Saturday, November 2, 2013

Cool methods used in the article, "Circulating C3 Necessary........in a Mouse Model"

To aid one’s understanding of this article, I would like to briefly introduce the method that researchers used to determine whether C3 synthesized within the synovium in joints is important in promoting inflammation, specifically for antibody-mediated arthritis in a mouse model.

First, they made four bone marrow chimeras between normal and non-C3 mice. Wild type bone marrow was injected into non-C3 mouse (C3 only in bone marrow); non-C3 bone marrow was injected into non-C3 mouse (control); non-C3 bone marrow was injected into wild type mouse (C3 only in circulation); . Wild type bone marrow was injected into wild type mouse (control). *Refer to Figure 1 (A) for visual aid.

Because antibodies can be shared between animals, serum that contains antibody from sever arthritic mouse was introduced into those chimeras’ ankle joint to elicit arthritic reaction. Immunofluorescence staining against C3 was used to detect C3 deposition in the joint (collagen). Enzyme-linked immunosorbent assay (ELISA) was used to measure the amount of circulating C3.

They also made parabiotic mice (Parabiosis is a surgical union of two organisms allowing 
sharing of the blood circulation.) to see whether C3 had to be produced locally within the joint, or whether it could be made at distant sites and transported to the joint via the circulation. Non-C3 and WT mice (no C3 in bone marrow) were sutured together to share the circulation. WT mouse was radiated to induce C3 production via parenchymal cells. The goal here was to introduce C3 to non-C3 mouse via circulation from distant site and compare those two mice’s reaction elicited by antibodies from arthritic mouse. *Refer to Figure 3 and 4 for visual aid.

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