Neuronal differentiation elicited by glial cell line‐derived neurotrophic factor and ciliary neurotrophic factor in adrenal chromaffin cell line tsAM5D immortalized with temperature‐sensitive SV40 T‐antigen

Abstract

To understand the characteristics of tsAM5D cells immortalized with the temperature-sensitive simian virus 40 large T-antigen, we first examined the responsiveness of the cells to ligands of the glial cell line-derived neurotrophic factor (GDNF) family. tsAM5D cells proliferated at the permissive temperature of 33 degrees C in response to either GDNF or neurturin, but not persephin or artemin. At the nonpermissive temperature of 39 degrees C, GDNF or neurturin caused tsAM5D cells to differentiate into neuron-like cells; however, the differentiated cells died in a time-dependent manner. Interestingly, ciliary neurotrophic factor (CNTF) did not affect the GDNF-mediated cell proliferation at 33 degrees C but promoted the survival and differentiation of GDNF-treated cells at 39 degrees C. In the presence of GDNF plus CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of various neuronal marker genes, indicating that the cells had undergone neuronal differentiation. In addition, tsAM5D cells caused to differentiate by GDNF plus CNTF at 39 degrees C became dependent solely on nerve growth factor (NGF) for their survival and neurite outgrowth. Moreover, upon treatment with GDNF plus CNTF, the dopaminergic phenotype was suppressed by the temperature shift. Thus, we demonstrated that tsAM5D cells had the capacity to differentiate terminally into neuron-like cells in response to GDNF plus CNTF when the oncogene was inactivated by the temperature shift. This cell line provides a useful model system for studying the role of a variety of signaling molecules for GDNF/CNTF-induced neuronal differentiation.

A sparse differential clustering algorithm for tracing cell type changes via single-cell RNA-sequencing data

“…Fxyd6 was also detected as a downregulated condition-specific gene for the naïve cells in cluster 1 and has previously been shown to respond to hypoxia ( 72 ). Condition-specific genes for the naïve cells in cluster 1 include genes involved in differentiation, Cntfr ( 73 ), targets of Notch signaling, Fjx1 ( 74 ) and genes involved in warding off neuronal disorders, Tpp1 ( 75 , 76 ). Some condition-dependent genes for the naïve cells in cluster 1 have also been shown to play a role in the functioning and differentiation of NSCs such as Fgfr3 ( 77 ), Sparcl1 ( 78–80 ) and Aqp4 ( 81 ), while Gpc5 has been shown to activate Hedgehog signaling ( 82 ), which plays a role in determining stem cell positional identity ( 83 ).…”

Paracrine influence of human perivascular cells on the proliferation of adenocarcinoma alveolar epithelial cells

“…PSPN was first identified as a glial cell line-derived neurotrophic factor (GDNF) that had neurotrophic effects on some neuronal cell types. For example, PSPN enhanced the morphological differentiation and phenotypic induction of dopaminergic neurons and could be considered as a cell therapy for Parkinson's disease [24252627]. However, very little is known about the role of PSPN in tumor progression.…”

Past, Present and Future of Human Chromaffin Cells: Role in Physiology and Therapeutics

“…In Spain, the first chromaffin cell transplant was performed in 1987 (López-Lozano et al 1989), and the researchers emphasized the need to carefully dissect the adrenal medulla to avoid including adrenocortical cells, which could impair the acquisition of a neuronal phenotype by glucocorticoid secretion (Unsicker et al 1978). On the other hand, this process is facilitated by trophic factors such as NGF, FGFb, and GDNF (López-Lozano et al 1999;Murata et al 2008a;Murata et al 2008b;Olson et al 1991). In effect, simultaneous grafting with other tissues seems to promote the survival of implanted cells (Date 1996;López-Lozano et al 1999;Olson et al 1991).…”