原发性开角型青光眼的遗传学调查与OPTN基因突变的研究
摘要
目的：
本课题对重庆地区一个POAG家系进行了详尽的遗传学调查以及系谱分析，拟阐明此家系POAG的发病与遗传的关系，并分析其遗传特征。进而通过分子生物学方法对家系中POAG患者、一级亲属的OPTN基因的编码外显子进行PCR扩增，以期发现是否存在OPTN基因的突变，并分析突变可能造成的影响。
研究方法：
1. 收集POAG家系，并根据孟德尔原则进行详尽的遗传学调查分析。
2. 提取所有样本的基因组DNA，PCR 扩增的13个外显子，部分外显子周围的内含子，共4350bp。
3. 对家系中所有POAG患者及一级亲属OPTN基因编码外显子直接测序。
4. 根据测序结果，选择特异性内切酶在散发性正常对照组中进行限制性长度片段多态性分析。
5. 统计分析家系POAG组和对照组中OPTN基因突变分布差异性。
6. 对检测到的OPTN基因突变进行生物信息学分析。
结果：
1. 该家系可追踪到的有五代共54人，男性29人，女性25人。其中已确诊的POAG患者17 例，男性患者11人，女性患者6人， 6人已经死亡。家系中连续四代发病。POAG的患病率为31％。
2. 患者IV-5、III-5、 III-4以及一级亲属IV-7中第4外显子412G>A.。患者II-5的第5外显子603T>A。患者III-5第10外显子1267-1268ins C/1271-1272ins C。III-3患者第12外显子1537-1538ins C。一级亲属V-3出现15外显子1878-1879ins A。
3. 32例正常对照组中有1例出现412G>A突变，其他突变型在对照组中均未发现。
4. 在不计算412G>A突变情况下，POAG家族发生OPTN基因编码区的突变概率为20%，而对照组中未发现。经X2分析，X2=6.93> X20.016.63，P<0.01, POAG家族与对照组相差显著。
生物信息学分析表明除第4外显子突变不改变OPTN蛋白的结构外，其他外
5. 显子突变均将导致结构的变化，影响OPTN的重要功能域。
结论：
1. 重庆忠县李某POAG家系遗传方式属常染色体显性遗传。在家系中存在导致POAG发病的突变的基因。
2. OPTN基因突变与本家系POAG发病相关。
3. 基因突变造成OPTN蛋白一二级结构和重要功能域异常，导致OPTN蛋白功能的异常可能是POAG发病的重要原因之一。
4. 本研究中共发现5种类型的OPTN基因突变（412G>A、 603T>A 1267-1268ins C/1271-1272ins C、 1537-1538ins C、 1878-1879ins A）。除412G>A、 603T>A两种与报道相同外，其余均为本次研究中第一次发现。
Glaucoma is the second leading cause of blindness which affect over 70 million people worldwide. Primary open angle glaucoma (POAG) is the most common subtype and the first reason of blindness of black man in America. Moreover, There are many subjects suffering from POAG in China. POAG is a kind of complex latent disease characterized by chronic progressive optic ganglion cells apoptosis and visual field injury. Thus, It is difficult to diagnoses and treat in early period. Although it is not yet wholly understood about the mechanism of glaucoma, more and more evidences suggest that there is an association between heredity and gene mutations with pathogenesis of glaucoma. In spite of great progress of glaucoma genetics, the hereditary form of POAG is not clear. on the other hand, about ten genes loci were found to associate with POAG after the discovery that TIGR gene links to POAG based on molecular genetics assay. The optineurin（optic neuropathy inducing, OPTN）, a novel gene linked to POAG, was found by Reiaze and his colleagues in 2002. Mutations in the OPTN gene were found in 16.7% of 54 families with autosomal dominant adult-onset POAG, and in 13.6% of the sporadic glaucoma cases. However, there is no report about mutations of OPTN gene in Chinese POAG cases.
Purpose:
To investigate the relationship between POAG and the mutations of OPTN gene in China, a POAG family in ChongQings was followed up, and the blood specimens of POAG individuals and controls were collected and processed by molecular biological strategy.
Methods:
1. To analyze the genetic form of the pedigree according to Mandalian hereditary rules after we collect a great family with POAG.
2. The genomic DNA was extracted from the peripheral blood of all samples. A total of 4350bp of 13 encoding exons and introns beside exons were amplified by PCR using the sequence-specific primers.
3. Automated sequencing method was used to screen and identify mutations of OPTN
gene of both POAG subjects and first degree relatives in this family.
4. Restriction endonuclease analysis was used to detect the same mutation in the controls samples.
5. The frequency of genetypes between POAG group and controls were compared by statistics.
6. The sequence alterations were analyzed by bioinfomatics.
Results:
1. This pedigree could be traced to 54 members in 5 generations, Including 29 males and 25 females. There are 17 patients with POAG, including 11 males and 6 females. Six patients had died in this family. The four continuous generations exist patients with POAG. The prevalence rate is 31%.
2. The encoding exons of OPTN gene of all alive patients and collected first degree relatives were directly automatic- sequenced, it is found that 3 patients ( IV-5、III-5、 III-4) and one first degree relative(IV-7) exist the 412G>A mutation in 4th exon. II-5 patient has a 603T>A alteration. III-5 was found that two C pyrimidine insertions between 1267-1268 site and 1271-1272 of 10th exon, respectively. III-3 exist 1537-1538ins (insert) C alteration. 1878-1879ins A had been found in a first degree relative (V-3).
3. No same mutations of OPTN gene were detected in 32 controls, except one person has a 412G>A mutation.
4. Rate of observed mutations in POAG family is 20%, no mutations were found in controls. The mutation rate of POAG family group is obviously higher than that in the controls group( X2=6.93> X20.016.63，P<0.01).
5. Bioinformatics analysis indicated that all mutations will result in the alteration of OPTN protein structure which may underlies its abnormal functional domain except for 412G>A mutation.
Conclusions:
1. The form of heredity in this POAG family in Zhong county of Chongqing is autosomal dominant trait, and there must be a abnormal gene which result in POAG in this family.
2. The mutation of OPTN gene is responsible for onset of POAG in this pedigree.
3. Function abnormality of OPTN protein, which are induced by change of the structure and functional domains caused by mutation of OPTN, may be an important m
本课题对重庆地区一个POAG家系进行了详尽的遗传学调查以及系谱分析，拟阐明此家系POAG的发病与遗传的关系，并分析其遗传特征。进而通过分子生物学方法对家系中POAG患者、一级亲属的OPTN基因的编码外显子进行PCR扩增，以期发现是否存在OPTN基因的突变，并分析突变可能造成的影响。
研究方法：
1. 收集POAG家系，并根据孟德尔原则进行详尽的遗传学调查分析。
2. 提取所有样本的基因组DNA，PCR 扩增的13个外显子，部分外显子周围的内含子，共4350bp。
3. 对家系中所有POAG患者及一级亲属OPTN基因编码外显子直接测序。
4. 根据测序结果，选择特异性内切酶在散发性正常对照组中进行限制性长度片段多态性分析。
5. 统计分析家系POAG组和对照组中OPTN基因突变分布差异性。
6. 对检测到的OPTN基因突变进行生物信息学分析。
结果：
1. 该家系可追踪到的有五代共54人，男性29人，女性25人。其中已确诊的POAG患者17 例，男性患者11人，女性患者6人， 6人已经死亡。家系中连续四代发病。POAG的患病率为31％。
2. 患者IV-5、III-5、 III-4以及一级亲属IV-7中第4外显子412G>A.。患者II-5的第5外显子603T>A。患者III-5第10外显子1267-1268ins C/1271-1272ins C。III-3患者第12外显子1537-1538ins C。一级亲属V-3出现15外显子1878-1879ins A。
3. 32例正常对照组中有1例出现412G>A突变，其他突变型在对照组中均未发现。
4. 在不计算412G>A突变情况下，POAG家族发生OPTN基因编码区的突变概率为20%，而对照组中未发现。经X2分析，X2=6.93> X20.016.63，P<0.01, POAG家族与对照组相差显著。
生物信息学分析表明除第4外显子突变不改变OPTN蛋白的结构外，其他外
5. 显子突变均将导致结构的变化，影响OPTN的重要功能域。
结论：
1. 重庆忠县李某POAG家系遗传方式属常染色体显性遗传。在家系中存在导致POAG发病的突变的基因。
2. OPTN基因突变与本家系POAG发病相关。
3. 基因突变造成OPTN蛋白一二级结构和重要功能域异常，导致OPTN蛋白功能的异常可能是POAG发病的重要原因之一。
4. 本研究中共发现5种类型的OPTN基因突变（412G>A、 603T>A 1267-1268ins C/1271-1272ins C、 1537-1538ins C、 1878-1879ins A）。除412G>A、 603T>A两种与报道相同外，其余均为本次研究中第一次发现。
Glaucoma is the second leading cause of blindness which affect over 70 million people worldwide. Primary open angle glaucoma (POAG) is the most common subtype and the first reason of blindness of black man in America. Moreover, There are many subjects suffering from POAG in China. POAG is a kind of complex latent disease characterized by chronic progressive optic ganglion cells apoptosis and visual field injury. Thus, It is difficult to diagnoses and treat in early period. Although it is not yet wholly understood about the mechanism of glaucoma, more and more evidences suggest that there is an association between heredity and gene mutations with pathogenesis of glaucoma. In spite of great progress of glaucoma genetics, the hereditary form of POAG is not clear. on the other hand, about ten genes loci were found to associate with POAG after the discovery that TIGR gene links to POAG based on molecular genetics assay. The optineurin（optic neuropathy inducing, OPTN）, a novel gene linked to POAG, was found by Reiaze and his colleagues in 2002. Mutations in the OPTN gene were found in 16.7% of 54 families with autosomal dominant adult-onset POAG, and in 13.6% of the sporadic glaucoma cases. However, there is no report about mutations of OPTN gene in Chinese POAG cases.
Purpose:
To investigate the relationship between POAG and the mutations of OPTN gene in China, a POAG family in ChongQings was followed up, and the blood specimens of POAG individuals and controls were collected and processed by molecular biological strategy.
Methods:
1. To analyze the genetic form of the pedigree according to Mandalian hereditary rules after we collect a great family with POAG.
2. The genomic DNA was extracted from the peripheral blood of all samples. A total of 4350bp of 13 encoding exons and introns beside exons were amplified by PCR using the sequence-specific primers.
3. Automated sequencing method was used to screen and identify mutations of OPTN
gene of both POAG subjects and first degree relatives in this family.
4. Restriction endonuclease analysis was used to detect the same mutation in the controls samples.
5. The frequency of genetypes between POAG group and controls were compared by statistics.
6. The sequence alterations were analyzed by bioinfomatics.
Results:
1. This pedigree could be traced to 54 members in 5 generations, Including 29 males and 25 females. There are 17 patients with POAG, including 11 males and 6 females. Six patients had died in this family. The four continuous generations exist patients with POAG. The prevalence rate is 31%.
2. The encoding exons of OPTN gene of all alive patients and collected first degree relatives were directly automatic- sequenced, it is found that 3 patients ( IV-5、III-5、 III-4) and one first degree relative(IV-7) exist the 412G>A mutation in 4th exon. II-5 patient has a 603T>A alteration. III-5 was found that two C pyrimidine insertions between 1267-1268 site and 1271-1272 of 10th exon, respectively. III-3 exist 1537-1538ins (insert) C alteration. 1878-1879ins A had been found in a first degree relative (V-3).
3. No same mutations of OPTN gene were detected in 32 controls, except one person has a 412G>A mutation.
4. Rate of observed mutations in POAG family is 20%, no mutations were found in controls. The mutation rate of POAG family group is obviously higher than that in the controls group( X2=6.93> X20.016.63，P<0.01).
5. Bioinformatics analysis indicated that all mutations will result in the alteration of OPTN protein structure which may underlies its abnormal functional domain except for 412G>A mutation.
Conclusions:
1. The form of heredity in this POAG family in Zhong county of Chongqing is autosomal dominant trait, and there must be a abnormal gene which result in POAG in this family.
2. The mutation of OPTN gene is responsible for onset of POAG in this pedigree.
3. Function abnormality of OPTN protein, which are induced by change of the structure and functional domains caused by mutation of OPTN, may be an important m
引文
1. 眼科全书 李凤鸣主编 北京人民卫生出版社 1996年第一版
2. H.A.Quigley. Number of people with glaucoma worldwide. Br J Ophthalmol. 1996, 80: 389-393
3. H.A.Quigley. Open angle glaucoma. N Engl J Med. 1993,328: 1097-1106
4. Duke-Elder S(ed). System of ophthalmology. Vol 3. London:Kimpton,1969:398-402
5. Biro I. Notes upon the guestion of hereditary glaucoma. Ophthalmologica 1951,122: 228-234
6. Stone EM, Fingert JH, Alward WLM et al. Identification of a gene that causes primary open angle glaucoma. Science 1997, 275: 668-670
7. Rezaize T, Child A, Hitchings R et al, Adult-onset primary open-angle glaucoma caused by mutations in optineurin. Science 2002, 295: 1077-1079
8. Gelehrter TD, Collins FS, Ginshburg D et al. 医学遗传学原理（中译版）第二版
9. Anderson DR .Automated static perimetry.St. Louis:Mosby, 1992 ,12
10. Thompson JD, Gibson TJ, Plewniak F, et al. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 1997, 24: 4876-4882
11. Mulder NJ, Apweiler, Zdobnov EM, et al. The InterPro Database, 2003 brings increased coverage and new features. Nucl Acids Res. 2003, 31: 315-318.
12. Armaly MF. Inheritance of dexamethasone hypertension and glaucoma. Arch Ophthalmol 1967 , 77: 747-765
13. Shin DH. Family history in primary open angle glaucoma . Arch Ophthalmol 1977, 94: 598-608
14. Jay B, Paterson G. The genetics of simple glaucoma . Trans Opehalmo Soc UK 1972 , 94 : 161-168
15. 朱启仲，胡铮。原发性开角型青光眼遗传方式探讨。中华眼科杂志 1988，24（1）：6－9
16. Finger JH , Heon E , Liebmann JM , et al. Analysis of myocilin mutations in 1703 glaucoma patients from five different populations. Hum Mol Gen , 1999 , 8:899-905.
17. 范宝剑,梁旭辉,彭智培等原发性开角型青光眼小梁网糖皮质激素诱导反应蛋白基因单核苷酸多态性研究,中华医学杂志 2002,82(11):743-747
卓业鸿，葛坚，郭彦，等。我国原发性开角型青光眼患者TIGR基因突变的筛选、
18. 克隆及序列分析。中华眼科杂志，2000，36：416－419
19. Francois J , Heintz De, Bree C. Personal research on the heredity of chronic simple glaucoma. Am J Ophthalmol 1996, 62(6): 1067-71
20. Crombie Al . Heredity glaucoma. Br J Ophthalmol 1964,48:143-154
21. Martin JP. Familial glaucoma. Br J Ophthalmol , 1974, 58:536-543
22. Yehong Zhou , Jian Ge , Mei Wang , et al. The analysis of pedigree GZ .1 with primary open angle glaucoma. Eye science, 2000,16:53-55
23. Sarfarazi M , Child A , Stoilova D , et al. Locallization of the fourth locus (GLC1E) for adult-onset primary open angle glaucoma to the 10p15-p14 region. Am J Hum Genet 1998 , 62: 641-652
24. Y Li, J Kang, Horwitz MS, et al. Interaction of an adenovirus E3-14.7 Da protein with a novel tumor necrosis factor alpha-inducible cellular protein containing leucine zipper domains.Mol Cell Biol 1998, 18:1601-1610
25. Schwamborn K, Weil R, Courtois G, et al. , A Israel , Phorbol esters and cytokines regulate the expression the NEMO-related protein , a molecule involved in a NF-kappa B-independent pathway , J Biol Chem 2000, 275: 22780-22789
26. Hattula K, Biol J, Peranen. FIP-2 , a coiled-coil protein , links Huntingtin to Rab8 and modulates cellular morphogenesis. Curr Biol. 2000,10:1603-1606
27. Jason L. Vittiow , Teresa . B Expression of optineurin , a glaucoma-linked gene , is influenced by elevated intraocular pressure . B.B.R.C 2002, 298:67-74
28. L Yuan , A. H. Neufeld . Tumor necrosis factor-alpha: a potentially neurodestructive cytokine produced by glia in the human glaucomatous optic nerve head , Glia 2000, 32:42-50
29. G Tezel , M. B. Wax.Increased production of tumor necrosisi factor-alpha by glial cells exposed to simulated ischemia or elevated hydrostatic pressure induces apoptosisi in coculturee retinal ganglion cells , J neurosci. 2000 , 20:8693-8700
30. A Cotinet , O Goureau , D Hicks , et al. Tumor necrosis factor and nitric wide production by retinal muller glial cells from rats exhibiting inherited retinal dystrophy , Glia 1997, 20:59-69
31. Alexander JP, Acott TS. Involvement of protein kinase C in TNF alphal regulation of trabecular matrix metalloproteinases and TIMPs , Invest Ophthalmol Vis Sci , 2001 , 42:2831-2838
Rezaie T , Child A , Hitchings R et al . supplementary Web material is available on
32. science online at WWW. sciencemag. Org /cgi /conten t/full / 295 / 5557 / 1077 / DC1
33. Hurst HC ， Transcription factors 1: bZIP proteins. Protein Profile 1995;2(2):101-68
34. Lavigne P, Crump MP, Gagne SM, et al. Insights into the mechanism of heterodimerization from the 1H-NMR solution structure of the c-Myc-Max heterodimeric leucine zipper. J Mol Biol 1998 , 281(1): 165-81
35. Evans RM, Hollenberg SM. Zinc fingers: gilt by association. Cell 1988，52(5):783-789
36. Vincent AL, Billingsler G , Buys Y , et al. Digenic inheritance of early-onset glaucoma: CYP1B1 , a potential modifier gene , Am J Hum Genet 2002 , 70:448-460
37. Lam DSC, Leung YF, Chua JHK, et al . Trunctions in the TIGR in individuals with and without primary open angle glaucoma . Invest Ophthalmol Vis Sci, 2000, 41 : 1386-1391
38. J Peranen, P. Auvine, H. Virta, et al. Rab8 promotes polarized membrane transport through reorganization of actin and microtubules in fibroblasts, J Cell Biol. 1996, 135: 153-167
39. K Hattula, H. Peranen , Fip-2, a coiled-coil protein, links Huntingtin to Rab8 and modulates cellular morphogenesis, Curr. Biol, 2000,10: 1603-1606
40. 萨姆布鲁克 J，弗里奇 EF，曼尼阿蒂斯 T. 分子克隆实验指南。1992年第二版，科学出版社，919－939
41. Sheffield VC, Stone EM, Alward WLM, et al. Genetic linkage of familial open angle glaucoma to chromosome 1q21-31. Nature Cenet 1993, 4: 47-50
42. Stone EM, Finger JH, Aleard WLM, et al. Indentification of a gene that causes primary open angle glaucoma. Science 1997, 275: 668-670
2. H.A.Quigley. Number of people with glaucoma worldwide. Br J Ophthalmol. 1996, 80: 389-393
3. H.A.Quigley. Open angle glaucoma. N Engl J Med. 1993,328: 1097-1106
4. Duke-Elder S(ed). System of ophthalmology. Vol 3. London:Kimpton,1969:398-402
5. Biro I. Notes upon the guestion of hereditary glaucoma. Ophthalmologica 1951,122: 228-234
6. Stone EM, Fingert JH, Alward WLM et al. Identification of a gene that causes primary open angle glaucoma. Science 1997, 275: 668-670
7. Rezaize T, Child A, Hitchings R et al, Adult-onset primary open-angle glaucoma caused by mutations in optineurin. Science 2002, 295: 1077-1079
8. Gelehrter TD, Collins FS, Ginshburg D et al. 医学遗传学原理（中译版）第二版
9. Anderson DR .Automated static perimetry.St. Louis:Mosby, 1992 ,12
10. Thompson JD, Gibson TJ, Plewniak F, et al. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 1997, 24: 4876-4882
11. Mulder NJ, Apweiler, Zdobnov EM, et al. The InterPro Database, 2003 brings increased coverage and new features. Nucl Acids Res. 2003, 31: 315-318.
12. Armaly MF. Inheritance of dexamethasone hypertension and glaucoma. Arch Ophthalmol 1967 , 77: 747-765
13. Shin DH. Family history in primary open angle glaucoma . Arch Ophthalmol 1977, 94: 598-608
14. Jay B, Paterson G. The genetics of simple glaucoma . Trans Opehalmo Soc UK 1972 , 94 : 161-168
15. 朱启仲，胡铮。原发性开角型青光眼遗传方式探讨。中华眼科杂志 1988，24（1）：6－9
16. Finger JH , Heon E , Liebmann JM , et al. Analysis of myocilin mutations in 1703 glaucoma patients from five different populations. Hum Mol Gen , 1999 , 8:899-905.
17. 范宝剑,梁旭辉,彭智培等原发性开角型青光眼小梁网糖皮质激素诱导反应蛋白基因单核苷酸多态性研究,中华医学杂志 2002,82(11):743-747
卓业鸿，葛坚，郭彦，等。我国原发性开角型青光眼患者TIGR基因突变的筛选、
18. 克隆及序列分析。中华眼科杂志，2000，36：416－419
19. Francois J , Heintz De, Bree C. Personal research on the heredity of chronic simple glaucoma. Am J Ophthalmol 1996, 62(6): 1067-71
20. Crombie Al . Heredity glaucoma. Br J Ophthalmol 1964,48:143-154
21. Martin JP. Familial glaucoma. Br J Ophthalmol , 1974, 58:536-543
22. Yehong Zhou , Jian Ge , Mei Wang , et al. The analysis of pedigree GZ .1 with primary open angle glaucoma. Eye science, 2000,16:53-55
23. Sarfarazi M , Child A , Stoilova D , et al. Locallization of the fourth locus (GLC1E) for adult-onset primary open angle glaucoma to the 10p15-p14 region. Am J Hum Genet 1998 , 62: 641-652
24. Y Li, J Kang, Horwitz MS, et al. Interaction of an adenovirus E3-14.7 Da protein with a novel tumor necrosis factor alpha-inducible cellular protein containing leucine zipper domains.Mol Cell Biol 1998, 18:1601-1610
25. Schwamborn K, Weil R, Courtois G, et al. , A Israel , Phorbol esters and cytokines regulate the expression the NEMO-related protein , a molecule involved in a NF-kappa B-independent pathway , J Biol Chem 2000, 275: 22780-22789
26. Hattula K, Biol J, Peranen. FIP-2 , a coiled-coil protein , links Huntingtin to Rab8 and modulates cellular morphogenesis. Curr Biol. 2000,10:1603-1606
27. Jason L. Vittiow , Teresa . B Expression of optineurin , a glaucoma-linked gene , is influenced by elevated intraocular pressure . B.B.R.C 2002, 298:67-74
28. L Yuan , A. H. Neufeld . Tumor necrosis factor-alpha: a potentially neurodestructive cytokine produced by glia in the human glaucomatous optic nerve head , Glia 2000, 32:42-50
29. G Tezel , M. B. Wax.Increased production of tumor necrosisi factor-alpha by glial cells exposed to simulated ischemia or elevated hydrostatic pressure induces apoptosisi in coculturee retinal ganglion cells , J neurosci. 2000 , 20:8693-8700
30. A Cotinet , O Goureau , D Hicks , et al. Tumor necrosis factor and nitric wide production by retinal muller glial cells from rats exhibiting inherited retinal dystrophy , Glia 1997, 20:59-69
31. Alexander JP, Acott TS. Involvement of protein kinase C in TNF alphal regulation of trabecular matrix metalloproteinases and TIMPs , Invest Ophthalmol Vis Sci , 2001 , 42:2831-2838
Rezaie T , Child A , Hitchings R et al . supplementary Web material is available on
32. science online at WWW. sciencemag. Org /cgi /conten t/full / 295 / 5557 / 1077 / DC1
33. Hurst HC ， Transcription factors 1: bZIP proteins. Protein Profile 1995;2(2):101-68
34. Lavigne P, Crump MP, Gagne SM, et al. Insights into the mechanism of heterodimerization from the 1H-NMR solution structure of the c-Myc-Max heterodimeric leucine zipper. J Mol Biol 1998 , 281(1): 165-81
35. Evans RM, Hollenberg SM. Zinc fingers: gilt by association. Cell 1988，52(5):783-789
36. Vincent AL, Billingsler G , Buys Y , et al. Digenic inheritance of early-onset glaucoma: CYP1B1 , a potential modifier gene , Am J Hum Genet 2002 , 70:448-460
37. Lam DSC, Leung YF, Chua JHK, et al . Trunctions in the TIGR in individuals with and without primary open angle glaucoma . Invest Ophthalmol Vis Sci, 2000, 41 : 1386-1391
38. J Peranen, P. Auvine, H. Virta, et al. Rab8 promotes polarized membrane transport through reorganization of actin and microtubules in fibroblasts, J Cell Biol. 1996, 135: 153-167
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