Biochemical polymorphisms in South African freshwater fish. Isoenzyme patterns in fish of the families Cyprinidae and Salmonidae

Isoenzyme patterns were studied in local populations of the carp ( Cyprinus carpio ) and the bream ( Sarotherodon mossambicus ) of the Cyprinid family. and in the trout ( Sabno giardneri ) of the family Salmonidae. Homogenates of heart muscle extracts were used in the identification of PGI. NADP-IDH. PGM and 6-PGD.
Polymorphisms found and gene frequencies obtained are discussed separately for each enzyme and species or population.
In the bream, variation was found only at the PGI locus. In the carp only the PGM locus was polymorphic whereas both PGM and IDH showed variation in the trout. The variation at the PGM locus in the trout cannot support the three locus model suggested elsewhere and clearly indicates a single locus for the mo-nomeric enzyme.
The large variety of NADP-IDH types in trout not only illucidates the complexity of this locus but confirms a disomic mode of inheritance. Genetic differences in the trout populations could be related to possible advantages for management purposes.     

Biochemical polymorphisms in South African freshwater fish. Isoenzyme patterns in fish of the families Cyprinidae and Salmonidae

Isoenzyme patterns were studied in local populations of the carp (Cyprinus carpio) and the bream (Sarotherodon mossambicus) of the Cyprinid family, and in the trout (Salmo giardneri) of the family Salmonidae. Homogenates of heart muscle extracts were used in the identification of PGI, NADP-IDH, PGM and 6-PGD. Polymorphisms found and gene frequencies obtained are discussed separately for each enzyme and species or population. In the bream, variation was found only at the PGI locus. In the carp only the PGM locus was polymorphic whereas both PGM and IDH showed variation in the trout. The variation at the PGM locus in the trout cannot support the three locus model suggested elsewhere and clearly indicates a single locus for the monomeric enzyme. The large variety of NADP-IDH types in trout not only illucidates the complexity of this locus but confirms a disomic mode of inheritance. Genetic differences in the trout populations could be related to possible advantages for management purposes.     

Effects of prolactin and androgens on enzymes of carbohydrate metabolism in prostate of castrated bonnet monkeys Macaca radiata (Geoffroy).

Effects of prolactin (PRL), bromocriptine (Br), testosterone propionate (TP), dihydrotestosterone (DHT) and the combinations of these androgens with PRL/Br on the specific activities of caudal and cranial prostatic cellular enzymes involved in carbohydrate metabolism in castrated mature bonnet monkeys have been studied. Castration decreased all the enzymes studied such as hexokinase (HK), 6-phosphofructokinase (6-PFK), glyceraldehyde-3-phosphate dehydrogenase (G-3-PD), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) in the cranial and caudal prostates. PRL elevated the activities of all the enzymes above normal except G-3-PD of cranial lobe. In the caudal lobe, PRL brought back the activities of HK, PFK, PK, G-6-PD to normal and 6-PGD above normal except G-3-PD. TP/DHT treatment increased all the enzymes in both the lobes. PRL given along with TP/DHT further enhanced the androgen action with regard to HK, PK, G-6-PD and 6-PGD of cranial and PFK, G-3-PD, PK, G-6-PD and 6-PGD of caudal lobe. Br treatment did not produce any alteration of these enzymes in both the lobes. In the cranial lobe, during Br+TP/DHT treatment, the stimulating effects of androgen were unaffected on all the enzymes except PK. On the other hand in the caudal, the stimulatory effects of androgens were affected and the activities of HK, PFK, PK and 6-PGD were significantly decreased. The present results suggest that PRL has a direct as well as a synergistic action with androgens on enzymes of EMP and HMP shunt in the prostates of monkeys.     

Regulation and rate of the hexose monophosphate shunt in Rana ridibunda erythrocytes

1. Resting rates of Rana ridibunda erythrocyte glucose consumption and 14CO2 production from 1-14C-glucose were found to be significantly lower than the respective values in human erythrocytes. 2. In the presence of 1-14C-glucose Methylene Blue stimulated 14CO2 production 7-fold, while in the presence of 6-14C-glucose Methylene Blue stimulated 14CO2 production 1.2-fold. 3. The Km of G-6-PD for G-6-P and NADP were 29 and 12 microM, respectively while the Km of 6-PGD for 6-PG and NADP were 83 and 32 microM, respectively. The Ki of G-6-PD and 6-PGD for NADPH were 80 and 12 microM, respectively. 4. Excess amounts of NADP resulted in a significant decrease of 14CO2 production from 1-14C-glucose in total haemolysates. 5. ATP, ADP and fructose diphosphate inhibited both G-6-PD and 6-PGD, the latter being more sensitive than G-6-PD to their inhibitory effect, 2,3-DPG and reduced and oxidized glutathione showed a marked inhibitory effect on 6-PGD, while the phosphorylated trioses inhibited only G-6-PD. 6. Physiological concentrations of oxidized glutathione decreased the inhibition exercised by NADPH on G-6-PD. 7. The possible role of the two dehydrogenases in the regulation of the HMS is discussed.     

Polymorphism and genetic control of some isozyme systems in mutant sunflower lines

Polymorphism of six isozyme systems (ME, EST, 6-PGD, AAT, ACP, IDH) of sunflower mutant inbred lines has been studied. It was shown that only three of them were polymorphic (ME, EST, 6-PGD). Two isoforms were observed for each polymorphic enzymic zone. Genetic control of malic-enzyme has been studied and it was determined that the identified polymorphic zone of the enzyme was controlled by one gene. The genes of malic-enzyme synthesis, anodal esterase and 6-phosphogluconat-dehydrogenase are inherited independently.     

Studies of enzymes connected with erythrocyte glutathione metabolism in a rural tropical population

Summary The activities of the erythrocyte enzymes hexokinase (HK), glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconate dehydrogenase (6-PGD), glutathione reductase (GR) and glutathione peroxidase (GSH-PO) were determined in a group of 12 Europeans and in a group of 103 male Thai subjects in northern Thailand. In the Thai group there were 16 subjects with G-6-PD deficiency and 28 subjects with abnormally low levels of GR activity. A comparison of the enzyme activities in the different subgroups indicated that HK and 6-PGD are not influenced by G-6-PD deficiency whereas GR and GSH-PO activities are significantly higher in G-6-PD deficient subjects. In the group with low GR activity G-6-PD and GSH-PO showed a tendency to an elevation of activity when compared with the normal control group. Significant positive correlations exist between G-6-PD and 6-PGD in the normal group and between GR and GSH-PO in the G-6-PD deficient group. A negative correlation between GR and GSH-PO was present in the group with low GR activities. A study of the families of subjects with low activity of GR did not yield evidence for the existence of a deficiency polymorphism.

---

Zusammenfassung Bei 12 Europäern und einer Gruppe von 103 männlichen thailändischen Versuchspersonen wurden die Aktivitäten der Erythrocytenenzyme Hexokinase (HK), Glucose-6-Phosphat-Dehydrogenase (G-6-PD), 6-Phosphogluconat-Dehydrogenase (6-PGD), Glutathion-Reduktase (GR) und Glutathion-Peroxidase (GSH-PO) bestimmt. In der Thai-Gruppe waren 16 Personen mit G-6-PD-Mangel und 28 Personen mit abnormal niedrigen Aktivitäten der GR. Ein Vergleich der Enzymaktivitäten in verschiedenen Untergruppen zeigte, daß HK und 6-PGD durch G-6-PD-Mangel nicht beeinflußt werden. Im Gegensatz hierzu sind die Aktivitäten der GR und der GSH-PO bei G-6-PD-Mangel signifikant erhöht. In der Gruppe mit erniedrigter GR-Aktivität bestand eine Tendenz zu erhöhten Werten für G-6-PD und GSH-PO. Die Korrelationen zwischen G-6-PD und 6-PGD in der Gruppe mit normaler G-6-PD und die zwischen GR und GSH-PO in der Gruppe mit G-6-PD-Mangel waren signifikant. In der Gruppe mit erniedrigter GR-Aktivität fand sich eine negative Korrelation zwischen GR und GSH-PO. Die Untersuchungen in Familien von Personen mit niedriger GR-Aktivität ergaben keinen sicheren Hinweis auf das Vorliegen eines GR-Mangel-Polymorphismus in der untersuchten Bevölkerung.

---

---

Established and supported by Stiftung Volkswagenwerk, Hannover.     

Isozyme diversity in sour,sweet, and ground cherry

Thirty-six sour (Prunus cerasus L.), sweet (P. avium L.), and ground cherry (P. fruticosa Pall.) selections were evaluated for seven enzyme systems and principal coordinate analysis was used to examine isozyme divergence among these cherry species. The enzyme systems studied were phosphoglucose isomerase (PGI), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphogluconate dehydrogenase (6-PGD), leucine aminopeptidase (LAP), shikimate dehydrogenase (SKDH), and malate dehydrogenase (MDH). The first principal coordinate, which accounted for 41% of the total variation, separated the diploid sweet cherry selections from the sour, ground, and sour x ground cherry tetraploids. An additional 86 selections were evaluated for up to six of the enzyme systems to determine the polymorphisms at the enzyme loci and the level of heterozygosity between the diploid sweet cherry and the tetraploid species and interspecific hybrids. 6-PGD was the most polymorphic enzyme exhibiting 16 patterns. The tetraploid cherry species were more heterozygous than the diploid sweet cherry with an average heterozygosity of 78% compared to 19% for the diploids.     

Contribution to the genetics of serum β-lipoproteins in man

Summary Blood and serum from 17 Macaca mulatta were analysed for haptoglobins, transferrins, 6-phosphogluconate dehydrogenase, adenylate kinase, adenosine deaminase, phosphoglucomutase, acid phosphatase, glucose-6-phosphate dehydrogenase and phosphohexose isomerase. Compared to the human pattern, the AcPh and the ADA components of macaques are fast moving; AK, PGM, 6-PGD and G-6-PD have almost uniform and similar electrophoretic mobilities; and the Hp, Tf and PHI show differential mobilities. All these macaques possess similar karyotypes.Aided by UGC Junior Research Fellowship.     

Genetic polymorphisms in juvenile-onset diabetes

Nine genetic polymorphic systems (ACP1, PGM1, ADA, AK, G-6-PD, Hp, ABO, Rh, MN), were studied in a series of 138 subjects affected by JOD. Differences between diabetic patients and controls were observed in the distribution of phenotypes of the red cell acid phosphatase (ACP1), and the ABO and MN blood groups.     

Enzyme activities during imbibition and germination of seeds of tamarack (Larix laricina)

Activities of six enzymes from extracts of separated embryos and gametophytes of tamarack [ Larix laricina (Du Roi) K. Koch] seeds were assayed at various stages of imbibition and germination. On a per seed part basis, activities of 6-phosphogluconate dehydrogenase (6-PGD, EC 1.1.1.44), glucose-6-phosphate dehydrogenase (G-6-PD, EC 1.1.1.49), malate dehydrogenase (NAD⁺–MDH, EC 1.1.1.37), isocitrate dehydrogenase (NADP⁺–IDH, EC 1.1.1.42), soluble peroxidase (PER, EC 1.11.1.7), and acid phosphatase (ACP, EC 3.1.3.2) from both the embryo and gametophyte tissues generally increased slowly, following cold stratification for 30 days and imbibition under germinating conditions for 5 days, but then increased at a faster rate with emergence of the radicle and subsequent growth of the seedling. The rate of increase of enzyme activity was highest for PER. Soluble protein levels also increased with imbibition and germination, with about 3 times greater levels present in the gametophyte than in the embryo. Heat inactivation experiments showed that, except for G-6-PD, activities were stable up to 40°C. Inactivation occurred at lower temperatures for G-6-PD, while higher temperatures were required for PER. Incubation of extracts for 7 days at 4°C indicated that loss of enzyme activity was greatest for G-6-PD (3.9% remaining) and least for PER and ACP (94 and 95% remaining, respectively).     

Genetic structure of the populations of native inhabitants in the northeastern USSR. V. The Chukot Evens

The genetic structure of two Chukot Evens subpopulations (314 individuals) for electrophoretic protein systems and taste sensitivity to PTC was studied. 17 of the 39 loci were polymorphic (43.59%). The following systems were completely monomorphic: diaphorase NAD H (Dia); glucose-6-phosphate dehydrogenase (G-6-PD); glutamatoxalate transaminase (GOT); carbonic anhydrase (Ca-1); catalase (Ct), lactate dehydrogenase (loci LDH-A and LDH-B); leucine aminopeptidase (Lap); malate dehydrogenase (MDH); purine nucleoside phosphorylase (PNP); superoxide phosphorylase (PNP); superoxide dismutase (SOD); phosphoglucomutase-2 (PGM2); cholinesterase (locus E1); red cell esterase (4 loci); albumin (Alb); hemoglobin (Hb A and B); ceruloplasmin (Cp); and blood, gren, using the standard method. The following systems were polymorphic: red cell acid phosphatase (AcP); phosphoglucomutase-1 (PGM1); 6-phosphogluconate dehydrogenase (PGD); glutamatepyruvate transaminase (GPT); glyoxalase-1 (GLO-1); esterase (EsD); adenilatkinase (AK); alkaline phosphatase (Pp); cholinesterase (locus E2); haptoglobin (Hp); transferrin (Tf); group-specific component (Gc) and ABO, MN, Lewis, P blood groups and taste sensitivity to PTC. The following allele frequencies for polymorphic loci have been detected: AKI = 0.994; GLO = 1I = 0.082; GPT1 = 0.653; AcPA = 0.400; AcPB = 0.599; AcPC = 0.001; PGDA = 0.944; PGM1(1) = 0.906; EsD1 = 0.897; E2+ = 0.048; HpI = 0.394; GcI = 0,919; Tfc = 0.987; r(O) = 0.669; p(A) = 0.184; q(B) = 0.146; M = 0.711; Le = 0.411; P1+ = 0.521; t = 0.295. The genetic structure of Chukot Evens population is significantly nearer to that of the other ethnic groups of the North-East, in comparison with the genetic structure of Evenks of the Middle Siberia.     

Activity of some glycolytic and pentose phosphate pathway enzymes during the development of adventitious roots

Activities of phosphofructokinase (PFK, EC 2.7.1.11), glyceraldehyde 3-phosphate (NAD) dehydrogenase [G-3-PD(NAD), EC 1.2.1.12], glucose 6-phosphate dehydrogenase (G-6-PD, EC 1.1.1.49), and 6-phosphogluconate dehydrogenase (6-PGD, EC 1.1.1.44) were determined in bean cuttings (Phaseolus vulgaris L. cv. Top Crop) over 4 days, encompassing adventitious root primordium initiation and development. Effects of applied auxin and “endogenous root-forming stimulus”(ERS) on enzyme activities, concentrations of reducing sugars, and primordium development were also determined during the first 4 days of propagation. Effects of auxin were determined through use of applied indole-3-acetic acid (IAA) or 2,3,5-triiodobenzoic acid. Effects of ERS were evaluated by means of decapitation of cuttings. Increased basipetal transport and increased metabolism of reducing sugars occurred in leafy cuttings in response to applied IAA and to ERS. Primordium development and activities of the four enzymes increased in leafy cuttings under conditions that simultaneously increased basipetal transport and metabolism of reducing sugars. Three types of enzyme activity response were found: (i) activity increased over time by ERS and by applied IAA [G-3-PD(NAD)], (ii) activity increased over time by ERS but not by applied IAA (PFK, G-6-PD), (iii) activity increased over time but not by ERS or applied IAA (6-PGD). Increases in G-3-PD(NAD), G-6-PD, and PFK activity in leafy cuttings were positively related to primordium development. 6-PGD activity increased in leafy cuttings during primordium development and may have supported it. However, equal increases occurred in decapitated cuttings, in which the long-term development of primordia was supressed. Results for G-3-PD(NAD) that were obtained in an experiment with jack pine (Pinus banksiana Lamb.) seedling cuttings were similar to results for the same enzyme in bean cuttings. G-3-PD(NAD) activity in naphthaleneacetic acid-treated jack pine cuttings increased with time, in comparison with untreated cuttings, before root emergence.     

Red cell antigen, serum protein and red cell enzyme polymorphisms in Karkar Islanders and inhabitants of the adjacent North Coast of New Guinea

Blood samples from the Waskia and Takia populations of Karkar Island, Papua New Guinea, and other nearby mainland populations, were tested for genetic variation in blood group, serum protein and red cell enzyme systems. Polymorphic variation was present in the ABO, P, MNS, Rh, Lewis, Duffy, Kidd and Gerbich blood group systems, in the Hp and Tf serum protein systems, and in the acid phosphatase, 6-PGD, ADA, PGM, MDH, and G-6-PD enzyme systems. A small number of variants was found in other systems: there were 4 Lu(a+), 1 Kp(a+), 2 C variants in the acid phosphatase system, 6 LDH variants, 1 ADA3-1 and 1 AK2-1 sample. All samples were negative for the red cell antigens Cw, Vw, He, K, Jsa, Dia, Wra, Rd and Marriott, and no variation was observed in the PHI enzyme system. The results are discussed in relation to those obtained on other Papua New Guinea populations.     

Erythrocyte G-6-PD and 6-PGD genetic polymorphisms in South African Negroes,with a note on G-6-PD and the malaria hypothesis

Summary Sample of 981 and 998 South African Negroes belonging to seven different ethnic groups were screned for G-6-PD and 6-PGD phenotypes, respectively. The results are discussed in terms of the interethnic variability and the possible adaptive values of these genetic polymorphisms. Particular attention is paid to the geographic co-distribution and interrelationship of G-6-PD deficiency and the occurrence of malaria in South Africa.     

Electrophoretic patterns of glucose metabolizing enzymes and acid phosphatase in mouse and human neuroblastoma cells

The electrophoretic patterns of glucose metabolizing enzymes and acid phosphatase in mouse and human neuroblastoma cells were investigated. Mouse neuroblastoma cells had one band of lactate dehydrogenase (LDH) and two bands of acid phosphatase, whereas human neuroblastoma cells had five bands of LDH and one band of acid phosphatase. Glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) were expressed as a single band in both mouse and human neuroblastoma cells. The electrophoretic pattern of LDH was similar in mouse neuroblastoma cells grown in culture or in vivo. The electrophoretic band of G-6-PD in mouse neuroblastoma cells grown in vivo appeared to be less dense than that observed in cells grown in culture; however, the reverse was true for 6-PGD. Among all enzymes examined, only the electrophoretic pattern of G-6-PD in cAMP-induced “differentiated” mouse neuroblastoma was different in comparison to control cells.     

Glucose-6-?hosphate dehydrogenase deficiency,haptoglobin and hemoglobin variants in dogs

Blood samples from 31 male and 34 female, adult, healthy dogs of different breeds were studied for erythrocytic G-6-PD activity. The hemolysates were also studied electrophoretically for G-6-PD, 6-PGD, and hemoglobin variants. Most of the plasma samples revealed a human Hp 1–1 type of band while three samples had an additional fast-moving band that disappeared on addition of an excess of hemoglobin and two samples were ahaptoglobinemic. G-6-PD deficiency was detected in eight samples, and it was more frequent in males than in females. The implications of G-6-PD deficiency with no difference in the electrophoretic pattern and of ahaptoglobinemia are discussed with respect to different genetic and clinical possibilities.     

Genetic structure of the populations of native inhabitants of the northeastern USSR. IV. The Koryaks of Kamchatka

Genetic structure of four Kamchatka subpopulations (675 individuals) was estimated for 25 erytrocyte and serum systems, some blood groups and for taste sensitivity to PTC. 23 of 38 loci examined are completely monomorphic. These are: AK, Ca-1, Cat, Dia, Est1-4, GOT, G-6-PD, LDH A and B, MDH, PGM2, SoD, Hb alpha and beta, ChE1, Lap, Alb, Cp, Tf, Rh. Following allele frequencies were found for polymorphic loci: AcPA = = 0.616; AcPB = 0.383; AcPC = 0.0015; EsD1 = 0.882; GLO - I1 = 0.156; GPT1 = 0.611; PGDA = = 0.959; PGM1(1) = 0.953; ChE2+ = 0.039; Gc1 = 0.888; Hp1 = 0.173; r(0) = 0.620; P(A) = 0.201; q(B) = 0.179; le = 0.192; M = 0.397; P1+ = 0.585; t = 0.371. According to monomorphic and polymorphic loci set, Kamchatka Koryaks are rather similar to other ethnic North-East Asiatic groups, being the most approximate to Reindeer Chuckchies and the most remote from Alaskan and Asiatic Eskimos. In other words, the extent of genetic differences between Kamchatka Koryaks and North-East populations corresponds to the geographic distribution and the degree of ecological differences in these populations. Analysis of interpopulation heterogeneity permitted to reveal the extent of contribution of individual loci to "differentiation" of North-East ethnic groups. The possible influence of ecological factors on interpopulation and intersubpopulation heterogeneity of the loci analysed is discussed.     

Genetic Variability in Scotch Pine in the Southeastern Part of Its Range

The main parameters of genetic variability have been determined in an isolated natural Scotch pine population from Chita oblast (Tsasuchei Forest) by analysis of 19 genes coding for nine enzymes: GDH, IDH, LAP, PGM, AAT, ADH, MDH, 6-PGD, and DIA. Polymorphic genes constituted 63.2% of all structural genes studied in the population at the 99% polymorphism criterion. The mean number of alleles per locus was 1.63. The observed and expected heterozygosities were 0.237 and 0.251, respectively. These estimates are close to the corresponding mean values for Scotch pine according to the data on 18 or more structural genes.     

Genetic variability in Scotch pine in the southeastern part of its range

The main parameters of genetic variability have been determined in an isolated natural Scotch pine population from Chita oblast' (Tsasuchei Forest) by analysis of 19 genes coding for nine enzymes: GDH, IDH, LAP, PGM, AAT, ADH, MDH, 6-PGD, and DIA. Polymorphic genes constituted 63.2% of all structural genes studied in the population at the 99% polymorphism criterion. The mean number of alleles per locus was 1.63. The observed and expected heterozygosities were 0.237 and 0.251, respectively. These estimates are close to the corresponding mean values for Scotch pine according to the data on 18 or more structural genes.     

Interstrain differences in red cell enzyme activities in mice and rats.

1. Interstrain differences in red blood cell enzyme activities were studied in mice (BALB/c, C57BL/6, C3H/He, DBA/2 and ddY) and rats (Donryu, F344/N, SD, Wistar and Wistar/ST), and were also compared with hamster, guinea-pig and rabbit. 2. The enzyme activities measured were: glutathione S-transferase (GST), glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconate dehydrogenase (6-PGD), NADPH-diaphorase (ND), hexokinase (Hx), glutamate oxaloacetate transaminase (GOT), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE). 3. There were marked variations in the activities of some red cell enzymes (e.g. GST, Hx, ND), while others (e.g. G-6-PD, 6-PGD) were much less variable both within different strains and species.