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Sumit Tewari

on 10 January 2013

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Transcript of Apoptosis

Murder of a CELL CELLS Cell Signalling APOPTOSIS Biochemical events lead to characteristic cell changes (morphology) and death. Apoptosis is the process of programmed cell death (PCD) that may occur in multicellular organisms. Intrinsic cell programme. The cell is “told” to die. Kill the defective cell (ex: Cancer cells) by sending appropriate signals Death Ligands

Death receptors (eg. DR4) Receptors are protein molecules to which one or more specific kinds of signaling molecules may attach.

Receives chemical signals originating externally from the cell.

Numerous receptor types are found within a typical cell and each type is linked to a specific biochemical pathway. (Ex: Death receptors)

Receptor Clustering Cell STORY RECEPTOR CLUSTERING Protein A Sulpho SMCC
crosslinker DR4 Antibody In vitro Studies Average Distance between nano particle = 60nm APOPTOSIS Ab-MNP
Death Receptor APOPTOSIS Adaptor proteins APOPTOSIS Demolition phase DISC
Death-inducing signalling complex Procasepase 8 Active Caspase 8 Caspase 3 Ab-MNP
Death Receptors
Adaptor Proteins Active caspase-8 and caspase-3 are immunostained with Alexa-594-labelled secondary antibodies (Red flourescence). Growing signal cascade Apoptosis signalling through caspase activation does not occur when each component of the magnetic switch, such as magnetic field, antibody or MNP, is applied alone Membrane Inversion + Blebbing + Morphological Changes FITC (fluorescein isothiocyanate)–annexin V and propidium iodide staining Interference Contrast Micrographs Defective cells Dead cells Using Cell counting kit-8 (CCK-8) Spatial Controllability Caspase 3 In-vivo Complexity of Live Biological systems Zebra Fish embryo TNF receptor (OTR) for apoptosis signalling
closeness to the human DR4 Zebrafish OTR Antibody on MNP (zAb-MNP) Zebrafish Embryo Cycle Bright Field Microscopy Morphological Alteration Fluorescence study

Immunostaining Conclusions drawn Successful turning On of the Magnetic Switch observed in both in-vitro and in-vivo (Zebrafish) studies.
Spatially and temporally controlled receptor clustering has been observed. Applications It can work with any surface membrane receptors.
Can be extended to other clinically useful membrane receptors like - vascular endothelial growth factor receptors, Toll -like receptors. Who killed the Cell ? Apoptosis One cell stage zAb-MNP NdFeB (Neodymium) magnets DLD-1 Cancer cells "There is plenty of room in the bottom "
- Richard Feynman A magnetic switch for the control of cell death signalling in in vitro and in vivo systems Sumit Tewari
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