Human Molecular Genetics

Human Molecular Genetics Advance Access originally published online on January 27, 2005
Human Molecular Genetics 2005 14(6):725-733; doi:10.1093/hmg/ddi068

This Article Abstract FREE Full Text (PDF) All Versions of this Article: 14/6/725    most recent ddi068v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (110) Request Permissions Google Scholar Articles by Monemi, S. Articles by Sarfarazi, M. Search for Related Content PubMed PubMed Citation Articles by Monemi, S. Articles by Sarfarazi, M. Social Bookmarking          What's this?

Human Molecular Genetics, Vol. 14, No. 6 © Oxford University Press 2005; all rights reserved

Identification of a novel adult-onset primary open-angle glaucoma (POAG) gene on 5q22.1

Sharareh Monemi¹, George Spaeth², Alexander DaSilva¹, Samuel Popinchalk¹, Elena Ilitchev³, Jeffrey Liebmann³, Robert Ritch³, Elise Héon⁴, Ronald Pitts Crick⁵, Anne Child⁶ and Mansoor Sarfarazi^1,*

¹Molecular Ophthalmic Genetics Laboratory, Surgical Research Center, Department of Surgery, University of Connecticut Health Center, Farmington, CT, USA, ²Glaucoma Service, Wills Eye Hospital, Philadelphia, PA, USA, ³Glaucoma Service, New York Eye and Ear Infirmary, New York, NY, USA, ⁴Department of Ophthalmology and Vision Sciences, The Hospital for Sick Children, Toronto, Ontario, Canada, ⁵International Glaucoma Association, London, UK and ⁶Department of Cardiological Sciences, St George's Hospital Medical School, London, UK

^* To whom correspondence should be addressed. Tel: +1 8606793629; Fax: +1 8606797524; Email: mansoor{at}neuron.uchc.edu

Received December 7, 2004; Accepted January 18, 2005

	ABSTRACT

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 
Glaucoma is a leading cause of blindness in virtually every country. Development of an accurate diagnostic test for presymptomatic detection of individuals at risk is an urgent requisition for this condition. Herein, we report mapping of a new adult-onset primary open-angle glaucoma (POAG) locus on 5q22.1 (GLC1G) and identification of its defective gene. Mutation screening of seven candidate genes from the GLC1G critical region (2 Mb between D5S1466 and D5S2051) identified only one significant alteration in the WDR36 (WD40-repeat 36) gene. This mutation (i.e. D658G) was segregated in all affected members of our first GLC1G-linked family but it was absent in 476 normal control chromosomes. Further screening of WDR36 in a total of 130 POAG families revealed 24 DNA variations. Overall, four mutations (N355S, A449T, R529Q and D658G) were identified in 17 (5.02–6.92%) unrelated POAG subjects, 11 with high-pressure and six with low-pressure glaucoma. These mutations were absent in a minimum of 200 normal control chromosomes and, further they were conserved between WDR36 orthologues in mouse, rat, dog, chimp and human. WDR36 is a novel gene with 23 exons, which encodes for 951 amino acids and a protein with multiple G-beta WD40 repeats. By northern blotting, two distinct mRNA transcripts of 5.9 and 2.5 kb were observed in human heart, placenta, liver, skeletal muscle, kidney and pancreas. WDR36 gene expression in lens, iris, sclera, ciliary muscles, ciliary body, trabecular meshwork, retina and optic nerve were established by RT–PCR. In mouse, two transcripts of 3.5 and 2.9 kb showed analogous expression patterns to human. mRNA expressions were detected in 7-, 11-, 15- and 17-day-old developing mouse embryos. In summary, WDR36 is a novel causative gene for adult-onset POAG at the GLC1G locus. Specific ocular expressions and observed mutations are consistent with WDR36 role in etiology of both high- and low-pressure glaucoma.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 
Glaucoma is a group of ocular disorders characterized by a specific pattern of optic nerve and visual field defects. This condition is one of the two leading causes of blindness, affecting over 67 million people worldwide (1). Open-angle glaucoma is usually asymptomatic until the late stages of the disease, by which time significant and irreversible optic nerve damage has already taken place (2,3). As the sensitivity of current diagnostic techniques is suboptimal, the diagnosis of glaucoma is usually made once an irreversible damage has already occurred. As glaucoma related visual loss is preventable in many cases, there is an urgent need to diagnose glaucoma at its early stages and to institute appropriate neuroprotective management of the ganglion cells (4,5). Mapping, cloning and identification of novel mutations involved in the etiology of glaucoma provide a significant opportunity for presymptomatic diagnosis, improved prognosis and better understanding of the etiology of this blinding condition.

Adult-onset primary open-angle glaucoma (POAG; MIM no. 137760) is the commonest form of this ocular group, usually manifesting itself after the age of 40 years (6,7). The prevalence of POAG is 1–2% over age 50 in white populations (8) and four to five times greater in black populations of the same age (9). Clinical diagnosis of all groups is based on characteristic changes of the optic nerve head and visual field, which are usually accompanied by increased intraocular pressure (IOP) (10). Family history is an important risk factor for POAG, and genetic study of families shows that dominant genes are predominantly involved in this condition (7,11,12).

During the last decade, a total of six genetic loci (13–18) and two genes have been reported for POAG: myocilin (MYOC) is primarily mutated in juvenile-onset subjects (19), whereas optineurin (OPTN) is mainly mutated in low pressure POAG individuals (20,21). Here, we report chromosomal mapping of a new POAG locus on 5q22.1 (designated as GLC1G) and identification of its causative gene, WDR36 (WD40-repeat 36), from within this region. We further present the first expression analysis of this gene in a variety of human ocular and non-ocular tissues. For an evolutionary comparative analysis, we further determined expression of the homologue of this gene (Wdr36) in various stages of mouse embryonic developments as well as a group of tissues from the adult animals.

	RESULTS

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 
Positional mapping of a new locus (GLC1G) for adult-onset POAG
Initially, genome scans of two large POAG families (i.e. POAG-002 and POAG-527) with 20 affected and nine glaucoma suspects were used to obtain a provisional hint of linkage to the 5q33–q35 region (22). Extended saturation mapping established critical recombination in two of the affected individuals in POAG-002 and, therefore, excluded genetic linkage of this family to this region of 5q. However, additional genotyping in POAG-527 revealed consistent sharing of an extended affected haplotype from 5q21.3 (D5S1466) to 5q35.2 (D5S498) in all seven affected members of this family (23). Therefore, in light of this new linkage information and due to a discrepancy that currently exists between genetic linkage and physical map position of several DNA markers from this region of chromosome 5 (including D5S1466), the upper physical boundary for this new POAG locus is now 5q21.3 and not 5q33 as initially envisaged (22).

After a hint of linkage was observed with the 5q region, seven additional POAG families with a total of 194 individuals and 31 living affected subjects were genotyped. Only two of these families were consistent with linkage to the same 5q region. Further saturation mapping in these families confined this new linkage to an interval of 35 Mb (Fig. 1) that is flanked by D5S1466 (5q21.3) and D5S1480 (5q31.3). Additional recombination in a subject with ocular hypertension (OH) and for DNA marker of D5S404 (5q23.1) suggested that the identified genetic linkage may be limited to 7 Mb (Fig. 1). In a parallel study, a group of 638 individuals (including more than 400 affected subjects) in 139 POAG families were being screened in our laboratory for the entire genome. As this screening also covered this newly identified region on 5q, a number of new families showed potential linkage to this region. Altogether, a total of seven families were consistent with linkage to this region of 5q. Table 1 shows individual and multi-point LOD score values for three closely linked DNA markers of D5S1462, D5S2501 and D5S1505 (Fig. 1). This newly identified POAG locus has now been designated as GLC1G by HUGO Gene Nomenclature Committee (HGNC). Recently, Samples et al. (24) and Kramer et al. (25) reported linkage in a single adult-onset POAG family and, within a 6.6 Mb region (Fig. 1) that is flanked by D5S1721 (5q21.2) and D5S2051 (5q22.1). Taken together, these two overlapping linkage data defined the critical region of the GLC1G locus between D5S1466 (5q21.3) and D5S2051 (5q22.1) and within a region of 2 Mb (Fig. 1). This newly identified GLC1G locus provided an opportunity for rapid mutation screening and identification of the defective gene at this locus.

View larger version (18K): [in this window] [in a new window]   Figure 1. Partial genetic and physical map of chromosome 5q. Cytogenetic banding and map positions (in Megabases) for a selected group of DNA markers are shown. This study identified two flanking markers of D5S1466 (109 Mb from the top of chromosome 5) and D5S1480 (144 Mb) and a further cross over with D5S404 (116 Mb) in a subject with OH. Another study by Samples et al. (24) and Kramer et al. (25) identified a region between D5S1721 (104 Mb) and D5S2051 (111 Mb). An overlapping region of 2 Mb between D5S1466 and D5S2051 defined the GLC1G locus. Order of seven known genes in this overlapping region that were fully sequenced in our study are also shown. D5S2501 maps between AK125070 and BC017169.

 

View this table: [in this window] [in a new window]   Table 1. Two-point and multi-point LOD score values for three closely GLC1G-linked markers

 
GLC1G candidate genes screening
The critical region of the GLC1G locus contains seven known genes (MAN2A1, AK125070, BC017169, TSLP, WDR36, CAMK4 and STARD4) and at least three other predicted genes (LOC153778, LOC441101 and LOC441102). For each of the seven known genes (Fig. 1), a series of genomic primers were made and their respective coding exons were PCR amplified and screened for mutation by direct DNA sequencing. Analysis of genomic DNA from at least two affected subjects of the original GLC1G-linked family of POAG-527 revealed four sequence alterations in only two genes. Three of these variations (i.e. IVS6–5A>G, S337S and A413V in CAMK4) were polymorphisms and only one sequence change in WDR36 proven to be significant. The observed variation in this gene was a single heterozygous DNA alteration in exon 17 (c.1973A>G; GAC>GGC) that is predicted to change aspartic acid (acidic) to glycine (neutral and polar) at amino acid 658 (D658G). This mutation (Fig. 2D) was observed in seven affected (with mean age of 63.7 years) and nine asymptomatic gene carriers (with mean age of 40.4 years) but it was absent in nine normal members of this family (with mean age of 45.3 years) and another six married-in normal spouses (Fig. 2E). Screening of a total of 476 normal control chromosomes by restriction enzyme digestion (BglI) showed that this mutation is absent in all the normal subjects tested. Direct sequencing of WDR36 in another 129 unrelated affected glaucoma probands (including the six GLC1G-linked families) identified additional 23 DNA alterations (Table 2). Three of these alterations (N355S, A449T and R529Q) (Fig. 2A–C) were considered disease causing, as they were not present in any of the public databases and in a group of at least 200 tested chromosomes from normal control subjects. We directly sequenced the genomic DNA of at least one affected subject from each of the original seven linked families. However, two of these families did not show any DNA variations in the coding regions of the WDR36 gene. Therefore, it is possible that other mutations interfering with the mRNA splicing machinery, mutations within the promoter of this gene or presence of other DNA rearrangements such as insertion, deletion or duplication is responsible for the POAG phenotype in these two families. Alternatively, it is possible that another POAG locus is located within our identified genetic linkage and, therefore, mutations in another gene are responsible for these two families.

View larger version (38K): [in this window] [in a new window]   Figure 2. Predicted POAG-causing mutations in the WDR36 gene. (A, B) Partial reverse sequence for N355S (AAT>AGT) and A449T (GCA>ACA); (C, D) Partial forward electropherogram for R529Q (CGA>CAA) and D658G (GAC>GGC). Vertical line identifies the intron–exon boundaries for A449T and D658G sequences. (E) Segregation of D658G mutation in our original GLC1G-linked family of POAG-527. BglI restriction digestion of a 389 bp PCR fragment produces two products of 272 and 117 bp only in presence of the mutant allele. For simplicity, only seven affected (filled-in), nine gene carriers (dot inside their symbols), two normal subjects and four available spouses are shown. All other normal subjects with no D658G mutation in the third and fourth generations were omitted.

 

View this table: [in this window] [in a new window]   Table 2. Distribution of 24 sequence variations identified in the WDR36 gene

 
Expression study of WDR36 at RNA level
As a first step in determining the role of WDR36 in glaucoma pathogenesis, we studied expression of this gene in different human ocular and non-ocular tissues by northern blot and reverse transcriptase–polymerase chain reaction (RT–PCR). Northern blotting of several human non-ocular mRNA samples revealed two different transcripts (i.e. 5.9 and 2.5 kb) that are highly expressed in heart, placenta, liver, skeletal muscle and pancreas (Fig. 3A). Using RT–PCR, similar WDR36 expressions were also detected in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas (Fig. 3B). WDR36 expression was also detected in a number of ocular tissues including lens, iris, sclera, ciliary muscles, ciliary body, trabecular meshwork, retina and optic nerve (Fig. 3C). As a comparative study, we also determined mRNA expression patterns for the orthologue of this gene (Wdr36) in mouse. Northern blotting of Wdr36 in a panel of adult mouse tissues revealed analogous expression patterns to human with only one exception; in skeletal muscle, the expression was stronger in human (Fig. 4A). Using RT–PCR, high levels of Wdr36 expression were detected in mouse heart, brain, liver, skeletal muscle, kidney, testis and lower expressions in spleen and lung (Fig. 4B). Study of Wdr36 during various stages of mouse embryonic developments showed that this gene is detectable in 7-day-old embryos (Fig. 4C).

View larger version (76K): [in this window] [in a new window]   Figure 3. WDR36 gene expression in various human tissues. (A) Northern blotting of non-ocular tissues revealed two transcripts of 5.9 and 2.5 kb (top panel). Beta-actin used as a standard control. Heart, placental, liver, smooth muscle and pancreas exhibited the strongest signals. (B) mRNA expression patterns of WDR36 in various non-ocular tissues by RT–PCR. Placental, lung, liver, kidney and pancreas produced the strongest signals. (C) RT–PCR of WDR36 in a group of human ocular tissues showing a strong gene expression in iris, sclera, ciliary muscle, ciliary body, retina and optic nerve.

 

View larger version (64K): [in this window] [in a new window]   Figure 4. Wdr36 gene expression in mouse. (A) Northern blot analysis exhibited two transcripts of 3.5 and 2.9 kb. Strongest signals observed in heart, liver, kidney and testis. (B) mRNA gene expression in various adult mouse tissues. Similar to northern blotting, spleen and lung had the weakest signals. (C) RT–PCR of Wdr36 in mouse embryo at different gestational age. The earliest gene expression was at the embryonic day 7.

 

	DISCUSSION

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 
This article describes mapping of a new locus for POAG (GLC1G) on the 5q22.1 region and identification of its disease-causing gene, WDR36 (GenBank accession no. NM_139281). As WDR36 is a novel gene, using northern blot analysis we determined the transcript size of this gene in two commercially available human and mouse mRNA blots, and using RT–PCR, we established expression of this gene in human ocular and non-ocular as well as in embryonic and adult mouse tissues.

WDR36 has recently been identified as one of the genes that is uniquely involved in T-cell activation and highly co-regulated with interleukin 2 (IL2). This gene encodes for a T-cell activation protein with a minimum of eight WD40 repeats (26) and, therefore, it is also recognized as T-cell activation WD repeat protein (TA-WDRP). This gene contains 23 exons and encodes for a 951 amino acid protein. The encoded protein consists of at least four known motifs: a guanine nucleotide binding protein (or G)-beta WD40 repeat, an AMP-dependent synthetase and ligase, a cytochrome cd1-nitrite reductase-like (C-terminal heme d1) and an Utp21-specific WD40 associated putative domain.

Mutation analysis of WDR36 in 130 unrelated affected individuals revealed a total of 24 allelic variants, of which 12 were amino acid coding and 12 involved intronic alterations (Table 2). Comparative sequence alignments of this gene among human, chimp, dog, rat and mouse revealed that 11 of these amino acids (except V727) have been fully conserved among these species (Fig. 5). Evolutionary conservation of the four identified disease-causing mutations (N355S, A449T, R529Q and D658G) in at least five different species and their absence in a minimum of 200 human normal control chromosomes imply that these four amino acids perform a fundamentally important role in the organization of WDR36 protein.

View larger version (22K): [in this window] [in a new window]   Figure 5. Partial amino acid alignment and evolutionary conservation of 12 WDR36 variations in human and four other species.

 
Although one of the WDR36 disease-causing mutations identified in this study (A449T) is not part of a known protein motif, the other three are located in separate G-beta WD40 repeats (Fig. 5). The N355S mutation in exon 8 maps to the fourth WD40 repeat (covering amino acids 321–361), the R529Q in exon 13 maps to the sixth WD40 repeat (amino acids 525–567) and D658G in exon 17 maps to the eighth WD40 repeat (amino acids 653–692). The G proteins are a family of membrane-associated proteins that act as intermediaries in transduction of the signals generated by transmembrane receptors. The G-beta subunit is required for membrane anchoring and receptor recognition. Structurally, the G-beta consists of eight tandem repeats of about 40 residues, each containing a central Trp-Asp dipeptide (thus named WD40 repeats). Therefore, mutations affecting the structure of these WD40 repeats may interfere with interaction of WDR36 with other proteins. This repetitive WD40 segment is also present in over 250 proteins that are encoded by a variety of genes mapping to every one of the human chromosomes. None of the WDR36 detected amino acid alterations in this study was located in the AMP-binding domain (amino acids 553–564) and only one single nucleotide polymorphism or SNP (V727V) was identified from within the Utp21 domain (amino acids 724–948).

The D658G mutation (Figs 2D and Fig. 5) that is located within the eighth WD40 repeat also maps to the C-terminal part of the cytochrome heme cd1 (cyt cd1) domain (amino acids 109–318 and 537–686). This ‘cyt cd1’ is part of a bi-functional enzyme with cytochrome oxidase activity (27). It is interesting that we have previously identified another member of the cytochrome P450 family (CYP1B1) that is involved in the etiology of primary congenital glaucoma (28,29). Therefore, there might be a functional association between WDR36 and CYP1B1, as mutations in these two genes are responsible for two different forms of glaucoma.

As D658G mutation was originally observed in five out of 130 unrelated and familial POAG subjects, we further tested for the presence of this one particular mutation in an additional 540 unrelated affected individuals from different POAG subgroups. This mutation creates a new recognition site (BglI) thus providing a rapid method for its screening (Fig. 2E). In total, 13 out of 670 (1.94%) subjects tested were found to be heterozygous for D658G (seven with high- and six with low-pressure glaucoma). Originally, we sequenced the entire of WDR36 gene in a total of 130 familial POAG cases and identified three mutations of N355S, A449T and R529Q in four families (3.08%) and the common mutation of D658G in five families. Altogether, nine out of 130 (6.92%) families that were fully sequenced showed mutations in the WDR36 gene. When additional 540 unrelated affected subjects (268 familial and 272 sporadic) were only tested for presence of D658G, this mutation was observed in another five familial and three sporadic cases. Altogether, D658G was observed in 2.51% of familial (10 out of 398) and 1.10% of sporadic (three out of 272) cases, thus providing a combined frequency of 1.94% (13 out of 670) for this one mutation alone. When this is added to the frequency of 3.08% obtained from the other three mutations, a minimum mutation rate of 5.02% is obtained for this gene. After the entire of WDR36 gene in the earlier-mentioned 540 familial and sporadic cases is fully sequenced, it is anticipated that additional mutations will be identified thus, altering the mutation frequency of 5.02–6.92% as presented here. In summary, we identified four different WDR36 mutations in 17 subjects with either high-pressure (65% of subjects) or low-pressure (35% of subjects) POAG. This observation indicates that the WDR36 gene is involved in etiology of both types of glaucoma and IOP, as traditionally used to group these into two separate clinical entities, may not be supported by this study and by recent molecular delineation of this group of optic neuropathies (20,21).

We also identified three other amino acid alterations of L25P, A163V and Y216P in a group of 15 POAG subjects (Table 2). As these three alterations were absent in all of the public databases, we screened for their presence in 428, 422 and 200 normal control chromosomes, respectively. As the ratio of their presence in the POAG group was significantly higher than that obtained for the normal group (11.55 versus 2.21%, respectively), we considered these as potential disease-susceptibility alterations (Tables 2 and Fig. 5). The role of these amino acid variations in the etiology of POAG requires further investigation.

We also observed three amino acid polymorphisms in our POAG or normal control subjects (I264V, M671V and V727V). In addition to these, the Ensembl database lists two other polymorphisms (S90I and A149A) that were not observed in our study. The two other silent amino acid polymorphisms (G134G and V714V) identified in this study were also present in the public EST databases. It is interesting that during this study (Table 2), we also observed 100% linkage disequilibrium between G134G and Y216P, and 95% between V714V and IVS18+216C>T. Of the 12 intronic alterations that were identified in this study, only four have previously been deposited in the SNP databases (Table 2). Therefore, we screened a total of 90 normal control chromosomes for the presence of these previously unreported intronic alterations. As shown in Table 2, only two of these changes (i.e. IVS7–39T>G and IVS8+92G>A) were absent in the normal control chromosomes but the significance of their presence in the POAG subjects is not clear at this point and requires further investigation.

As there is limited in silico expression data for WDR36, we studied the mRNA transcripts of this gene in various mouse and human tissues. Using northern analysis two different transcript sizes were detected in human and mouse (Figs 3 and 4). However, it is not clear at this point whether these two transcripts were produced by alternative splicing or by use of two different promoters. In humans, intense hybridization was observed in heart, placenta, liver, skeletal muscle, kidney and pancreas (Fig. 3A and B). In mouse, heart, liver and kidney produced the highest signal (Fig. 4A and B). Using RT–PCR, we determined expression of WDR36 in lens, iris, sclera, ciliary muscles, ciliary body, trabecular meshwork, retina and optic nerve (Fig. 3C). In mouse, expressions were detected in 7-, 11-, 15- and 17-day-old developing embryos (Fig. 4C).

Although the in vivo function of WDR36 is yet unknown, identification of this gene as an adult-onset POAG gene provides an opportunity to find and to study biochemical pathways that are expected to be involved in the pathogenesis of this group of optic neuropathies. WDR36 contains multiple G-beta WD40 repeats that are also present in a large family of proteins with diverse functions. As this WD40 repeat motif is also involved in protein–protein interaction and as WDR36 is now implicated in the etiology of POAG, this finding provides an opportunity to search for WDR36-interacting proteins aiming to identify other proteins/genes that have direct functional effect on the pathophysiology of this blinding condition. Additionally, although different mutations in WDR36 were observed in both high- and low-pressure glaucoma, molecular screening of this gene may provide a useful tool for presymptomatic detection of individuals at risk in families or in the elderly population.

	MATERIALS AND METHODS

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 
Family ascertainment
The clinical status of all affected POAG members of studied families have previously been confirmed by their respective caring ophthalmologists. The diagnoses of glaucoma agreed with the standard criteria and included abnormal cup/disc ratio and glaucomatous visual field loss with or without increased IOP. The original GLC1G-linked family of POAG-527 has a total of 108 members in five generations, of whom 54 (seven affected, five glaucoma suspect and 42 unaffected) participated in this study. The other large family of POAG-002 that was originally used in our genome scan but subsequently proven to be unlinked to the GLC1G locus has a total of 99 members in four generations, 40 of whom have been sampled previously (13 affected, four OH and 23 unaffected). The clinical status of all other unrelated familial and sporadic POAG subjects screened during this study have previously been confirmed. The University of Connecticut Health Center Institutional Review Board has approved this study for inclusion of Human Subjects in Research.

Genome-wide scan
A genome-wide scan was carried out with two different sets of fluorescently end-labeled primers (ABI and CHLC-Weber linkage mapping sets). The amplified PCR products were loaded onto a 96-well gel and run on an ABI-377 automated DNA sequencer. Computer assisted data collection and genotyping were performed with the ABI GENESCAN analysis and ABI GENOTYPER computer software. The obtained genotypic data were analyzed and further used for additional linkage analysis and saturation mapping.

Saturation mapping and refinement of the GLC1G critical region
All genotypes either from the ABI or from the CHLC-Weber florescent linkage mapping sets were automatically transferred into an in-house database management system (DMS) program. Pedigrees and their genotypic data for each of the 22 chromosomes were subsequently exported to the CYRILLIC program and haplotypes were constructed for each pedigree and for each chromosome. Co-inheritance of affected-bearing haplotypes with the POAG phenotype was inspected manually for each of the studied families. Using additional short tandem repeat polymorphism (STRP) markers potential regions of interest were saturated in these families. After a hint of linkage was observed on chromosome 5q, fluorescent genotyping and saturation mapping of other families with new STRP markers were carried out in order to reduce the identified interval even further. The two-point and multi-point LOD scores were calculated with the MLINK and LINKMAP components of the LINKAGE program, respectively (30).

Mutation screening
After linkage to the GLC1G locus was established and its candidate region was reduced to a region of 2 Mb, all the seven known genes mapping to the critical region (Fig. 1) were selected for mutation screening. Primers were designed to flank intron–exon boundaries of the selected genes and PCR amplification performed using genomic DNA of affected individuals (primer sequences are available on request). Direct sequencing carried out with ABI-Big Dye Terminator Cycle sequencing kit and run on an ABI-3100 Genetic Analyzer and DNA Sequencer.

RNA analysis
Under a dissecting microscope, the lens, iris, ciliary body, ciliary muscle, trabecular meshwork, sclera, retina and optic nerve were micro-dissected and immediately used for RNA extraction. The TRIzol reagent method (Invitrogen) was used to isolate RNA from these ocular tissues. For RT–PCR, cDNA were made from purified RNAs, using random hexamer primers and Superscript III reverse transcriptase (Invitrogen). Furthermore, expression of the WDR36 gene in human and mouse tissues were investigated by northern hybridization. Radiolabeled probes were prepared from a 691 and 693 bp PCR fragment amplified from human and mouse cDNA, respectively. Expression of this gene was studied by use of the same probe and a premade blot (Clontech) containing poly(A)+ mRNA (2 µg/lane) from heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas in human and heart, brain, spleen, lung, liver, skeletal muscle, kidney and testis in mouse.

	ACKNOWLEDGEMENTS

 
We are indebted to our patients and their families for their participation in this study. We specially would like to thank Dr Ordan Lehmann for his help with initial genetic ascertainment and blood sampling and Dr Sushma Rai for additional sampling and cell transformation of the first GLC1G-linked family of POAG-527, Dr Nilanjana Maulik for her help with northern blotting, Dr Ivaylo Stoilov for his help with eye dissection, Dr Jeff Coppin and Jose Aragon-Martin for their technical assistance. This work was funded by grants from the National Eye Institute (EY-09947), International Glaucoma Association and University of Connecticut General Clinical Research Center (M01RR-06192). A.C. would like to thank the Bluff Field Charitable Fund, St George's Hospital NHS Trust and Medical School for their financial support.

	REFERENCES

TOP ABSTRACT INTRODUCTION RESULTS DISCUSSION MATERIALS AND METHODS REFERENCES

 

1. Quigley, H.A. (1996) Number of people with glaucoma worldwide. Br. J. Ophthalmol., 80, 389–393.[Abstract/Free Full Text]

2. Crick, R.P. and Khaw, P.T. (2003) A Text Book of Clinical Ophthalmology. 3rd edn. World Scientific, Singapore.

3. Fechtner, R.D. and Weinreb, R.N. (1994) Mechanisms of optic nerve damage in primary open angle glaucoma. Surv. Ophthalmol., 39, 23–42.[CrossRef][Web of Science][Medline]

4. Ritch, R. (2000) Neuroprotection: is it already applicable to glaucoma therapy? Curr. Opin. Ophthalmol., 11, 78–84.[CrossRef][Medline]

5. Hitchings, R.A. (1999) Glaucoma in the new millennium. Ophthalmol. Clin. North Am., 12, 519–531.[CrossRef]

6. Shields M., Ritch R. and Krupin, T. (1996) Classification of the Glaucomas. Mosby, St Louis.

7. Sarfarazi, M. (1997) Recent advances in molecular genetics of glaucomas. Hum. Mol. Genet., 6, 1667–1677.[Abstract/Free Full Text]

8. Tuck, M.W. and Crick, R.P. (1998) The age distribution of primary open angle glaucoma. [Erratum (1999) Ophthalm. Epidemiol., 6, 84] Ophthalmic Epidemiol., 5, 173–183.[CrossRef][Medline]

9. Tielsch, J.M., Katz, J., Singh, K., Quigley, H.A., Gottsch, J.D., Javitt, J. and Sommer, A. (1991) A population-based evaluation of glaucoma screening: the Baltimore eye survey. Am. J. Epidemiol., 134, 1102–1110.[Abstract/Free Full Text]

10. Realini, T. and Fechtner, R.D. (2000) Target intraocular pressure in glaucoma management. Ophthalmol. Clin. North Am., 13, 407–415.[CrossRef]

11. Lee, D.A., Brubaker, R.F. and Hruska, L. (1985) Hereditary glaucoma: a report of two pedigrees. Ann. Ophthalmol., 17, 739–741.[Web of Science][Medline]

12. Bennett, S.R., Alward, W.L. and Folberg, R. (1989) An autosomal dominant form of low tension glaucoma. Am. J. Ophthalmol., 108, 238–244.[Web of Science][Medline]

13. Sheffield, V.C., Stone, E.M., Alward, W.L., Drack, A.V., Johnson, A.T., Streb, L.M. and Nichols, B.E. (1993) Genetic linkage of familial open angle glaucoma to chromosome 1q21–q31. Nat. Genet., 4, 47–50.[CrossRef][Web of Science][Medline]

14. Stoilova, D., Child, A., Trifan, O.C., Crick, R.P., Coakes, R.L. and Sarfarazi, M. (1996) Localization of a locus (GLC1B) for adult-onset primary open angle glaucoma to the 2cen–q13 region. Genomics, 36, 142–150.[CrossRef][Web of Science][Medline]

15. Wirtz, M.K., Samples, J.R., Kramer, P.L., Rust, K., Topinka, J.R., Yount, J., Koler, R.D. and Acott, T.S. (1997) Mapping a gene for adult-onset primary open-angle glaucoma to chromosome 3q. Am. J. Hum. Genet., 60, 296–304.[Web of Science][Medline]

16. Trifan, O.C., Traboulsi, E.I., Stoilova, D., Alozie, I., Nguyen, R., Raja, S. and Sarfarazi, M. (1998) A third locus (GLC1D) for adult-onset primary open-angle glaucoma maps to the 8q23 region. Am. J. Ophthalmol., 126, 17–28.[CrossRef][Web of Science][Medline]

17. Sarfarazi, M., Child, A., Stoilova, D., Brice, G., Desai, T., Trifan, O.C., Poinoosawmy, D. and Crick, R.P. (1998) Localization of the fourth locus (GLC1E) for adult-onset primary open-angle glaucoma to the 10p15–p14 region. Am. J. Hum. Genet., 62, 641–652.[CrossRef][Web of Science][Medline]

18. Wirtz, M.K., Samples, J.R., Rust, K., Lie, J., Nordling, L., Schilling, K., Acott, T.S. and Kramer, P.L. (1999) GLC1F, a new primary open-angle glaucoma locus, maps to 7q35–q36. Arch. Ophthalmol., 117, 237–241.[Abstract/Free Full Text]

19. Stone, E.M., Fingert, J.H., Alward, W.L., Nguyen, T.D., Polansky, J.R., Sunden, S.L., Nishimura, D., Clark, A.F., Nystuen, A., Nichols, B.E. et al. (1997) Identification of a gene that causes primary open angle glaucoma. Science, 275, 668–670.[Abstract/Free Full Text]

20. Rezaie, T., Child, A., Hitchings, R., Brice, G., Miller, L., Coca-Prados, M., Heon, E., Krupin, T., Ritch, R., Kreutzer, D. et al. (2002) Adult-onset primary open-angle glaucoma caused by mutations in optineurin. Science, 295, 1077–1079.[Abstract/Free Full Text]

21. Sarfarazi, M. and Rezaie, T. (2003) Optineurin in primary open angle glaucoma. Ophthalmol. Clin. North Am., 16, 529–541.[CrossRef][Medline]

22. Monemi, S., Child, A., Lehmann, O., Spaeth, G.L., Crick, R.P. and Sarfarazi, M. (2003) Genome scan of two large families with adult-onset primary open angle glaucoma (POAG) suggests a probable locus on 5q33–q35. The Association for Research in Vision and Ophthalmology (ARVO) Annual Meeting, Fort Lauderdale, FL. (Abstract no. 1128). Available from: http://abstracts.IOVS.org/cgi/content/abstract/44/5/1128.

23. Monemi, S., Child, A., Dasilva, A., Lehmann, O., Spaeth, G.L., Crick, R.P. and Sarfarazi, M. (2003) Provisional mapping of an adult-onset primary open angle glaucoma (POAG) locus to 5q33–q35. Am. J. Hum. Genet., 73, 534. Available from: http://www.ashg.org/genetics/abstracts/abso3/f2139.htm.

24. Samples, J.R., Sykes, R., Man, J., Rust, K., Kramer, P.L. and Wirtz, M.K. (2004) GLC1G: Mapping a new POAG locus on chromosome 5. The Association for Research in Vision and Ophthalmology (ARVO) Annual Meeting, Fort Lauderdale, FL. (Abstract no. 4622). Available from: http://abstracts.IOVS.org/cgi/content/abstract/45/5/4622.

25. Kramer, P.L., Samples, J.R., Schilling, K., Sykes, R.L., Man, J., Rust, K. and Wirtz, M.K. (2004) Mapping the GLC1G locus for primary open-angle glaucoma (POAG) in an Oregon family of Dutch origin. Am. J. Hum. Genet., 75. Available from: http://www.ashg.org/genetics/abstracts/abso4/f1914.htm.

26. Mao, M., Biery, M.C., Kobayashi, S.V., Ward, T., Schimmack, G., Burchard, J., Schelter, J.M., Dai, H., He, Y.D. and Linsley, P.S. (2004) T lymphocyte activation gene identification by coregulated expression on DNA microarrays. Genomics, 83, 989–999.[CrossRef][Web of Science][Medline]

27. Hajdu, J., Neutze, R., Sjogren, T., Edman, K., Szoke, A., Wilmouth, R.C. and Wilmot, C.M. (2000) Analyzing protein functions in four dimensions. Nat. Struct. Biol., 7, 1006–1012.[CrossRef][Web of Science][Medline]

28. Stoilov, I., Akarsu, A.N. and Sarfarazi, M. (1997) Identification of three different truncating mutations in cytochrome P4501B1 (CYP1B1) as the principal cause of primary congenital glaucoma (buphthalmos) in families linked to the GLC3A locus on chromosome 2p21. Hum. Mol. Genet., 6, 641–647.[Abstract/Free Full Text]

29. Sarfarazi, M., Stoilov, I. and Schenkman, J.B. (2003) Genetics and biochemistry of primary congenital glaucoma. Ophthalmol. Clin. North Am., 16, 543–554.[CrossRef][Medline]

30. Lathrop, G.M. and Lalouel, J.M. (1984) Easy calculations of lod scores and genetic risks on small computers. Am. J. Hum. Genet., 36, 460–465.[Web of Science][Medline]

CiteULike    Connotea    Del.icio.us    What's this?

This article has been cited by other articles:

				F. Carbonaro, T. Andrew, D. A. Mackey, T. L. Young, T. D. Spector, and C. J. Hammond Repeated Measures of Intraocular Pressure Result in Higher Heritability and Greater Power in Genetic Linkage Studies Invest. Ophthalmol. Vis. Sci., November 1, 2009; 50(11): 5115 - 5119. [Abstract] [Full Text] [PDF]

				L. M. E. van Koolwijk, P. R. Healey, R. A. Hitchings, P. Mitchell, P. C. Sham, P. McGuffin, and A. C. Viswanathan Major Genetic Effects in Glaucoma: Commingling Analysis of Optic Disc Parameters in an Older Australian Population Invest. Ophthalmol. Vis. Sci., November 1, 2009; 50(11): 5275 - 5280. [Abstract] [Full Text] [PDF]

				X. Jiao, Z. Yang, X. Yang, Y. Chen, Z. Tong, C. Zhao, J. Zeng, H. Chen, D. Gibbs, X. Sun, et al. Common variants on chromosome 2 and risk of primary open-angle glaucoma in the Afro-Caribbean population of Barbados PNAS, October 6, 2009; 106(40): 17105 - 17110. [Abstract] [Full Text] [PDF]

				M. Nakano, Y. Ikeda, T. Taniguchi, T. Yagi, M. Fuwa, N. Omi, Y. Tokuda, M. Tanaka, K. Yoshii, M. Kageyama, et al. Three susceptible loci associated with primary open-angle glaucoma identified by genome-wide association study in a Japanese population PNAS, August 4, 2009; 106(31): 12838 - 12842. [Abstract] [Full Text] [PDF]

				M. M. Walsh, H. Yi, J. Friedman, K.-i. Cho, N. Tserentsoodol, S. McKinnon, K. Searle, A. Yeh, and P. A. Ferreira Gene and Protein Expression Pilot Profiling and Biomarkers in an Experimental Mouse Model of Hypertensive Glaucoma Experimental Biology and Medicine, August 1, 2009; 234(8): 918 - 930. [Abstract] [Full Text] [PDF]

				N. Comes and T. Borras Individual molecular response to elevated intraocular pressure in perfused postmortem human eyes Physiol Genomics, July 9, 2009; 38(2): 205 - 225. [Abstract] [Full Text] [PDF]

				T. K. Footz, J. L. Johnson, S. Dubois, N. Boivin, V. Raymond, and M. A. Walter Glaucoma-associated WDR36 variants encode functional defects in a yeast model system Hum. Mol. Genet., April 1, 2009; 18(7): 1276 - 1287. [Abstract] [Full Text] [PDF]

				S. Park, Y. Jamshidi, D. Vaideanu, M. Bitner-Glindzicz, S. Fraser, and J. C. Sowden Genetic Risk for Primary Open-Angle Glaucoma Determined by LMX1B Haplotypes Invest. Ophthalmol. Vis. Sci., April 1, 2009; 50(4): 1522 - 1530. [Abstract] [Full Text] [PDF]

				Y. H. Kwon, J. H. Fingert, M. H. Kuehn, and W. L.M. Alward Primary Open-Angle Glaucoma N. Engl. J. Med., March 12, 2009; 360(11): 1113 - 1124. [Full Text] [PDF]

				A. X. C. N. Valente, S. B. Roberts, G. A. Buck, and Y. Gao Functional organization of the yeast proteome by a yeast interactome map PNAS, February 3, 2009; 106(5): 1490 - 1495. [Abstract] [Full Text] [PDF]

				Y. Zheng, J. Ge, G. Huang, J. Zhang, B. Liu, Y.-M. Hur, and M. He Heritability of Central Corneal Thickness in Chinese: The Guangzhou Twin Eye Study Invest. Ophthalmol. Vis. Sci., October 1, 2008; 49(10): 4303 - 4307. [Abstract] [Full Text] [PDF]

				M Akiyama, K Yatsu, M Ota, Y Katsuyama, K Kashiwagi, F Mabuchi, H Iijima, K Kawase, T Yamamoto, M Nakamura, et al. Microsatellite analysis of the GLC1B locus on chromosome 2 points to NCK2 as a new candidate gene for normal tension glaucoma Br J Ophthalmol, September 1, 2008; 92(9): 1293 - 1296. [Abstract] [Full Text] [PDF]

				J. M. Skarie and B. A. Link The Primary open-angle glaucoma gene WDR36 functions in ribosomal RNA processing and interacts with the p53 stress-response pathway Hum. Mol. Genet., August 15, 2008; 17(16): 2474 - 2485. [Abstract] [Full Text] [PDF]

				Y. Liu, S. Schmidt, X. Qin, J. Gibson, K. Hutchins, C. Santiago-Turla, J. L. Wiggs, D. L. Budenz, S. Akafo, P. Challa, et al. Lack of Association between LOXL1 Variants and Primary Open-Angle Glaucoma in Three Different Populations Invest. Ophthalmol. Vis. Sci., August 1, 2008; 49(8): 3465 - 3468. [Abstract] [Full Text] [PDF]

				F Carbonaro, T Andrew, D A Mackey, T D Spector, and C J Hammond Heritability of intraocular pressure: a classical twin study Br J Ophthalmol, August 1, 2008; 92(8): 1125 - 1128. [Abstract] [Full Text] [PDF]

				S. Chakrabarti, K. N. Rao, I. Kaur, R. S. Parikh, A. K. Mandal, G. Chandrasekhar, and R. Thomas The LOXL1 Gene Variations Are Not Associated with Primary Open-Angle and Primary Angle-Closure Glaucomas Invest. Ophthalmol. Vis. Sci., June 1, 2008; 49(6): 2343 - 2347. [Abstract] [Full Text] [PDF]

				M. He, B. Liu, W. Huang, J. Zhang, Q. Yin, Y. Zheng, D. Wang, and J. Ge Heritability of Optic Disc and Cup Measured by the Heidelberg Retinal Tomography in Chinese: The Guangzhou Twin Eye Study Invest. Ophthalmol. Vis. Sci., April 1, 2008; 49(4): 1350 - 1355. [Abstract] [Full Text] [PDF]

				P. Healey, F. Carbonaro, B. Taylor, T. D. Spector, P. Mitchell, and C. J. Hammond The Heritability of Optic Disc Parameters: A Classic Twin Study Invest. Ophthalmol. Vis. Sci., January 1, 2008; 49(1): 77 - 80. [Abstract] [Full Text] [PDF]

				F. Pasutto, C. Y. Mardin, K. Michels-Rautenstrauss, B. H. F. Weber, H. Sticht, G. Chavarria-Soley, B. Rautenstrauss, F. Kruse, and A. Reis Profiling of WDR36 Missense Variants in German Patients with Glaucoma Invest. Ophthalmol. Vis. Sci., January 1, 2008; 49(1): 270 - 274. [Abstract] [Full Text] [PDF]

				F. W. Rozsa, K. Scott, H. Pawar, S. Moroi, and J. E. Richards Effects of Timolol on MYOC, OPTN, and WDR36 RNA Levels Arch Ophthalmol, January 1, 2008; 126(1): 86 - 93. [Abstract] [Full Text] [PDF]

				S. de Voogd, R. C. W. Wolfs, N. M. Jansonius, A. G. Uitterlinden, H. A. P. Pols, A. Hofman, and P. T. V. M. de Jong Estrogen Receptors Alpha and Beta and the Risk of Open-angle Glaucoma: The Rotterdam Study Arch Ophthalmol, January 1, 2008; 126(1): 110 - 114. [Abstract] [Full Text] [PDF]

				S. Chakrabarti, K. R. Devi, S. Komatireddy, K. Kaur, R. S. Parikh, A. K. Mandal, G. Chandrasekhar, and R. Thomas Glaucoma-Associated CYP1B1 Mutations Share Similar Haplotype Backgrounds in POAG and PACG Phenotypes Invest. Ophthalmol. Vis. Sci., December 1, 2007; 48(12): 5439 - 5444. [Abstract] [Full Text] [PDF]

				K. K. Abu-Amero, J. Morales, M. N. Osman, and T. M. Bosley Nuclear and Mitochondrial Analysis of Patients with Primary Angle-Closure Glaucoma Invest. Ophthalmol. Vis. Sci., December 1, 2007; 48(12): 5591 - 5596. [Abstract] [Full Text] [PDF]

				G. Thorleifsson, K. P. Magnusson, P. Sulem, G. B. Walters, D. F. Gudbjartsson, H. Stefansson, T. Jonsson, A. Jonasdottir, A. Jonasdottir, G. Stefansdottir, et al. Common Sequence Variants in the LOXL1 Gene Confer Susceptibility to Exfoliation Glaucoma Science, September 7, 2007; 317(5843): 1397 - 1400. [Abstract] [Full Text] [PDF]

				L. M. E. van Koolwijk, D. D. G. Despriet, C. M. van Duijn, L. M. Pardo Cortes, J. R. Vingerling, Y. S. Aulchenko, B. A. Oostra, C. C. W. Klaver, and H. G. Lemij Genetic Contributions to Glaucoma: Heritability of Intraocular Pressure, Retinal Nerve Fiber Layer Thickness, and Optic Disc Morphology Invest. Ophthalmol. Vis. Sci., August 1, 2007; 48(8): 3669 - 3676. [Abstract] [Full Text] [PDF]

				B.-C. Park, M. Tibudan, M. Samaraweera, X. Shen, and B. Y.J.T. Yue Interaction between two glaucoma genes, optineurin and myocilin Genes Cells, August 1, 2007; 12(8): 969 - 979. [Abstract] [Full Text] [PDF]

				J. H. Fingert, W. L. M. Alward, Y. H. Kwon, S. P. Shankar, J. L. Andorf, D. A. Mackey, V. C. Sheffield, and E. M. Stone No Association Between Variations in the WDR36 Gene and Primary Open-Angle Glaucoma Arch Ophthalmol, March 1, 2007; 125(3): 434 - 436. [Full Text] [PDF]

				J. L. Wiggs Genetic Etiologies of Glaucoma Arch Ophthalmol, January 1, 2007; 125(1): 30 - 37. [Abstract] [Full Text] [PDF]

				S. P. Suriyapperuma, A. Child, T. Desai, G. Brice, A. Kerr, R. P. Crick, and M. Sarfarazi A New Locus (GLC1H) for Adult-Onset Primary Open-angle Glaucoma Maps to the 2p15-p16 Region Arch Ophthalmol, January 1, 2007; 125(1): 86 - 92. [Abstract] [Full Text] [PDF]

				F. W. Rozsa, K. M. Scott, H. Pawar, J. R. Samples, M. K. Wirtz, and J. E. Richards Differential Expression Profile Prioritization of Positional Candidate Glaucoma Genes: The GLC1C Locus Arch Ophthalmol, January 1, 2007; 125(1): 117 - 127. [Abstract] [Full Text] [PDF]

				D. Y. Wang, B. J. Fan, J. K. H. Chua, P. O. S. Tam, C. K. S. Leung, D. S. C. Lam, and C. P. Pang A Genome-wide Scan Maps a Novel Juvenile-Onset Primary Open-Angle Glaucoma Locus to 15q Invest. Ophthalmol. Vis. Sci., December 1, 2006; 47(12): 5315 - 5321. [Abstract] [Full Text] [PDF]

				B.-C. Park, X. Shen, M. Samaraweera, and B. Y.J.T. Yue Studies of Optineurin, a Glaucoma Gene: Golgi Fragmentation and Cell Death from Overexpression of Wild-Type and Mutant Optineurin in Two Ocular Cell Types Am. J. Pathol., December 1, 2006; 169(6): 1976 - 1989. [Abstract] [Full Text] [PDF]

				P. L. Kramer, J. R. Samples, S. Monemi, R. Sykes, M. Sarfarazi, and M. K. Wirtz The Role of the WDR36 Gene on Chromosome 5q22.1 in a Large Family With Primary Open-Angle Glaucoma Mapped to This Region. Arch Ophthalmol, September 1, 2006; 124(9): 1328 - 1331. [Abstract] [Full Text] [PDF]

				B. Liu, H. Chen, T. G. Johns, and A. H. Neufeld Epidermal growth factor receptor activation: an upstream signal for transition of quiescent astrocytes into reactive astrocytes after neural injury. J. Neurosci., July 12, 2006; 26(28): 7532 - 7540. [Abstract] [Full Text] [PDF]

				J. Wu, A. W. Hewitt, C. M. Green, M. A. Ring, P. J. McCartney, J. E. Craig, and D. A. Mackey Disease Severity of Familial Glaucoma Compared With Sporadic Glaucoma Arch Ophthalmol, July 1, 2006; 124(7): 950 - 954. [Abstract] [Full Text] [PDF]

				M. A. Hauser, R. R. Allingham, K. Linkroum, J. Wang, K. LaRocque-Abramson, D. Figueiredo, C. Santiago-Turla, E. A. del Bono, J. L. Haines, M. A. Pericak-Vance, et al. Distribution of WDR36 DNA Sequence Variants in Patients with Primary Open-Angle Glaucoma. Invest. Ophthalmol. Vis. Sci., June 1, 2006; 47(6): 2542 - 2546. [Abstract] [Full Text] [PDF]

				M. B. Petersen, G. Kitsos, J. R. Samples, N. D. Gaudette, E. Economou-Petersen, R. Sykes, K. Rust, M. Grigoriadou, G. Aperis, D. Choi, et al. A Large GLC1C Greek Family with a Myocilin T377M Mutation: Inheritance and Phenotypic Variability Invest. Ophthalmol. Vis. Sci., February 1, 2006; 47(2): 620 - 625. [Abstract] [Full Text] [PDF]

				T. Toh, S. H. M. Liew, J. R. MacKinnon, A. W. Hewitt, J. L. Poulsen, T. D. Spector, C. E. Gilbert, J. E Craig, C. J. Hammond, and D. A. Mackey Central Corneal Thickness Is Highly Heritable: The Twin Eye Studies Invest. Ophthalmol. Vis. Sci., October 1, 2005; 46(10): 3718 - 3722. [Abstract] [Full Text] [PDF]

				J. C. Charlesworth, T. D. Dyer, J. M. Stankovich, J. Blangero, D. A. Mackey, J. E. Craig, C. M. Green, S. J. Foote, P. N. Baird, and M. M. Sale Linkage to 10q22 for Maximum Intraocular Pressure and 1p32 for Maximum Cup-to-Disc Ratio in an Extended Primary Open-Angle Glaucoma Pedigree Invest. Ophthalmol. Vis. Sci., October 1, 2005; 46(10): 3723 - 3729. [Abstract] [Full Text] [PDF]

				D P Dimasi, A W Hewitt, C M Green, D A Mackey, and J E Craig Lack of association of p53 polymorphisms and haplotypes in high and normal tension open angle glaucoma J. Med. Genet., September 1, 2005; 42(9): e55 - e55. [Abstract] [Full Text] [PDF]

				S. W. M. John Mechanistic Insights into Glaucoma Provided by Experimental Genetics The Cogan Lecture Invest. Ophthalmol. Vis. Sci., August 1, 2005; 46(8): 2650 - 2661. [Full Text] [PDF]

				V. Vittal, A. Rose, K. E. Gregory, M. J. Kelley, and T. S. Acott Changes in Gene Expression by Trabecular Meshwork Cells in Response to Mechanical Stretching Invest. Ophthalmol. Vis. Sci., August 1, 2005; 46(8): 2857 - 2868. [Abstract] [Full Text] [PDF]

				T. Rezaie, D. M. Waitzman, J. L. Seeman, P. L. Kaufman, and M. Sarfarazi Molecular Cloning and Expression Profiling of Optineurin in the Rhesus Monkey Invest. Ophthalmol. Vis. Sci., July 1, 2005; 46(7): 2404 - 2410. [Abstract] [Full Text] [PDF]

				R. R. Allingham, J. L. Wiggs, E. R. Hauser, K. R. Larocque-Abramson, C. Santiago-Turla, B. Broomer, E. A. Del Bono, F. L. Graham, J. L. Haines, M. A. Pericak-Vance, et al. Early Adult-Onset POAG Linked to 15q11-13 Using Ordered Subset Analysis Invest. Ophthalmol. Vis. Sci., June 1, 2005; 46(6): 2002 - 2005. [Abstract] [Full Text] [PDF]

This Article Abstract FREE Full Text (PDF) All Versions of this Article: 14/6/725    most recent ddi068v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (110) Request Permissions Google Scholar Articles by Monemi, S. Articles by Sarfarazi, M. Search for Related Content PubMed PubMed Citation Articles by Monemi, S. Articles by Sarfarazi, M. Social Bookmarking          What's this?

Online ISSN 1460-2083 - Print ISSN 0964-6906

Copyright © 2009 Oxford University Press

Oxford Journals Oxford University Press

• Site Map
• Privacy Policy
• Frequently Asked Questions

Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan American National Biography Booksellers' Information Service Children's Fiction and Poetry Children's Reference Corporate & Special Sales Dictionaries Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks Humanities International Education Unit Law Medicine Music Online Products Oxford English Dictionary Reference Rights and Permissions Science School Books Social Sciences Very Short Introductions World's Classics