초록 ▼

MsrB3 유전자 결손 마우스의 청각 및 전정기능검사를 수행한 결과, 심도난청소견을 확인하였으며, 주사전자현미경을 이용해 내이 상피세포의 손상과정을 확인하였음. 면역형광조직검사로 내이에서 Msrb3의 발현양상을 규명하였으며, quantitative RT-PCR을 이용해 발달시기별 Msrb3 mRNA의 발현량을 조사하였음. 투과전자현미경을 이용한 내이의 초미세구조의 변화관찰 및 손상구조물 규명과 FM1-43을 이용한 내이유모세포 MET 채널기능검사를 통해 MsrB3가 hair cell의 발생에는 영향을 주지 않으며 발생 후 stere

MsrB3 유전자 결손 마우스의 청각 및 전정기능검사를 수행한 결과, 심도난청소견을 확인하였으며, 주사전자현미경을 이용해 내이 상피세포의 손상과정을 확인하였음. 면역형광조직검사로 내이에서 Msrb3의 발현양상을 규명하였으며, quantitative RT-PCR을 이용해 발달시기별 Msrb3 mRNA의 발현량을 조사하였음. 투과전자현미경을 이용한 내이의 초미세구조의 변화관찰 및 손상구조물 규명과 FM1-43을 이용한 내이유모세포 MET 채널기능검사를 통해 MsrB3가 hair cell의 발생에는 영향을 주지 않으며 발생 후 stereocilia의 유지에 문제를 야기함을 밝힘. Msrb3 유전자 결손 마우스와 정상 마우스 내이의 시기별, 조직별 RNA 정제 및 microarray 수행을 통해 MsrB3의 연관 단백질 후보를 발굴하였음. 내이 발달 시기별 ROS 정량 및 형태적 변화의 비교 분석 결과, MsrB3의 기능이 조직 전반적 수준에서는 ROS와 직접적 관련이 없음을 밝힘. 산화환원 관련 내이 손상 약물 치료법 개발을 위해 Cisplatin에 대한 alpha lipoic acid의 효과를 검증하였으며, 국소적 약물 전달 시스템을 위한 실험 조건을 확립하였음.

Abstract ▼

Ⅰ.Purpose
The purpose of this research aims using the MsrB3 knock out (KO) mouse model to understand the morphological function of the MsrB3 in the inner ear which is related with redox and to investigate factors which causes the hearing loss and to investigate a new effective drug delivery syste

Ⅰ.Purpose
The purpose of this research aims using the MsrB3 knock out (KO) mouse model to understand the morphological function of the MsrB3 in the inner ear which is related with redox and to investigate factors which causes the hearing loss and to investigate a new effective drug delivery system into the inner ear to treat the hearing loss.
Ⅱ.Contents
1.Pathologic physiological identification of the MsrB3 KO mouse
Hearing test of the MsrB3 KO mouse
Morphological analysis of inner ear of the MsrB3 KO mouse
Functional analysis of mechanoelectrical transduction (MET) of the hair cells of the MsrB3 KO mouse
2.Investigating the hearing loss caused by MsrB3 dysfunction
Functional analysis of the MsrB3 related genes in the inner ear
Quantification of ROS in the developmental stages of the inner ear of the MsrB3 KO mouse
3.Development of the drug therapy on the damaged inner ear related with redox
Development a new drug delivery system into the inner ear for treatment of hearing loss
Ⅲ.Results
1.Pathologic physiological identification of the MsrB3 KO mouse
The MsrB3 KO mouse showed the severe hearing loss
The result of the morphological test on the inner ear of the MsrB3 KO mouse showed phenomenon of stereocilia disorder
The result of the MET of the hair cells in the MsrB3 KO mouse showed the activation of the MET channel was measured
2.Investigating the hearing loss caused by MsrB3 dysfunction
The protein which showed different expression compared with the wild type mouse was detected by the microarray
The result of the ROS quantity change of the MsrB3 KO mouse by the developmental stages showed no differences with the wild type mouse
3.Development of the drug therapy on the damaged inner ear related with redox
The ototoxicity analysis of the cisplatin showed that when the concentration increased the number of the HEI-OC1 cells drastically decreased
The effects of ALA on the ototoxicity of the cisplatin was confirmed by the survival rate of the ALA treated cells reached almost up to the normal rate compared to the control group with only treated with the cisplatin
A drug delivery system by using the cochlear explant was prepared and for the long term usage of the drug treatment, the tissues were cultured with the drug delivery system for 14 days and the possibility of maintenance was confirmed
Ⅳ.Applicable planning
The study on the early diagnosis which is the need in most to the hereditary hearing loss patients will be done. The study will be focused on the gene therapy and further studies using drugs and viruses will be done.