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4 protocols using c57br cdj

1

Mouse Diversity Panel Protocol

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Thirty-six male inbred mouse strains (129S1/SvImJ, 129X1/SvJ, A/J, AKR/J, BALB/cByJ, BTBR T+Itpr3tf/J, BUB/BnJ, C3H/HeJ, C57BLKS/J, C57BL/6J, C57BR/cdJ, C58/J, CBA/J, CZECHII/EiJ, DBA/2J, FVB/NJ, I/LnJ, KK/HiJ, LG/J, LP/J, MA/MyJ, NOD/LtJ, NON/LtJ, NZB/BINJ, NZO/HiLtJ, NZW/LacJ, PERA/EiJ, PL/J, PWD/PhJ, PWK/PhJ, RIIIS/J, SEA/GnJ, SJL/J, SM/J, SWR/J, and WSB/EiJ), aged 10–12 weeks, were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). This panel of isogenic mice was chosen based on priority strains from the Mouse Diversity Panel.26 (link) Four mice were used per strain. Male mice were housed four per cage in polycarbonate cages on a 12-hour light/dark cycle (lights on at 7 am), with access to food and water ad libitum. All procedures were approved by the Institutional Animal Care and Use Committee and followed the guidelines set forth by the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals.
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2

Histologic Evaluation of Mouse Hearts

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Ten female and 10 male mice were obtained from The Jackson Laboratory (Bar Harbor, ME) as retired breeders (i.e., 5–10 months of age depending on the rotation time for the colony)(Flurky et al. 2009 ) from the following 16 inbred strains: A/J, BALB/cByJ, BALB/cJ, C3H/HeJ, C57BL/6J, C57BL/10J, C57BR/cdJ, C57L/J, DBA/2J, FVB/NJ, KK/HlJ, LP/J, PWD/PhJ, SWR/J, MRL/MpJ-Faslpr/J, and SJL/J. Mice were euthanized by CO2 asphyxiation, weighed, and the hearts were removed and processed for histologic evaluation.
All protocols were reviewed and approved by The Jackson Laboratory Animal Care and Use Committee. Mouse handling and care were followed according to the Public Health Service animal welfare policies using IACUC approved methods.
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3

Genetic Diversity of Mouse Strains

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129S1/SvImJ, A/J, AKR/J, BALB/cByJ, BTBRT+tf/J, BUB/BnJ, C3H/HeJ, C57BL/10J, C57BL/6J, C57BLKS/J, C57BR/cdJ, C57L/J, CAST/EiJ, CBA/J, DBA/2J, FVB/NJ, KK/HlJ, LP/J, MRL/MpJ, NOD.B10Sn-H2b/J (a congenic strain with the NOD genetic background but with a histocompatibility locus from a diabetes-resistant strain), NON/ShiLtJ, NZO/HlLtJ, NZW/LacJ, P/J, PL/J, PWD/PhJ, RIIIS/J, SJL/J, SM/J, SWR/J, and WSB/J were obtained from The Jackson Laboratory (Bar Harbor, ME). Three of the strains were not evaluated at all time points because of lymphoma development (AKR/J), lack of sufficient mice for analysis (CAST/EiJ), or self-mutilations resulting in euthanasia for humane purposes (SJL/J). (9 , 10 (link)) Taken together the strains selected were genetically diverse, which was estimated by SNP genotyping of over 100 strains into 7 distinct genetic groups, so that there were 3–7 strains representing each of the groups.(11 (link))
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4

Characterization of Mlph ln/ln Mice

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Leaden (C57J/L) and C57BR/cdJ mice were purchased from The Jackson Laboratory. To minimize potential effects of spontaneous mutations occurring after separation of these inbred strains, C57L/J were crossed with C57BR/cdJ, and the resultant heterozygous mice were intercrossed to generate Mlph ln /Mlph ln mice, which were used in the current study. Animal experiments were performed according to the rules and regulations of the Animal Care and Experimentation Committees of Gunma University (Gunma, Japan) and the University of Tokyo (Tokyo, Japan). Only male mice and their tissues and cells were phenotypically characterized in this study. Blood glucose levels were determined by a glucose oxidase method using Glutest Pro GT-1660 (Sanwa Kagaku Kenkyujyo). Insulin was measured by an AlphaLISA insulin kit (PerkinElmer). Pancreatic islet isolation, perifusion secretion assays, and morphometric electron microscopic analysis of granule distribution were performed as described previously (13, 14) .
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