초록 ▼

먼저, 제브라피쉬 유모세포 재생과정에서 발현되는 마이크로 RNA의 screening을 시행하였음. 측선 유모 세포 및 지지세포에 형광단백질 발현이 되는 teansgenic 제브라피쉬의 embryo에 neomycin등의 ototoxic drug 투여 후 FACS를 이용하여 재생되는 hair cell을 collection 함. 재생되는 cell을 microarray 분석을 시행함. 측선의 감각유모세포의 재생과정에서 증가 또는 억제되는 microRNA를 detection 하였음.
Screening하여 선택된 마이크로 RNA의 역할을

먼저, 제브라피쉬 유모세포 재생과정에서 발현되는 마이크로 RNA의 screening을 시행하였음. 측선 유모 세포 및 지지세포에 형광단백질 발현이 되는 teansgenic 제브라피쉬의 embryo에 neomycin등의 ototoxic drug 투여 후 FACS를 이용하여 재생되는 hair cell을 collection 함. 재생되는 cell을 microarray 분석을 시행함. 측선의 감각유모세포의 재생과정에서 증가 또는 억제되는 microRNA를 detection 하였음.
Screening하여 선택된 마이크로 RNA의 역할을 규명함.
제브라피쉬에서 유모세포 재생과정에서만 변화가 있는 마이크로 RNA를 제브라피쉬에서 과발현 또는 억제시켰을때 유모세포의 생성 및 재생에 미치는 영향을 분석하였음.
발견된 마이크로 RNA 183 family의 경우 development를 방해할 뿐만 아니라 ototoxic drug 노출 후 regeneration 과정에도 영향을 끼쳐 regeneration을 방해함을 밝힘. 주된 기전은 transdifferentiation inhibition으로 분석하고 있음.

Abstract ▼

The critical structural and functional roles of distinct collagens have been observed on heritable and acquired human diseases. An example of such a situation is type XVII collagen (also known as the 180-kDa bullous pemphigoid antigen or bullous pemphigoid antigen 2:BP180 and BPAG2, respectively). T

The critical structural and functional roles of distinct collagens have been observed on heritable and acquired human diseases. An example of such a situation is type XVII collagen (also known as the 180-kDa bullous pemphigoid antigen or bullous pemphigoid antigen 2:BP180 and BPAG2, respectively). The corresponding human gene. COL17A1, which consists of 56 exons, has been cloned and mapped to the long arm of chromorome 10, region 10q24.3. Type XVII collagen is a structural component of dermal-epidermal junction, forming hemidesmosomal protein compiexes together with a6b4 integrin, plectin, and the 230-kD bullous pemphigoid antigen (BPAG1). This conciusion is based early observation demonstrating that type XVII collagen serves as an autoantigen in an acquired blistering skin disease, bullous pemphigoid.
During the process of characterizing the type XVII collagen (col17a1a) in the zebrafish (Danio rerio) which clearly demonstrates ultrastructural features similar to those in human basement membrane zones of the skin, we found zebrafish COL17A1 orthologues (col17a1b) which related to mechanosensory hair cell. As noted above, zebrafish has two COL17A1-related gene sequences, col17a1 and col17a1b, as discovered by analysis of the online gene database. col17a1a and col17a1b showed considerable homology with each other, and were encoding proteins structurally related to human type XVII collagen. In situ hybridization of the developing zebrafish revealed that col17a1a was expressed in the skin in a punctate arrangement, while the expression of col17a1b was limited to the lateral line which consists of neuromasts of the developing animal.
To characterize the expression of col17a1b, we performed in situ hybridization with a probe recognizing col17a1b sequence. Subsequent injection of a morpholino corresponding to 5' UTR of col17a1b sis not reveal any change in morphology when examined at 75hpf to and followed up to 5dpf. However, staining for acetylated a-tubulin, a market for hair cells, showed decreased immunosignal.
col17a is primarily expressed in the cutaneous basement membrane zone and "knock-down underlying dermis accompanying the loss of hemidesmosomes.
In contrast, col17a1b is expressed in neural tissues, and "knock-down" of its expression results in reduction of hair cells. It is of interest that type XVII collagen was recently shown to be present in neurons of the cerebral cortex, hippocampus and amygdale of the human brain. Animal studies have also demonstrated the presence of type XVII collagen in the retina, cerebellum and the olfactory bulb. It has been postulated that in analogy to hemidesmosomes at the dermal-epidermal junction, type XVII collagen may participate in providing anchorage to the neural elements to the surrounding extracellular matrix. It has also been suggested that type XVII collagen might function as a synaptic stabilizer in the central nervous system, influencing synaptic plasticity. In this context it is of interest to note that an association between bullous pemphigoid, an autoimmune blistering disorder with circulating anti-type XVII collagen antibodies, and neurological disorders, such as age-related neurodegenerative disorders and multiple sclerosis, has been proposed.
In summary, we have identified two distinct COL17A1 orthologues, col17a1a and col17a1b, which are expressed exclusively in the skin or the lateral of zebrafish, respectively. col17a1a zebrafish model system may potentially serve as a screening tool for pharmacologic interventions to counteract. col17a1b in zebrafish appears to be required for auditory hair cell development and it differs from mammalian auditory hair cell. col17a1b can be a candidate gene which make hair cell regenerate and must be analyzed further.

목차 Contents

• 표지 ... 1
• I. 총괄현황 ... 2
• 표지 ... 3
• 제출문 ... 4
• 보고서 요약서 ... 5
• 요약문 ... 6
• SUMMARY ... 7
• 연구 성과 실적 및 향후 계획 ... 9
• 참여연구원 현황표 ... 13
• II. 총괄연구과제 연구결과 ... 14
• 1. 연구개발과제의 배경 및 필요성 ... 15
• 2. 국내외 기술개발 현황 ... 20
• 3. 연구개발과제의 추진체계 ... 21
• 4. 연구개발수행 내용 및 결과 ... 21
• 5. 목표달성도 및 관련분야 기여도 ... 26
• 6. 향후 연구성과 추진 계획 ... 28
• 7. 연구개발 결과의 파급효과 ... 28
• 8. 연구개발결과의 활용계획 ... 28
• 9. 연구개발과정에서 수집한 해외과학기술정보 ... 28
• 10. 참고문헌 ... 29
• III. 첨부서류 ... 31
• 1. 자체평가의견서 ... 32
• 2. 실적 증빙자료 ... 32
• 표지 ... 33
• 끝페이지 ... 33