Characterization of CoCl2-induced reactive oxygen species (ROS): Inductions of neurite outgrowth and endothelin-2/vasoactive intestinal contractor in PC12 cells by CoCl2 are ROS dependent, but those by MnCl2 are not.
CoCl(2) and MnCl(2) are hypoxic mimetic agents. We previously found that expression of ET-2/VIC, one of hypoxia-related factors, and the induction of neurite outgrowth in PC12 cells through ROS induced by CoCl(2). MnCl(2) also are known to induce neurite outgrowth in PC12 cells. However, it is unclear whether the mechanism of the effect induced by these metals is same. In the present study, we evaluated biological effects induced by MnCl(2) and compared with those induced by CoCl(2). Furthermore, we analyzed sources of CoCl(2)-induced ROS generation. MnCl(2) up-regulated ET-2/VIC gene expression and ET-2/VIC peptide production as CoCl(2) did, but not affect ET-1 gene expression, in the neurite outgrowth of PC12 cells. NAC did not at all inhibit the effects induced by MnCl(2). Furthermore, addition of MnCl(2) to the culture medium did not generate ROS as CoCl(2) did. These results indicate that ET-2/VIC expression is a common pathway in neurite outgrowth induced by CoCl(2) and MnCl(2), but the effects induced by CoCl(2) are ROS dependent, whereas the effects induced by MnCl(2) are ROS independent. Taken together, the mechanism for the effects by CoCl(2) was different from that by MnCl(2). The ROS, were not decomposed by catalase or SOD, were rapidly generated by reaction of CoCl(2) mainly with components of HS rather than with FBS or DMEM. Some ROS generated by reaction of CoCl(2) with components of HS may participate in the observed neurite outgrowth of PC12 cells.
Kotake-Nara
Eiichi
E
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Ibaraki 305-8566, Japan. CQX05272@nifty.com
Saida
Kaname
K
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002712:Chlorides / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology*; D019333:Endothelin-2 / Q000096:biosynthesis* / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D036341:Intercellular Signaling Peptides and Proteins; D017895:Manganese Compounds / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects*; D016716:PC12 Cells; D010455:Peptides / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*
D002712:Chlorides; D019333:Endothelin-2; D036341:Intercellular Signaling Peptides and Proteins; D017895:Manganese Compounds; D010455:Peptides; D017382:Reactive Oxygen Species; C059568:vasoactive intestinal constrictor; D003035:Cobalt; C018021:cobaltous chloride; C025340:manganese chloride