Protective Effects of Neurotrophic Factors on Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL)-mediated Apoptosis of Murine Adrenal Chromaffin Cell Line tsAM5D

Abstract

We previously established the murine adrenal chromaffin cell line tsAM5D, which was immortalized with the temperaturesensitive simian virus 40 large T-antigen. tsAM5D cells have the capacity to differentiate into neuron-like cells in response to neurotrophic factors when the culture temperature is shifted from 33 to 39°C. In this model system, the temperature shift in the absence of neurotrophic factors led to cell death. Hoechst staining analysis revealed that typical apoptotic nuclei appeared in a time-dependent manner after the temperature shift. Upon shifting to 39°C, the degradation of T-antigen was accompanied by the transcriptional activation of p53 protein. Among the p53 target genes, death receptor 5 (DR5), which is the receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), showed the highest level of induction. Interestingly, TRAIL-neutralizing antibody protected tsAM5D cells from the temperature shift-induced apoptotic cell death by blocking the activation of caspase-8 and -3, indicating the involvement of TRAIL-mediated death signaling in the temperature shift-induced apoptosis. Glial cell line-derived neurotrophic factor (GDNF) inhibited the TRAIL-mediated activation of caspase-8 in tsAM5D cells exposed to 39°C and cooperated with basic fibroblast growth factor and ciliary neurotrophic factor. Interestingly, the temperature shift induced oligomerization of DR5, which is the earliest process necessary for transduction of the death signal. This oligomerization was inhibited by treatment with GDNF plus ciliary neurotrophic factor but not by that with GDNF alone or GDNF plus basic fibroblast growth factor. These results are discussed with respect to the intracellular mechanism underlying the protective function of neurotrophic factors against TRAIL-mediated death signaling.The temperature-sensitive tsA58 mutant of the simian virus 40 large T-antigen (tsSV40T) 2 (1) has been utilized for development of mammalian cell lines displaying differentiated characteristics. Cells immortalized with the tsSV40T oncogene proliferate under the permissive temperature of 33°C, and they differentiate at the nonpermissive temperature of 39°C (2, 3). An advantage of tsSV40T is the ability to study the differentiation function of the cell without the interference of the transforming oncogene at the nonpermissive temperature. In fact, several tsSV40T-immortalized neuronal and non-neuronal cell lines have been shown to retain their capacity to differentiate even when the immortalizing oncogene has been inactivated (4 -11). Therefore, the tsSV40T-transformed cell line can be useful for studying the regulatory mechanism by which target cells differentiate in response to extracellular stimuli. Tyrosine hydroxylase is the rate-limiting enzyme in the biosynthesis of the catecholamines dopamine, norepinephrine, and epinephrine (12, 13). Tyrosine hydroxylase is selectively expressed in the central and peripheral catecholaminergic neurons and in the adrenal medullary chromaffin cells. Previously, we demonstrat...

Catalytically active Yersinia outer protein P induces cleavage of RIP and caspase-8 at the level of the DISC independently of death receptors in dendritic cells

“…The expression of CD95 on DC is dependent on DC maturation as it was shown that 14% of mouse BMDC expressed CD95 at day 8 of cultivation [57] which is in accordance with the CD95 expression (13%) of DC used in this study. The TNF-related apoptosis-inducing ligand (TRAIL) mainly induces apoptosis in transformed (tumor) cells [58,59] sparing healthy, non-transformed cells [60][61][62]. However, some reports indicate that TRAIL may also trigger cell death in primary cells, i.e., mouse thymocytes [63,64].…”

Establishment and characterization of a noradrenergic adrenal chromaffin cell line, tsAM5NE, immortalized with the temperature-sensitive SV40 T-antigen

“…In fact, it was earlier reported that, in some of tsSV40T-immortalized cell lines, the upshift to the nonpermissive temperature not only induces growth arrest but also causes apoptotic cell death Yanai and Obinata, 1994;Zheng et al, 1994;Guenal et al, 1997;Walther et al, 1999;Tavelin et al, 1999;Wade et al, 1999). Because p53 has the ability to induce apoptotic cell death (Evan et al, 1998;Gottlieb et al, 1998), it has been suggested that the apoptotic death of tsSV40T-immortalized cells upon temperature upshift reflects the biological activity of p53 released from its complex with the thermolabile T-antigen (Murata et al, 2006;Yanai and Obinata, 1994;Zheng et al, 1994). GDNF and neurturin individually promoted the tsAM5NE cell survival and acted to cause the cells to differentiate into neuron-like cells.…”

Complex Changes in the Apoptotic and Cell Differentiation Programs during Initiation of the Hair Follicle Response to Chemotherapy

“…TRAIL-neutralizing antibody has been shown previously to effectively inhibit apoptosis in different models (Cantarella et al , 2010; Murata et al , 2006). Treatment of the follicles with TRAIL-neutralizing antibody alone did not result in any significant changes in keratinocyte proliferation or apoptosis, while expression of TRAIL in the hair matrix was markedly decreased suggesting for inhibition of TRAIL activity (data not shown).…”