Fluorescent peptide-PNA chimeras for imaging monoamine oxidase A mRNA in neuronal cells.
Bioconjug Chem
; 23(2): 158-63, 2012 Feb 15.
Article
em En
| MEDLINE
| ID: mdl-22239616
ABSTRACT
Monoamine oxidases (MAO) catalyze the oxidative deamination of many biogenic amines and are integral proteins found in the mitochondrial outer membrane. Changes in MAO-A levels are associated with depression, trait aggression, and addiction. Here we report the synthesis, characterization, and in vitro evaluation of novel fluorescent peptide-peptide nucleic acid (PNA) chimeras for MAOA mRNA imaging in live neuronal cells. The probes were designed to include MAOA-specific PNA dodecamers, separated by an N-terminal spacer to a µ-opioid receptor targeting peptide (DAMGO), with a spacer and a fluorophore on the C-terminus. The probe was successfully delivered into human SH-SY5Y neuroblastoma cells through µ-opioid receptor-mediated endocytosis. The K(d) by flow cytometry was 11.6 ± 0.8 nM. Uptake studies by fluorescence microscopy showed â¼5-fold higher signal in human SH-SY5Y neuroblastoma cells than in negative control CHO-K1 cells that lack µ-opioid receptors. Moreover, a peptide-mismatch control sequence showed no significant uptake in SH-SY5Y cells. Such mRNA imaging agents with near-infrared fluorophores might enable real time imaging and quantitation of neuronal mRNAs in live animal models.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
RNA Mensageiro
/
Ácidos Nucleicos Peptídicos
/
Imagem Molecular
/
Corantes Fluorescentes
/
Monoaminoxidase
/
Neurônios
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article