Isolation of specific peptides that home to dorsal root ganglion neurons in mice.
We isolated peptides that home to mouse dorsal root ganglion (DRG) from a phage library expressing random 7-mer peptides fused to a minor coat protein (pIII) of the M13 phage. An in vitro biopanning procedure yielded 113 phage plaques after five cycles of enrichment by incubation with isolated DRG neurons and two cycles of subtraction by exposure to irrelevant cell lines. Analyses of the sequences of this collection identified three peptide clones that occurred repeatedly during the biopanning procedure. Phage-antibody staining revealed that the three peptides bound to DRG neurons of different sizes. To determine if the peptides would recognize neuronal cells in vivo, we injected individual GST-peptide-fusion proteins into the subarachnoid space of mice and observed the appearance of immunoreactive GST in the cytosol of DRG neurons with a similar size distribution as that observed in vitro, indicating that the GST-peptide-fusion proteins were recognized and taken up by different DRG neurons in vivo. The identification of homing peptide sequences provides a powerful tool for future studies on DRG neuronal function in vitro and in vivo, and opens up the possibility of neuron-specific drug and gene delivery in the treatment of diseases affecting DRG neurons.
Oi
Jiro
J
Department of Medicine, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan.
Terashima
Tomoya
T
Kojima
Hideto
H
Fujimiya
Mineko
M
Maeda
Kengo
K
Arai
Ryohachi
R
Chan
Lawrence
L
Yasuda
Hitoshi
H
Kashiwagi
Atsunori
A
Kimura
Hiroshi
H
D000595:Amino Acid Sequence / Q000502:physiology; D000818:Animals; D017104:Bacteriophage M13; D036022:Capsid Proteins / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D048429:Cell Size / Q000187:drug effects; D003001:Cloning, Molecular / Q000379:methods; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005822:Genetic Vectors / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009475:Neurons, Afferent / Q000187:drug effects* / Q000378:metabolism; D009619:Nociceptors / Q000187:drug effects / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism* / Q000494:pharmacology
D036022:Capsid Proteins; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins