Effective delivery of Pep-1-cargo protein into ischemic neurons and long-term neuroprotection of Pep-1-SOD1 against ischemic injury in the gerbil hippocampus.
We examined the intracellular delivery of Pep-1-cargo protein against transient ischemic damage in the hippocampal CA1 region in gerbils. For this study, we introduced green fluorescent protein (GFP) and constructed Pep-1-GFP protein. At 12h after Pep-1-GFP treatment, GFP fluorescence was shown in almost CA1 pyramidal neurons in ischemic animals; in the sham-operated group, GFP fluorescence was shown in a few pyramidal neurons. Next, we confirmed the long-term effects of Pep-1-Cu,Zn-superoxide dismutase 1 (SOD1) against ischemic damage. In behavioral test, locomotor activity was significantly increased in Pep-1- and Pep-1-SOD1-treated groups 1 day after ischemia/reperfusion; the locomotor activity in the Pep-1-treated group was higher than that of the Pep-1-SOD1-treated group. Thereafter, the locomotor activity in both groups was decreased with time. Four days after ischemia/reperfusion, the locomotor activity in the Pep-1-SOD1-treated group was similar to that of the sham group; in the Pep-1-treated group, the activity was lower than that of the sham group. In the histochemical study, the cresyl violet positive neurons in the Pep-1-SOD1-treated group were abundantly detected in the hippocampal CA1 region 5 days after ischemia/reperfusion. In biochemical study, SOD1 protein level and activity in all Pep-1-treated ischemic groups were significantly lower than that of the Pep-1-SOD1-treated group. Our results indicate that Pep-1-cargo fusion proteins can be efficiently delivered into neurons in the ischemic hippocampus, and that Pep-1-SOD1 treatment in ischemic animals show a neuroprotection in the ischemic hippocampus for a long time.
Cho
Jun Hwi
JH
Department of Emergency Medicine, College of Medicine, Kangwon National University, Chuncheon 200-701, South Korea.
Hwang
In Koo
IK
Yoo
Ki-Yeon
KY
Kim
So Young
SY
Kim
Dae Won
DW
Kwon
Young-Guen
YG
Choi
Soo Young
SY
Won
Moo-Ho
MH
D000818:Animals; D001921:Brain / Q000473:pathology; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology; D016923:Cell Death; D003543:Cysteamine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D016503:Drug Delivery Systems; D005849:Gerbillinae; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006624:Hippocampus / Q000473:pathology*; D002546:Ischemic Attack, Transient / Q000188:drug therapy / Q000473:pathology; D008297:Male; D009043:Motor Activity / Q000187:drug effects; D009474:Neurons / Q000473:pathology*; D018696:Neuroprotective Agents; D010455:Peptides / Q000008:administration & dosage / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000008:administration & dosage / Q000494:pharmacology*; D000072105:Superoxide Dismutase-1
D018696:Neuroprotective Agents; C503924:Pep-1 peptide; D010455:Peptides; D049452:Green Fluorescent Proteins; D003543:Cysteamine; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1