Animal Core

COBRE in Lipidomics and Pathobiology

 

Core Services

The mission of the core is to assist investigators perform experimentation with genetically modified mice, particularly mice bearing mutations in sphingolipid pathway genes).

Consultation on mouse experimentation

·       Planning of all phases of mouse experimentation

·       Assistance with preparation of IACUC animal protocols.

·       Assistance with preparing information for the Vertebrate Animals section in NIH grant applications.

Supplying mice for experimentation

·       The core will provide required genetically modified mice (e.g., mice bearing mutations in sphingolipid pathway genes) for experimentation.  The services include mouse husbandry, genotyping necessary to provide mice of specified age and sex.

The core also maintains a number of genetically modified mouse strains (Table 1), but will consider raising other types of mice that would be of broad interest.

 

Table 1. Strains of genetically modified mice currently housed by the core.

Targeted Gene	Gene Name	Background
SK1	Sphingosine kinase 1	BL6
SK2	Sphingosine kinase 2	BL6 and FVB
S1P2	S1P receptor 2	BL6
S1P3	S1P receptor 3	BL6
ASM	Acid sphingomyelinase	BL6
NCdase	Neutral ceramidase	BL6
CerS4	Ceramide synthase 4	BL6
CerS5	Ceramide synthase 6*	BL6
LDLR	Low density lipoprotein receptor*	BL6
SK1-/-;SK2-/+;LDLR-/-*	Compound mutant	BL6

*Currently a helicobacter positive colony.

 

Special procedures

The core will conduct special procedures including: 

·       Special diet feeding (e.g., atherogenic diet)

·       Metabolic cage housing (e.g., urine collection)

·       Mini pump implantation

Full service tissue procurement 

·       On request the core will perform isolation and fixation of tissues.  Isolated tissue can be provided fresh, frozen, fixed or preserved in RNAlater.

Full service isolation of primary cells from genetically modified and wildtype mice

·       On request the core will prepare primary cell lines from genetically modified and wildtype mice. For example the core can isolate mouse embryonic fibroblasts (MEFs) or endothelial cells.

·       Currently, the core has stocks of MEFs including SK1 null MEFs (BL6) and wildtype MEFs (BL6).

Full service isolation of tissue RNA from genetically modified and wildtype mice

·       Core personnel will isolate RNA from tissues of genetically modified and wildtype mice.  Isolated RNA will be quality controlled using an Agilent Bioanalyzer.

·       Upon request we can also prepare cDNA from isolated RNA.         

Genotyping

·       The core offers full service genotyping.  This service includes mouse-tail clipping, extraction and purification of DNA and PCR. The core will also offer assistance with designing a PCR-based genotyping protocol for a given transgenic mouse.  This includes PCR primer design and optimizing PCR conditions. 

Procedure Training

The core offers training in a range of techniques including:

·       Intravenous (IV) injection: Tail vein injection of agents (e.g., BrdU, drugs, siRNA, Evans blue dye).

·       Blood Isolation: Isolation of blood by sub-mandibular bleeding or heart puncture.

·       Euthanasia: (including perfusion fixation if required).           

·       Immunohistochemistry:  Core personnel have extensive experience with performing immunohistological staining (confocal microscopy) and histochemical staining e.g., DAB, H&E, alizarin red, oil Red O staining.

All animal procedures are performed according MUSC IACUC approved animal protocol.

Fees

Service fees are currently being determined.