Biochemical and Biophysical Research Communications
Functional inhibition of the p75 receptor using a small interfering RNA
Section snippets
Materials and methods
Isolation of Schwann cells. Sciatic nerves were dissected from mice at postnatal day 4, treated with 0.1% collagenase type I (Sigma) and 0.25% trypsin (Gibco-BRL), and incubated with 200 U/ml DNase1 (Takara) for 30 min at 37 °C. The cells were triturated and plated on poly-l-lysine-coated Lab-Tek four-chamber tissue culture slides (Nunc) cultured in DMEM containing 10% fetal calf serum (FCS).
Isolation of DRG neurons. DRGs were removed from mice at postnatal day 9 and dissociated into single cells
siRNA construct
Several groups showed evidence for sequence-specific inhibition of the target gene when the appropriate siRNAs were transfected with somatic mammalian cell lines [18], [19]. siRNAs have been shown to act as guides for the enzymatic complex required for the degradation of the target mRNA [20]. Specific repression of target gene is caused by duplexes of 20–23 bp and the 21 nt duplexes with 2 nt 3′ overhangs were shown to be efficient in mediating RNAi in Drosophila[21]. In a recent study in
References (29)
- et al.
Neurotrophin signal transduction in the nervous system
Curr. Opin. Neurobol.
(2000) - et al.
The uniqueness of being a neurotrophin receptor
Curr. Opin. Neurobiol.
(2001) - et al.
Neurotrophin binding to the p75 receptor modulates Rho activity and axonal outgrowth
Neuron
(1999) - et al.
Neurotrophin stimulate the release of dopamine from rat mesencephalic neurons via Trk and p75Lntr receptors
J. Biol. Chem.
(1996) - et al.
Nerve growth factor (NGF)-induced calcium influx and intracellular calcium mobilization in 3T3 cells expressing NGF receptors
J. Biol. Chem.
(1999) - et al.
Targeted mutation of the gene encoding the low affinity NGF receptor p75 leads to deficits in the peripheral sensory nervous system
Cell
(1992) - et al.
Prior exposure to neurotrophins blocks inhibition of axonal regeneration by MAG and myelin via cAMP-dependent mechanism
Neuron
(1999) - et al.
RNAi: Double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals
Cell
(2000) - et al.
Biochemical and functional interactions between the neurotrophin receptors trk and p75NTR
EMBO J.
(1999) - et al.
The neurotrophin receptor p75NTR: novel functions and implications for disease of the nervous system
Nat. Neurosci.
(2002)
p75 is important for axon growth and Schwann cell migration during development
J. Neurosci.
Ceramide signaling downstream of the p75 neurotrophin receptor mediates the effects of nerve growth factor on outgrowth of cultured hippocampal neurons
J. Neurosci.
Nerve growth factor and its low-affinity receptor promote Schwann cell migration
Proc. Natl. Acad. Sci. USA
The low-affinity neurotrophin receptor p75 regulates the function but not the selective survival of specific subpopulations of sensory neurons
J. Neurosci.
Cited by (37)
Neurotropin® accelerates peripheral nerve regeneration in a rat sciatic nerve crush injury model
2024, Journal of Orthopaedic Science‘Nano–in–nano’ hybrid liposomes increase target specificity and gene silencing efficiency in breast cancer induced SCID mice
2017, European Journal of Pharmaceutics and BiopharmaceuticsCitation Excerpt :Among the various strategies employed in the treatment of diseases such as cancer, HIV infection, Parkinson’s disease, etc., gene therapy using siRNA has evolved as one of the prominent clinical modalities [1,2] siRNA regulates the gene expression through a mechanism known as RNA interference (RNAi), which is a natural cellular tool that is involved in the sequence–specific down regulation of gene expression levels [3].
Prokaryotic expression of recombinant human p75NTR-Fc fusion protein and its effect on the neurite outgrowth of dorsal root ganglia neuron
2009, Journal of Medical Colleges of PLAIL-1β promotes neurite outgrowth by deactivating RhoA via p38 MAPK pathway
2008, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Cell culture. DRG neurons were cultured as described previously [14]. DRGs from Wistar rats at postnatal day 9 or 10 were dissociated by incubation with 0.25% trypsin, 0.1% collagenase, and 200 U/ml DNase I.