Oxidative stress reveals heterogeneity of FMRP granules in PC12 cell neurites.
PC12 cells are a well-known model of parasympathetic neurons. They have also been used to study the dynamics of heterologously expressed fragile X mental retardation (FMRP) granule trafficking down neurites. Here, we demonstrate that undifferentiated and differentiated PC12 cells harbor endogenous FMRP-containing granules. These granules are not stress granules because they do not associate with an authentic stress granule marker protein T-cell internal antigen 1 (TIA-1). Treatment with sodium arsenite induces stress granule formation in undifferentiated and differentiated PC12 cells. In NGF-treated cells, FMRP-containing stress granules are observed in the soma, neurites and growth cones by co-immunostaining with anti-TIA-1 antibody. These data demonstrate that all three microdomains respond similarly to oxidative stress. Nevertheless, we find significantly less co-localization of FMRP and TIA-1 and FMRP and its homologs in the neurites of differentiated PC12 cells treated with sodium arsenite than in the soma or growth cones. The heterogeneity of these granules suggests that FMRP has multiple roles in neurites.
Dolzhanskaya
Natalia
N
Biochemical Molecular Neurobiology Laboratory, Department of Molecular Biology, New York State Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, NY 10314, USA.
Merz
George
G
Denman
Robert B
RB
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D004347:Drug Interactions; D005455:Fluorescent Antibody Technique / Q000379:methods; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology; D020932:Nerve Growth Factor / Q000737:chemistry / Q000494:pharmacology; D016501:Neurites / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D018384:Oxidative Stress; D016716:PC12 Cells / Q000187:drug effects; D021381:Protein Transport / Q000502:physiology; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats
D051860:Fragile X Messenger Ribonucleoprotein 1; D020932:Nerve Growth Factor; D016376:Oligonucleotides, Antisense; D016601:RNA-Binding Proteins; C489666:Tial1 protein, rat