中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2013年
1期
42-47
,共6页
朱光洁%马登滨%钱晓云%周函%陈杰%王芳%高下
硃光潔%馬登濱%錢曉雲%週函%陳傑%王芳%高下
주광길%마등빈%전효운%주함%진걸%왕방%고하
肌球蛋白轻链激酶%小鼠,基因敲除%毛细胞,内%诱发电位,听觉,脑干
肌毬蛋白輕鏈激酶%小鼠,基因敲除%毛細胞,內%誘髮電位,聽覺,腦榦
기구단백경련격매%소서,기인고제%모세포,내%유발전위,은각,뇌간
Myosin-light-chain Kinase%Mice,knockout%Hair cells,inner%Evoked potentials,auditory,brain stem
目的 建立特异性敲除内毛细胞肌球蛋白轻链激酶(myosin light chain kinase,MLCK)基因小鼠模型,初步评价敲除该基因对小鼠听功能的影响.方法 从DNA水平对Mlck基因敲除小鼠作基因型鉴定并验证敲除效率；以短声和8、16、32 kHz短纯音作为刺激音对基因敲除组及对照组小鼠行听性脑干反应(ABR)测试,分析两组小鼠反应阈值及波形差异.结果 根据鼠尾DNA PCR结果能够初步判定小鼠基因型,分离内毛细胞提取DNA,PCR结果证实目的基因被剔除.Mlck 基因敲除组小鼠短声及短纯音各频率ABR平均阈值较对照组均有升高,差异具有统计学意义(P值均＜0.05)；其中高频阈值升高更为显著,在32 kHz两组平均阈值相差＞20 dB.基因敲除组小鼠16 kHz短纯音60、50、40 dBSPL声强下Ⅰ波幅值明显小于对照组,差异具有统计学意义(P值均＜0.05).两组小鼠16 kHz短纯音在60 dBSPL下的Ⅰ/Ⅱ、Ⅰ/Ⅲ波幅比差异无统计学意义(P值均＞0.05).结论 内毛细胞特异性敲除Mlck小鼠成功将目的基因剔除,敲除后小鼠听力受损,尤以高频明显.
目的 建立特異性敲除內毛細胞肌毬蛋白輕鏈激酶(myosin light chain kinase,MLCK)基因小鼠模型,初步評價敲除該基因對小鼠聽功能的影響.方法 從DNA水平對Mlck基因敲除小鼠作基因型鑒定併驗證敲除效率；以短聲和8、16、32 kHz短純音作為刺激音對基因敲除組及對照組小鼠行聽性腦榦反應(ABR)測試,分析兩組小鼠反應閾值及波形差異.結果 根據鼠尾DNA PCR結果能夠初步判定小鼠基因型,分離內毛細胞提取DNA,PCR結果證實目的基因被剔除.Mlck 基因敲除組小鼠短聲及短純音各頻率ABR平均閾值較對照組均有升高,差異具有統計學意義(P值均＜0.05)；其中高頻閾值升高更為顯著,在32 kHz兩組平均閾值相差＞20 dB.基因敲除組小鼠16 kHz短純音60、50、40 dBSPL聲彊下Ⅰ波幅值明顯小于對照組,差異具有統計學意義(P值均＜0.05).兩組小鼠16 kHz短純音在60 dBSPL下的Ⅰ/Ⅱ、Ⅰ/Ⅲ波幅比差異無統計學意義(P值均＞0.05).結論 內毛細胞特異性敲除Mlck小鼠成功將目的基因剔除,敲除後小鼠聽力受損,尤以高頻明顯.
목적 건립특이성고제내모세포기구단백경련격매(myosin light chain kinase,MLCK)기인소서모형,초보평개고제해기인대소서은공능적영향.방법 종DNA수평대Mlck기인고제소서작기인형감정병험증고제효솔；이단성화8、16、32 kHz단순음작위자격음대기인고제조급대조조소서행은성뇌간반응(ABR)측시,분석량조소서반응역치급파형차이.결과 근거서미DNA PCR결과능구초보판정소서기인형,분리내모세포제취DNA,PCR결과증실목적기인피척제.Mlck 기인고제조소서단성급단순음각빈솔ABR평균역치교대조조균유승고,차이구유통계학의의(P치균＜0.05)；기중고빈역치승고경위현저,재32 kHz량조평균역치상차＞20 dB.기인고제조소서16 kHz단순음60、50、40 dBSPL성강하Ⅰ파폭치명현소우대조조,차이구유통계학의의(P치균＜0.05).량조소서16 kHz단순음재60 dBSPL하적Ⅰ/Ⅱ、Ⅰ/Ⅲ파폭비차이무통계학의의(P치균＞0.05).결론 내모세포특이성고제Mlck소서성공장목적기인척제,고제후소서은력수손,우이고빈명현.
Objective To investigate the function of myosin light chain kinase(MLCK) in hearing in mouse by generating inner hair cell-specific Mlck knockout mice and analyze the effect on their hearing.Methods Cross Mlck floxed mice with IHC-Cre mice,the genotype and knockout efficiency were confirmed by PCR.We used auditory brain stem response (ABR) to evaluate mice hearing function at different frequencies.Results Mlck konckout mice were selected by mice tail DNA genotyping and confirmed the deletion of the target gene by isolated inner hair cell DNA genotyping.Mlck-deficient mice showed impaired hearing with a rise in ABR threshold response to click and three different pure tones (8 kHz,16 kHz,32 kHz),and the rise was over 20 dB at high-frequency(32 kHz).Further analyses of waveforms showed that wave-Ⅰ amplitudes on 60 dB SPL,50 dB SPL and 40 dBSPL in response to tone (16 kHz) were less than control group(P ＜ 0.05) on average,but the ratio of wave Ⅰ / Ⅱ and Ⅰ / Ⅲ1 were not difference (P ＞0.05).Conclusions Mlck is successfully deleted in inner hair cell-specific Mlck knockout mice.Mlck knockout mice display a significantly higher threshold in response to click and tones,especially in high-frequencies.
