Nerve Growth Factor (NGF)-induced Calcium Influx and Intracellular Calcium Mobilization in 3T3 Cells Expressing NGF Receptors

Abstract

The neurotrophins have been implicated in the acute regulation of synaptic plasticity. Neurotrophin-stimulated presynaptic calcium uptake appears to play a key role in this process. To understand the mechanism of neurotrophin-stimulated calcium uptake, the regulation of calcium uptake and intracellular mobilization by nerve growth factor (NGF) was investigated using NIH 3T3 cells stably transfected with either the high affinity NGF receptor p140 trk (3T3-Trk) or the low affinity NGF receptor p75 NGFR (3T3-p75). In 3T3-Trk cells, NGF increased both calcium uptake and intracellular calcium mobilization. In 3T3-p75 cells, NGF increased calcium uptake but not intracellular calcium mobilization. K-252a alone increased intracellular calcium in 3T3-Trk cells but not in 3T3-p75 cells. Nifedipine, an inhibitor of calcium uptake through L-type calcium channels, inhibited the action of NGF on both 3T3-Trk cells and 3T3-p75 cells, indicating that both p140trk and p75 NGFR receptors are linked to nifedipine-sensitive L-type calcium channels. These studies show that either NGF receptor will support increases in intracellular calcium but that p140 trk does so by increasing both uptake and mobilization, whereas p75 NGFR does so by increasing uptake only.

Stable release of BDNF from the fibroblast cell line NIH3T3 grown on silicone elastomers enhances survival of spiral ganglion cells in vitro and in vivo

“…However, no intrinsic expression of other neurotrophic factors as like CNTF, FGF1/2, GDNF, NGF, NT3, NT4 and NT5 has been reported in NIH3T3 cells. Instead, this fibroblast cell line seem to present the ideal platform for genetical modifications for functional and structural related studies, ie calcium signalling, neurotrophic factor receptor binding, neuronal survival and bioactivity of neurotrophic factors in dose dependent manner (Dazert et al, 1998;Fryer et al, 1997;Glass et al, 1991;Jerregard et al, 2001;Jiang et al, 1999;Li et al, 2002;Wissel et al, 2008). Thus, we assume that the changes in SGC survival and neurite sprout observed in co-cultivation assays with non-modified NIH3T3 cells result from additional endogenous BDNF and/or other growth factor secreted from bystanding cells, in particular glial and satellite cells that are present in primary SGC cultures.…”

Chopper, a New Death Domain of the p75 Neurotrophin Receptor That Mediates Rapid Neuronal Cell Death

“…It has recently been shown that p75 NTR can regulate cellular calcium levels, causing an influx of calcium into the cell after NGF stimulation (68), suggesting that p75 NTR may interact with an ion channel or channel regulator. Furthermore, many studies show that membrane localization of calpain is necessary for its activation (66), which would explain the requirement for membrane attachment for Chopper killing action.…”

The Atypical Protein Kinase C-interacting Protein p62 Is a Scaffold for NF-κB Activation by Nerve Growth Factor

“…3D). Likewise, p62 down-regulation in NIH-3T3 cells expressing either p75 or TrkA receptor (20) (Fig. 3D) also abrogated NGF-induced activation of NF-B.…”