Down-regulation of amyloid precursor protein by peptide nucleic acid oligomer in cultured rat primary neurons and astrocytes.
The amyloid precursor protein (APP) and its proteolytic cleavage products, the amyloid beta peptides, have been implicated as a cause of Alzheimer's disease. Peptide nucleic acids (PNA), the DNA mimics, have been shown to block the expression of specific proteins at both transcriptional and translational levels. Generally, the cellular uptake of PNA is low. However, recent studies have indicated that the effect of unmodified antisense PNA uptake is more pronounced in nervous tissue. In this study we have shown that biotinylated PNA directed to the initiator codon region of the APP mRNA (-4 - +11) was taken up into the cytoplasm of primary rat cerebellar granule cells and cortical astrocytes, using fluorescence and confocal microscopy studies. Uptake of PNA was faster in neurons than in astrocytes. Western blotting analysis showed that APP was strongly down-regulated in both neurons and astrocytes. Thus, unmodified PNA can be used for studies on the function of APP in neurons and astrocytes.
Adlerz
Linda
L
Department of Neurochemistry and Neurotoxicology, Stockholm University, SE-10691 Stockholm, Sweden.
Soomets
Ursel
U
Holmlund
Linda
L
Viirlaid
Säde
S
Langel
Ulo
U
Iverfeldt
Kerstin
K
D000199:Actins / Q000145:classification / Q000187:drug effects / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000187:drug effects / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D007150:Immunohistochemistry; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D020135:Peptide Nucleic Acids / Q000378:metabolism / Q000494:pharmacology*; D016372:RNA, Antisense / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D000199:Actins; D016564:Amyloid beta-Protein Precursor; D010446:Peptide Fragments; D020135:Peptide Nucleic Acids; D016372:RNA, Antisense