A novel peptide, colivelin, prevents alcohol-induced apoptosis in fetal brain of C57BL/6 mice: signaling pathway investigations.
Fetal alcohol exposure is known to induce cell death through apoptosis. We found that colivelin (CLN), a novel peptide with the sequence SALLRSIPAPAGASRLLLLTGEIDLP, prevents this apoptosis. Our initial experiment revealed that CLN enhanced the viability of primary cortical neurons exposed to alcohol. We then used a mouse model of fetal alcohol exposure to identify the intracellular mechanisms underlying these neuroprotective effects. On embryonic day 7 (E7), weight-matched pregnant females were assigned to the following groups: (1) ethanol liquid diet 25% (4.49% v/v) ethanol derived calories; (2) pair-fed control; (3) normal chow; (4) ethanol liquid diet combined with administration (i.p.) of CLN (20 microg/20 g body weight); and (5) pair-fed combined with administration (i.p.) of CLN (20 microg/20 g body weight). On E13, fetal brains were collected and assayed for TdT-mediated dUTP nick end labeling staining, caspase-3 colorimetric assay, enzyme-linked immunosorbent assay, and Meso scale discovery electrochemiluminescence. CLN blocked the alcohol-induced decline in brain weight and prevented alcohol-induced: apoptosis, activation of caspase-3 and increases of cytosolic cytochrome c, and decreases of mitochondrial cytochrome c Analysis of proteins in the upstream signaling pathway revealed that CLN down-regulated the phosphorylation of the c-Jun N-terminal kinase. Moreover, CLN prevented alcohol-induced reduction in phosphorylation of BAD protein. Thus, CLN appears to act directly on upstream signaling proteins to prevent alcohol-induced apoptosis. Further assessment of these proteins and their signaling mechanisms is likely to enhance development of neuroprotective therapies.
Sari
Y
Y
Program in Neuroscience, Indiana University Bloomington, IN 47405, USA. ysari@indiana.edu
Chiba
T
T
Yamada
M
M
Rebec
G V
GV
Aiso
S
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001921:Brain / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D045304:Cytochromes c / Q000378:metabolism; D004622:Embryo, Mammalian; D004789:Enzyme Activation; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D047908:Intracellular Signaling Peptides and Proteins / Q000493:pharmacokinetics / Q000494:pharmacology*; D020935:MAP Kinase Signaling System; D018811:Maternal Exposure; D008431:Maternal-Fetal Exchange; D051379:Mice; D008810:Mice, Inbred C57BL; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000493:pharmacokinetics / Q000494:pharmacology*; D009929:Organ Size; D010766:Phosphorylation; D011247:Pregnancy; D014018:Tissue Distribution
C508523:Colivelin; D047908:Intracellular Signaling Peptides and Proteins; D018696:Neuroprotective Agents; D000431:Ethanol; D045304:Cytochromes c; D020928:Mitogen-Activated Protein Kinases; D053148:Caspase 3