| |
 |
UNIVERSITY OF BUCHAREST
FACULTY OF PHYSICS Guest
2024-11-22 2:07 |
 |
|
|
|
Conference: Bucharest University Faculty of Physics 2007 Meeting
Section: Electricity and Biophysics
Title:
An in vitro model of NGF-induced apoptosis in rat primary sensory neurons
Authors:
Elfrida Pfeiffer1, Octavian Calborean2, Nicoleta Pascu2, Bogdan Amuzescu2
Affiliation:
1 National Institute of R&D for Optoelectronics, Bucharest
2 Department of Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest
E-mail
frida@inoe.inoe.ro
Keywords:
nerve growth factor, dorsal root ganglion, neuronal viability
Abstract:
Nerve Growth Factor (NGF) is a trophic factor required during development for the growth and survival of sympathetic neurons, sensory neurons, and neurons in the central nervous system (Nicol and Vasko, 2007). Beside classical high affinity tyrosine kinase receptors (trkA – half-activatory concentration 10^-11 M) which act via MAPK, protein kinase B/AKT and phospholipase C{gamma} to increase neuronal survival and neurite growth, low affinity non-specific receptors like neurotrophin 3 (p75-NTR3 – half-activatory concentration 10^-9 M) lead to JNK-mediated apoptosis. Within the present study we established an in vitro model of NGF-induced apoptosis that may be useful in the assessment of neuroprotective agents. NGF-7S of murine submaxillary gland origin was applied at different concentrations (2 ng/ml and 200 ng/ml) to primary sensory neurons obtained by enzymatic dissociation of dorsal root ganglia extracted from adult rats and maintained in culture for one day, in DMEM:F10 supplemented with 10% FBS. Neuronal viability was assessed by microspectrofluorimetry with propidium iodide. We noticed that there was a relationship between the NGF concentration and the extent of neuronal viability: at high NGF concentration (200 ng/ml) the proportion of dead cells (estimated by the integral of fluorescence intensity over a microscopic field using a low-magnification objective and a CCD camera for image acquisition) was almost significantly higher (one-tailed paired t test: t = 2.54, p = 0.06) compared to that of cells cultured in control conditions or exposed to a low NGF concentration (2 ng/ml).
|
|
|
|