This article describes a method for inducing central nervous system (CNS) infections in mouse pups using targeted viral injections. The procedure involves both intracerebral and intraperitoneal injections to facilitate localized and systemic viral spread.
Begin with two syringes filled with different volumes of media containing the viruses targeted for central nervous system (CNS) infection.
Keep the mouse pup in a prone position. Disinfect the head and inject the smaller volume of the virus solution intracerebrally through the lambda region.
This allows the viruses to bypass the blood-brain barrier and directly reach the brain tissue.
Next, hold the mouse pup in a supine position. Disinfect and inject the larger volume of the virus solution into the abdomen.
This facilitates the systemic spread of the virus through the blood vessels, infecting other parts of the CNS.
The injected viruses enter neuronal cells and use host machinery to replicate, leading to cell death.
The viruses also trigger the immune system, causing CNS inflammation and establishing a CNS infection model.
Before initiating the infection procedure, dilute non-adapted dengue virus 2 stock to a 1 times 10 to the fifth plaque forming units per 40 microliters of RPMI 1640 medium concentration. Next, load 10.3 milliliter syringe equipped with a 30-gauge needle with 10 microliters of diluted virus and 10.3 milliliters syringe equipped with a 30-gauge needle with 30 microliters of virus per animal.
For intracerebral delivery of the virus, press the oracle between the thumb and index finger to gently restrain a seven-day-old suckling Institute of Cancer Research Mouse in a prone position, and inject the 10 microliter volume of diluted virus into the lambda area at the intersection of the sagittal and lambdoid suture. After the intracerebral injection, use the thumb and index finger to hold the mouse in a supine position and gently intraperitoneally, inject the 30-microliter volume of diluted virus into the Murine abdomen.
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