Detection of caspase-9 activation in the cell death of the Bcl-x-deficient mouse embryo nervous system by cleavage sites-directed antisera

Fujita, Eriko; Urase, Koko; Egashira, Jun; Miho, Yasuko; Isahara, Kyoko; Uchiyama, Yasuo; Isoai, Atushi; Kumagai, Hiroshi; Kuida, Keisuke; Motoyama, Noboru; Momoi, Takashi

doi:10.1016/s0165-3806(00)00066-3

Cited by 25 publications

(17 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1). Moreover, the Xenopus pattern of caspase 9 activation is similar to that obtained using a mouse caspase 9 antibody on mouse tissue extracts, which also showed two bands corresponding to the progressive proteolytic cleaved form of the mouse active caspase 9 (Fujita et al, 2000). Thus, it seems probable that the two bands detected using the Xenopus antibody correspond to the procaspase to active caspase conversion for the Xenopus caspase 9.…”

Section: Fig 2 Expression Of Majorsupporting

confidence: 67%

“…Why there is a shift between the two bands of activated caspase 9 at stage 59/60 is not clear. However, the cleavage pattern of Xenopus caspase 9 shows a strong similarity with mouse caspase 9, which also shows two bands corresponding to the same Asp cleavage sites as found in Xenopus (Fujita et al, 2000). After stage 60, the decrease in the signal intensity for the two cleaved caspase 9 fragments can be explained by the fact that, from this stage, tail regression increases strongly up to the end of metamorphosis.…”

Section: Discussionmentioning

confidence: 78%

“…In mouse, the five C-terminal amino acids of the P15 fragment (i.e., LDQLD-330, Fig. 1) have been used previously to synthesize successfully an antibody against active mouse caspase 9 (Fujita et al, 2000).…”

Section: Domainmentioning

confidence: 99%

See 2 more Smart Citations

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

et al. 2005

View full text Add to dashboard Cite

The climax of amphibian metamorphosis is marked by thyroid hormone-dependent tadpole tail resorption, implicating apoptosis of multiple cell types, including epidermal cells, fibroblasts, nerve cells, and muscles. The molecular cascades leading to and coordinating the death of different cell types are not fully elucidated. It is known that the mitochondrial pathway, and in particular the Bax and XR11 genes, regulates the balance between apoptosis and survival in muscle. However, the down-stream factors modulated by changes in mitochondrial permeability have not been studied in a functional context. To investigate further the mitochondrial-dependent pathway, we analyzed the regulation and the role of caspase 9 in Xenopus tadpoles. We report that caspase 9 mRNA is expressed in the tail before metamorphosis and increases before and during climax. Similarly, at the protein level, the production of active forms of caspase 9 increases in muscle tissue as metamorphosis progresses. To assess the functional role of caspase 9, we designed a dominant-negative protein. Overexpression of this dominant-negative abrogates both Bax-induced cell death in vitro and muscle apoptosis in vivo during natural metamorphosis. These findings consolidate a model of metamorphic muscle death that directly implicates the mitochondrial pathway and the apoptosome. Developmental Dynamics 233:76 -87, 2005.

show abstract

Section: Fig 2 Expression Of Majorsupporting

confidence: 67%

Section: Discussionmentioning

confidence: 78%

See 1 more Smart Citation

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

et al. 2005

View full text Add to dashboard Cite

show abstract

“…FLAG-tagged processing fragments of other caspases such as rat caspase-2D169, mouse caspase-3D175, caspase-7D198, caspase-D8D387, and caspase-9D368 were prepared as described previously. 13 Preparation of antisera against cleavage sites of caspase-12…”

Section: Methodsmentioning

confidence: 99%

Caspase-12 processing and fragment translocation into nuclei of tunicamycin-treated cells

et al. 2002

View full text Add to dashboard Cite

Excess endoplasmic reticulum (ER) stress induces processing of caspase-12, which is located in the ER, and cell death. However, little is known about the relationship between caspase-12 processing and cell death. We prepared antisera against putative caspase-12 cleavage sites (anti-m12D318 and anti-m12D341) and showed that overexpression of caspase-12 induced autoprocessing at D 318 but did not induce cell death. Mutation analysis confirmed that D 318 was a unique autoprocessing site. In contrast, tunicamycin, one of the ER stress stimuli, induced caspase-12 processing at the Nterminal region and the C-terminal region (both at D 318 and D 341 ) and cell death. Anti-m12D318 and anti-m12D341 immunoreactivities were located in the ER of the tunicamycin-treated cells, and some immunoreactivities were located around and in the nuclei of the apoptotic cells. Thus, processing at the N-terminal region may be necessary for the translocation of processed caspase-12 into nuclei and cell death induced by ER stress. Some of the caspase-12 processed at the N-terminal and C-terminal regions may directly participate in the apoptotic events in nuclei.

show abstract

“…The decreased apoptosis causes brain malformations and perinatal lethality. A specific antibody that can recognize the mature form of caspase-9 in situ has recently been generated (38). With this antibody, processing of caspase-9 has been detected in the lamina terminalis, dorsal root ganglia, and ventral hindbrain of mouse embryos at E11.5 (38).…”

Section: Generation Of Cleavagementioning

confidence: 99%

Identification of a Caspase-9 Substrate and Detection of Its Cleavage in Programmed Cell Death during Mouse Development

Nakanishi¹,

Maruyama²,

Shibata³

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

The caspase family of proteases represents the main machinery by which apoptosis occurs. In vitro studies have revealed that upstream caspases are activated in response to apoptotic stimuli, and the active caspases in turn process downstream effector caspases that are involved in the destruction of cellular structure. Caspase-9 is an upstream caspase that can become active in response to cellular damage, including deprivation of growth factors and exposure to oxidative stress in vitro. Little is known, however, about how activation of caspase-9 is temporally and spatially regulated in vivo, e.g. during development. We have identified vimentin as the first example of a caspase-9 substrate that is not a downstream procaspase. Immunohistochemical analysis, using a specific antibody against the vimentin fragments generated by caspase-9, showed that caspase-9 cleaves vimentin in apoptotic cells in the embryonic nervous system and the interdigital regions. This result is consistent with observations that gene knockouts of caspase-9 and its activator, Apaf-1, result in developmental defects in these tissues. Our results show that the specific antibody is useful for in situ detection of caspase-9 activation in programmed cell death.

show abstract

Detection of caspase-9 activation in the cell death of the Bcl-x-deficient mouse embryo nervous system by cleavage sites-directed antisera

Cited by 25 publications

References 31 publications

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

Caspase-12 processing and fragment translocation into nuclei of tunicamycin-treated cells

Identification of a Caspase-9 Substrate and Detection of Its Cleavage in Programmed Cell Death during Mouse Development

Contact Info

Product

Resources

About