Study of Lesion Sizes in Osmotically Stressed RBCs

Datta, Sanchari (2015) Study of Lesion Sizes in Osmotically Stressed RBCs. Masters thesis, Indian Institute of Science Education and Research Kolkata.

PDF (MS dissertation of Sanchari Datta (10MS35)) Sanchari_Datta_10MS35_MS_Thesis_2015.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

As some of the membranes of animal cells inhabit mechanically stressful environments, they are highly susceptible to membrane rupture and hence undergo membrane repair. It is generally accepted that the mechanism of repair is linked to the size of the lesion formed. In the lab, it has been established that global stress like hypo-osmotic shock does rupture cells however observed only under cold shock. My thesis aims to readdress the issue of membrane lesion size in global mechanical stress in RBCs since RBCs lack endomembrane, have tighter size distribution and most importantly, rupture even without “cold” hypo-osmotic shock. I have quantified the rupture dynamics, showing clearly the presence of a single rupture point as well that the rupture size is independent of the shock strength. I have also observed that perturbing the cholesterol content (cholesterol depletion by MβCD) immensely affects the rupture propensities and increases lesion sizes. It was shown earlier in the lab that HeLa and CHO (cells with endomembrane and nucleus) when ruptured by cold hypo-osmotic shock show no differences in lesion sizes even when treated with actin perturbing drugs, or depleted of ATP. Learning from RBC experiments, I am also looking at the effect of MβCD (cholesterol depleting agent) on the rupture charecteristics in HeLa cells. Future investigations using reflection interference contrast microscopy will aim to characterize local heterogeneities in the membrane that result in varied rupture propensities under a global stress mechanism.

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