KISEP Otology Korean J Otolaryngol 1998;418:971-975 기니픽와우에서성장인자수용체의분포 이병돈 박영진 안희영 장혁순 강주원 Expression of Receptor Tyrosine Kinases in the Guinea Pig Cochlea Byung Don Lee, MD, Young Jin Park, MD, Hyuck Soon Chang, MD, Hee Young Ahn, MD and Ju Won Kang, MD Department of Otolaryngology, College of Medicine, Soonchunhyang University, Seoul, Korea ABSTRACT Background and ObjectivesDamage in the auditory sensory epithelia is irreversible and the resulting hearing loss is permanent in mammals. Nevertheless, mammals like birds can recover their auditory function anatomically and physiologically after the sensorineural hearing loss. One of the functions of a receptor tyrosine kinase is to controll the cellular metabolisms like motility, growth, differentiation, regeneration, and ion transport via various pathways. There are no reports on the distribution of receptor tyrosine kinases in the guinea pig cochlea. The purpose of this study is to evaluate the expression of the fibroblast growth factor receptor FGFR and transforming growth factor receptor TGFR-, type II in the cochlear sensory epithelia of guinea pigs. Materials and MethodsWe investigated the immunoreactivity and functional roles of the receptor tyrosine kinases-fgfr and TGFR-type II - by using surface preparation technique in the cochlea of the Preyer s positive, pigmented guinea pig. ResultsThe results showed that receptor tyrosine kinases were expressed in the cytoplasm of Deiter s and Hensen s cells to FGFR and TGFR- type II, and the stereocillia of inner and outer hair cells to FGFR by surface preparation technique. ConclusionThe findings suggest that receptor tyrosine kinases are involved in the cellular growth, differentiation and regeneration in the cochlear supporting cells, and ion transport in the hair cells of the mammals. We found that receptor tyrone kinases are not related to outer hair cells participating in the PLC1-IP3 second messenger system. Korean J Otolaryngol 1998;418:971-975 KEY WORDSReceptor tyrosine kinase Signal transduction Auditory regeneration. 971
성장인자수용체의 발현 ered solution(ph 7.5, 이하 PBS)과 4% paraform-alde- the first 28 N-Terminal residues of the mature human hyde를 각각 100 ml를 좌심실을 통해 관류 고정하였다. 단 TGF-β receptor, Upstate Biotechnology. Inc., U.S.A)를 두 후 otic capsule을 열고 등골을 적출하고 정원창과 와우의 각각 1%의 BSA로 1 50과 1 5로 희석하여 4 에서 12 apical turn의 골편을 제거하여 24시간 동안 4% paraformaldehyde로 와우를 재고정하였다. 와우 고정 후 면역조직 화학 염색을 위해 실체 줌현미경과 드릴을 사용하여 와우의 외측벽과 혈관조를 박리하였다. 비특이적 결합을 방지하기 위 해 8% bovine serum albumin(이하 BSA)으로 2시간 반응 시킨 후, 일차 항체는 다클론성 IgG rabbit anti-chicken 섬 유아세포 성장인자 수용체 항체(rabbit polyclonal IgG, An ti-chicken FGF Receptor, residues 119-114 of the deduced cdna sequence of the extracellular domain of the chicken FGF receptor, Upstate Biotechnology. Inc., U.S. A) 및 rabbit anti-human 형질변환인자 수용체 항체(rabbit polyclonal IgG, Anti-Human TGF-β Receptor, type II, Fig. 1. Control. Surface preparation of cochlear sensory epithelia. OHC outer hair cell, SC supporting cell (mid-turn, ABC method, 1,000). Fig. 2. Surface preparation of cochlear sensory epithelia stained with Anti-chicken FGF receptor. Strong immunoreactive staining was seen in Deiter s cells (arrow), not outer hair cells. IHC inner hair cell, OHC outer hair cell (mid-turn, ABC method, 1,000). 972 Fig. 3. Surface preparation of cochlear sensory epithelia stained with Anti-chicken FGF receptor. Strong immunoreactive staini-g was seen in Hensen s cells (arrow) (mid-turn, ABC method, 1,000). Fig. 4. Surface preparation of cochlear sensory epithelia stained with Anti-human TGF receptor. Strong immunoreactive staining was seen in Deiter s cells (arrow), not outer hair cells. OHC outer hair cell (mid-turn, ABC method, 1,000). Fig. 5. Surface preparation of cochlear sensory epithelia stained with Anti-human TGF receptor. Strong immunoreactive staining wa s seen in Hensen s cells (arrow) (mid-turn, ABC method, 1,000). Korean J Otolaryngol 1998;41(8):971-975
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