Hum. Hered. 25 : 161 -171 (1975)
Further Data on the Incidence and Segregation of Genetically Determined Electrophoretic Variants of Human Red Cell NADH Diaphorase L. W illiams and D. A. Hopkinson Department of Anatomy, University of Queensland, Brisbane, and MRC Human Biochemical Genetics Unit, Gallon Laboratory, University College London
Key Words. NADH diaphorase variants • Isozymes • Australian population Abstract. Human red cell NADH diaphorase isozyme patterns have been examined in 3,060 unrelated Australians of European origin, by starch gel electrophoresis. 26 people with variant isozyme patterns were encountered: 12 were phenotype Dia 2- I and 13 were Dia 4-1. A new variant isozyme pattern (Dia 7 1) was identified. No variants were identi fied in 100 Melanesians and 70 Australian Aborigines.
Introduction Several electrophoretic variants of human red cell NADH diaphorase have been identified and shown to be determined by variant alleles at an auto somal locus. Some of these variants were discovered in patients with niethaemoglobinaemia associated with diaphorase deficiency [2, 6, 7]: others were found in random surveys of normal healthy individuals [3-5, 8, 10. II]. This paper presents further data on the frequency of electrophoretic variation in healthy individuals and is principally concerned with Australian population samples.
Blood samples from healthy unrelated individuals were collected into ACD anti coagulant tubes and lysates were prepared as soon as possible by freezing and thawing thrice washed red cells. The NADH diaphorase isozyme patterns were then examined by starch gel electrophoresis using the method of H opkinson et ol. (5).
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Materials and Methods
162
Wll LIAMS Hohkinson Table I. Frequency o f NADU diaphorase variants in dilferent populations
Population group
Total number tested
Europeans Adults 2,933 127 Newborns 100 Melanesians Australian Aborigines 70 Chinese 15 9 Others Totals
3,254
Diaphorase phenotype 1 4 1 2 1
2,908 126 100 70 15 9 3.228
11 1
13 -
-
12
7 1
1 -
-
13
1
Total number of variants
25 1 26
Results and Discussion
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Population Data 2.933 unrelated adult Australians of European origin were tested and 25 individuals with variant electrophoretic patterns were encountered (table I). Of these. 11 were identical with the Dia 2 1 phenotype and 13 were identical with the Dia 4-1 phenotype. So in this population sample the frequency of the Dia 2-1 phenotype is 0.38% and that of the Dia 4-1 phenotype is 0.44%. No Dia 3-1. Dia 5-1 or Dia 6-1 variants were encountered. One other variant isozyme pattern not previously described was identified in one individual: it has been designed Dia 7 - 1. It is shown diagrammatically. together with the other variant isozyme patterns, in figure I, and in a photo graph of a starch gel in figure 2. This variant pattern was found to be a con stant individual characteristic by repeatedly checking fresh blood samples from this individual. As in the other variant patterns, two isozymes are present in Dia 7 I. one corresponding with the main isozyme of the Dia I phenotype: the other has a mobility slower than the Dia 1 isozyme, but faster than the variant isozyme of Dia 4-1. The variant isozyme of Dia 7-1 has about the same staining intensity as the Dia I isozyme; it does not appear to be associated with any enzyme deficiency. From comparison with other variants, phenotype Dia 7 1 would appear to represent the heterozygous combination of the common DIA' allele and a new variant allele designated DIA'. 127 cord blood samples from babies of Australian parents were also screened electrophoretically. These specimens were generally rather less
NA DH Diaphorasc Variants
163
HD A
Origin________________________________________________________________ Dia 1 2-1 3-1 4-1 5-1 6-1 7-1
~
Fig. I. Diagram showing red cell NADH diaphorasc isozyme patterns in individuals of the common phenotype Dia I and six rare phenotypes.
Dia 1
7-1
4-1
7-1
4-1
• - SS ¡Si? — 1
Fig. 2. Photograph of a starch gel showing the patterns of NADH diaphorase isozymes in phenotypes Dia I, Dia 4-1 and Dia 7-1.
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Origin
£
Table II. NADH diaphorase: incidence of variant phenotypes in unrelated, healthy individuals of various population groups
Australian Greek Cypriot Californian Michigan Seattle Total
2 1
1,975 725 1,008 989
8 (3) 5 16
-
-
12 1
-
13 -
-
-
-
3,060 101 2(X) several hundred1 175 8,233
Diaphorase phenotype
(2) 45
3-1
4-1
1
8 (1) 5
-
1
27
5-1
1 -
6-1
7-1
2 -
-
7
-
-
-
-
-
-
-
-
-
-
-
8
2
1
1
Total Number Incidence number o f o f different of variants variants variants/ 1,000 population
20 4 17 16
5 2 3 1
10.1 5.5 16.9 16.2
26 1 0
3 1 0
8.6 9.9 0
0 2
0 1
0 11.4
86
6
10.5
H opkinson el a/., 1970 TARIVIRDlANCtfl/., 1970 Potrakki elal., 1972 Santachiara-Benerecetti eta/., 1972
W i l lJAMS H o P K I N S O N
European English German West German Italian
Total number tested
Hopkinscn et al., 1970 K aplan and Beutler, 1967 Brewer et ai. 1967 D oti er et al., 1970
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Population group
Table II. (Continued) Indian London India Lebanese Sydney Negro London Michigan
-
7
-
1
-
-
147
1
-
-
-
-
1
1
1
-
-
-
(1) -
78
-
4 8
1 2
10.2 16.3
H opkinson el a!.. 1970 Blake and K irk , 1972
-
1
i
6.8
Blake and K irk , 1972
3
3
11.8
Hopkinson el a!.. 1970
-
-
-
1 0
1 0
0
Brewer
Further Data on the Incidence and Segregation of Genetically Determined Electrophoretic Variants of Human Red Cell NADH Diaphorase L. W illiams and D. A. Hopkinson Department of Anatomy, University of Queensland, Brisbane, and MRC Human Biochemical Genetics Unit, Gallon Laboratory, University College London
Key Words. NADH diaphorase variants • Isozymes • Australian population Abstract. Human red cell NADH diaphorase isozyme patterns have been examined in 3,060 unrelated Australians of European origin, by starch gel electrophoresis. 26 people with variant isozyme patterns were encountered: 12 were phenotype Dia 2- I and 13 were Dia 4-1. A new variant isozyme pattern (Dia 7 1) was identified. No variants were identi fied in 100 Melanesians and 70 Australian Aborigines.
Introduction Several electrophoretic variants of human red cell NADH diaphorase have been identified and shown to be determined by variant alleles at an auto somal locus. Some of these variants were discovered in patients with niethaemoglobinaemia associated with diaphorase deficiency [2, 6, 7]: others were found in random surveys of normal healthy individuals [3-5, 8, 10. II]. This paper presents further data on the frequency of electrophoretic variation in healthy individuals and is principally concerned with Australian population samples.
Blood samples from healthy unrelated individuals were collected into ACD anti coagulant tubes and lysates were prepared as soon as possible by freezing and thawing thrice washed red cells. The NADH diaphorase isozyme patterns were then examined by starch gel electrophoresis using the method of H opkinson et ol. (5).
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 6:33:34 PM
Materials and Methods
162
Wll LIAMS Hohkinson Table I. Frequency o f NADU diaphorase variants in dilferent populations
Population group
Total number tested
Europeans Adults 2,933 127 Newborns 100 Melanesians Australian Aborigines 70 Chinese 15 9 Others Totals
3,254
Diaphorase phenotype 1 4 1 2 1
2,908 126 100 70 15 9 3.228
11 1
13 -
-
12
7 1
1 -
-
13
1
Total number of variants
25 1 26
Results and Discussion
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 6:33:34 PM
Population Data 2.933 unrelated adult Australians of European origin were tested and 25 individuals with variant electrophoretic patterns were encountered (table I). Of these. 11 were identical with the Dia 2 1 phenotype and 13 were identical with the Dia 4-1 phenotype. So in this population sample the frequency of the Dia 2-1 phenotype is 0.38% and that of the Dia 4-1 phenotype is 0.44%. No Dia 3-1. Dia 5-1 or Dia 6-1 variants were encountered. One other variant isozyme pattern not previously described was identified in one individual: it has been designed Dia 7 - 1. It is shown diagrammatically. together with the other variant isozyme patterns, in figure I, and in a photo graph of a starch gel in figure 2. This variant pattern was found to be a con stant individual characteristic by repeatedly checking fresh blood samples from this individual. As in the other variant patterns, two isozymes are present in Dia 7 I. one corresponding with the main isozyme of the Dia I phenotype: the other has a mobility slower than the Dia 1 isozyme, but faster than the variant isozyme of Dia 4-1. The variant isozyme of Dia 7-1 has about the same staining intensity as the Dia I isozyme; it does not appear to be associated with any enzyme deficiency. From comparison with other variants, phenotype Dia 7 1 would appear to represent the heterozygous combination of the common DIA' allele and a new variant allele designated DIA'. 127 cord blood samples from babies of Australian parents were also screened electrophoretically. These specimens were generally rather less
NA DH Diaphorasc Variants
163
HD A
Origin________________________________________________________________ Dia 1 2-1 3-1 4-1 5-1 6-1 7-1
~
Fig. I. Diagram showing red cell NADH diaphorasc isozyme patterns in individuals of the common phenotype Dia I and six rare phenotypes.
Dia 1
7-1
4-1
7-1
4-1
• - SS ¡Si? — 1
Fig. 2. Photograph of a starch gel showing the patterns of NADH diaphorase isozymes in phenotypes Dia I, Dia 4-1 and Dia 7-1.
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 6:33:34 PM
Origin
£
Table II. NADH diaphorase: incidence of variant phenotypes in unrelated, healthy individuals of various population groups
Australian Greek Cypriot Californian Michigan Seattle Total
2 1
1,975 725 1,008 989
8 (3) 5 16
-
-
12 1
-
13 -
-
-
-
3,060 101 2(X) several hundred1 175 8,233
Diaphorase phenotype
(2) 45
3-1
4-1
1
8 (1) 5
-
1
27
5-1
1 -
6-1
7-1
2 -
-
7
-
-
-
-
-
-
-
-
-
-
-
8
2
1
1
Total Number Incidence number o f o f different of variants variants variants/ 1,000 population
20 4 17 16
5 2 3 1
10.1 5.5 16.9 16.2
26 1 0
3 1 0
8.6 9.9 0
0 2
0 1
0 11.4
86
6
10.5
H opkinson el a/., 1970 TARIVIRDlANCtfl/., 1970 Potrakki elal., 1972 Santachiara-Benerecetti eta/., 1972
W i l lJAMS H o P K I N S O N
European English German West German Italian
Total number tested
Hopkinscn et al., 1970 K aplan and Beutler, 1967 Brewer et ai. 1967 D oti er et al., 1970
Downloaded by: King's College London 137.73.144.138 - 1/16/2019 6:33:34 PM
Population group
Table II. (Continued) Indian London India Lebanese Sydney Negro London Michigan
-
7
-
1
-
-
147
1
-
-
-
-
1
1
1
-
-
-
(1) -
78
-
4 8
1 2
10.2 16.3
H opkinson el a!.. 1970 Blake and K irk , 1972
-
1
i
6.8
Blake and K irk , 1972
3
3
11.8
Hopkinson el a!.. 1970
-
-
-
1 0
1 0
0
Brewer
