Detection of activated Caspase-3 by a cleavage site-directed antiserum during naturally occurring DRG neurons apoptosis.
We prepared a cleavage site-directed antiserum against Caspase-3 (anti-p20/17), which reacts with the p20/17 fragment (p20/17) activated by cleavage but not proCaspase-3 (p32), and examined the relationship between the activation of Caspase-3 and apoptosis. We identified p20/17-positive cells where cell death occurs naturally: interdigits of the forelimbs, small intestine epithelium, thymus, trigeminal ganglia, and dorsal root ganglia of mouse embryos. Withdrawal of nerve growth factor induced the appearance of p20/17-positive cells with DNA fragmentation in the culture of dorsal root ganglia neurons, while DNA fragmentation was detected in both p20/17-positive and -negative neurons in dorsal root ganglia of mouse embryos. These results suggest that not only activation of Caspase-3 but also other molecular mechanism play a role in the naturally occurring dorsal root ganglia apoptosis. Cleavage site-directed antisera against Caspases will be useful for the analysis of the molecular mechanism of naturally occurring apoptosis during development.
Kouroku
Y
Y
Division of Development and Differentiation, National Institute of Neuroscience, NCNP, Tokyo, Japan.
Urase
K
K
Fujita
E
E
Isahara
K
K
Ohsawa
Y
Y
Uchiyama
Y
Y
Momoi
M Y
MY
Momoi
T
T
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000276:immunology / Q000378:metabolism*; D053938:DNA Fragmentation; D004622:Embryo, Mammalian / Q000201:enzymology; D005314:Embryonic and Fetal Development; D004789:Enzyme Activation / Q000502:physiology; D005727:Ganglia, Spinal / Q000502:physiology; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D051381:Rats
D009414:Nerve Growth Factors; C505409:Casp3 protein, mouse; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases