大鼠胚胎期嗅上皮中嗅细胞发育及细胞凋亡的研究
|
摘要
[研究背景]
嗅觉是哺乳动物的重要感觉器官之一,它同视觉、听觉一样,是人体捕获外界信息的特殊装置。人类的嗅觉具有辨别气味、增进食欲、识别环境、报警等作用。嗅觉还通过中枢神经系统影响人的情绪、调节生命周期[1]。对一些特殊职业,如香精师、美食家、侦察员、医师、公安消防人员等,灵敏的嗅觉更是必不可少。嗅神经元是唯一暴露在外界环境中的神经元容易受到外界的损伤。FrankME[2]等报道人类永久性嗅觉障碍发病率约1.2%,暂时性嗅觉障碍发病率约62.4%,由于缺乏客观检查方法和发生机制不明等原因,嗅觉障碍一直缺乏有效的治疗方法。随着人们生活水平的提高,生活质量有更高的要求,嗅觉越来越引起重视。本实验通过对大鼠嗅感觉上皮发育过程的组织化学及大鼠嗅上皮细胞凋亡的研究以期对嗅觉障碍的发生及治疗提供一定的理论依据。
[研究目的2
免疫组化方法检测神经特异性烯醇酶(NSE)及嗅细胞标记蛋白(OMP)在胎鼠嗅上皮发育过程中的表达,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记技术(TUNEL)检测胎鼠嗅上皮发育过程中的细胞凋亡,以揭示哺乳动物胚胎期嗅上皮的发生、发育、成熟及凋亡过程。
[研究方法]
取大鼠不同孕期(E13-E21)胚胎鼻部标本,免疫组化方法检测神经特异性烯醇酶(NSE)及嗅细胞标记蛋白(OMP)在不同胎龄胎鼠嗅上皮中的表达及其规律。TUNEL方法检测其嗅上皮细胞凋亡,并计算细胞凋亡指数(AI)。统计学方法数据以x±s表示,采用spss10.0统计软件处理,各组间平均数比较采用非参数秩和检验,P＜0.05为具有统计学意义。
[结果]
大鼠E13天胎鼠嗅上皮中即有神经特异性烯醇酶表达(NSE),细胞数量多,细胞排列尚不规律,没有形成典型的柱状上皮形态。随胎龄增加阳性细胞数量增加,排列规律,细胞形态更接近双极细胞。E14-E21天胎鼠,在嗅上皮中均可见大量阳性细胞。E13天胎鼠嗅上皮中未见OMP阳性表达细胞。在大鼠E14天胎鼠,嗅上皮中开始出现嗅细胞标记蛋白(OMP)阳性细胞,数量少散在分布,随胎龄增加阳性细胞数量增多。至E17天达高峰,以后渐趋稳定。在E13天胎鼠嗅上皮中可见细胞凋亡,细胞凋亡数量少,E13-E15天胎鼠嗅上皮中细胞凋亡数量较稳定,至E16天胎鼠嗅上皮中凋亡细胞数量明显增加,达到高峰,E18天-E21天胎鼠嗅上皮中凋亡细胞数量逐渐减少渐趋于稳定。E16天胎鼠嗅上皮中凋亡指数与其他天数之间P＜0.05,差异有显著性。其他组间P＞0.05。
[结论]
嗅细胞标记蛋白(OMP)与神经特异性烯醇化酶(NSE)检测的联合应用可反应嗅上皮中嗅细胞发生、发育及成熟过程。大鼠胚胎发育过程中,胚胎中期嗅上皮中已有发育成熟的嗅神经元,胚胎后期嗅上皮已基本发育成熟,大鼠在出生时已具有一定的嗅功能。在嗅上皮发育过程中存在细胞凋亡,细胞凋亡在嗅上皮发生伊始(E13天)即已出现,是一生理过程,E16天出现一高峰,与嗅上皮和嗅球建立突触联系的时间相吻合,可能是两者相互作用的结果,提示细胞凋亡在嗅觉发育过程中对嗅细胞数量的稳定起重要作用。
Background:The olfactory is the important sensory organ for mammalian,as well as the visual sense,the auditory sense.It is a special structure for human to capture surrounding message.It can indentify the odour,discern circumstance and promote appetite and so on.Otherwise,it can affect human emotion and adjust life cycle by central nervous system.The sensitive olfactory is absolutely necessary to special occupation,such as the gastrologist,doctor,fire-fighter etc.Then,the olfactory recetor neurns are uniqueness neurns which expose to surrounding and facility to be hurt.Frank ME reported that the morbility of the permance dysosmia is 1.2%.The temporality dysosmia is 62.4%.As the objectively examination is deficiency and the genesis mechanism of dysosmia is not clear.The dysosmia is not cured effectively.This research will utilize immunohistochemical method and the Tdt-mediated dUTP nick end-labeling techniques to detect the development of the mouse olfactory epithelium.It perhaps provides a new path to treat the dysosmia.
Object:
This research is expected to detect and compare the expression and the value of the neuro-special endolase(NSE),the olfactory marker protein(OMP)and the apoptotic cells in the mouse olfactory epithelium of the different embryomic stages. The immunohistochemical method and the Tdt-mediated dUTP nick end-labeling techniques(TUNEL)were used.we expect to explore the developmental process of the mammal olfactory epithelium.
Method:
Embryonic mice were sacrificed by decapitation in defferent embryomic stages.The embryonic tissues were dissected out and fixed by immersion in 4% paraformaldehyde.The expression condition of NSE,OMP were examined by immunohistochemical method in the different embryomic stages from 13th day to the 21th day.TUNEL was applied to detect apoptotic cells of olfactory epithelium in the different embryomic stages,the apoptotic index was expressed for x±s.The statistic data of the apoptotic index was analyzed by Kruskal-Wallis test(P＜0.05)using the software systat version 10.0.
Result:
In embryonic 13th day,There were positive cells of NSE in the olfactory epitheliu.From E13 to E21,The positive cell of NSE increased and gradually stabilized.Otherwise,the positive cells of OMP were observed since E14 and the quantity was few.In E13 there was no the OMP-positive cell in olfactory epithelium.The noticeable increasing in number of OMP-positive neurons was observed in the olfactory epithelium with embryonic development.In E17,the quantity of OMP-positive neurons reached the peak.Afterward,quantity of OMP-positive neurons reached stabilization.The apoptotic process in the olfactory epithelium was observed form E13 to E21.In E13,the number of apoptotic cells was few and the AI was low.In E14,the number of apoptotic cells increased.A massive wave of cell death was evidenced in E16 and finally decreased to low levels at E19.Afterward,few apoptotic figures were observed and the apoptotic index in E20,E21 was very low.The apoptotic index of E16 was obviously defferentance from others by Kruskal-Wallis test(P＜0.05).The defferentance of the AI between the others was not obvious(P＜0.05).
Conclusion:
Olfactory receptor neurns of mousefetuses has begun since E13.But in this differentiated stage,the olfactory receptor neurns are immature.In E14,the mature Olfactory receptor neurns emerge and the quantity is few.With the increasing of embryonic days,the number of the mature Olfactory receptor neurns increases.To E21,the olfactory epithelium has been mature.The apoptotic process takes part in the development of the mouse olfactory epithelium.The apoptosis is a active process in the development of the mouse olfactory epithelium and the effect of the apoptosis is important.
嗅觉是哺乳动物的重要感觉器官之一,它同视觉、听觉一样,是人体捕获外界信息的特殊装置。人类的嗅觉具有辨别气味、增进食欲、识别环境、报警等作用。嗅觉还通过中枢神经系统影响人的情绪、调节生命周期[1]。对一些特殊职业,如香精师、美食家、侦察员、医师、公安消防人员等,灵敏的嗅觉更是必不可少。嗅神经元是唯一暴露在外界环境中的神经元容易受到外界的损伤。FrankME[2]等报道人类永久性嗅觉障碍发病率约1.2%,暂时性嗅觉障碍发病率约62.4%,由于缺乏客观检查方法和发生机制不明等原因,嗅觉障碍一直缺乏有效的治疗方法。随着人们生活水平的提高,生活质量有更高的要求,嗅觉越来越引起重视。本实验通过对大鼠嗅感觉上皮发育过程的组织化学及大鼠嗅上皮细胞凋亡的研究以期对嗅觉障碍的发生及治疗提供一定的理论依据。
[研究目的2
免疫组化方法检测神经特异性烯醇酶(NSE)及嗅细胞标记蛋白(OMP)在胎鼠嗅上皮发育过程中的表达,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记技术(TUNEL)检测胎鼠嗅上皮发育过程中的细胞凋亡,以揭示哺乳动物胚胎期嗅上皮的发生、发育、成熟及凋亡过程。
[研究方法]
取大鼠不同孕期(E13-E21)胚胎鼻部标本,免疫组化方法检测神经特异性烯醇酶(NSE)及嗅细胞标记蛋白(OMP)在不同胎龄胎鼠嗅上皮中的表达及其规律。TUNEL方法检测其嗅上皮细胞凋亡,并计算细胞凋亡指数(AI)。统计学方法数据以x±s表示,采用spss10.0统计软件处理,各组间平均数比较采用非参数秩和检验,P＜0.05为具有统计学意义。
[结果]
大鼠E13天胎鼠嗅上皮中即有神经特异性烯醇酶表达(NSE),细胞数量多,细胞排列尚不规律,没有形成典型的柱状上皮形态。随胎龄增加阳性细胞数量增加,排列规律,细胞形态更接近双极细胞。E14-E21天胎鼠,在嗅上皮中均可见大量阳性细胞。E13天胎鼠嗅上皮中未见OMP阳性表达细胞。在大鼠E14天胎鼠,嗅上皮中开始出现嗅细胞标记蛋白(OMP)阳性细胞,数量少散在分布,随胎龄增加阳性细胞数量增多。至E17天达高峰,以后渐趋稳定。在E13天胎鼠嗅上皮中可见细胞凋亡,细胞凋亡数量少,E13-E15天胎鼠嗅上皮中细胞凋亡数量较稳定,至E16天胎鼠嗅上皮中凋亡细胞数量明显增加,达到高峰,E18天-E21天胎鼠嗅上皮中凋亡细胞数量逐渐减少渐趋于稳定。E16天胎鼠嗅上皮中凋亡指数与其他天数之间P＜0.05,差异有显著性。其他组间P＞0.05。
[结论]
嗅细胞标记蛋白(OMP)与神经特异性烯醇化酶(NSE)检测的联合应用可反应嗅上皮中嗅细胞发生、发育及成熟过程。大鼠胚胎发育过程中,胚胎中期嗅上皮中已有发育成熟的嗅神经元,胚胎后期嗅上皮已基本发育成熟,大鼠在出生时已具有一定的嗅功能。在嗅上皮发育过程中存在细胞凋亡,细胞凋亡在嗅上皮发生伊始(E13天)即已出现,是一生理过程,E16天出现一高峰,与嗅上皮和嗅球建立突触联系的时间相吻合,可能是两者相互作用的结果,提示细胞凋亡在嗅觉发育过程中对嗅细胞数量的稳定起重要作用。
Background:The olfactory is the important sensory organ for mammalian,as well as the visual sense,the auditory sense.It is a special structure for human to capture surrounding message.It can indentify the odour,discern circumstance and promote appetite and so on.Otherwise,it can affect human emotion and adjust life cycle by central nervous system.The sensitive olfactory is absolutely necessary to special occupation,such as the gastrologist,doctor,fire-fighter etc.Then,the olfactory recetor neurns are uniqueness neurns which expose to surrounding and facility to be hurt.Frank ME reported that the morbility of the permance dysosmia is 1.2%.The temporality dysosmia is 62.4%.As the objectively examination is deficiency and the genesis mechanism of dysosmia is not clear.The dysosmia is not cured effectively.This research will utilize immunohistochemical method and the Tdt-mediated dUTP nick end-labeling techniques to detect the development of the mouse olfactory epithelium.It perhaps provides a new path to treat the dysosmia.
Object:
This research is expected to detect and compare the expression and the value of the neuro-special endolase(NSE),the olfactory marker protein(OMP)and the apoptotic cells in the mouse olfactory epithelium of the different embryomic stages. The immunohistochemical method and the Tdt-mediated dUTP nick end-labeling techniques(TUNEL)were used.we expect to explore the developmental process of the mammal olfactory epithelium.
Method:
Embryonic mice were sacrificed by decapitation in defferent embryomic stages.The embryonic tissues were dissected out and fixed by immersion in 4% paraformaldehyde.The expression condition of NSE,OMP were examined by immunohistochemical method in the different embryomic stages from 13th day to the 21th day.TUNEL was applied to detect apoptotic cells of olfactory epithelium in the different embryomic stages,the apoptotic index was expressed for x±s.The statistic data of the apoptotic index was analyzed by Kruskal-Wallis test(P＜0.05)using the software systat version 10.0.
Result:
In embryonic 13th day,There were positive cells of NSE in the olfactory epitheliu.From E13 to E21,The positive cell of NSE increased and gradually stabilized.Otherwise,the positive cells of OMP were observed since E14 and the quantity was few.In E13 there was no the OMP-positive cell in olfactory epithelium.The noticeable increasing in number of OMP-positive neurons was observed in the olfactory epithelium with embryonic development.In E17,the quantity of OMP-positive neurons reached the peak.Afterward,quantity of OMP-positive neurons reached stabilization.The apoptotic process in the olfactory epithelium was observed form E13 to E21.In E13,the number of apoptotic cells was few and the AI was low.In E14,the number of apoptotic cells increased.A massive wave of cell death was evidenced in E16 and finally decreased to low levels at E19.Afterward,few apoptotic figures were observed and the apoptotic index in E20,E21 was very low.The apoptotic index of E16 was obviously defferentance from others by Kruskal-Wallis test(P＜0.05).The defferentance of the AI between the others was not obvious(P＜0.05).
Conclusion:
Olfactory receptor neurns of mousefetuses has begun since E13.But in this differentiated stage,the olfactory receptor neurns are immature.In E14,the mature Olfactory receptor neurns emerge and the quantity is few.With the increasing of embryonic days,the number of the mature Olfactory receptor neurns increases.To E21,the olfactory epithelium has been mature.The apoptotic process takes part in the development of the mouse olfactory epithelium.The apoptosis is a active process in the development of the mouse olfactory epithelium and the effect of the apoptosis is important.
引文
1、 Sullivan SL,Dryer L,Information processing in mammalian olfactory system[J],Neurobion.1996 May,30(1):20-36.
2、 Allergic rhinitis and olfactory loss Ann Allergy Asthma Immunnl.1995 Oct 75(4)311-316
3、 Holbrook EH,Leopold DA.Anosmia:diagnosis and management[J].Curr Opin Otolaryngol Head Neck Surg.2003 Feb;11(1):54-60,
4、 黄选兆,汪吉宝.实用耳鼻咽喉科学[M].北京:人民卫生出版社,2006.96-97:
5、 Matthew H,Kaufman.The atlas of mouse development[M].San Deigo:Elsevier,2003:215-217.
6、 Marshall CT,Lu C,Winstead W,et al,The therapeutic potential of human olfactory-derived stem cells.Histol Histopathol[J].2006 Jun;21(6):633-43
7、 Reden J,Mueller A,Mueller C,et al.Recovery of Olfactory Function Following Closed Head Injury or Infections of the Upper Respiratory Tract.Arch Otolaryngol Head Neck Surg,2006,132:265-269.
8、 胡金玮,黄维国,姜鸿彦,等.胚胎期豚鼠前庭终器细胞凋亡的检测[J].第四军医大学学报,2002:23(16):1514-1517.
9、 Biffo S,Sassoe'M Pognetto,Verdun L,et al,Bulbectomy enhances neurogenesis and cell turnover of primary olfactory neurons but does not abolish carnosine expression[J],Eur.J.Neurosci.1992,4:1398 - 1406.
10、 Schwob J.E,Mielesko K.e.,Stasky A.A.,Olfactory sensory neurons are trophically dependent on the olfactory bulb for their prolonged survival,[J].Neurosci.1992.12:3896 - 3919.
11、 Amoore JE,Steinle S 1991.A graphic history of specific anosmia.In Chemical Senses.Vol.3.C.J.Lancet D,Vertebrate olfactory reception.Annual Review of Neuroscience.1986,9:329-355.
12、 Beets J,Olfactory response and molecular structure.Handbook of sensory physiology,1971,4:257-321。
13 、 Graziadei P P C, Okano M ,Neuronal degeneration and regeneration in olfactory epithelium of pigeon following transaction of first cranial nerve. Acta anat. 1979:104,220-236.
14、 Yamagishi M,Nakamura H,Takahashi S,al et ,Olfactory receptor celkimmmunocytochemistry for nervous system-specific proteins and reevaluation of their precursor cells. Archs Histol.Cytol.52,Suppl.375-381.
15、 Osako Y, Otsuka T, Taniguchi M, et al. Oxytocin enhances presynaptic and postsynaptic glutamatergic transimision between rat olfactory bubl neurons in culture [J],ncurosci Lett,2001,299(1):65
16、 Bock E.Nervous system specific proteins[J].Neurochem 1978,30:7-14 17、 Schmechel DE,Marangos PJ,Brightman MW.Neuron-specific enolase is a molecular marker for peripheral and central neuoenddocrine cells[J].Nature 1979,276:834-846.
18、 Heilmann S, Hummel T,Margolis FL,et al. Immunhistochemical distribution ofgalectin-3,and olfactory marker protein in human olfactory epithelium.Histochem Cell Biol,2000,113:241-245.
19、 Nibu K,Li G,Zhang X,et al. Olfactory neuron-specific expression of NeuroD in mouse and human nasal mucosa.Cell Tissue Res,1999,298:405-414.
20、 Margolis FL.A brain protein uniquer to the olfactory bulb. Proc Natl Acad Sci USA, 1972,69:1221-1224.
21、 Farbman AI,Margolish FL,Olfactory marker protein during ontogeny:immunohistochemical localization.Dev Biol 1980,74:205-215.
22、 Menco BP. Electron-microscopic demonstration of olfactory-marker protein with protein G-gold in freeze-substituted, lowicryl KIIM-embedded rat olfactory receptor cells. Cell Tissue Res, 1989,256:275-281.
23、 Suzuki Y, Takeda M. Basal cells in the mouse olfactory epithelium after axotomy: immunohistochemical and electron microscopic studies. Cell Tissue Res, 1991, 266: 239-245. 24、 Farbman AI. Cell biology of olfaction. New York: Cambridge, 1992.
25、 Farbman AI.Developmental biology of olfactory sensory neurons.Semin Cell Biol,1994,5:3-10.
26、 Verhaagen J,Oestreicher AB,Grillo M,et al.Neuroplasticity in the olfactory system:differential effects of central and peripheral lesions of the primary olfactory pathway on the expression of B-50 / GAP43 and the olfactory marker protein.J Neurosci Res,1990,26:31-44.
27、王鸿杰,张志文。嗅觉之谜-2004年诺贝尔生理学或医学奖简介[J]。生理科学进展,2005,36(1):93-94。
28、 Winner B,Cooper-Kuhn CM,Aigner R,et al.Long-term survival and cell death of newly generated neurons in the adult rat olfactory bulb[J].Eur J Neurosci,2002;16(9):1681-1869.
29、 Kerr JF,Wyllie AH,Currie AR.Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics Br,J C ancer,1972,26(4):239-257.
30、 Haunstetter A,lzumo S.Apoptosis:basic mechanisms and implications for cardiov asculard iseases.CircR es,19 98,82:11- 2 9.
31、 Marion M.TRAIL induced signaling and a poptosis.Toxicol Lett,2 003,139:89-97。
32、 Cowan CM,Thai J,Krajewski S,et al.Caspases 3 and 9 send a pro - apoptotic signal from synapse to cell body in olfactory receptor neurons[J].J Neurosci,2001,21(18):7099 - 7109.
33、 Holcomb JD,Mumm JS,Calof AI.Apoptosis in the neuronal lineage of the mouse olfactory epithelium:regulation in vivo and vitro.Dev B iol,1995,172:307-323
34、 Cowan C M,Thai J,Krajewski S,et al.Caspase-3 and 9 send pro-apoptotic signalfrom synapse to cell body in olfactory receptor neurons.J Neurosci,2001,21(18):7099-7109.
35、 Holcomb JD,MummJS,Calof AL.Apoptosis in the neuronal lineage of the mouse olfactory epithelium:regulation in vivo and in vitro[J].Dev Biol,1995,172(1):307- 323.
36、 Mahalik TJ . Apparent apoptotic cell death in the olfactory epithelium of adult rodents : death occurs at different developmeanal stages [J ]. J Comp Neurol ,1996 ,372(3): 457 - 464.
37、 OppenheimRW. Cell death during development of the nervous system. A nnu Rev Neurosic, 1991,14:453.
38、 Mathormet M,Lalloue F,Petit B, et al. Differential responses of olfactory neurons to axotomy at embryonic and postnatal stages. Neuroscience. 2002;109(2):207-17.
39、 Halliwell B , Gall CM. Free radicals in biology and medicine.Oxofrd:Clarendon Press, 1989.
1. Minai C, Bracale A M, Moreschi C, et al. Post—traumatic anosmia: description of a clinical case, proposal of a standardized protocol and medico-legal comments[J]. Acta Otorhinolaryngol Hal, 2002, 22: 142— 149.
2. Temmel A F. Quint C, Schickinger F B, et al. Characteristics of olfactory disorders in relation to major causes of olfactory loss[J]. Arch Otolaryngol Head Neck Surg, 2002, 128: 635— 641.
3. KERN R C. QLJINN B, ROSSEAU G, et al. Post—traumatic olfactory dysfunction[J]. Laryngoscope, 2000, 110: 2106— 2109.
4. Mouly AM,Gervais R. Polysynaptic potentia—tion at different levels of rat olfactory pathways following learning[J]. Learn Mem, 2002,9: 66—75.
5. Burdach KJ ,Doty RL. The effects of mouth movement ,swallowing. and spitting on retronasal odor perception[J ]. Physiol Behav ,1987 ,41(4) :353 - 356.
6. Donald Leopold. A perplexing olfactory loss[J ]. Arch otolaryngol Head and Neck Surg ,2000 ,126 :803.
7. Thovnas Humme. Perspectives in olfactory loss following viral infections of the upper respiratory tract. Arch otolaryngol [J ] .Head and Neck Surg ,2000 ,126:802.
8. Damm M, Quante G, Jungehlsing M , et al. Impact of functional endoscopic sinus surgery on symptoms and quality of life in chronic rhino sinusitis [J]. Laryngoscop, 2002, 112:310-315.
9. Moll B, Klimek L, Eggers G. Comparison of olfactory, function in patients with seasonal and perennial allergic rhinitis[J]. Allergy, 1998, 53: 297—301.
10. Ph Rombaux, S Collet, Ph Eloy. Smell disorders in ENT clinic[J]. B. ENT, 2005, KSuppl 1): 97-109.
11. Beverly J. Cowart, Karin Flynn-Rodden, Stephen J McGeady. Hyposmia in allergic rhinitis[J . J Allergy Clin Immunol, 1993, 91: 747-751.
12.Malnic B,Godfrey PA,Buck LB.the human olfactory receptor gene family.Proc Natl Acad Sci USA,2004,101:2584-2589.
13.Hu Y,Tanriverdi F,MacColl GS,et al.Kallmann' s syndrome:molecular pathogenesis.Int J Biochem cell Biol,2003,35:1157-1162.
14.Abolmaali,Nasreddin D,Hietschold Volker,et aI.M R evaluation in patients with isolated anosmia since birth or early childho[J].NR Am J NeuroradioI,2002,23(1):157-64.
15.Yousem DM,Geckle RJ,Bilker W,et al.MR evaluation of patients with congenital hyposmia or anosmia.AJR Am J roentgenol,1996,166:439-443.
16.Jafek BW,Gordon AS,Moran DT,et al.CongenitaI anosmia[J].Ear Nose Throat J,1990,69:331-337.
17.Leopold DA,Hornung DE,Schwob JE.Congenital lack of olfactory ability.Ann Otol Rhinol Laryngol,1992,101:229-236.
18.Tisingh G,Berendse HW,Bergmans P,et al.Loss of olfaction in denovo and treated Parkinso n s disease:possible implica-tions for early diagnosis[J].Mov Disord,2001,16(1):41-46.
19.Seiden A M,Duncan H J.The diagnosis of a conductive olfactory loss.Laryngoscope,2001,111:9-14.
20.Braun D,Wagner W,Zenner H P,et al.Disabling disturbance of olfaction in a dental technician following exposure tO methyl ethacrylate.Int Arch Occup Environ Heahh,2002,75:73-74.
21.Sunderman F W.Nasal toxicity,carcinogenicity,and olfactory uptake of metals.Ann Clin Lab Sci,200 1,3 1:3-24.
22.Lee S H,Lim H H,Lee H M,et al.Olfactory mucosal findings in patients with persistent anosmia after endoscopic sinus surgery.Ann Otol Rhinol Laryngol,2000,109(8 Pt 1):720-725
23.史丽,陈瑛,任翔,等.神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达.中华耳鼻咽喉科杂志,2003,38(3):180-182.
24.关桂梅,董震,杨占泉,等.大鼠嗅球中凋亡相关基因蛋白和成纤维细胞因 子表达的相关性及其意义.中华耳鼻咽喉科杂志,2000,35(5):345-347.
25.倪道凤,嗅觉障碍及嗅功能检查{J} 临床耳鼻咽喉科杂志,2003,17(9):571-575。
26.Ishimaru T.Shimada T,M iwa T.et al.Electrically stimulated olfactory evoked potential in olfactory disturbance.Ann Otol Rhinol I aryngol。 2002,111:518-522.
27.Abolmaali ND,Hietschold V,Nogl TJ,et al.MR evaluation in patients with isolated anosmia since birth or early childhood.AJNR Am J Neuroradiol,2002 Jan;23(1):157-164.
28.KurahashiT,MeniniA。Mechanism of odorant adaptation in the olfactory receptor cell.Nature,1997,385:725-729.
29.Duncan HJ.Seiden AM.Long-term follow-up of olfactory loss secondary to head trauma an d upper respiratory tract infection.Arch Otolaryngol Head Neck Surg.1995:121:1183-1187.
30.Hummel T.Th erapy of olfactory loss.Laryngorhinootologie.2003:82:552-554.
31.赵红,魏永祥,苗旭涛等,嗅觉障碍与慢性鼻-鼻窦炎患者鼻腔鼻窦形态的相关性研究[J]临床耳鼻咽喉-头颈外科杂志,2007,21(17):771-774.
32.汪学勇,影响慢性鼻窦炎鼻息肉患者术后嗅觉改善的因素[J]临床耳鼻咽喉科杂志,2003,17(10):609-611。
33.周兵,韩德民,刘华超,等.少年儿童鼻内镜手术远期疗效及相关临床因素探讨.中华耳鼻咽喉科杂志.2003;38:255-258。
34.Leopold DA,Schuob JE,Youngentob SL,et al.SuccessfuItreatment of phantosmiawith preservation of olfaction[J].Arch OtoIaryngoI Head and Neck Surg,1991,1 17(12):1402-1406.
2、 Allergic rhinitis and olfactory loss Ann Allergy Asthma Immunnl.1995 Oct 75(4)311-316
3、 Holbrook EH,Leopold DA.Anosmia:diagnosis and management[J].Curr Opin Otolaryngol Head Neck Surg.2003 Feb;11(1):54-60,
4、 黄选兆,汪吉宝.实用耳鼻咽喉科学[M].北京:人民卫生出版社,2006.96-97:
5、 Matthew H,Kaufman.The atlas of mouse development[M].San Deigo:Elsevier,2003:215-217.
6、 Marshall CT,Lu C,Winstead W,et al,The therapeutic potential of human olfactory-derived stem cells.Histol Histopathol[J].2006 Jun;21(6):633-43
7、 Reden J,Mueller A,Mueller C,et al.Recovery of Olfactory Function Following Closed Head Injury or Infections of the Upper Respiratory Tract.Arch Otolaryngol Head Neck Surg,2006,132:265-269.
8、 胡金玮,黄维国,姜鸿彦,等.胚胎期豚鼠前庭终器细胞凋亡的检测[J].第四军医大学学报,2002:23(16):1514-1517.
9、 Biffo S,Sassoe'M Pognetto,Verdun L,et al,Bulbectomy enhances neurogenesis and cell turnover of primary olfactory neurons but does not abolish carnosine expression[J],Eur.J.Neurosci.1992,4:1398 - 1406.
10、 Schwob J.E,Mielesko K.e.,Stasky A.A.,Olfactory sensory neurons are trophically dependent on the olfactory bulb for their prolonged survival,[J].Neurosci.1992.12:3896 - 3919.
11、 Amoore JE,Steinle S 1991.A graphic history of specific anosmia.In Chemical Senses.Vol.3.C.J.Lancet D,Vertebrate olfactory reception.Annual Review of Neuroscience.1986,9:329-355.
12、 Beets J,Olfactory response and molecular structure.Handbook of sensory physiology,1971,4:257-321。
13 、 Graziadei P P C, Okano M ,Neuronal degeneration and regeneration in olfactory epithelium of pigeon following transaction of first cranial nerve. Acta anat. 1979:104,220-236.
14、 Yamagishi M,Nakamura H,Takahashi S,al et ,Olfactory receptor celkimmmunocytochemistry for nervous system-specific proteins and reevaluation of their precursor cells. Archs Histol.Cytol.52,Suppl.375-381.
15、 Osako Y, Otsuka T, Taniguchi M, et al. Oxytocin enhances presynaptic and postsynaptic glutamatergic transimision between rat olfactory bubl neurons in culture [J],ncurosci Lett,2001,299(1):65
16、 Bock E.Nervous system specific proteins[J].Neurochem 1978,30:7-14 17、 Schmechel DE,Marangos PJ,Brightman MW.Neuron-specific enolase is a molecular marker for peripheral and central neuoenddocrine cells[J].Nature 1979,276:834-846.
18、 Heilmann S, Hummel T,Margolis FL,et al. Immunhistochemical distribution ofgalectin-3,and olfactory marker protein in human olfactory epithelium.Histochem Cell Biol,2000,113:241-245.
19、 Nibu K,Li G,Zhang X,et al. Olfactory neuron-specific expression of NeuroD in mouse and human nasal mucosa.Cell Tissue Res,1999,298:405-414.
20、 Margolis FL.A brain protein uniquer to the olfactory bulb. Proc Natl Acad Sci USA, 1972,69:1221-1224.
21、 Farbman AI,Margolish FL,Olfactory marker protein during ontogeny:immunohistochemical localization.Dev Biol 1980,74:205-215.
22、 Menco BP. Electron-microscopic demonstration of olfactory-marker protein with protein G-gold in freeze-substituted, lowicryl KIIM-embedded rat olfactory receptor cells. Cell Tissue Res, 1989,256:275-281.
23、 Suzuki Y, Takeda M. Basal cells in the mouse olfactory epithelium after axotomy: immunohistochemical and electron microscopic studies. Cell Tissue Res, 1991, 266: 239-245. 24、 Farbman AI. Cell biology of olfaction. New York: Cambridge, 1992.
25、 Farbman AI.Developmental biology of olfactory sensory neurons.Semin Cell Biol,1994,5:3-10.
26、 Verhaagen J,Oestreicher AB,Grillo M,et al.Neuroplasticity in the olfactory system:differential effects of central and peripheral lesions of the primary olfactory pathway on the expression of B-50 / GAP43 and the olfactory marker protein.J Neurosci Res,1990,26:31-44.
27、王鸿杰,张志文。嗅觉之谜-2004年诺贝尔生理学或医学奖简介[J]。生理科学进展,2005,36(1):93-94。
28、 Winner B,Cooper-Kuhn CM,Aigner R,et al.Long-term survival and cell death of newly generated neurons in the adult rat olfactory bulb[J].Eur J Neurosci,2002;16(9):1681-1869.
29、 Kerr JF,Wyllie AH,Currie AR.Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics Br,J C ancer,1972,26(4):239-257.
30、 Haunstetter A,lzumo S.Apoptosis:basic mechanisms and implications for cardiov asculard iseases.CircR es,19 98,82:11- 2 9.
31、 Marion M.TRAIL induced signaling and a poptosis.Toxicol Lett,2 003,139:89-97。
32、 Cowan CM,Thai J,Krajewski S,et al.Caspases 3 and 9 send a pro - apoptotic signal from synapse to cell body in olfactory receptor neurons[J].J Neurosci,2001,21(18):7099 - 7109.
33、 Holcomb JD,Mumm JS,Calof AI.Apoptosis in the neuronal lineage of the mouse olfactory epithelium:regulation in vivo and vitro.Dev B iol,1995,172:307-323
34、 Cowan C M,Thai J,Krajewski S,et al.Caspase-3 and 9 send pro-apoptotic signalfrom synapse to cell body in olfactory receptor neurons.J Neurosci,2001,21(18):7099-7109.
35、 Holcomb JD,MummJS,Calof AL.Apoptosis in the neuronal lineage of the mouse olfactory epithelium:regulation in vivo and in vitro[J].Dev Biol,1995,172(1):307- 323.
36、 Mahalik TJ . Apparent apoptotic cell death in the olfactory epithelium of adult rodents : death occurs at different developmeanal stages [J ]. J Comp Neurol ,1996 ,372(3): 457 - 464.
37、 OppenheimRW. Cell death during development of the nervous system. A nnu Rev Neurosic, 1991,14:453.
38、 Mathormet M,Lalloue F,Petit B, et al. Differential responses of olfactory neurons to axotomy at embryonic and postnatal stages. Neuroscience. 2002;109(2):207-17.
39、 Halliwell B , Gall CM. Free radicals in biology and medicine.Oxofrd:Clarendon Press, 1989.
1. Minai C, Bracale A M, Moreschi C, et al. Post—traumatic anosmia: description of a clinical case, proposal of a standardized protocol and medico-legal comments[J]. Acta Otorhinolaryngol Hal, 2002, 22: 142— 149.
2. Temmel A F. Quint C, Schickinger F B, et al. Characteristics of olfactory disorders in relation to major causes of olfactory loss[J]. Arch Otolaryngol Head Neck Surg, 2002, 128: 635— 641.
3. KERN R C. QLJINN B, ROSSEAU G, et al. Post—traumatic olfactory dysfunction[J]. Laryngoscope, 2000, 110: 2106— 2109.
4. Mouly AM,Gervais R. Polysynaptic potentia—tion at different levels of rat olfactory pathways following learning[J]. Learn Mem, 2002,9: 66—75.
5. Burdach KJ ,Doty RL. The effects of mouth movement ,swallowing. and spitting on retronasal odor perception[J ]. Physiol Behav ,1987 ,41(4) :353 - 356.
6. Donald Leopold. A perplexing olfactory loss[J ]. Arch otolaryngol Head and Neck Surg ,2000 ,126 :803.
7. Thovnas Humme. Perspectives in olfactory loss following viral infections of the upper respiratory tract. Arch otolaryngol [J ] .Head and Neck Surg ,2000 ,126:802.
8. Damm M, Quante G, Jungehlsing M , et al. Impact of functional endoscopic sinus surgery on symptoms and quality of life in chronic rhino sinusitis [J]. Laryngoscop, 2002, 112:310-315.
9. Moll B, Klimek L, Eggers G. Comparison of olfactory, function in patients with seasonal and perennial allergic rhinitis[J]. Allergy, 1998, 53: 297—301.
10. Ph Rombaux, S Collet, Ph Eloy. Smell disorders in ENT clinic[J]. B. ENT, 2005, KSuppl 1): 97-109.
11. Beverly J. Cowart, Karin Flynn-Rodden, Stephen J McGeady. Hyposmia in allergic rhinitis[J . J Allergy Clin Immunol, 1993, 91: 747-751.
12.Malnic B,Godfrey PA,Buck LB.the human olfactory receptor gene family.Proc Natl Acad Sci USA,2004,101:2584-2589.
13.Hu Y,Tanriverdi F,MacColl GS,et al.Kallmann' s syndrome:molecular pathogenesis.Int J Biochem cell Biol,2003,35:1157-1162.
14.Abolmaali,Nasreddin D,Hietschold Volker,et aI.M R evaluation in patients with isolated anosmia since birth or early childho[J].NR Am J NeuroradioI,2002,23(1):157-64.
15.Yousem DM,Geckle RJ,Bilker W,et al.MR evaluation of patients with congenital hyposmia or anosmia.AJR Am J roentgenol,1996,166:439-443.
16.Jafek BW,Gordon AS,Moran DT,et al.CongenitaI anosmia[J].Ear Nose Throat J,1990,69:331-337.
17.Leopold DA,Hornung DE,Schwob JE.Congenital lack of olfactory ability.Ann Otol Rhinol Laryngol,1992,101:229-236.
18.Tisingh G,Berendse HW,Bergmans P,et al.Loss of olfaction in denovo and treated Parkinso n s disease:possible implica-tions for early diagnosis[J].Mov Disord,2001,16(1):41-46.
19.Seiden A M,Duncan H J.The diagnosis of a conductive olfactory loss.Laryngoscope,2001,111:9-14.
20.Braun D,Wagner W,Zenner H P,et al.Disabling disturbance of olfaction in a dental technician following exposure tO methyl ethacrylate.Int Arch Occup Environ Heahh,2002,75:73-74.
21.Sunderman F W.Nasal toxicity,carcinogenicity,and olfactory uptake of metals.Ann Clin Lab Sci,200 1,3 1:3-24.
22.Lee S H,Lim H H,Lee H M,et al.Olfactory mucosal findings in patients with persistent anosmia after endoscopic sinus surgery.Ann Otol Rhinol Laryngol,2000,109(8 Pt 1):720-725
23.史丽,陈瑛,任翔,等.神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达.中华耳鼻咽喉科杂志,2003,38(3):180-182.
24.关桂梅,董震,杨占泉,等.大鼠嗅球中凋亡相关基因蛋白和成纤维细胞因 子表达的相关性及其意义.中华耳鼻咽喉科杂志,2000,35(5):345-347.
25.倪道凤,嗅觉障碍及嗅功能检查{J} 临床耳鼻咽喉科杂志,2003,17(9):571-575。
26.Ishimaru T.Shimada T,M iwa T.et al.Electrically stimulated olfactory evoked potential in olfactory disturbance.Ann Otol Rhinol I aryngol。 2002,111:518-522.
27.Abolmaali ND,Hietschold V,Nogl TJ,et al.MR evaluation in patients with isolated anosmia since birth or early childhood.AJNR Am J Neuroradiol,2002 Jan;23(1):157-164.
28.KurahashiT,MeniniA。Mechanism of odorant adaptation in the olfactory receptor cell.Nature,1997,385:725-729.
29.Duncan HJ.Seiden AM.Long-term follow-up of olfactory loss secondary to head trauma an d upper respiratory tract infection.Arch Otolaryngol Head Neck Surg.1995:121:1183-1187.
30.Hummel T.Th erapy of olfactory loss.Laryngorhinootologie.2003:82:552-554.
31.赵红,魏永祥,苗旭涛等,嗅觉障碍与慢性鼻-鼻窦炎患者鼻腔鼻窦形态的相关性研究[J]临床耳鼻咽喉-头颈外科杂志,2007,21(17):771-774.
32.汪学勇,影响慢性鼻窦炎鼻息肉患者术后嗅觉改善的因素[J]临床耳鼻咽喉科杂志,2003,17(10):609-611。
33.周兵,韩德民,刘华超,等.少年儿童鼻内镜手术远期疗效及相关临床因素探讨.中华耳鼻咽喉科杂志.2003;38:255-258。
34.Leopold DA,Schuob JE,Youngentob SL,et al.SuccessfuItreatment of phantosmiawith preservation of olfaction[J].Arch OtoIaryngoI Head and Neck Surg,1991,1 17(12):1402-1406.