Inorganic pyrophosphatase: a new polymorphic allozyme locus in Pacific salmon

Genetic polymorphism of inorganic pyrophosphatase was investigated in 2799 individuals in four species of Pacific salmon: chinook salmon (Oncorhynchus tshawytscha), coho salmon (O. kisutch), kokanee (O. nerka), and steelhead (O. mykiss), using horizontal starch gel electrophoresis. This enzyme system appears to be an isolocus system with electrophoretically indistinguishable allozymes encoded by two loci (PP-1,2*) expressed in retinal tissue. Mendelian inheritance was observed with a breeding study in three family crosses. Population variability in four species was characterized in 44 populations from the U.S. Pacific coast. Three alleles were found in chinook salmon; two alleles each were found in coho salmon, kokanee, and steelhead. Chinook salmon and kokanee populations differed enough with respect to PP-1,2* frequencies that this isolocus is useful for genetic stock identification in these species.     

NADP-dependent isocitrate dehydrogenase of Astyanax scabripinnis (Pisces, Characidae) from three altitudes at Grande Stream, Campos do Jord?o, SP.

Electrokinetic, thermic, and kinetic properties of products of NADP-dependent isocitrate dehydrogenase (IDHP; EC 1.1.1.42) loci of Astyanax scabripinnis (Pisces, Characidae) collected at three different altitudes (700 m, 1,800 m, and 1,920 m) of Grande Stream at Campos do Jord?o, State of S?o Paulo, Brazil, were analyzed. Two IDHP bidirectionally divergent loci, a single skeletal muscle, the IDHP-A*, and a single liver IDHP-B*, both polymorphic, were detected in the three different altitude populations. The variant allele *128 at the IDHP-A* locus, had its highest frequency detected in the 1,920 m population (0.494). Among the nine variant alleles detected at the IDHP-B* locus (*37, *57, *69, *79, *85, *114, *119, *124, and *140), the *37 and 79 were detected only in 1,800 m population. Chi-square values showed that only the 700 m population is not in Hardy-Weinberg equilibrium for the IDHP-A* locus, while for the IDHP-B* locus, no population is. Homogeneity Chi2 test indicated that the populations are significantly different in their A and B phenotype frequencies. Wright's FST mean value (0.036 and 0.32, IDHP-A* and IDHP-B*, respectively) was 0.178 for the three altitude populations which means that 82% of total genetic diversity was found among individuals of each one of the populations. Stability at environmental temperatures (16 degrees to 21 degrees C), and apparent Km and Vmax values of each A-phenotype skeletal muscle crude extract suggest different roles of A-isoforms during the increased lipogenesis that occurs in fish at low temperatures.     

Silencing of duplicate genes: a null allele polymorphism for lactate dehydrogenase in brown trout (Salmo trutta)

A previously described isozyme polymorphism at one of two skeletal muscle LdhA loci in brown trout is due to a null allele, Ldh1(n), producing no detectable catalytic activity. Homozygotes for this allele have approximately only 56% of the LDH activity in skeletal muscle relative to homozygotes for the active allele. The remaining activity results from enzyme subunits produced by other LDH loci. The Ldh1(n) allele is common and widespread throughout brown trout populations in Sweden and is also found in populations from Ireland. The persistence of duplicate gene expression for the LdhA loci in almost all salmonid species is best explained by natural selection against individuals containing null alleles. However, there is no indication of natural selection against brown trout with the Ldh1(n) allele: We suggest that the selection against individuals containing null alleles that is apparently responsible for the persistence of duplicate LdhA loci in salmonids occurs only under certain environmental conditions.      

The human glutathione S-transferases: Developmental aspects of the GST1, GST2, and GST3 loci

The expression of the GST1, GST2, and GST3 loci in fetal, neonatal, and infant tissues has been studied using starch gel electrophoresis and chromatofocusing. Each locus demonstrated developmental changes in expression, some of which were specific to a single tissue while others occurred in several tissues. GST1 was not usually expressed in any of the tissues studied before 30 weeks of gestation but steadily increased thereafter until adult levels were reached in late infancy. In neonates and older infants the frequencies of the GST1*0, GST1*1, and GST1*2 alleles were 0.79, 0.07, and 0.14, respectively. GST2 was always expressed in liver and adrenal but was only weakly expressed in spleen, cardiac muscle, and diaphragm. In kidney this locus was not usually expressed until nearly 1 year after birth. The GST3 isoenzymes were present in all fetal, neonatal, and infant tissues, although their expression in liver decreased after 30 weeks of gestation. Other isoenzymes with fast anodal mobilities were also identified in several tissues; these are believed to be GST3 isoenzymes that have undergone posttranslational modification rather than products of the putative GST4 locus. No specifically fetal isoenzymes were detected.     

Genetic basis of creatine kinase isozymes in skeletal muscle of salmonid fishes

The genetic basis of isozyme phenotypes of creatine kinase (CK) from extracts of skeletal muscle of salmonids has been resolved through breeding data including double heterozygous crosses and backcrosses of rainbow trout (Salmo gairdneri), and backcrosses of coho salmon (Oncorhynchus kisutch). The two-, three-, or four-banded phenotypes of homozygous individuals and all heterozygous and hybrid phenotypes of ten salmonid species are readily explained by the following model: (1) there are no detectable heterodimers either between allelic products at a single locus or between loci; (2) each allele is represented electrophoretically by two bands, presumably a reflection of stable posttranslational modification of a single polypeptide unit; (3) CK of salmonid muscle is encoded by two loci—CK-1 and CK-2. The distance separating the paired bands reflecting each allele provides a basis for two groupings—a broad-spaced group (including all species of Oncorhynchus tested excepting O. masou) and a narrow-spaced group (including all species of Salmo tested and O. masou). The relationships among species suggested by the relative mobilities and spacings of these CK bands are consistent with taxonomic schemes inferred from morphological, cytogenetic, and other isozymic data.     

Immunochemical characterization of human red cell acid phosphatase isozymes

An immunological study was performed on human red cell acid phosphatase (ACP1) isozymes encoded by different alleles, each of which is expressed as an electrophoretically fast (f) isozyme and a slow (s) isozyme. These isozymes reacted as two immunochemically different groups. Allele-specific reactions were not detected between either the f isozymes or the s isozymes. Quantitation of ACP1 isozymes in red cells by crossed immunoelectrophoresis revealed a phenotype-dependent variation in the concentration of isozyme protein. A simple gene dosage effect was indicated and the ordering of the ACP1 alleles (ACP1*A < ACP1*B < ACP1*C < ACP1*E) was identical to that found for enzyme activity levels. Also, an allele effect on the proportion between s and f isozymes (s/f) was observed; the ordering here was ACP1* B < ACP1*A < ACP1*, which is the same as that reported for the susceptibility to modulation with purines. These variations in isozyme protein levels appear to account for the phenotypic differences in the intensity of the isozyme bands, when activity-stained after electrophoresis, and in the red cell enzyme activity levels. Investigation of two carriers of a Null allele showed no evidence of an aberrant protein product, and half-normal concentrations of enzyme protein were observed in the red cells of these individuals.     

Liver aldehyde oxidase and xanthine oxidase genetics in the mouse

A 'null' activity variant for the major liver isozyme of aldehyde oxidase (AOX-1) in adult male mice and an electrophoretically distinct, high activity variant of the second liver isozyme (AOX-2) were used to examine the segregation of the genetic loci encoding these enzymes (Aox-1 and Aox-2 respectively) in breeding studies. A single recombinant between these loci was observed among the 147 backcross progeny examined, which confirms a previous report (Holmes, 1979) for close linkage and genetic distinctness of the two loci. An activity variant for mouse liver xanthine oxidase (XOX) is also reported which behaved as though controlled by codominant alleles at a single locus (designated Xox-1 ). Genetic analyses showed that the Xox-1 locus segregated independently of the multiple- A ox loci.     

Geographic variation and temporal population differentiation of chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> from some regions of the Russian Far East

Allozyme variation of chum salmon Oncorhynchus keta was examined in the populations from nine rivers of the Russian Far East. A total of 12 loci were tested, and eight of these were shown to be polymorphic. The greatest contribution to the samples differentiation was made by the EST-D1* and sIDHP-2* loci. Most of the allele diversity was distributed among the individuals within the samples. The among-sample differences within races were statistically significant and accounted for slightly more than 7% of total variation. The differences between the races constituted 0.1% and were statistically nonsignificant. Genetic similarity of seasonal races within single locality allowed suggestion on their secondary formation after settling of founder population into a certain region.     

Genetic comparison of salmon from the White Sea and north-western Atlantic Ocean

Samples of salmon Salmo salar from the River Kachkovka and the River Nilma in northern Russia were analysed by starch gel electrophoresis and compared to three Norwegian stocks, the Neiden river in northern Norway and Øyreselv and Hopselv rivers on the west coast. The comparison included the following polymorphic loci: AAT-4 *, IDDH-2 *, IDHP-3 *, MDH- 3,4 *, MEP-2 *, ESTD * as well as the newly discovered polymorphic loci FBALD-3 * and TPI-3 *. Samples were run side by side on gels, and the alleles found in the Russian stocks were the same as those found in the Norwegian stocks, although the electrophoretic methods used lead to differences in designations of alleles. A polymorphism in ESTD * which involves a slow allele was commonly observed in the three northern populations of the Nilma, Kachkovka and Neiden rivers. This allele was absent in the other Norwegian stocks and in a major brood stock of farmed salmon in Norway. The IDHP-3 * 116 allele was found in unusually high frequencies in the northern populations. Thus, the variability observed at these two loci indicates a barrier to gene flow between the northern salmon stocks and the more southern stocks in the East Atlantic area.     

Enolase isozymes in Coho salmon.

1. Analysis of the enolase isozymic distribution has been performed in tissues of the Coho salmon, using electrophoretic separation on cellulose acetate strips followed by localization of enzymatic activity. 2. A total of six electrophoretically distinct forms are seen in Coho salmon in patterns that differ both qualitatively and quantitatively from one tissue to another. 3. The isozymes in skeletal muscle and liver are sufficiently similar to one another that a purification procedure previously developed for trout muscle enolase by Cory & Wold (1966) can be used to partially purify enolase from either of the above-mentioned Coho tissues. The main form of enolase in Coho muscle has an isoelectric point of 7.57. 4. Both liver and skeletal muscle enolases can be reversibly denatured in guanidine HCl and subsequently renatured. Liver enolase appeared to renature somewhat faster than muscle enolase under the same conditions. 5. While polyploidy among salmonids may contribute to the complexity of enolase patterns in fish, the differences in isozymic patterns seen from one tissue to another indicate the presence of distinct, nonallelic genes, probably arising through gene duplication.     

Y-chromosome-specific microsatellite variation in Australian aboriginals

The frequency distributions of 4 highly polymorphic Y-chromosome-specific microsatellites (DYS19, DYS390, DYS391, and DYS392) were determined in 79 unrelated Australian Aboriginal males from the Northern Territory. These results are compared with those observed in worldwide populations at both the locus and the haplotype level. Common alleles in Aboriginals are DYS19*15 (49%), DYS19*14 (28%), DYS390*19 (39%), DYS390*24 (20%), DYS391*10 (72%), DYS392*11 (63%), and DYS392*13 (28%). No evidence of reduced gene diversity was observed for these Y-chromosome alleles. DYS390 exhibits the most complex arrangement, displaying a bimodal distribution composed of common alleles (*22-*26), and rare short alleles (*18-*20), with an intermediate allele (*21) being absent. DYS390*20, previously reported only in Papuans and Samoans, is observed for the first time in Aboriginals. Compared with a recent study of Aboriginals, our sample exhibits considerable diversity in the haplotypes associated with the rare DYS390*19 allele, indicating that this allele is of considerable antiquity, if it arose as a single deletion event. Combining all 4 Y-chromosome-linked microsatellites produced 41 unique haplotypes, which were linked using a median-joining network. This network shows that most (78%) of our Aboriginal haplotypes fall into 2 distinct clusters, which likely represent 2 separate lineages. Seven haplotypes are shared with haplotypes found in a recent study of Aboriginals, and 7 are shared with a Spanish population. The cluster of Aboriginal haplotypes associated with the short DYS390 alleles does not share any haplotypes with the Spanish, indicating that this cluster of haplotypes is unique to Australian Aboriginals. Limited data from 4 worldwide populations used to construct haplotypes based on 3 loci (DYS19, DYS390, DYS392) show that only 4 of these haplotypes are seen in Australian Aboriginals. Shared haplotypes may be the result of admixture and/or recurrent mutation at these loci. Expanding the haplotype analysis to include biallelic markers on the Y chromosome will resolve this issue.     

Malate dehydrogenase in subtropical fish belonging to the orders characiformes,siluriformes and perciformes I. Duplicate gene expression and polymorphism

• 1.1. Electrophoretic analysis of the soluble malate dehydrogenase (sMDH) from 22 subtropical fish belonging to the orders Characiformes, Siluriformes and Perciformes, collected in 10 reservoirs of São Paulo State and in two lakes of Minas Gerais State, Brazil, indicates that at least two sMDH loci, MDH-A¹ and MDH-B¹, are active. In addition to this latter locus, in Hoplias malabaricus (Erythrinidae, Characiformes), a MDH-A 1,3¹ isoloci is proposed in order to explain the six-banded pattern detected in all the individuals screened.
• 2.2. In attempting to explain the multiplicity of compounds detected in 87% of the Geophagus brasiliensis (Cichlidae, Perciformes) specimens analyzed, three hypotheses are proposed: the event of duplication in processing the presence of three loci with a null allele within the MDH-B¹, and overdominance.
• 3.3. In 87% of the species here studied, a bidirectionally divergent pattern of expression of the sMDH loci was observed, in which the least anodal isozyme A₂ predominated in liver, and the most anodal isozyme B₂ predominated in skeletal muscle. In two siluriform species, Pimelodela gracilis and Hypostomus regani, and in one perciform, Tilapia rendalli, a unidirectionally divergent pattern, in which the isozyme A₂ predominated in every tissue analyzed, was observed.
• 4.4. Polymorphism in at least one of the sMDH loci was detected in 9% of the species studied here: Leporinus friderici (Characiformes) at the MDH-A¹ and P. gracilis at both sMDH loci. In L. friderici and Pimelodus maculatus (Siluriformes), rare alleles at the MDH-B¹ locus were detected. Polymorphism at the mitochondrial locus was detected in Tilapia rendalli.

     

Genetic characterization in four sciaenid species from the Arabian Sea

Four sciaenid species Johnieops dussumieri, Kathala axillaris, Pennahia macropthalmus and Otolithes ruber were analysed electrophoretically for genetic variation at 18 loci (16 in P. macropthalmus and O. ruber ). Twelve loci were polymorphic in J. dussumieri , 10 in K. axillaris , three in P. macropthalmus and 12 in O. ruber ( P <0·99). Average heterozygosity ranged from 0·033 ± 0·100 to 0·070 ± 0·122. The allele frequencies of 14 loci were used to estimate Nei's genetic distance (). The values ranged from 0·334 to 0·612. Three isozyme loci ( LDH-B*, MDH-2* and G3PDH-1* ) were found to be the most reliable species-specific markers.     

Genetic variability inCebus appella paraguayanus: Biochemical analysis of seven loci and variation in Glyoxalase I (E.C.4.4.1.5)

An analysis of seven loci inCebus apella paraguayanus showed that Glyoxalase I was polymorphic due the appearance of two alleles (GLO*2 andGLO*3) with frequencies of 0.955 and 0.045, respectively. Of the two alleles,GLO*2 was electrophoretically similar to the most common allele found in the human andAotus. These results confirmed our previous findings in the same population sample showing that this subspecies has a very low genetic variation among New World primates.     

A comparative electrophoretic analysis of mammalian carbonic anhydrase isozymes: evidence for a third isozyme in red skeletal muscles.

1. Starch gel electrophoretic patterns of carbonic anhydrase (CA) isozymes were examined from tissue extracts of cats, sheep, rabbits and mice. 2. In addition to the widely distributed and extensively studied B and C isozymes, an additional isozyme (called CA-A) was observed. 3. Tissue distribution studies showed the A isozyme to be predominantly localized in red skeletal muscles, although this activity was also observed in white and "mixed" skeletal muscles of the cat, sheep and rabbit, as well as sheep lung and rabbit liver. 4. A, B and C isozymes of carbonic anhydrase from cat, sheep and mice exhibited independent variations in nett surface charge. In terms of decreasing anodal migration, the following results are reported: cat A greater than C greater than B; sheep C greater than B greater than A; and mouse B greater than C greater than A. 5. These results are consistent with the existence of 3 genetic loci encoding carbonic anhydrase in mammalian tissues.     

A genetic analysis of the London strain of rainbow trout

The London strain of rainbow trout (Oncorhynchus mykiss) was created by interbreeding three other strains of rainbow trout and therefore was expected to have higher levels of genetic variation than other strains of rainbow trout. We examined 129 London strain rainbow trout from Indiana by allozyme electrophoresis to assess levels of genetic variation and to examine the relationship between the London strain and other hatchery strains. When using the same loci to compare with other hatchery strains the London strain showed levels of genetic variation within the range of other hatchery strains: mean heterozygosity of 0.053 (0.031-0.099), 1.27 (1.20-1.60) alleles per locus and 20.0% (20.0-40.0%) of the loci were polymorphic. The London strain is somewhat distinct from other hatchery strains (D=0.009-0.072), in part because of the high frequency of the sIDHP*40 allele.     

Evolution of isozyme loci and their differential tissue expression

Summary The phylogeny of the creatine kinase (CK, EC 2.7.3.2) isozyme loci and their differential tissue expressions were determined for representatives of 65 families of vertebrates, with emphasis on the fishes. The transition from the single creatine kinase locus, characteristic of certain echinoderms, to the two creatine kinase loci which are orthologous to those present in all vertebrates, occurred early in the chordate line. The majority of pre-teleostean fishes possesses only these two CK loci (A and C). These loci are relatively generalized in their tissue expressions which are variable among species of primitive fishes. The third and fourth creatine kinase loci (B and D) arose separately in the ancestors of the bony fishes and appear to be the result of regional genome duplications. Concomitant with the increase in the number of isozyme loci has been an increase in the specificity of their tissue expression. In the advanced teleost fishes the four CK loci are differentially expressed in a characteristic manner. The A₂ isozyme predominates in skeletal muscle, the B₂ isozyme in eye and brain, the C₂ isozyme in stomach muscle, and the D₂ isozyme is found exclusively in testis. We propose a phylogeny of the creatine kinase genes in the lower chordates based on the time of appearance of new CK loci, the sequence in which the loci achieve a tissue restricted expression, and the immunochemical relatedness of the orthologous and paralogous gene products.     

Discrimination of walleye, Stizostedion vitreum vitreum (Mitchill), stocks by isozyme analysis with cellulose acetate

Summary. Five walleye, Stizosfedion vitreum vitreum (Mitchill), populations from within the state of Minnesota were examined for isozyme gene frequencies. Of 26 loci identified and scored, nine polymorphic loci were found. The ADH*, MDH-3* and PROT-4* loci were found to be the most informative for distinguishing the populations. A north-south cline in the frequency of PROT-4* alleles was found, with a higher frequency of the fast allele being found in the southern population. This is probably a reflection of genetic isolation caused by glaciation. Cellulose acetate was used to resolve the isozymes. The advantages of cellulose acetate over starch in terms of decreased run time, smaller sample and stain volumes and superior resolution advocate the use of cellulose acetate for fish isozyme analyses. Comparisons between starch and cellulose acetate separations showed similar results with some exceptions. Cellulose acetate resolved FH* and PROT-2* polymorphisms that were not resolved on starch, but failed to resolve the MDH-1 locus. These differences did not affect the results.     

Haplotype analysis of the apolipoprotein E and apolipoprotein C1 loci in Portugal and São Tomé e Príncipe (Gulf of Guinea): linkage disequilibrium evidence that APOE*4 is the ancestral APOE allele

The joint distributions of phenotypes from the apolipoprotein E gene (APOE) and from a closely linked restriction site polymorphism at the apolipoprotein C1 locus (APOC1) were studied in population samples from Portugal and S?o Tomé e Príncipe (Gulf of Guinea), a former Portuguese colony that was originally populated by slaves imported from the African mainland. The frequencies of the APOE alleles (*2, *3, and *4) in Portugal and S?o Tomé fitted the ranges of variation generally observed in European and African populations, respectively. Haplotype analysis showed that in both populations the strength of linkage disequilibrium was highest for the APOE*2 allele and lowest for the APOE*4 allele, suggesting that the origin of the APOE alleles followed a 4-->3-->2 pathway and thus providing independent confirmation of the results from sequence homology studies with nonhuman primates. In accordance with global trends in the distribution of human genetic variation, the European sample from Portugal presented more intense linkage disequilibrium between APOE and APOC1 than the African sample from S?o Tomé where, despite the short 4-kb distance that separates the 2 loci, the level of association between the APOC1 alleles and APOE*4 was nonsignificant.     

Protein loci in the Arctic charr, Salvelinus alpinus L.: electrophoretic expression and genetic variability patterns

In the search for electrophoretically detectable protein loci in the Arctic charr, Salvelinus alpinus L ., tissue samples of eye, liver, and muscle from a total of 934 specimens collected at 10 Swedish localities were analysed. General protein staining and specific staining for 33 enzymes revealed 52 detectable loci; 37 of which were considered usable in population surveys. Variability was observed at four loci coding for esterase ( est-2 ), the liver-specific form of lactate dehydrogenase ( ldh-3 ), and the skeletal muscle form of malate dehydrogenase ( mdh-4, 5 ); genetic variation at loci coding for ldh-3 and mdh-4, 5 has not previously been described in the Arctic charr. Relating our results to the multiple locus studies presented in the literature including Arctic charr from Ireland and North America reveals polymorphism at approximately one-third of the loci studied in the Arctic charr, and the fraction of variable loci does not appear lower in this species than in other salmonids. There were highly significant allele frequency differences between samples. Nevertheless, there was a high degree of genetic similarity among all the populations sampled indicating that they were derived from a relatively recent common ancestor. The results are discussed in relation to the current controversy concerning the number of major evolutionary lines in Scandinavian Arctic charr.