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MB Sample ID: SA164694
| Local Sample ID: | StrainsMetabs_B6_1_8 |
| Subject ID: | SU001852 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6 ; PWK/PhJ ; WSB/EiJ ; NZO/HILtJ ; A/J ; 129S1/SvImJ ; CAST/EiJ ; NOD/ShiLtJ |
| Age Or Age Range: | 8-12 weeks |
| Gender: | Female |
| Animal Animal Supplier: | JAX (except C57BL/6, who came from Charles River) |
| Animal Housing: | standard |
| Animal Light Cycle: | standard |
| Animal Feed: | TEKLAD global |
| Animal Water: | standard |
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Treatment:
| Treatment ID: | TR001865 |
| Treatment Summary: | See protocol document for full study details. 2 female C57BL/6 mice were given intraperitoneal (i.p.) injections of 100uL frozen stock of P. chabaudi-infected red blood cells (iRBCs). When parasitemia reached 10-20% at 8-10 days post-infection, mice were euthanized and blood obtained via cardiac puncture. Blood was diluted to 105 iRBC / 100uL in Kreb’s saline with glucose (KSG) and administered i.p. to experimental animals at a dose of 105 iRBCs. https://www.nature.com/articles/nprot.2011.313 Control animals received 100uL of vehicle i.p. Only female mice 8-12 weeks of age were used for P. chabaudi experiments. Experiments were performed in multiple cohorts. Parasitemia was quantified via thin blood smear, methanol fixation, KaryoMAX Giemsa (GIBCO) staining, and manual microscope counting at 100X magnification. RBCs were quantified using a BD Accuri C6 Plus cytometer (see Longitudinal Infection monitoring). Longitudinal monitoring was performed as described previously (Torres et al. 2016, PLOS Biol, “Tracking Resilience to Infections by Mapping Disease Space”). For each mouse, baseline RBC, weight, body temperature, and blood glucose measurements were collected between 1 and 5 days prior to infection. In some cases, blood glucose was collected only at baseline sampling and on the day of sacrifice. Mice were restrained during sample collection using tail-access rodent restrainers (Stoelting Co.). Blood was collected from the tail vein by nicking the end of the tail with disinfected surgical scissors, and depositing the blood into EDTA-coated capillary tubes to prevent clotting. For total RBC quantitation, 2uL of blood was diluted in 1mL of cold 1x Hank’s Balanced Salt Solution (HBSS) and kept on ice until absolute RBC counts were obtained using forward and side scatter gates on a BD Accuri C6 Plus flow cytometer. To record body temperature, mice in the metabolic screen experiments were implanted with subcutaneous electronic temperature and ID transponders (IPTT-300 transponders, Bio Medic Data System, Inc) one week prior to infection. Mice were locally anesthetized using a 2% lidocaine solution (100 ug delivered per dose) prior to implantation. Temperature data was recorded using a DAS-7006/7 s reader (Bio Medic Data System, Inc). Subsequent to metabolic screen experiments, body temperatures were measured using a thermocouple thermometer and mouse rectal probe (World Precision Instruments, RET-3). Blood glucose measurements were obtained with 2uL of tail vein blood analyzed with a Bayer CONTOUR Blood Glucose Monitor and Test Strips. Post-infection sampling began on day 4 or 5 post-infection. Parasitemia values were obtained as detailed above. Parasite density is the number of iRBCs per microliter of blood, and is calculated by multiplying parasitemia by the number of total RBCs. |
| Treatment Protocol Filename: | strainsmetabs_methods_for_mw.docx |
