RESCUE-ESE are shown above the genomic sequence of exon 3 for each .... bull's-eye maculopathy. Epiretinal. Central macular atrophy. Centra l well define d.
Gardner et al., Human Mutation
Supp. Figure S1. Cone dystrophy and colour blindness segregating with different cone opsin array haplotypes. Pedigree of Family 22 with segregation of X-linked cone dystrophy (filled symbols) and protanopia (shaded symbols). Segregation of the cone opsin array haplotypes is shown beneath each individual in the pedigree. Xlinked cone dystrophy segregates with haplotype p.C203R on one X-chromosome and protanopia segregates with a single M- gene on a different X-chromosome. Individual IV-7, unusually for a female, is colour blind and is a compound heterozygote for p.C203R and a single M- gene haplotype.
1
Gardner et al., Human Mutation
S10
S12
S2
S5
S17
S19
S20
S23
S21
S26
N
Supp. Figure S2. Electroretinogram traces for adult subjects. Subject ID is given on the left for each trace. DA refers to Dark Adapted; LA to Light Adapted; PERG to Pattern ERG. Supp. Table S1 provides further details.
2
Gardner et al., Human Mutation
3
Supp. Figure S3. Distribution of electroretinogram amplitudes by mutation class and patient. Full-field ERG amplitudes in 10 patients. Amplitudes are expressed as a percentage of the age-matched lower limit of normal and are shown for dark-adapted ERGs (DA0.01; DA010.0 a-wave) and light adapted ERGs (LA 3.0 30Hz; LA 3.0 b-wave). The genotype is stated for each subject. Lower limit of normal is defined as the minimum normal value minus 5% of the reference interval (maximum-minimum normal value in a group of healthy subjects).
Gardner et al., Human Mutation
4
150 ERG amplitude (% of age-matched normal)
LA 3.0 30Hz ERG
100
50
0
0
20
40 age (years)
60
80
Supp. Figure S4. Electroretinogram (ERG) amplitudes and patient age. Comparison of full-field ERG amplitudes (DA 10.0 ERG a-wave; LA 3.0 30Hz ERG) with age in 6 individuals with exon 3 interchange haplotypes. There is a trend for ERG amplitudes (corrected for age) to decrease with advancing age.
Gardner et al., Human Mutation
5
Supp. Figure S5. Serial electroretinograms (ERG) of patients S21 and S26. Comparison of fullfield ERG parameters at baseline with those at follow-up in S21 and S26. In S21 DA 0.01 and DA 10.0 a-wave amplitudes showed marked reduction in amplitude after 12 years. In S26 all amplitudes were reasonably stable but DA0.01 ERG showed an increase in peak time of 15ms bilaterally after an interval of 5 years.
Gardner et al., Human Mutation
6
Supp. Figure S6. Splicing motif prediction comparison for wild-type and disease associated L- and M- exon 3 haplotypes. (A) Splice enhancer motifs (yellow type) predicted by RESCUE-ESE are shown above the genomic sequence of exon 3 for each haplotype (shown to left of figure). The SNP c.532A>G results in loss of a splice enhancer motif (white arrow) in all 3 disease associated haplotypes LIAVA, LVAVA and MIAVA. (B) Splice silencer motifs (red type) predicted by FAS-ESS are shown above exon 3 genomic sequence. SNP c.532A>G results in the gain of a splice silencer motif (black arrow) in all three disease associated haplotypes while other SNPs in the haplotype have no predicted effect on splicing.
Gardner et al., Human Mutation
7
Supp. Table S1. Summary of clinical findings Mutation
LCR DEL
Subject Age (S)yrs Family (F) ID
S1 F1
10
Age of onset
Clinical
Refract ERG ion
LCR DEL
S2 F2
S3 F3
14
16
OCT
FAF
Visual Acuity
Poor vision from 5 yrs
HRR/Ishih ara plates Nystagmus
Poor vision from 3 mths
M +++
Cone ++++ Rod technically poor Mac ++++
Myopic
M +++
Cone++++ Rod + Mac ++++ S-cone normal
Myopic
Slight granular appearance to ellipsoid line centrally
ND
Seen once
M+
Cone ++++ Rod normal Mac ++++ S-cone absent
ND
ND
ND
No 5 yrs
6/30 BE
Nystagmus 6/36 BE Tri-3/4 Pro/deusome (abn deutan) Nystagmus
Poor vision from 18 6/38 RE mths 6/48 LE
Tri-4/4 Pro/deu - 0
Progression Test Interval
Tri -4/4 Pro/deu – 0 LCR DEL
Fundus appearance
Normal
ND
No 5 yrs
Gardner et al., Human Mutation Mutation
LCR DEL
LCR DEL
Subject Age (S)yrs Family (F) ID
S4 F4
S5 F5
17
58
Age of onset
8 Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
Poor vision from infancy
Poor vision from infancy
HRR/Ishih ara plates Nystagmus
Test Interval
M+
ND
ND
ND
ND
6/30 RE 6/38 LE Trit-1/4 Pro/deu - 0 No documented Nystagmus
Progression
No 2 yrs
M +++
Cone +++ Peripapillary Rod normal atrophy Mac ++++
Disruption of central outer retinal layer
ND
ND
ND
ND
No 8 yrs
6/60 RE 6/48 LE
p.C203R
S6 F6
4
Poor vision from early infancy
Tri-3/4 Pro/deu - 0 Nystagmus 6/60 RE 6/48 LE
Colour testing ND
M ++
ND
No 4 yrs
Gardner et al., Human Mutation Mutation
p.C203R
p.C203R
Subject Age (S)yrs Family (F) ID
S7 F7
S8 F8
5
8
Age of onset
9 Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
Poor vision from 6 mths
Poor vision from 5 yrs
HRR/Ishih ara plates Nystagmus 6/48 RE 6/38 LE Tritan-1/4 Pro/deu - 0 No documented Nystagmus
Progression Test Interval
Myopic astigma tism
Cone +++Rod normal Mac ND S-cone normal
Blond fundus
Normal
Norm al
No
M ++
Cone +++ Rod normal Mac ++++
ND
Myopic Astigm atism
Cone +++ Rod normal Mac ND
Normal
ND
ND
No 1 yr
Myopic Astigm atism
Cone ++++ Rod normal Mac ++++
Normal
Granular appearance to ellipsoid
ND
No
3 yrs
ND
ND
No 2 yrs
6/38 BE
p.C203R
p.C203R
S9 F9
S10 F10
11
14
Poor vision from early infancy Poor vision from
Tri-4/4 Pro/deusome Nystagmus 6/60 BE Tri-2/4 Pro/deu - 0 Nystagmus 6/19 RE
10 yrs
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
S11 F11
16
Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
early infancy
p.C203R
10
ND
Test Interval
HRR/Ishih ara plates 6/24 LE Tritan-3/4 Pro/deu9/20 Nystagmus
Progression
S-cone normal
line centrally
M+
Cone ++++ Rod normal
Peripheral Normal hyperpigment ation
ND
No 1 yr
M+
Cone +++ Rod + Mac +++ S-cone normal
Normal
Norm al
No
Cone +++ Rod normal
Bull’s- eye maculopathy, pigmentary changes,
6/60 BE Ishihara - 0
p.C203R
S12 F12
19
Poor vision from infancy
Nystagmus 6/36 BE Tri-4/4 Passed Berson Pro/deu-0
p.C203R
S13 F22
27
ND
Nystagmus 6/60 BE
M +++
Normal
2 yrs
ND
ND
No 7 yrs
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
11 Clinical
Refract ERG ion
S14 F22
18
OCT
FAF
Visual Acuity
ND
Nystagmus
Progression Test Interval
HRR/Ishih ara plates Colour testing ND
p.C203R
Fundus appearance
thinning of retinal vessels M+++
Cone ++++ Rod +
Pale discs, mottled macular pigmentation
ND
ND
No
M+
Cone ++++ Rod ND
Macular atrophy
ND
ND
No
M +++
Cone++++ Rod normal
Peripapillary atrophy
Normal
ND
No
6/18 BE at 1 yr 6/36 BE at 5yrs Colour testing ND p.C203R
S15 F22
19
ND
Nystagmus 2/60 BE - at 3 yrs 3/60 RE 6/60 LE - at 4yrs
LIAVA LIAVA
S16 F14
10
ND
Pro/deu-0 No documented
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
12 Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
Test Interval
HRR/Ishih ara plates Nystagmus
MVVVA
Progression
Mac ++++
2 yrs
6/18 BE
LIAVA LIAVA MVVVA
S17 F15
18
Poor vision from early infancy
Tri - 4/4 Pro/deu some No documented Nystagmus
M +++
Cone ++ Rod normal Mac ++++ S-cone normal
Mild macular hyperpigmentation
Normal
Cone +++ Rod normal Mac +++ S-cones normal
Blond fundus
ND
Norm al
No 3 yrs
6/12 RE 6/18 LE
LVAVA
S18 F16
10
Poor vision from 9 mths
Tri-all Pro/deusome No documented Nystagmus 6/15 RE 6/12 LE
M ++
ND
No 1 yr
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
13 Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
Progression Test Interval
HRR/Ishih ara plates
LVAVA
S19 F13
40
Poor vision from 2 yrs
Tri-all Pro/deu- 0 No documented Nystagmus
M +++
6/24 BE
LVAVA
LIAVA
S20 F17
S21 F18
49
73
Poor vision from early infancy
Poor vision from early infancy
Tri-3/4 Pro/deu – 0 No documented Nystagmus
M +++
6/18 BE Tri-4/4 Pro/deu-0 No documented Nystagmus 6/60 BE
M +++
Cone +++ Rod ++ Mac ++++ S-cone normal
Moderate central macular atrophy, peripapillary atrophy
Central outer retinal atrophy
Norm al
Cone ++ Rod + Mac ++++ S cone normal
Peripapillary atrophy, mild macular hyperpigmention, pale optic discs
ND
Age 59 Cone ++++ Rod Normal Mac ++++ S-cone normal
Bilateral bull’s-eye maculopathy
Subfoveal lucency between ellipsoid and retinal pigment epithelial layers Central macular atrophy
Epiretinal
No 3 yrs
No 5 years
Centra l well define d macul ar
Yes 20 years Progression on ERG with
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
14 Clinical
Refract ERG ion
LIAVA MIAVA* MIAVA*
S22 F19
S23 F19
9
68
OCT
FAF
Visual Acuity
Poor vision from infancy
Poor vision from infancy
Nystagmus
Progression Test Interval
HRR/Ishih ara plates Tri-0/4 Ishihara -0 Pro/deu-0
LIAVA MIAVA* MIAVA*
Fundus appearance
membranes
M +++
Age 71 Cone ++++ Rod ++ S-cone normal Cone ++++ Rod +
Peripapillary atrophy Myopic fundus
hypo- increasing autofl rod uoresc dysfunction ence
Myopic thinning
ND
Stable over 6 years
ND
Centra ND l hypoa utofluore scent region with hyper autofluore scent rim
6/36 BE Color testing ND Nystagmus 6/60 BE Tri-4/4 Pro/deusome
Mac and Scone ND M+
Cone ++++ Rod normal Mac ++++ S-cone absent
Bilateral macular atrophy, peripapillary atrophy
Gardner et al., Human Mutation Mutation
MIAVA MIAVA MVVVA
MIAVA MIAVA MVVVA
LVAVA LVAVA MVVVA
Subject Age (S)yrs Family (F) ID
S24 F20
S25 F20
S26 F21
34
51
80
Age of onset
15 Clinical
Refract ERG ion
Fundus appearance
OCT
FAF
Visual Acuity
Poor vision from 6 mths
HRR/Ishih ara plates Nystagmus
Poor vision in late 20s
Test Interval
M +++
6/24 BE
Colour testing ND Poor Nystagmus vision Bilateral from 12 ptosis mths convergent squint
M +++
-18.0 D BE at age 13
Colour testing ND No documented Nystagmus
M+
Colour testing ND
Cone ++ Rod technically poor
Peripapillary atrophy, mild inferior retinal pigmentation
ND
Cone +++ Rod +++
Severe myopic degeneration
ND
Progres sive
6/18 BE
6/36 BE
Progression
Cone +++ Rod + Mac ++++ S cones Normal
Normal
ND
No 2 yrs
ND
No 13 yrs
Disruption of outer retinal layer
Norm al
Maternal Grandfather (I-1) registered blind at 58yrs Yes Progression on ERGs, over 5 yrs, Rod peak time prolonged, cone system
Gardner et al., Human Mutation Mutation
Subject Age (S)yrs Family (F) ID
Age of onset
16 Clinical Visual Acuity
Refract ERG ion
Fundus appearance
OCT
FAF
Progression Test Interval
HRR/Ishih ara plates function unchanged LCR DEL = locus control region deletion, LIAVA represents haplotype (GCG/AT/c/G/G), MIAVA represents haplotype (AAC/AT/c/G/G) MIAVA* represents (AAG/AT/c/G/G), LVAVA represents haplotype (GCG/GG/c/G/G), MVVVA represents haplotype (AAC/GG/t/G/G). Visual acuity refers to Snellen visual acuity, RE is right eye, LE is left eye, BE both eyes; HRR refers to Hardy-Rand-Rittler colour plates that were seen; ERG is electroretinogram; OCT is Optical Coherence Tomography; FAF is fundus autofluorescence; ND, not determined. Myopia; + low (6 Diopters). Mac +, ++, +++ and ++++ refer to mild (70-99% of normal amplitude), moderate (30-69% of normal), severe (1-29% of normal) and undetectable macular dysfunction, respectively. Cone +, ++, +++ and ++++ refer to mild (70-99% of normal amplitude), moderate (30-69% of normal), severe (1-29% of normal) and undetectable cone system dysfunction, respectively. Rod +, ++, +++ and ++++ refer to mild (70-99% of normal amplitude), moderate (30-69% of normal), severe (1-29% of normal) and undetectable rod system dysfunction, respectively.
Gardner et al., Human Mutation
17
Supp. Table S2. Electroretinogram results in paediatric patients (surface recordings and paediatric protocol) Mutation
Subject Age at ERG (yrs)
Stimulus Scotopic bright flash ERG
Photopic 30Hz ERG
Pattern ERG (PERG)
LCR
S1
5
Strobe
Unreliable
Undetectable
Undetectable
LCR
S3
10
Ganzfeld
Normal
Undetectable
Undetectable
p.C203R
S7
3
Strobe
Normal
Severe
Not Done
p.C203R
S8
5
Ganzfeld
Normal
Severe
Undetectable
p.C203R
S11
14
Strobe
Normal
Undetectable
Not done
p.C203R
S14
2
Strobe
Mildly reduced
Undetectable
Not done
p.C203R
S15
4
Strobe
Not done
Undetectable
Not done
Interchange
S16
6
Ganzfeld
Normal
Undetectable
Undetectable
Interchange
S22
5
Strobe
Mildly reduced
Undetectable
Not done
Gardner et al., Human Mutation
18
Supp. Table S3. ISCEV Electroretinograms (ERG), pattern ERGs and S-cone ERGs in adults Mutation
Subject
Age (at ERG)
ISCEV ERG
ISCEV ERG
ISCEV ERG
(DA10.0)
(LA 3.0 30Hz)
S-cone ERG
ISCEV PERG
(DA 0.01)
LCR
S2
10
Mild
Mild
Undetectable
Normal
Undetectable
LCR
S5
52
Normal
Normal
Severe
Not available
Undetectable
p.C203R
S10
10
Normal
Normal
Undetectable
Normal
Undetectable
p.C203R
S12
16
Mild
Mild
Severe
Normal
Severe
Interchange
S17
16
Normal
Mild
Moderate
Normal
Undetectable
Interchange
S19
38
Moderate
Mild
Severe
Normal
Undetectable
Interchange
S20
44
Mild
Normal
Moderate
Normal
Undetectable
Interchange
S21
59
Normal
Normal
Undetectable
Normal
Undetectable
Interchange
S23
63
Normal
Normal
Undetectable
Undetectable
Undetectable
Interchange
S26
69
Mild
Moderate
Severe
Normal
Undetectable
ERG measures of dysfunction; Mild: 70%-99% of normal amplitude, Moderate 30-69% of normal, Severe 1-29% of normal, Undetectable. *6mm pupils ISCEV refers to International Society for Clinical Electrophysiology of Vision; DA to Dark Adapted; LA to Light Adapted.
