Phosphorylation and cytoplasmic localization of MAPK p38 during apoptosis signaling in bone marrow granulocytes of mice irradiated in vivo and the role of amifostine in reducing these effects
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2011-01-01
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Segreto, Helena Regina Comodo [UNIFESP]
Oshima, Celina Tizuko Fujiyama [UNIFESP]
Franco, Marcello Fabiano de [UNIFESP]
Silva, Maria Regina Regis [UNIFESP]
Egami, Mizue Imoto [UNIFESP]
Teixeira, Vicente de Paulo Castro [UNIFESP]
Segreto, Roberto Araujo [UNIFESP]
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We studied p38 phosphorylation and its intracellular localization during p53 and Puma (a p53 upregulated modulator of apoptosis) apoptotic signaling pathway in bone marrow granulocytes in mice irradiated in vivo and the role of the radioprotector amifostine in ameliorating these responses. Sixty-four C578L mice were randomly assigned in two non-irradiated (Ami-/rad - and Ami +/rad -) and two irradiated (Ami - /rad + and Ami +/rad +) groups. Animals received 400 mg/kg of amifostine i.p. 30 min prior to a single whole body radiation dose of 7 Gy. the experiments were performed using immunohistochemistry for caspase-3, cleaved caspase-3, p53, p-p53 (Ser 15), Puma, p38 and p-p38 (Thr 180/Tyr 182) protein expression. in addition transmission electron microscopy was used for ultrastructural characterization of apoptosis. Data showed that: (i) amifostine significantly reduced the number of apoptotic cells, (ii) p-p53 and Puma proteins were strongly immunostained in granulocytes after irradiation (Ami - /rad +), (iii) amifostine decreased the immunostaining of the proteins (Ami +/rad +), (iv) p38 was immunolocalized in physiological conditions in the nucleus and cytoplasm of granulocytes and neither radiation nor amifostine changed the protein immunostaining or its subcellular distribution, but influenced its activation, (v) radiation-induced p38 phosphorylation and its cytoplasmic accumulation during apoptosis signaling in granulocytes after whole body high radiation dose and amifostine markedly reduced these effects. (C) 2010 Elsevier GmbH. All rights reserved.
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Acta Histochemica. Jena: Elsevier Gmbh, Urban & Fischer Verlag, v. 113, n. 3, p. 300-307, 2011.