【《遗传性听力损失疾病的研究文献综述》3500字】

遗传性听力损失疾病的研究文献综述根据不同的特点，听力损失有不同的分类方式。根据致病原因，听力损失可分为遗传性和非遗传性两大类，前者指的是由于基因突变导致的听力下降，而后者则是由于细菌感染、耳毒性药物、外伤和噪声暴露等环境因素造成的听力损失。据统计，超过一半的耳聋病人是由于遗传因素导致的ADDINEN.CITE<EndNote><Cite><Author>Howell</Author><Year>2019</Year><RecNum>238</RecNum><DisplayText>(Howell,2019)</DisplayText><record><rec-number>238</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1582683317">238</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Howell,R.R.</author></authors></contributors><auth-address>UniversityofMiamiMillerSchoolofMedicine,Miami,FL,USA.rhowell@.</auth-address><titles><title>Wemustnowputinplaceanupdated,comprehensivenewbornscreeningprogramfordeafandhard-of-hearinginfants</title><secondary-title>GenetMed</secondary-title></titles><periodical><full-title>GenetMed</full-title></periodical><pages>2439-2441</pages><volume>21</volume><number>11</number><edition>2019/08/16</edition><dates><year>2019</year><pub-dates><date>Nov</date></pub-dates></dates><isbn>1530-0366(Electronic) 1098-3600(Linking)</isbn><accession-num>31413371</accession-num><urls><related-urls><url>/pubmed/31413371</url></related-urls></urls><electronic-resource-num>10.1038/s41436-019-0638-3</electronic-resource-num></record></Cite></EndNote>(\o"Howell,2019#238"Howell,2019)，因此对遗传性听力损失的致病突变和致聋机制的研究十分重要。1遗传性听力损失疾病的研究现状1988年，Wallis等人通过定位克隆的方法，首次报道了NSHL的致聋基因POU3F4ADDINEN.CITE<EndNote><Cite><Author>Wallis</Author><Year>1988</Year><RecNum>2010</RecNum><DisplayText>(Wallisetal.,1988)</DisplayText><record><rec-number>2010</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1614655512">2010</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Wallis,C.</author><author>Ballo,R.</author><author>Wallis,G.</author><author>Beighton,P.</author><author>Goldblatt,J.</author></authors></contributors><auth-address>DepartmentofHumanGenetics,MedicalSchool,UniversityofCapeTown,SouthAfrica.</auth-address><titles><title>X-linkedmixeddeafnesswithstapesfixationinaMauritiankindred:linkagetoXqprobepDP34</title><secondary-title>Genomics</secondary-title><alt-title>Genomics</alt-title></titles><periodical><full-title>Genomics</full-title><abbr-1>Genomics</abbr-1></periodical><alt-periodical><full-title>Genomics</full-title><abbr-1>Genomics</abbr-1></alt-periodical><pages>299-301</pages><volume>3</volume><number>4</number><dates><year>1988</year></dates><isbn>0888-7543</isbn><accession-num>3243543</accession-num><urls><related-urls><url>/3243543</url></related-urls></urls><remote-database-name>PubMed</remote-database-name><language>eng</language></record></Cite></EndNote>(\o"Wallis,1988#2010"Wallisetal.,1988)，开启了通过全基因组遗传连锁分析和定位克隆的手段寻找致聋突变位点的新纪元。但是，由于定位区间错误或者突变基因未知等原因，仍然有部分经典的NSHL致病位点没有找到ADDINEN.CITEADDINEN.CITE.DATA(\o"Vona,2015#2009"Vonaetal.,2015)。21世纪初，随着人类全基因组测序工作完成ADDINEN.CITEADDINEN.CITE.DATA(\o"Lander,2001#2128"Landeretal.,2001)，结合定位克隆确定致病突变的区间，研究人员进一步发现了更多的NSHL致聋位点。同时，全基因组二代测序技术出现并迅速发展，为寻找遗传性疾病的突变提供了更加快速有效的手段ADDINEN.CITEADDINEN.CITE.DATA(\o"Vona,2015#2009"Vonaetal.,2015)。近年来，研究人员发现，大片段插入基因外显子或内含子区域的结构变异（structuralvariation，SV）同样也会造成小鼠耳聋的表型，表明致聋突变位点和类型有待进一步挖掘ADDINEN.CITE<EndNote><Cite><Author>Gagnier</Author><Year>2019</Year><RecNum>2053</RecNum><DisplayText>(Gagnieretal.,2019)</DisplayText><record><rec-number>2053</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1614741915">2053</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Gagnier,Liane</author><author>Belancio,VictoriaP.</author><author>Mager,DixieL.</author></authors></contributors><auth-address>1TerryFoxLaboratory,BCCancerandDepartmentofMedicalGenetics,UniversityofBritishColumbia,V5Z1L3,Vancouver,BCCanada. 2DepartmentofStructuralandCellularBiology,TulaneUniversitySchoolofMedicine,TulaneCancerCenter,TulaneCenterforAging,NewOrleans,LA70112USA.</auth-address><titles><title>Mousegermlinemutationsduetoretrotransposoninsertions</title><secondary-title>MobileDNA</secondary-title><alt-title>MobDNA</alt-title></titles><periodical><full-title>MobileDNA</full-title></periodical><alt-periodical><full-title>MobDNA</full-title></alt-periodical><pages>15</pages><volume>10</volume><dates><year>2019</year></dates><isbn>1759-8753</isbn><accession-num>31011371</accession-num><urls><related-urls><url>/31011371</url></related-urls></urls><electronic-resource-num>10.1186/s13100-019-0157-4</electronic-resource-num><remote-database-name>PubMed</remote-database-name><language>eng</language></record></Cite></EndNote>(\o"Gagnier,2019#2053"Gagnieretal.,2019)。随着高通量测序技术的进一步发展，长读段全基因组测序等技术为研究遗传性听力损失致聋突变的提供了新的检测方法ADDINEN.CITE<EndNote><Cite><Author>Rhoads</Author><Year>2015</Year><RecNum>1866</RecNum><DisplayText>(RhoadsandAu,2015)</DisplayText><record><rec-number>1866</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1608619871">1866</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Rhoads,A.</author><author>Au,K.F.</author></authors></contributors><auth-address>DepartmentofBiostatistics,UniversityofIowa,IowaCity,IA52242,USA. DepartmentofBiostatistics,UniversityofIowa,IowaCity,IA52242,USA;DepartmentofInternalMedicine,UniversityofIowa,IowaCity,IA52242,USA.Electronicaddress:kinfai-au@.</auth-address><titles><title>PacBioSequencingandItsApplications</title><secondary-title>GenomicsProteomicsBioinformatics</secondary-title></titles><periodical><full-title>GenomicsProteomicsBioinformatics</full-title></periodical><pages>278-89</pages><volume>13</volume><number>5</number><edition>2015/11/07</edition><keywords><keyword>DNAMethylation/genetics</keyword><keyword>Genome/*genetics</keyword><keyword>MolecularSequenceAnnotation/*methods</keyword><keyword>ProteinIsoforms/genetics</keyword><keyword>RepetitiveSequences,NucleicAcid/genetics</keyword><keyword>SequenceAnalysis,DNA/*instrumentation/*methods</keyword><keyword>Transcriptome/*genetics</keyword><keyword>Denovoassembly</keyword><keyword>Geneisoformdetection</keyword><keyword>Hybridsequencing</keyword><keyword>Methylation</keyword><keyword>Third-generationsequencing</keyword></keywords><dates><year>2015</year><pub-dates><date>Oct</date></pub-dates></dates><isbn>2210-3244(Electronic) 1672-0229(Linking)</isbn><accession-num>26542840</accession-num><urls><related-urls><url>/pubmed/26542840</url></related-urls></urls><custom2>PMC4678779</custom2><electronic-resource-num>10.1016/j.gpb.2015.08.002</electronic-resource-num></record></Cite></EndNote>(\o"Rhoads,2015#1866"RhoadsandAu,2015)。2遗传性听力损失致病的分子机制遗传性听力损失的致病变异绝大多数都是基因突变直接导致蛋白功能异常或无法正常合成蛋白质ADDINEN.CITE<EndNote><Cite><Author>Karousis</Author><Year>2019</Year><RecNum>1953</RecNum><DisplayText>(KarousisandMuhlemann,2019)</DisplayText><record><rec-number>1953</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1613013328">1953</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Karousis,E.D.</author><author>Muhlemann,O.</author></authors></contributors><auth-address>DepartmentofChemistryandBiochemistry,UniversityofBern,CH-3012Bern,Switzerland.</auth-address><titles><title>Nonsense-MediatedmRNADecayBeginsWhereTranslationEnds</title><secondary-title>ColdSpringHarbPerspectBiol</secondary-title></titles><periodical><full-title>ColdSpringHarbPerspectBiol</full-title></periodical><volume>11</volume><number>2</number><edition>2018/06/13</edition><keywords><keyword>Codon,Nonsense</keyword><keyword>*Eukaryota</keyword><keyword>*NonsenseMediatedmRNADecay</keyword><keyword>*PeptideChainTermination,Translational</keyword><keyword>ProteinBiosynthesis</keyword><keyword>RNA,Messenger/genetics/*metabolism</keyword></keywords><dates><year>2019</year><pub-dates><date>Feb1</date></pub-dates></dates><isbn>1943-0264(Electronic) 1943-0264(Linking)</isbn><accession-num>29891560</accession-num><urls><related-urls><url>/pubmed/29891560</url></related-urls></urls><custom2>PMC6360860</custom2><electronic-resource-num>10.1101/cshperspect.a032862</electronic-resource-num></record></Cite></EndNote>(\o"Karousis,2019#1953"KarousisandMuhlemann,2019)。迄今为止，已发现170多个基因突变与人类遗传性听力损失相关ADDINEN.CITEADDINEN.CITE.DATA(\o"Angeli,2012#1932"Angelietal.,2012;\o"Kremer,2019#230"Kremer,2019;\o"Shearer,2017#1933"Sheareretal.,2017)。根据基因翻译产生蛋白的功能可以大致分为四类：（1）细胞骨架基因，细胞骨架是生命体中每个细胞都需要的结构，但听觉系统中有许多特化结构因而也存在该系统所特有的细胞骨架蛋白，其中很多都与静纤毛的排列、长度以及相互间的连接相关，例如Myo7a、Espin等ADDINEN.CITEADDINEN.CITE.DATA(\o"Rhodes,2004#29"Rhodesetal.,2004;\o"Zheng,2000#2034"Zhengetal.,2000)；（2）细胞间连接蛋白基因，内耳各个细胞之间都存在着各种复杂的细胞间连接。位于毛细胞顶端的静纤毛之间通过顶连接相连。在血管纹的三层上皮细胞间广泛存在着紧密连接，对于维持内外淋巴液成分的动态平衡和离子浓度起到至关重要的作用。这类基因包括Pcdh15、Cdh23、Gjb2等ADDINEN.CITEADDINEN.CITE.DATA(\o"Alagramam,1999#2038"Alagramametal.,1999;\o"Bryda,2001#2039"Brydaetal.,2001;\o"DiPalma,2001#2040"DiPalmaetal.,2001;\o"Estivill,1998#2035"Estivilletal.,1998;\o"Kelley,1998#2036"Kelleyetal.,1998;\o"Wada,2001#2037"Wadaetal.,2001)；（3）转运体和离子通道基因，负责运输内外淋巴液中的各类离子，产生中阶蜗内电位，甚至直接参与毛细胞机械电转导过程，例如与毛细胞囊泡释放以及外毛细胞电致运动相关的基因。这类基因有Kcnq1、Tmc1/2、Otoferlin，Prestin等ADDINEN.CITEADDINEN.CITE.DATA(\o"Casimiro,2001#1951"Casimiroetal.,2001;\o"Kurima,2002#2046"Kurimaetal.,2002;\o"Lee,2000#251"Leeetal.,2000;\o"Liberman,2002#2044"Libermanetal.,2002;\o"Rodríguez-Ballesteros,2003#2043"Rodríguez-Ballesterosetal.,2003;\o"Vreugde,2002#2041"Vreugdeetal.,2002;\o"Wu,2004#2045"Wuetal.,2004)；（4）细胞外基质蛋白基因，主要包括编码盖膜组成成分的基因,这些基因突变会导致盖膜结构异常，这类基因有Tecta、Col2a1等ADDINEN.CITEADDINEN.CITE.DATA(\o"Donahue,2003#2049"Donahueetal.,2003;\o"Legan,2000#2047"Leganetal.,2000;\o"Legan,2005#2048"Leganetal.,2005)。目前，随着测序技术日趋完善，除了编码蛋白的基因上发生突变，研究人员发现了越来越多不同类型的致病变异ADDINEN.CITE<EndNote><Cite><Author>Kremer</Author><Year>2019</Year><RecNum>230</RecNum><DisplayText>(Kremer,2019)</DisplayText><record><rec-number>230</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1582683231">230</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Kremer,H.</author></authors></contributors><auth-address>Hearing&GenesDivision,DepartmentofOtorhinolaryngologyandDepartmentofHumanGenetics,DondersInstituteofBrain,CognitionandBehaviour,RadboudUniversityMedicalCenter,Nijmegen,theNetherlands.Electronicaddress:hannie.kremer@radboudumc.nl.</auth-address><titles><title>Hereditaryhearingloss;abouttheknownandtheunknown</title><secondary-title>HearRes</secondary-title></titles><periodical><full-title>HearRes</full-title></periodical><pages>58-68</pages><volume>376</volume><edition>2019/01/23</edition><keywords><keyword>*Genotype-phenotypecorrelation</keyword><keyword>*Hereditarydeafness</keyword><keyword>*wes</keyword><keyword>*wgs</keyword></keywords><dates><year>2019</year><pub-dates><date>May</date></pub-dates></dates><isbn>1878-5891(Electronic) 0378-5955(Linking)</isbn><accession-num>30665849</accession-num><urls><related-urls><url>/pubmed/30665849</url></related-urls></urls><electronic-resource-num>10.1016/j.heares.2019.01.003</electronic-resource-num></record></Cite></EndNote>(\o"Kremer,2019#230"Kremer,2019)，包括（1）致病变异发生在已知耳聋基因“未知”的外显子或剪接位点ADDINEN.CITEADDINEN.CITE.DATA(\o"Bowl,2017#239"Bowletal.,2017)。Rehman等人发现，在Myo15a基因第二个内含子中存在一个之前未注释的由218个核苷酸组成的外显子，并且包含新的转录起始位点ADDINEN.CITE<EndNote><Cite><Author>Rehman</Author><Year>2016</Year><RecNum>1493</RecNum><DisplayText>(Rehmanetal.,2016)</DisplayText><record><rec-number>1493</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1588147746">1493</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Rehman,AtteeqU.</author><author>Bird,JonathanE.</author><author>Faridi,Rabia</author><author>Shahzad,Mohsin</author><author>Shah,Sujay</author><author>Lee,Kwanghyuk</author><author>Khan,ShaheenN.</author><author>Imtiaz,Ayesha</author><author>Ahmed,ZubairM.</author><author>Riazuddin,Saima</author><author>Santos-Cortez,RegieLynP.</author><author>Ahmad,Wasim</author><author>Leal,SuzanneM.</author><author>Riazuddin,Sheikh</author><author>Friedman,ThomasB.</author></authors></contributors><titles><title>MutationalSpectrumofMYO15AandtheMolecularMechanismsofDFNB3HumanDeafness</title><secondary-title>HumanMutation</secondary-title></titles><periodical><full-title>HumanMutation</full-title></periodical><pages>991-1003</pages><volume>37</volume><number>10</number><section>991</section><dates><year>2016</year></dates><isbn>10597794</isbn><urls></urls><electronic-resource-num>10.1002/humu.23042</electronic-resource-num></record></Cite></EndNote>(\o"Rehman,2016#1493"Rehmanetal.,2016)；（2）致病变异位于已知的耳聋基因内含子、启动子、增强子、抑制子或者阻隔子等非编码区域ADDINEN.CITE<EndNote><Cite><Author>Bonev</Author><Year>2016</Year><RecNum>2129</RecNum><DisplayText>(BonevandCavalli,2016)</DisplayText><record><rec-number>2129</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1616158728">2129</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Bonev,Boyan</author><author>Cavalli,Giacomo</author></authors></contributors><auth-address>InstituteofHumanGenetics,UPR1142NationalCentreforScientificResearch(CNRS);andUniversityofMontpellier,141RuedelaCardonille,34396MontpellierCedex5,France.</auth-address><titles><title>Organizationandfunctionofthe3Dgenome</title><secondary-title>Naturereviews.Genetics</secondary-title><alt-title>NatRevGenet</alt-title></titles><alt-periodical><full-title>NatRevGenet</full-title></alt-periodical><pages>661-678</pages><volume>17</volume><number>11</number><dates><year>2016</year></dates><isbn>1471-0064</isbn><accession-num>27739532</accession-num><urls><related-urls><url>/27739532</url></related-urls></urls><electronic-resource-num>10.1038/nrg.2016.112</electronic-resource-num><remote-database-name>PubMed</remote-database-name><language>eng</language></record></Cite></EndNote>(\o"Bonev,2016#2129"BonevandCavalli,2016)。例如，在DFNB1中GJB2基因的启动子区域的致病变异造成该基因的转录水平降低ADDINEN.CITE<EndNote><Cite><Author>Matos</Author><Year>2007</Year><RecNum>2130</RecNum><DisplayText>(Matosetal.,2007)</DisplayText><record><rec-number>2130</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1616159802">2130</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Matos,T.D.</author><author>Caria,H.</author><author>Simões-Teixeira,H.</author><author>Aasen,T.</author><author>Nickel,R.</author><author>Jagger,D.J.</author><author>O'Neill,A.</author><author>Kelsell,D.P.</author><author>Fialho,G.</author></authors></contributors><titles><title>Anovelhearing-loss-relatedmutationoccurringintheGJB2basalpromoter</title><secondary-title>Journalofmedicalgenetics</secondary-title><alt-title>JMedGenet</alt-title></titles><alt-periodical><full-title>JMedGenet</full-title></alt-periodical><pages>721-725</pages><volume>44</volume><number>11</number><dates><year>2007</year></dates><isbn>1468-6244</isbn><accession-num>17660464</accession-num><urls><related-urls><url>/17660464</url><url>/pmc/articles/PMC2752183/pdf/721.pdf</url></related-urls></urls><remote-database-name>PubMed</remote-database-name><language>eng</language></record></Cite></EndNote>(\o"Matos,2007#2130"Matosetal.,2007)。而在DFNX2的研究中发现对于内耳发育必不可少的一个远距离顺式作用元件（cis-regulator）的缺失会导致POU3F4基因功能异常ADDINEN.CITEADDINEN.CITE.DATA(\o"Naranjo,2010#1498"Naranjoetal.,2010)；（3）致病变异影响调控子而非编码蛋白的基因，如小RNA（microRNA）和长非编码RNA。2009年，Lewis和Mencia等在人和小鼠中发现miRNA-96的功能缺失会导致听力损失ADDINEN.CITEADDINEN.CITE.DATA(\o"Lewis,2009#245"Lewisetal.,2009;\o"Mencia,2009#244"Menciaetal.,2009)。3遗传性听力损失小鼠的研究方法对于遗传性耳聋致病变异的分析多起始于针对病人家系和耳聋小鼠品系的连锁分析及定位克隆，并进一步通过测序分析等实验手段寻找致病突变ADDINEN.CITE<EndNote><Cite><Author>Angeli</Author><Year>2012</Year><RecNum>1932</RecNum><DisplayText>(Angelietal.,2012)</DisplayText><record><rec-number>1932</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1612835461">1932</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Angeli,S.</author><author>Lin,X.</author><author>Liu,X.Z.</author></authors></contributors><auth-address>DepartmentofOtolaryngology,UniversityofMiami,Miami,Florida33136,USA.</auth-address><titles><title>Geneticsofhearinganddeafness</title><secondary-title>AnatRec(Hoboken)</secondary-title></titles><periodical><full-title>AnatRec(Hoboken)</full-title></periodical><pages>1812-29</pages><volume>295</volume><number>11</number><edition>2012/10/10</edition><keywords><keyword>Animals</keyword><keyword>Deafness/*genetics</keyword><keyword>*GeneticDiseases,Inborn</keyword><keyword>Hearing/*genetics</keyword><keyword>Humans</keyword><keyword>Mice</keyword></keywords><dates><year>2012</year><pub-dates><date>Nov</date></pub-dates></dates><isbn>1932-8494(Electronic) 1932-8486(Linking)</isbn><accession-num>23044516</accession-num><urls><related-urls><url>/pubmed/23044516</url></related-urls></urls><custom2>PMC4523052</custom2><electronic-resource-num>10.1002/ar.22579</electronic-resource-num></record></Cite></EndNote>(\o"Angeli,2012#1932"Angelietal.,2012)。2007年，Sedigheh等人在研究耳聋病人家系的致聋突变时，发现其中4个独立的家系的突变位点都在DFNB59基因中，该基因编码Pejvakin蛋白。随后，他们通过构建其中一个家系的致病突变R183W敲入的纯合小鼠，发现基因敲入的小鼠听神经传导功能异常导致耳聋ADDINEN.CITEADDINEN.CITE.DATA(\o"Delmaghani,2006#33"Delmaghanietal.,2006)。同年，Martin等人对一株正向遗传学筛选获得的耳聋小鼠Sirtaki品系进行研究，发现Pejvakin基因突变造成小鼠外毛细胞异常，出现渐进性耳聋的表型ADDINEN.CITEADDINEN.CITE.DATA(\o"Schwander,2007#34"Schwanderetal.,2007)。但是由于人类家系有限以及逐年减少的家系规模，我们需要更多的动物模型来寻找致聋基因。小鼠和人的听觉系统具有较高的保守性，包括耳蜗的结构、听觉形成的机制以及与听觉功能相关的基因等方面ADDINEN.CITEADDINEN.CITE.DATA(\o"Fritzsch,2003#2131"FritzschandBeisel,2003;\o"Kawamoto,2001#2051"Kawamotoetal.,2001)。小鼠在实验室环境下近亲繁育过程中，会出现少量携带自发突变的个体，同时为了获得更多疾病动物模型，研究人员通常会进行大规模的ENU诱变，促使小鼠产生全基因组范围内不同类型的突变。结合相应的筛选手段，可以从自发突变和ENU诱变产生的动物中找到平衡或（和）听力异常的可遗传小鼠品系进行后续研究ADDINEN.CITEADDINEN.CITE.DATA(\o"HrabédeAngelis,2000#2132"HrabédeAngelisetal.,2000)。CochleaCochleaABRwaves图1.SEQ图1.\*ARABIC1ABR测听原理及波形示意图ABR测听原理示意图。当小鼠受到声音刺激后，耳蜗中产生的电信号经听神经（auditorynerve，AN）传至耳蜗核（cochlearnucleus，CN），到达对侧上橄榄核复合体（superiorolivarycomplex，SOC）后，再由外侧丘系传到下丘脑（inferiorcolliculus，IC），最终到达听皮层（Auditorycortex，AC）。经典的ABR波形如图所示，其中波I-IV分别记录听神经、耳蜗核、上橄榄核和下丘脑的信号。ADDINEN.CITEADDINEN.CITE.DATA(\o"Möhrle,2016#2103"Möhrleetal.,2016)为了从自发突变或ENU诱变产生的子代中筛选到听力损失的小鼠，常用的检测方法是ABR和DPOAEADDINEN.CITEADDINEN.CITE.DATA(\o"Eggermont,2019#1888"Eggermont,2019;\o"Moulin,1994#2012"Moulinetal.,1994)。ABR是评估小鼠听力状况的指标之一，通过检测小鼠对于声音刺激的同步化神经发应，评估听觉的灵敏度。其生物学基础是来自耳蜗的I型传入神经先投射到耳蜗核，随后大部分神经冲动经斜方体交叉到对外侧的上橄榄核，再由外侧丘系传到下丘脑，最终到达大脑皮层的听皮层区域ADDINEN.CITEADDINEN.CITE.DATA(\o"Möhrle,2016#2103"Möhrleetal.,2016)。测定小鼠的ABR信号，并通过信号大小、波形的形状以及最大幅值等综合评估小鼠的听力状况（REF_Ref67127703\h图1.11）。正常小鼠ABR的阈值一般在20~30dBSPL，若超过该阈值则认为该小鼠听力受损，若在最大声强90dBSPL仍检测不到III波，则认为该小鼠重度耳聋。哺乳动物耳蜗中基底膜能够分辨不同频率的声音刺激。20世纪50年代，George提出行波理论，该理论指出当耳蜗受到外界声音刺激时，首先会在基底膜靠近卵圆窗的位置（底端）产生振动并向顶端逐渐传递。基底膜产生的波动的振幅从底回开始逐渐增大，并在某一特定位置达到最大值，随后迅速降低。通常高频声音刺激在基底膜的底回产生最大振幅，而低频声音产生的最大振幅则对应基地膜的顶回位置。耳蜗能够分辨频率差别细微的两个声音，仅靠基底膜对不同频率声音的响应无法实现如此精细的区分。20世纪90年代末，Johnstone和Boyle发现，在活体动物中外毛细胞对于声音的刺激还存在主动放大机制，能够将不同频率的信号进行非线性放大，从而大大提高的了频率的分辨率（REF_Ref67127661\h图1.12）ADDINEN.CITE<EndNote><Cite><Author>Fettiplace</Author><Year>2017</Year><RecNum>1452</RecNum><DisplayText>(Fettiplace,2017)</DisplayText><record><rec-number>1452</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1587805608">1452</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Fettiplace,R.</author></authors></contributors><auth-address>DepartmentofNeuroscience,UniversityofWisconsinSchoolofMedicineandPublicHealth,Madison,Wisconsin,USA.</auth-address><titles><title>HairCellTransduction,Tuning,andSynapticTransmissionintheMammalianCochlea</title><secondary-title>ComprPhysiol</secondary-title></titles><periodical><full-title>ComprPhysiol</full-title></periodical><pages>1197-1227</pages><volume>7</volume><number>4</number><edition>2017/09/16</edition><keywords><keyword>Animals</keyword><keyword>Calcium/metabolism</keyword><keyword>HairCells,Auditory/metabolism/*physiology</keyword><keyword>Humans</keyword><keyword>IonChannels/genetics/metabolism</keyword><keyword>*Mechanotransduction,Cellular</keyword><keyword>*SynapticTransmission</keyword></keywords><dates><year>2017</year><pub-dates><date>Sep12</date></pub-dates></dates><isbn>2040-4603(Electronic) 2040-4603(Linking)</isbn><accession-num>28915323</accession-num><urls><related-urls><url>/pubmed/28915323</url></related-urls></urls><custom2>PMC5658794</custom2><electronic-resource-num>10.1002/cphy.c160049</electronic-resource-num></record></Cite></EndNote>(\o"Fettiplace,2017#1452"Fettiplace,2017)。图1.SEQ图1.\*ARABIC2外毛细胞主动放大机制示意图对于纯音刺激，在基底膜自身的频率选择性的基础上，其对应频率非常狭窄的区域内外毛细胞会产生振动，将信号放大约50dBSPL，从而提高耳蜗感知不同频率纯音的灵敏度。图片引自http://www.cochlea.eu针对外毛细胞这一特点，研究人员开发了一系列测定耳声发射的技术，可以经外耳道记录内耳在外界声音刺激下产生的微弱信号，DPOAE即为其中一种。DPOAE是外毛细胞对于特定频率的两个纯音f1和f2（f1<f2）在基底膜上进行放大后产生的信号，通常用来初步评估小鼠的听力水平以及耳蜗基底膜上不同频率外毛细胞的存活情况。记录DPOAE时，两个纯音信号的频率有特定的关系，并分别独立传递到外耳道，经外毛细胞放大后，在外耳道记录2f1-f2信号的强度并判断DPOAE的阈值（REF_Ref67130076\h图1.13）ADDINEN.CITE<EndNote><Cite><Author>Moulin</Author><Year>1994</Year><RecNum>2012</RecNum><DisplayText>(Moulinetal.,1994)</DisplayText><record><rec-number>2012</rec-number><foreign-keys><keyapp="EN"db-id="0sfs5dvpdfxs2ketx0jvdwt2xdeaafrx2saw"timestamp="1614669369">2012</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Moulin,A.</author><author>Bera,J.C.</author><author>Collet,L.</author></authors></contributors><auth-address>PhysiologieSensorielleAuditionetVoix,UnitéassociéeauCNRS1447,HôpitalEdouard-Herriot,Lyon,France.</auth-address><titles><title>Distortionproductotoacousticemissionsandsensorineuralhearingloss</title><secondary-title>Audiology:officialorganoftheInternationalSocietyofAudiology</secondary-title><alt-title>Audiology</alt-title></titles><periodical><full-title>Audiology:officialorganoftheInternationalSocietyofAudiology</full-title><abbr-1>Audiology</abbr-1></periodical><alt-periodical><full-title>Audiology:officialorganoftheInternationalSocietyofAudiology</full-title><abbr-1>Audiology</abbr-1></alt-periodical><pages>305-326</pages><volume>33</volume><number>6</number><dates><year>1994</year></dates><isbn>0020-6091</isbn><accession-num>7741665</accession-num><urls><related-urls><url>/7741665</url></related-urls></urls><remote-database-name>PubMed</remote-database-name><language>eng</language></record></Cite></EndNote>(\o"Moulin,1994#2012"Moulinetal.,1994)。图1.SEQ图1.\*ARABIC3DPOAE测定原理示意图由外耳道给小鼠两个有特定关系的不同频率纯音（f1<f2）刺激，并经外耳道记录在基底膜上产生的2f1-f2的畸变耳声发射产物。图片引自/参考文献Acevedo-Arozena,A.,Wells,S.,Potter,P.,Kelly,M.,Cox,R.D.,andBrown,S.D.M.(2008).ENUMutagenesis,aWayForwardtoUnderstandGeneFunction.AnnualReviewofGenomicsandHumanGenetics9,49-69.Alagramam,K.N.,Kwon,H.Y.,Cacheiro,N.L.,Stubbs,L.,Wright,C.G.,Erway,L.C.,andWoychik,R.P.(1999).Anewmouseinsertionalmutationthatcausessensorineuraldeafnessandvestibulardefects.Genetics152,1691-1699.Alkan,C.,Coe,B.P.,andEichler,E.E.(2011).Genomestructuralvariationdiscoveryandgenotyping.NatRevGenet12,363-376.Angeli,S.,Lin,X.,andLiu,X.Z.(2012).Geneticsofhearinganddeafness.AnatRec(Hoboken)295,1812-1829.Arnold,C.N.,Xia,Y.,Lin,P.,Ross,C.,Schwander,M.,Smart,N.G.,Muller,U.,andBeutler,B.(2011).Rapididentificationofadiseasealleleinmousethroughwholegenomesequencingandbulksegregationanalysis.Genetics187,633-641.Artus,J.,andHadjantonakis,A.-K.(2011).Generationofchimerasbyaggregationofembryonicstemcellswithdiploidortetraploidmouseembryos.Methodsinmolecularbiology(Clifton,NJ)693,37-56.Ashby,J.,Gorelick,N.J.,andShelby,M.D.(1997).Mutationassaysinmalegermcellsfromtransgenicmice:overviewofstudyandconclusions.Mutationresearch388,111-122.Azcoitia,V.,Aracil,M.,Martínez-A,C.,andTorres,M.(2005).ThehomeodomainproteinMeis1isessentialfordefinitivehematopoiesisandvascularpatterninginthemouseembryo.Developmentalbiology280,307-320.Barhanin,J.,Lesage,F.,Guillemare,E.,Fink,M.,Lazdunski,M.,andRomey,G.(19