The effect of neurotrophic factors on morphology, TRPV1 expression and capsaicin responses of cultured human DRG sensory neurons.
We have studied the effect of key neurotrophic factors (NTFs) on morphology, levels of the vanilloid receptor-1 (TRPV1) and responses to capsaicin in adult human sensory neurons in vitro. Avulsed dorsal root ganglia (DRG, n = 5) were cultured with or without a combination of nerve growth factor (NGF), glial cell (line)-derived growth factor (GDNF) and neurotrophin3 (NT3) for 5 days. In the absence of NTFs, the diameter of neurons ranged from 20 to 100 microm (mean 42 +/- 4 microm). Adding NTFs caused a significant increase in neuronal sizes, up to 120 microm (mean diameter 62 +/- 5 microm, P < 0.01, t-test), an overall 35% increase of TRPV1-positive neurons (P < 0.003), and notably of large TRPV1-positive neurons > 80 microm (P < 0.05). Responses to capsaicin were significantly enhanced with calcium ratiometry (P < 0.0001). Short duration (1h) exposure of dissociated sensory neurons to NTFs increased numbers of TRPV1-positive neurons, but not of TRPV3, Nav 1.8 and IK1 and the morphological size-distribution remained similar to intact post-mortem DRG neurons. NTFs thus increase size, elevate TRPV1 levels and enhance capsaicin responses in cultured human DRG neurons; these changes may relate to pathophysiology in disease states, and provide an in vitro model to study novel analgesics.
Peripheral Neuropathy Unit, Imperial College, Area A, Ground Floor, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK.
U
U
Anand
W R
WR
Otto
M A
MA
Casula
N C
NC
Day
J B
JB
Davis
C
C
Bountra
R
R
Birch
P
P
Anand
D000328:Adult; D002211:Capsaicin / Q000494:pharmacology*; D048429:Cell Size; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000166:cytology*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism; D006801:Humans; D008297:Male; D062559:NAV1.8 Voltage-Gated Sodium Channel; D020932:Nerve Growth Factor / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism*; D009475:Neurons, Afferent / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020933:Neurotrophin 3 / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated / Q000096:biosynthesis; D015222:Sodium Channels / Q000096:biosynthesis; D050916:TRPV Cation Channels / Q000096:biosynthesis*
C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D062559:NAV1.8 Voltage-Gated Sodium Channel; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D024681:Potassium Channels, Calcium-Activated; C568317:SCN10A protein, human; D015222:Sodium Channels; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; C459445:TRPV3 protein, human; D020932:Nerve Growth Factor; D002211:Capsaicin