Mice overexpressing human caspase 3 appear phenotypically normal but exhibit increased apoptosis and larger lesion volumes in response to transient focal cerebral ischaemia

Kerr, Lorraine E.; McGregor, Ailsa L.; Amet, L; Asada, Tomohisa; Spratt, Christopher; Allsopp, Timothy E.; Harmar, Anthony J.; Shen, Sanbing; Carlson, G; Logan, Nicola; Kelly, John; Sharkey, John

doi:10.1038/sj.cdd.4401449

Cited by 28 publications

(18 citation statements)

References 72 publications

(91 reference statements)

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“…In transgenic mice, overexpression of the human caspase-3 gene did not induce morphogenetic defects during normal development but leads to greater apoptosis under stress conditions (Kerr et al, 2004). The transgenic mice showed significantly larger lesions when they were subjected to focal cerebral ischaemia-reperfusion injury.…”

Section: Discussionmentioning

confidence: 92%

Extensive apoptosis and abnormal morphogenesis in pro-caspase-3 transgenic zebrafish during development

Yamashita

Mizusawa

Hojo

et al. 2008

Journal of Experimental Biology

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SUMMARYThe pro-apoptotic caspase-3 gene has been shown to have key functions in the execution of apoptosis (programmed cell death) in vertebrate cells. However, the central role of caspase-3 in morphogenesis during development remains unclear. In this study, transgenic zebrafish that overexpress full-length pro-caspase-3 were generated to determine the effects of caspase genes on vertebrate morphogenesis and stress tolerance. The enhanced expression of the full-length pro-caspase-3 cDNA induced extremely high levels of caspase activity and extensive apoptosis in the transgenic embryos, and 33-46% of F2 embyos in the transgenic lines exhibited some form of morphological abnormality. Pro-caspase-3 transgenic zebrafish exhibited abnormal morphogenesis in the eyes, notochord, heart and yolk sac, suggesting that enhanced processing of pro-caspase-3 triggers significant apoptotic responses in the specific target tissues that are undergoing morphogenesis during development. The transgenic fish had reduced eye size and showed degeneration of the retina, including the photoreceptor cell layers, whereas pigmentation and lens formation were not affected. In addition, heart failure due to a weakened heartbeat and reduced circulation was noted in the pro-caspase-3 transgenic embryos. The transgenic embryos were markedly sensitive to stress conditions, such as UV irradiation at 2 or 5 mJ cm -2 . On the other hand, caspase-3 deficiency through injection of antisense morpholino oligo into embryos repressed apoptosis and enhanced stress tolerance after UV irradiation. Therefore, the caspase-3-mediated proapoptotic signalling pathway and its activation play critical roles in the induction of apoptosis and stress tolerance during zebrafish embryogenesis.

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Section: Discussionmentioning

confidence: 92%

Extensive apoptosis and abnormal morphogenesis in pro-caspase-3 transgenic zebrafish during development

Yamashita

Mizusawa

Hojo

et al. 2008

Journal of Experimental Biology

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“…We therefore examined the role of nucleophosmin in vivo using a mouse model of stroke. Previous data from our laboratory (Kerr et al, 2004b) and many others (Matsushita et al, 1998;Cao et al, 2001) have demonstrated Bax involvement in the apoptotic cell death that occurs following experimental transient occlusion of the MCA. Bax translocation has also been reported in both rats (Cao et al, 2001) and mice (Gao et al, 2005) using this model.…”

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confidence: 73%

“…Monofilament occlusion of the MCA was performed in adult male mice (25-30 g, approximately 10-12 weeks) as described previously (Kerr et al, 2004b). Animals were subjected to 30 min of ischaemia and killed at 3, 6, 12 and 24 h by trans-cardiac perfusion with 4% paraformaldehyde under deep anaesthesia (pentobarbitone, 60 mg/kg intraperitoneally).…”

Section: Proximity Assaymentioning

confidence: 99%

Nucleophosmin is a novel Bax chaperone that regulates apoptotic cell death

et al. 2006

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The proapoptotic B-cell lymphoma-2 family protein Bax is a key regulatory point in the intrinsic apoptotic pathway. However, the factors controlling the process of Bax activation and translocation to mitochondria have yet to be fully identified and characterized. We performed affinity chromatography using peptides corresponding to the mitochondrial-targeting region of Bax, which is normally sequestered within the inactive structure. The molecular chaperone nucleophosmin was identified as a novel Bax-binding protein by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Reciprocal co-immunoprecipitation and proximity assays confirmed the Bax-nucleophosmin protein-protein interaction and verified that nucleophosmin only bound to activated conformationally altered Bax. Confocal microscopy in a cell-based apoptosis model, demonstrated that nucleophosmin translocation from nucleolus to cytosol preceded Bax movement. Specific knockdown of nucleophosmin expression using RNAi attenuated apoptosis as measured by mitochondrial cytochrome c release and activation of the caspase cascade. In a mouse model of ischaemic stroke, subcellular fractionation studies verified that nucleophosmin translocation occurred within 3 h, at a time before Bax translocation but after Bax conformational changes have occurred. Thus, we have elucidated a novel molecular mechanism whereby Bax becomes activated and translocates to the mitochondria to orchestrate mitochondrial dysfunction and apoptotic cell death, which opens new avenues for therapeutic intervention.

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“…Furthermore, serum withdrawal, which induced the repression of Dicer expression, indeed increased the expression of caspase-3. It has been reported that mice overexpressing caspase-3 are phenotypically normal under physiological conditions, while exhibiting increased apoptosis in response to ischemia-reperfusion injury, implying the role of an increased expression of caspase-3 in the increased susceptibility to degenerative insults (23). These results indicated the causative role of Dicer repression in caspase-3 upregulation in response to serum withdrawal and suggested that the upregulation of caspase-3 in the settings of Dicer downregulation involves an increased sensitivity to apoptosis.…”

Section: Discussionmentioning

confidence: 99%

Downregulation of Dicer expression by serum withdrawal sensitizes human endothelial cells to apoptosis

Asada¹,

Takahashi

Isodono

et al. 2008

American Journal of Physiology-Heart and Circulatory Physiology

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though the modulated expression of Dicer is documented upon neoplastic transformation, little is known of the regulation of Dicer expression by environmental stimuli and its roles in the regulation of cellular functions in primary cells. In this study, we found that Dicer expression was downregulated upon serum withdrawal in human umbilical vein endothelial cells (HUVECs). Serum withdrawal induced a time-dependent repression of Dicer expression, which was specifically rescued by vascular endothelial cell growth factor or sphingosine-1-phosphate. When Dicer expression was silenced by short-hairpin RNA against Dicer, the cells were more prone to apoptosis under serum withdrawal, whereas the rate of apoptosis was comparable with control cells in the serum-containing condition. Real-time PCR-based gene expression profiling identified several genes, the expression of which was modulated by Dicer silencing, including adhesion and matrix-related molecules, caspase-3, and nitric oxide synthase 3 (NOS3). Dicer silencing markedly impaired migratory functions without affecting cell adhesion and repressed phosphorylation of focal adhesion kinase and proline-rich tyrosine kinase 2 in adherent HUVECs. Dicer knockdown upregulated caspase-3 and downregulated NOS3 expression, and serum withdrawal indeed increased caspase-3 and decreased NOS3 expression. Furthermore, the overexpression of Dicer in HUVECs resulted in a marked reduction in apoptosis upon serum withdrawal and a decreased caspase-3 and increased NOS3 expression. The inhibition of NOS activity by N -nitro-L-arginine methyl ester abrogated the effect of Dicer overexpression to rescue the cells from serum withdrawal-induced apoptosis. These results indicated that serum withdrawal decreases Dicer expression, leading to an increased susceptibility to apoptosis through the regulation of caspase-3 and NOS3 expression. caspase 3; nitric oxide synthase 3 APOPTOSIS IS A PROCESS OF innate cellular death, controlled by complex and diverse molecular mechanisms with considerable cell-type specificity. Apoptosis plays important roles in various aspects of biology from the development to a wide range of diseases such as cancers and cardiovascular diseases. In the vasculature, the integrity of the endothelial lining is essential for vascular homeostasis and for normal organ function, and endothelial cell apoptosis has been implicated not only in normal physiological events such as blood vessel development, homeostasis, and remodeling but also in pathological conditions associated with endothelial dysfunction such as inflammatory and immune disorders, tumor growth, and atherosclerosis (31, 32, 46). Although apoptotic processes are tightly regulated by extracellular factors and intracellular signalings, the precise molecular mechanisms governing endothelial cell apoptosis have not been fully elucidated, and understanding the regulation of apoptosis is of great importance for the advancement of endothelial biology and for developing novel therapeutic strategies.Dicer is a cytoplasmic RN...

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Mice overexpressing human caspase 3 appear phenotypically normal but exhibit increased apoptosis and larger lesion volumes in response to transient focal cerebral ischaemia

Cited by 28 publications

References 72 publications

Extensive apoptosis and abnormal morphogenesis in pro-caspase-3 transgenic zebrafish during development

Extensive apoptosis and abnormal morphogenesis in pro-caspase-3 transgenic zebrafish during development

Nucleophosmin is a novel Bax chaperone that regulates apoptotic cell death

Downregulation of Dicer expression by serum withdrawal sensitizes human endothelial cells to apoptosis

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