Effects of Reg-2 on survival of spinal cord neurons in vitro.
Regeneration gene protein 2 (Reg-2) is a small secreted protein expressed in motor and sensory neurons of spinal cord during developmental stages and following injury of peripheral nerves. Reg-2 appears to act as a neurotrophic factor and protects injured neurons from death during regeneration. To illustrate these potential protective effects in vitro, we investigated the blocking effects of Reg-2 antibodies on the survival of primary cultured spinal cord neurons and astrocytes, as well as on neurite outgrowth. In addition, the effects of Reg-2 in neuron injury models induced by peroxide and mitochondrial poisoning were assessed. Our results showed that Reg-2 antibody markedly reduced survival and neurite outgrowth from neurons, whereas astrocyte survival was unaffected. Addition of Reg-2 into the culture medium had no effect on neuron survival or neurite outgrowth. However, the addition of the Reg-2 into culture media after peroxide treatment or cellular hypoxia insult induced by mitochondrial poisoning can reduce lactate dehydrogenase release levels and cell death. Thus, the data suggests that Reg-2 is essential for the survival and neurite outgrowth of developing spinal cord neurons but not the survival of glial cells, and that Reg-2 plays protective effects on spinal cord neurons against injury in vitro.
Fang
Marong
M
Institute of Anatomy and Cell Biology, Zhejiang University, China.
Huang
Jian-Ying
JY
Ling
Shu-Cai
SC
Rudd
John A
JA
Yew
David T
DT
Han
Shu
S
D000818:Animals; D000951:Antigens, Neoplasm / Q000494:pharmacology / Q000502:physiology*; D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects; D014408:Biomarkers, Tumor / Q000494:pharmacology / Q000502:physiology*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D037181:Lectins, C-Type / Q000502:physiology*; D008928:Mitochondria / Q000187:drug effects; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D016877:Oxidants / Q000494:pharmacology; D000073718:Pancreatitis-Associated Proteins; D010545:Peroxides / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000196:embryology*
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D037181:Lectins, C-Type; D016877:Oxidants; D000073718:Pancreatitis-Associated Proteins; D010545:Peroxides; D012333:RNA, Messenger; C000612142:Reg2 protein, rat; D007770:L-Lactate Dehydrogenase