Secreted neural apoptosis inhibiting proteins

An apoptosis inhibitor protein, secreted technology, applied in the field of molecular biology

Inactive Publication Date: 2007-01-10
AVENTIS PHARMA INC
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Moreover, the observed apoptosis after exposure to ceramide may not involve the action of NMDA receptors (Centeno et al., Neuroreport (1998) 9(18): 4199-4203; Moore et al., Br.J.Pharmacol.( 2002) 135(4): 1069-1077)

Method used

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Embodiment Construction

[0175] RNA extraction: The cultured cells or tissue homogenate were lysed in 1.5 ml Trizol (Gibco, Cat. No. 15596) per 10 cm culture dish or 50 mg tissue homogenate, respectively. The lysate was pipetted multiple times to homogenize the lysate (and subsequently transfer the cell lysate to a tube). After homogenization, the lysates were incubated at 30°C for 5 minutes to completely dissociate the nucleoprotein complexes. After incubation, chloroform was added to the lysate at a ratio of 0.2 ml chloroform (Sigma, Cat. No. C53 12) per 1 ml Trizol reagent, and the tube was shaken vigorously for 15 seconds. The lysates were then incubated at 30°C for 3 minutes. After incubation, the lysates were centrifuged at 12,000 xg for 15 minutes at 4°C. After centrifugation, remove the supernatant and rinse the remaining RNA pellet with 70% ethanol. The rinsed samples were then centrifuged at 7500 xg for 10 minutes at 4°C, and the resulting supernatant was discarded. The remaining RNA pel...

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Abstract

A novel neuroprotectant was identified by microarray analysis that is differentially expressed between the ventricular zone and the cortex of human adult and fetal brain. The secreted protein antagonizes Wnt action in Xenopus embryos. Methods are described for modulating free radical neurotoxicity by contacting cells with the protein, treating neuronal diseases associated with free radical-mediated cell death by administering the protein, determining neuroprotective genomic targets associated with select free radical toxicity pathways by screening with the protein and using the protein to identify other compounds that modulate the biological activity of the secreted protein and the cell machinery that reacts to the secreted protein.

Description

field of invention [0001] The invention belongs to the field of molecular biology, and in particular relates to the identification of a novel neuroprotective agent capable of regulating the cell death effect mediated by free radicals. Background of the invention [0002] Wnt and Frizzled [0003] Extracellular signaling molecules play a fundamental role as inducers of cell proliferation, migration, differentiation, and tissue morphogenesis during normal development (Finch et al., Proc. Natl. Acad. Sci. USA (1997) 94:6770-75) . In addition, such molecules function as regulators of apoptosis, the programmed cell death that plays an important role in the normal development and function of multicellular organisms. When dysfunctional, signaling molecules and apoptosis are involved in the pathogenesis of numerous diseases, see eg Thompson, Science (1995) 267:1456-1462. [0004] Apoptosis is involved in various normal and pathogenic biological events and can be induced by many u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47A61K38/17C12N5/07C12N5/073C12N5/0793
CPCA61K31/727G01N33/5017C12Q1/26G01N33/5011A61K38/1709G01N33/5047A61P21/04A61P25/00A61P25/16A61P25/18A61P25/28A61P39/06A61P43/00A61P9/00A61P9/10C07K14/47
Inventor J·梅里尔姚正斌W·佩特克O·霍克瓦G·基斯勒汪敏
Owner AVENTIS PHARMA INC
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