Method for alleviating phenotype of degenerative disease <i>Drosophila </i>model by using low-dose radiation

Abstract

The present invention relates to a degenerative disease model Drosophila. More particularly, a technique is disclosed for alleviating the phenotypes of a degenerative disorder symptom of Drosophila. Using this technique, illumination with low dose radiation on a degenerative disease model Drosophila alleviates symptoms of the degenerative disease.

Claims

1. A method of alleviating a phenotype of an Alzheimer's disease Drosophila model, the method comprising applying low-dose radiation, at 0.05 Gy, to a Drosophila embryo, wherein the Alzheimer's disease Drosophila model is GMR>Aβ42.

2. The method of claim 1, wherein the phenotype of the Alzheimer's disease is decreased mobility.

3. The method of claim 1, wherein the phenotype of the Alzheimer's disease is induction of apoptosis.

4. A method of alleviating a phenotype of an Alzheimer's disease Drosophila model, the method comprising: applying low-dose radiation, at 0.05 Gy, to a Drosophila embryo, wherein the Alzheimer's disease Drosophila model is elav>Aβ42.

5. The method of claim 4, wherein the phenotype of the Alzheimer's disease is a decreased mobility or induction of apoptosis.

6. The method of claim 1, wherein the phenotype of the Alzheimer's disease is induction of apoptosis.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows the effect of alleviating the rough eye phenotypem of an Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation;

(2) FIG. 2 shows the effect of reducing apoptosis of the Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation;

(3) FIG. 3 shows the effect of alleviating the decreased climbing ability phenotype of an Alzheimer's disease Drosophila model (elav>Aβ42) using low-dose radiation; and

(4) FIG. 4 shows the effect of reducing apoptosis of the Alzheimer's disease Drosophila model (elav>Aβ42) using low-dose radiation.

DETAILED DESCRIPTION OF THE EMBODIMENTS

(5) Hereinafter, a detailed description will be given of the present invention.

(6) An embodiment of the present invention pertains to a method of alleviating the phenotypes of a degenerative disease of Drosophila using low-dose radiation, the method comprising collecting a Drosophila embryo and treating low-dose radiation to the Drosophila embryo.

(7) Here, Drosophila is an Alzheimer's disease Drosophila model. The Alzheimer's disease Drosophila model is GMR>Aβ42 or elav>Aβ42. GMR>Aβ42 and elav>Aβ42 are Alzheimer's disease Drosophila models and are Drosophila models able to induce tissue-specific overexpression of Aβ42, a representative protein of Alzheimer's disease pathology, in the neurons and eyes using the UAS-GAL4 system.

(8) The low-dose radiation is preferably applied so that the cumulative dose thereof is 0.05 to 0.1 Gy (.sup.137Cs, 0.8 Gy/min), and more preferably 0.05 Gy.

(9) The degenerative disease is Alzheimer's disease. Alzheimer's disease is one of the main causes of senile dementia. The cause of Alzheimer's disease has been found to be highly associated with beta-amyloid protein. When beta amyloid is excessively produced in the body and accumulates in the brain, the function of the neurons deteriorates and thus Alzheimer's disease develops. Beta amyloid paralyzes or deteriorates the function of mitochondria in the neurons, thereby increasing the amount of reactive oxygen species released from the mitochondria. The reactive oxygen species increased makes a fatal wound on intracellular protein or DNA, which in turn results in damage or apoptosis of neurons in brain.

(10) The phenotypes of the degenerative disease include decreased climbing ability and induction of apoptosis.

(11) Another embodiment of the present invention pertains to a neurodegenerative disease Drosophila model, in which the phenotypes of the degenerative disease are alleviated by treating low-dose radiation to a Drosophila embryo.

(12) Here, Drosophila is an Alzheimer's disease Drosophila model. Also, the Alzheimer's disease Drosophila model is GMR>Aβ42 or elav>Aβ42.

(13) The low-dose radiation is preferably applied so that a cumulative dose thereof is 0.05 to 0.1 Gy (.sup.137Cs, 0.8 Gy/min), and more preferably 0.05 Gy.

(14) The degenerative disease is Alzheimer's disease.

(15) The phenotypes of the neurodegenerative disease include decreased climbing ability and induction of apoptosis.

(16) A better understanding of the present invention will be given through the following examples. These examples are merely set forth to illustrate the present invention but are not to be construed as limiting the scope of the present invention, as is apparent to those skilled in the art.

EXAMPLE 1

(17) Evaluation of effect of alleviating rough eye phenotype of Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation

(18) In order to evaluate the effect of alleviating the phenotype of Alzheimer's disease using low-dose radiation, an Alzheimer's disease Drosophila model (GMR>Aβ42) (Crowther et al., 2003, Neuroscience; Finelli et al., 2004, Mol. Cell. Neurosci.), exhibiting a rough eye phenotype by overexpressing Aβ42, a causative protein of Alzheimer's disease, specifically in the eyes through GMR-GAL4, was used. As a control, Drosophila having only GMR-GAL4 was used, and all Drosophila samples were cultured in an incubator at a temperature of 25° C. and a humidity of 60%.

(19) Respective embryos were collected from control Drosophila (GMR-GAL4) and Alzheimer's disease Drosophila model (GMR>Aβ42) for 6 hr. To evaluate the phenotype alleviation effect of cumulative doses, the embryos of the Alzheimer's disease Drosophila model (GMR>Aβ42) were divided into five groups, four groups of which were irradiated respectively at doses of 0.05 Gy, 0.1 Gy, 0.2 Gy, and 4 Gy (.sup.137Cs, 0.8 Gy/min). After irradiation of the low-dose radiation, these embryos were grown in an incubator until adulthood, and the eyes of individual adult Drosophila groups were observed using a microscope.

(20) The Drosophila eye microscopy images for confirming the effect of alleviating the rough eye phenotype of the Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation are shown in FIG. 1. Based on this result, the rough eye phenotype of the Alzheimer's disease Drosophila model (GMR>Aβ42) was confirmed to be alleviated at cumulative doses of 0.05 Gy and 0.1 Gy, and the phenotype alleviation effect was insignificant at 0.2 Gy.

EXAMPLE 2

(21) Evaluation of effect of reducing apoptosis of Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation

(22) Respective embryos were collected from control Drosophila (GMR-GAL) and the Alzheimer's disease Drosophila model (GMR>Aβ42) for 6 hr in the same manner as in Example 1. The embryos of the Alzheimer's disease Drosophila model (GMR>Aβ42) were divided into three groups, two groups of which were irradiated respectively at doses of 0.05 Gy and 4 Gy (.sup.137Cs, 0.8 Gy/min). After irradiation of the low-dose radiation, the embryos were grown in an incubator until they became 3.sup.rd instar larvae, and the larval eye discs were dissected, stained with acridine orange (AO), and then observed with a fluorescence microscope.

(23) The results of observation with a fluorescence microscope for confirming the effect of reducing the apoptosis of Alzheimer's disease Drosophila model (GMR>Aβ42) using low-dose radiation are shown in FIG. 2. Based on this result, apoptosis (the portion stained black in FIG. 2), known to be induced due to overexpression of Aβ42 in the Alzheimer's disease Drosophila model (GMR>Aβ42), was confirmed to be reduced by low-dose radiation of 0.05 Gy, whereas high-dose radiation of 4 Gy was found to increase apoptosis.

EXAMPLE 3

(24) Evaluation of effect of alleviating decreased climbing ability phenotype of Alzheimer's disease Drosophila model (elav-Aβ42) using low-dose radiation

(25) In order to evaluate the effect of alleviating the phenotype of Alzheimer's disease using low-dose radiation, an Alzheimer's disease Drosophila model (elav>Aβ42), exhibiting a decreased climbing ability phenotype through overexpression of the Aβ42 protein specifically in neurons through elav-GAL4, was used. As a control, Drosophila having only elav-GAL4 was used, and all Drosophila samples were cultured under incubation conditions of a temperature of 25° C. and a humidity of 60%.

(26) Respective embryos were collected from control Drosophila (elav-GAL4) and the Alzheimer's disease Drosophila model (elav>Aβ42) for 6 hr. The embryos of the Alzheimer's disease Drosophila model (elav>Aβ42) were divided into three groups, two groups of which were irradiated respectively at cumulative doses of 0.05 Gy and 4 Gy (.sup.137Cs, 0.8 Gy/min). After irradiation of the low-dose radiation, these embryos were grown in an incubator until adulthood, and male Drosophila samples were collected on the third day of adulthood, and the climbing ability of Drosophila groups was tested. The climbing ability test was performed in the same manner as in Hwang et al., (2013) PLoS Genetics.

(27) The results of evaluation of the effect of alleviating the decreased climbing ability phenotype of the Alzheimer's disease Drosophila model (elav>Aβ42) using low-dose radiation are shown in FIG. 3. Based on this result, the climbing ability was improved in the Alzheimer's disease Drosophila model (elav>Aβ42) at a cumulative dose of 0.05 Gy, and was significantly degraded by high-dose radiation of 4 Gy.

EXAMPLE 4

(28) Evaluation of effect of reducing apoptosis of Alzheimer's disease Drosophila model (elav>Aβ42) using low-dose radiation

(29) Respective embryos were collected from control Drosophila (elav-GAL4) and the Alzheimer's disease Drosophila model (elav>Aβ42) for 6 hr in the same manner as in Example 3, and the embryos of the Alzheimer's disease Drosophila model (elav>Aβ42) were divided into three groups, two groups of which were irradiated respectively at cumulative doses of 0.05 Gy and 4 Gy (.sup.137Gy, 0.8 Gy/min). After irradiation of the low-dose radiation, the embryos were grown in an incubator until they became larvae, and the brains of the larvae were dissected, stained with acridine orange (AO), and then observed with a fluorescence microscope.

(30) The results of observation with a fluorescence microscope for confirming the effect of reducing apoptosis of the Alzheimer's disease Drosophila model (elav>Aβ42) using low-dose radiation are shown in FIG. 4. Based on this result, apoptosis (the portion stained black in the image) induced due to overexpression of Aβ42 in the Alzheimer's disease Drosophila model (elav>Aβ42) was confirmed to be reduced by low-dose radiation of 0.05 Gy, and high-dose radiation of 4 Gy was found to further induce apoptosis.

(31) Although specific embodiments of the present invention have been disclosed in detail as described above, it will be obvious to those skilled in the art that such description is merely of preferable exemplary embodiments and is not to be construed to limit the scope of the present invention. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.