[Preliminary isolation and identification of neurotrophic factors sensory neurons in spinal dorsal horn of embryonic chicken].
Our previous studies have demonstrated that the extract from dorsal horn of embryonic chicken can promote apparently the neurite growth of spinal ganglion during development. In this study, we investigated progressively the neurotrophic factors for primary sensory neurons in spinal dorsal horn. The tissues of spinal dorsal horn from five hundred Hamburger Stage-40 embryonic chickens were taken. After homogenization and centrifugation, the supernatant was subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) for analyzing the protein in dorsal horn. The simple protein band fishing by cell (SPBFC) technique was employed in finding the protein bands with neurons attached, and then such protein bands, were taken and transferred to nitric cellulose paper for Western hybridization analysis by antibody for NGF, BDNF, NT-3 and GDNF respectively. The results of SPBFC showed that some neurons were attached to gel within the scope of mobility relative front (Rf) 0.10-0.19, 0.30-0.35, 0.50-0.55, 0.60-0.67 and 0.90-0.94 respectively, indicating there were some neurotrophic factors for maintaining neuron survival in the above Rf scope. Moreover, the result of Western hybridization showed that the positive hybridization bands in each scope were that of NGF, BDNF, GDNF; NGF, BDNF, NT-3; NGF, BDNF, NT-3; NGF, NT-3 and BDNF, NT-3 respectively; indicating that the neurotrophic effect of spinal dorsal horn tissue on primary sensory neurons involves NGF, BDNF, NT-3 and GDNF. These also suggest that the neuronal survival effect in each Rf scope involves at least two or more neurotrophic factors.
Wang
T
T
Center for Research on Neuroscience, Kunming Medical College, Kunmin 650031.
Feng
Z
Z
Zou
X
X
Li
L
L
Feng
Y
Y
D000818:Animals; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel; D005727:Ganglia, Spinal / Q000737:chemistry*; D009414:Nerve Growth Factors / Q000032:analysis / Q000302:isolation & purification*; D009475:Neurons, Afferent / Q000737:chemistry
D009414:Nerve Growth Factors