse Lines for Neuroscience

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Snap25-GCaMP6s (JAX#025111), Thy1-YFP-H (JAX#003782), and Cx3cr1-GFP (JAX#005582) mouse lines were purchased from The Jackson Laboratory and maintained on the C57BL/6J background. Mice were group-housed in the UCSC animal facility, with 12 h light-dark cycle and access to food and water ad libitum. Mice of both sexes were used in the study. All animal studies were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee (IACUC) of UCSC.
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10

Mouse and Zebrafish Experiments Protocol

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All animal experiments were conducted according to the National Institutes of Health guidelines for animal research. Procedures and protocols on mice and zebrafish were approved by the Institutional Animal Care and Use Committee at Janelia Research Campus, Howard Hughes Medical Institute; and the Animal Care and Use Committee at the University of California, Berkeley. Mice were housed in cages in groups of 1–5, under a normal light cycle, at a temperature of about 73 °F and humidity of 37%. Both males and females were used in this study, with ages ranging from 5 to > 30 weeks old, and included (Jackson Laboratories): Thy1-YFP-H, Thy1-GFP-M, Gad2-IRES-Cre X Ai14, C57BL/6J. Details on animal preparations are available below.
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