T. Yoshida et al., BOTH HUMAN FAS AND CHIMERIC FAS TNFR-1 MEDIATE COMPLETE CELL-DEATH INMURINE NIH/3T3 CELLS WITH LESS DISTINCT NUCLEAR FRAGMENTATION/, Proceedings of the Japan Academy. Series B Physical and biological sciences, 72(1), 1996, pp. 16-21
Necrotic and apoptotic changes have been rather conceptually defined,
and have shown varying morphological and biochemical features dependin
g on signaling pathways and cell lineages. In an attempt to investigat
e apoptotic and necrotic signals, we constructed a chimer ic gene enco
ding an extracellular human Fas and intracytoplasmic human TNFR-1 (TNF
receptor p55) and introduced it into murine NIH/3T3 cells for their h
eterotopic expression. Upon stimulation with an agonistic anti-human F
as monoclonal antibody those NIH/3T3 cell clones with the chimeric rec
eptor showed cytopathic changes quite similar to those with the wild t
ype Fas, although both receptors possess 'death' domains with differen
t binding proteins. In this forced induction of death of the fibroblas
tic cells, we found thus that: 1) Fas signal induced complete cell dea
th, 2) cytopathic changes in both NIH/3T3 derivatives were quite simil
ar with irregular nuclear fragmentation and less distinct DNA fragment
ation, and 3) intracytoplasmic domains were interchangeable between Fa
s and TNFR-1. This system may contribute to further analyses of the ce
ll death mechanisms, including downstream Fas/TNFR-1 signaling.