Effect of Mouse Hepatitis Virus (MHV) infection in alteration of Connexin 43 in Astrocytes, Meningeal fibroblastoid and Microglia

Banerjee, Kaveri (2014) Effect of Mouse Hepatitis Virus (MHV) infection in alteration of Connexin 43 in Astrocytes, Meningeal fibroblastoid and Microglia. Masters thesis, Indian Institute of Science Education and Research Kolkata.

PDF (MS dissertation of Kaveri Banerjee) Kaveri_Banerjee_Thesis.pdf - Submitted Version Restricted to Repository staff only Download (16MB)

Abstract

We observed that astrocytes, fibroblastoid cells and microglia can be enriched from mixed glial cultures. Astrocytes and fibroblastoid cells express Cx43 gap junction protein. These cells upon infection show syncytium formation. The Lucifer dye transfer experiment makes it evident that the gap junction channels are altered in infected cells. As a result of this the dye transfer is hindered in the syncytium forming regions. This same observation was seen in case of infected astrocytes. Thus we can conclude that MHV infection brings upon similar effect on fibroblastoid cells like astrocytes. Further investigation at mRNA and protein levels by Quantitative-PCR is required to comment on regulation at transcriptional or translational level. Confirmation of fibroblastoid type of cells further by using CD-34 antibody is also required. N9 microglial cell lines have shown a down regulation of Cx43 in MHV infected cells compared to uninfected ones. Literature reveals an up-regulation of Cx43 in primary microglia under wound stab conditions. Whether Cx43 is expressed by N9 is still a topic of research. Effect of MHV infection upon N9 is also not well documented. Thus, from the experiments we performed, it can be said that Cx43 is expressed in N9 cells. Further investigation of Cx43 regulation at mRNA levels and gap junction channel formation is yet to be done. In conclusion, it can be revealed that both astrocytes and fibroblastoids express Cx43.As they both express Cx43 they may be physiologically connected through gap junction in maintaining the homeostasis in CNS and BBB (Blood Brain Barrier). N9 also express Cx43 at protein level and altered due to MHV infection which can signify that expression of Cx43 may have a role in MHV infection.

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