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Multiple alphaII-spectrin breakdown products distinguish calpain and caspase dominated necrotic and apoptotic cell death pathways

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Abstract

Apoptosis and oncotic necrosis in neuronal and glial cells have been documented in many neurological diseases. Distinguishing between these two major types of cell death in different neurological diseases is needed in order to better reveal the injury mechanisms so as to open up opportunities for therapy development. Accumulating evidence suggests apoptosis and oncosis epitomize the extreme ends of a broad spectrum of morphological and biochemical events. Biochemical markers that can distinguish between the calpain and caspase dominated types of cell death would help in this process. In this study, three chemical agents, maitotoxin (MTX), staurosporine (STS) and thylenediaminetetraacetic acid (EDTA), were used to induce different types of cell death in PC12 neuronal-like cells. MTX-induced necrosis, as determined by the increased levels of calpain-specific cleaved fragments of spectrin by antibodies specific to the calpain-cleaved 150 kDa αII-spectrin breakdown product (SBDP150) and 145 kDa αII-spectrin breakdown product (SBDP145). In this paradigm, there were no detectable SBDP150i and SBDP120 fragments as determined by antibodies specific to the caspase-cleaved specific fragments similar to those seen in the EDTA-mediated apoptotic PC-12 cells. In contrast to the calpain specific MTX necrosis treatment and the caspase EDTA apoptotic treatment is the STS treatment which induced both proteases as shown by the increase in all the SBDP fragments. Furthermore, compared to SBDP150, SBDP145 appears to be a more specific and sensitive biomarker for calpain activation. Taken together, our results suggested calpains and caspases which dominate the two major types of cell death could be independently discriminated by specifically examining the multiple αII-spectrin cleavage breakdown products.

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Abbreviations

SBDP:

Spectrin breakdown product

MTX:

Maitotoxin

STS:

Staurosporine

EDTA:

Ethylenediaminetetraacetic acid

TBI:

Traumatic brain injury

PCD:

Programmed cell death

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Acknowledgements

We wish to thank Meghan O’Donoghue for her technical assistance. This work was supported by the Department of Defense grant DAMMED-03-1-0066, National Institutes of Health grants R01 NS049175-01-A1 and R01 NS051431. K.K.W. Wang and R.L. Hayes own stock of Banyan Biomarkers Inc., and may benefit financially as a result of the outcome of this research or the work reported in this publication.

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  1. Center of Innovative Research, Banyan Biomarkers Inc., 12085 Research Drive, Alachua, FL, 32615, USA

    Zhiqun Zhang, Stephen F. Larner, Ming Cheng Liu, Wenrong Zheng, Ronald L. Hayes & Kevin K. W. Wang

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  1. Zhiqun Zhang
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  2. Stephen F. Larner
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  3. Ming Cheng Liu
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Correspondence to Zhiqun Zhang or Kevin K. W. Wang.

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Zhiqun Zhang and Stephen F. Larner contributed equally to this work.

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Zhang, Z., Larner, S.F., Liu, M.C. et al. Multiple alphaII-spectrin breakdown products distinguish calpain and caspase dominated necrotic and apoptotic cell death pathways. Apoptosis 14, 1289–1298 (2009). https://doi.org/10.1007/s10495-009-0405-z

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