Differences in isoenzyme patterns of axenically and monoxenically grown Acanthamoeba and Hartmannella

Axenically and monoxenically grown Acanthamoeba castellanii, Acanthamoeba polyphaga and different isolates of Hartmannella vermiformis strains were examined by polyacrylamide isoelectric focusing in the pH range 3–10. Isoenzyme patterns of acid phosphatase (AP), propionyl esterase (PE), malate dehydrogenase (MDH), alcohol dehydrogenase (ADH), glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) were compared. Zymograms were used to reveal differences in typical isoenzyme patterns between axenically and monoxenically grown amoebae and to compare axenically grown A. castellanii, A. polyphaga and H. vermiformis. Comparison of zymograms for AP, PE and MDH between axenically grown Acanthamoeba and Hartmannella strains revealed different isoenzyme patterns. Acanthamoeba showed strong bands for ADH and extremely weak bands for GPI and PGM, while Hartmannella lacked ADH but possessed bands for GPI and PGM.\par Comparison of zymograms from axenically and monoxenically grown amoebae revealed a lower intensity and even lack of typical isoenzyme bands in lysates from monoxenic cultures. The observed changes in typical isoenzyme patterns induced by the bacterial substrate can influence the correct isoenzymatic typing of different strains in clinical and phylogenetic studies.     

Developmental rate and viability of rainbow trout with a null allele at a lactate dehydrogenase locus

We show that a previously described isozyme polymorphism in rainbow trout(Salmo gairdneri) is the result of an enzymatically inactive (i.e., null) allele(n). Ldh3 null homozygotes(n/n) and heterozygotes(100/n) have reductions of about 20 and 12% in total lactate dehydrogenase (LDH) activity at hatching, respectively. As juveniles,(100/n) fish have reductions in LDH activity of 15, 37, and 21% in brain, heart, and white muscle, respectively. Embryos with differntLdh3 phenotypes from 11 families do not differ significantly in either survival or hatching time. However, a second measure of developmental rate, the amount of malate dehydrogenase (MDH) and phosphoglucomutase (PGM) activity in 33-day-old embryos, suggests that(100/n) embryos develop more slowly than(100/100) embryos. In three of four families examined,(100/n) embryos have significantly lower amounts of total MDH activity (8–10%). In one of these,(100/n) embryos also have significantly lower total PGM activity (15%). These data suggest that the reduction in total LDH activity is associated with small but detectable delays in developmental rate but nondetectable differences in survival to hatching.This research was supported by NSF Grant BSR-8300039 to F. W. Allendorf and a postgraduate scholarship from the NSERC (Canada) to M. M. Ferguson.     

Genetic variability in Hoplias malabaricus (Osteichthyes: Erythrinidae) in fluvial and lacustrine environments in the Upper Paraná river floodplain (Paraná State,Brazil)

Genetic variability in Hoplias malabaricus, from two localities in the upper Paraná River floodplain, was investigated by starch and polyacrylamide gel electrophoresis. A total of 52 specimens were analyzed for 14 enzymatic systems. Twenty-three gene loci of 13 enzymatic systems (AAT, ACP, ADH, GDH, G6PDH, GPI, IDH, LDH, MDH, MEP, PGM, PER, and SOD) were analyzed by starch gel electrophoresis (Penetrose-30). The EST system was analyzed by polyacrylamide gel electrophoresis, and one polymorphic locus was found (EST-1). Twenty-four loci were detected. The proportion of polymorphic loci was 37.5% in the lagoon and 33.3% in the river. Significant differences in allele frequencies of five loci were found between specimens from the two environments. Expected mean heterozygosity (H _e = 0.14) is the same in the river and lagoon, however, Nei's genetic distance (D) between the population of the two locations was 0.049.     

棉铃虫地理种群的等位酶变异

利用聚丙烯酰胺梯度凝胶电泳检测了棉铃虫Helicoverpa armigera的13种等位酶：α-磷酸甘油脱氢酶(α-GPDH)、酸性磷酸酯酶(ACPH)、碱性磷酸酯酶(ALP)、醛氧化酶(AO)、酯酶(EST)、谷氨酸草酰乙酸转氨酶(GOT)、己糖激酶(HEX)、亮氨酸氨肽酶(LAP)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、苹果酸酶(ME)、磷酸葡萄糖变位酶(PGM)和黄嘌呤脱氢酶(XDH),染色采用双染法。对其中9种等位酶的遗传变异进行了分析,包括13个位点,6个位点表现出多态性,7个位点是单态的,其中多态性位点比例为46.15%。AO、GOT、LAP、LDH、ME和XDH计算出棉铃虫的平均杂合度为0.1160,南京、成都、武穴、衡阳和哈密5个种群的平均遗传距离为0.0008～0.0293,平均遗传相似度为0.9707～0.992。棉铃虫种群内存在很高的遗传多态性,而已测定的种群间遗传分化程度较小,种群间没有基因交流的障碍。迁飞阻碍了不同地理种群间的遗传分化。     

Comparison of Populations of Pratylenchus brachyurus Based on Isozyme Phenotypes

Enzymes from females of five Pratylenchus brachyurus populations and one P. scribneri population were analyzed by isoelectric focusing electrophoresis. Of the 18 enzyme systems investigated, only malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were detected from all five P. brachyurus populations and P. scribneri. Faint bands were detected for isocitrate dehydrogenase and phosphogluconate dehydrogenase from one P. brachyurus population. Three distinct phenotypic groups were found in the MDH and PGM systems for P. brachyurus populations, but only a single electromorph was detected for PGI. Multiple electromorphs for MDH, PGM, and PGI were detected for P. scribneri; there was no similarity among these patterns with those from P. brachyurus. No phenotypic differences in PGI were observed between females and mixed juveniles of one population of P. brachyurus.     

Allelic frequency variations at the MDH-1 locus within Schistosoma mansoni strains from Guadeloupe (French West Indies): ecological interpretation

1. An isoenzymatic study on Schistosoma mansoni from Guadeloupe has been carried out using isoelectric focusing in polyacrylamide gels. 2. Among the seven systems examined (LDH, MDH, G6PD, PGI, PGM, AcP, HK), only MDH showed variation at the MDH-1 locus and mdh-1a allele frequencies were used to characterise eight strains derived either from human or murine hosts, and representative of different transmission sites. 3. In comparison with criteria previously employed to type these strains (ecological context of their transmission sites, cercarial emergence patterns), mdh-1a frequency variations have been correlated to the degree of participation of the murine host reservoir in the parasite transmission dynamics within the different foci of Guadeloupe.     

Isozyme expression in F1 hybrids between carp and goldfish

Interspecific genetic differences in malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase (EST) isozymes in carp (Cyprinus carpio) and goldfish (Carassius auratus) were used to examine the allelic expressions in the hybrid between these species. A unique liver SOD and muscle LDH phenotype unambiguously identifies all presumed hybrid individuals. There was no evidence of F₂ or backcross phenotypes in hybrid individuals. Liver MDH and EST phenotypes in hybrids show a preferential expression of goldfish isozymes. Variation in the levels of carp liver MDH isozymes may result from the polymorphism of a regulatory mutation affecting isozyme expression, leading to gene silencing after duplication.This work was supported through NSERC (Canada) grants to James P. Bogart and John F. Leatherland.     

Interspecific hybridization between human and mouse somatic cells: Enzyme and linkage studies

Human-mouse somatic cell hybrids have been isolated and examined for enzyme and chromosome constitution. The enzymes assayed were lactate dehydrogenase (LDH), isocitrate dehydrogenase (IDH), NADP-dependent malate dehydrogenase (MDH), glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), phosphoglucomutase (PGM), and several esterases. Coexpression of mouse and human genomes and formation of heteropolymeric enzymes were observed in seven different hybrid populations for the enzymes LDH, IDH, MDH, and G6PD. Evidence predicated on the absence of chromosomal rearrangements is provided for the lack of genetic linkage in the human genome for these four enzymes, as well as for thymidine kinase.Supported in part by NIH Grants GM 09966 and 1-F1-GM-39,399 from the Institute of General Medical Sciences and by NIH Training Grant HD-32.     

Comparative study of red cell enzymes in Mirpur (a Portuguese-descendant community) and a neighbouring Hindu population of Gopalchak

Five enzyme systems (ACP1, PGM1, MDH, LDH and EsD) have been studied in Mirpur and Gopalchak. The allele PGM13 is present in polymorphic frequency in Mirpur. In respect of ACP1 and PGM1 loci, these two populations differ significantly.     

Temporal genetic variation of brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis> L. from Vorobiev creek (White sea) based on allozyme analysis

We performed an analysis of allozyme variation in brown trout from Vorobiev creek. Seventeen allozyme loci encoding glycerol-3-phosphate dehydrogenase (G3PDH), aspartate aminotransferase (AAT), malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase D (EST-D) were studied. We found statistically significant differences in allele frequencies for the AAT-1,2*, G3PDH-2,3*, LDH-5*, and MDH-2* loci between brown trout samples collected in 1981–1982 and/or 1992–1995. We suggest that temporal changes of allele frequencies in brown trout from Vorobiev Creek are associated with gene drift.     

Genetic Variation of Mantled Howler Monkeys (Alouatta palliata) from Costa Rica1

We examined genetic diversity of howler monkeys (Alouatta palliata) from Costa Rica. Blood samples of howler monkeys were collected at various locations in Costa Rica, and electrophoresis of total plasma proteins yielded no variation. We also conducted starch gel electrophoresis of red cell isozymes and did not find variation for any of the 14 loci analyzed (i.e., ACP, ADA, CA2, EST, GPI, IDH, LDH‐1, LDH‐2, MDH, PGD, PGM‐1, PGM‐2, SOD, and TPI). These findings were compared with the levels of genetic variation for A. seniculus and A. belzebul from one Brazilian population. Four of the 14 isozymes (ADA, GPI, PGD, and SOD) showed more than one allele for these species. Both A. seniculus and A. belzebul from Brazil showed similar levels of genetic variation. The potential causes of the low genetic variation in A. palliata from Costa Rica are discussed.     

Complex chromosome homologies between the rhesus monkey (Macaca mulatta) and man

The chromosome localization and gene synteny of soluble malate dehydrogenase (MDH1), soluble isocitrate dehydrogenase (IDH1), mitochondrial superoxide dismutase (SOD2), phosphoglucomutase-3 (PGM3), mitochondrial malate dehydrogenase (MDH2), beta-glucuronidase (GUSB), nucleoside phosphorylase (NP), pyruvate kinase M2 (PKM2), hexosaminidase A (HEXA), inosine triphosphatase (ITPA), and N-acetyl-alpha-D-galactosaminidase (NAGA) were determined in the rhesus monkey using somatic cell hybrids. Comparison with the human and Pongidae syntenic groups shows that chromosome banding homologies do not always correlate with gene mapping data.     

Evidence for two additional common alleles at the PGM1 locus (Phosphoglucomutase-E.C.:2.7.5.1)

Summary Lysates of erythrocytes, leukocytes, lymphocytes, and extracts of sperms were investigated for the PGM₁ isozymes by three techniques: starch gel electrophoresis, high voltage thin-layer agarose gel electrophoresis, and thinlayer isoelectric focusing on polyacrylamide gel. On starch, only the well known common phenotypes 1, 2-1, and 2 were demonstrable. On agarose, different distances of the two main cathodal bands (a, b) among the phenotypes 2-1 were noted. Furthermore, on agarose, some types considered as homozygous on starch gel had a single, sharp banded pattern, while others were broad and blurred. Optimal separation was achieved by isoelectric focusing on polyacrylamide gel. In 291 leukolysates, 10 different phenotypes were identified. These are considered as gene products of 4 different common alleles at the PGM₁ locus as suggested by preliminary family investigations. In a random population from Hessen these four alleles, had the following frequencies: PGM ₁ ^a1 0.6186, PGM ₁ ^a2 0.1718, PGM ₁ ^a3 0.1426, and PGM ₁ ^a4 0.067. The preliminary designation a1, a2, a3 and a4 was chosen as the assumed polymorphism was demonstrated on acrylamide and agarose. The sum of the frequencies PGM ₁ ^a1 and PGM ₁ ^a3 (the gene products of which have apparently the same electrophoretic mobility on starch) is similar to the frequency of the old PGM ₁ ¹ allele (0.757) in Caucasoids, PGM ₁ ^a2 and PGM ₁ ^a4 have a frequency of 0.2388 corresponding with the frequency of the old allele PGM ₁ ² .     

Null alleles at two lactate dehydrogenase loci in rainbow trout are associated with decreased developmental stability

Previous studies with rainbow trout (Oncorhynchus mykiss) have shown that allozymic heterozygotes have increased developmental stability, as measured by reduced fluctuating bilateral asymmetry. In this paper, we examine the phenotypic effects of null alleles at two lactate dehydrogenase (LDH) loci. If the association between allozymic heterozygosity and developmental stability is due largely to linked chromosomal segments, then we would expect null allele heterozygotes to have increased developmental stability. In contrast, heterozygotes for LDH null alleles in three populations have reduced developmental stability. This suggests that the reduction in enzyme activity at these loci is having a deleterious effect on development that is strong enough to mask any beneficial effects that may be associated with heterozygosity for these chromosomal segments. The LDH loci examined in this study are members of two different paralogous pairs of duplicate genes produced by the polyploidization of the ancestral salmonid genome. The apparent deleterious effects of these null alleles in heterozygotes could retard the possible loss of duplicate gene expression.     

Biochemical polymorphism of starry flounder, Platichthys stellatus, from the northwestern and northeastern Pacific Ocean.

Three enzyme systems, MDH, PGM and AGPDH, were investigated in starry flounder collected from off northwestern Hokkaido Island, Japan; western Kamchatka; USSR; Puget Sound, Washington, USA; and Yaquina Bay, Oregon, USA. Significant differences in MDH allele distribution were found between the Asian-Puget Sound and Yaquina Bay-Puget Sound samples suggesting that this system may be useful to separate different population units. The allele frequencies of MDH and PGM did not show significant variation with body length and handedness in starry flounder collected from Puget Sound, Washington, and did not support the hypothesis of differential viability between dextral and sinistrial forms of starry flounder.     

Origin of clonal diversity in triploid parthenogenetic Trichoniscus pusillus pusillus (Isopoda,Crustacea) based upon allozyme and nucleotide sequence data

Variation in PGM (phosphoglucomutase) and MDH (malate dehydrogenase) allozymes and in mitochondrial and nuclear (ribosomal) DNA gives evidence of at least three independent origins of triploid Trichoniscus pusillus pusillus. Much of the genetic variation found may reflect variation within the parental diploid population(s), but it is argued that some of the variation in PGM allozymes have accumulated within the parthenogenetic lines. Based upon the variation at this locus, 15 genetically distinct clones are distinguished.     

On the mechanism of action of tetracycline antibiotics

It was found that low oxytetracycline (OTC) concentrations inhibited malic dehydrogenase (MDH) and lactic dehydrogenase (LDH) inStaphylococcus aureus andEscherichia coli (1–5 μg/ml for MDH and 10 μg/ml for LDH). Inhibition of these enzymes occurred almost instantaneously and could be demonstrated after 3–4 minutes. No MDH activity was found in OTC-resistant variants of these microorganisms, but LDH activity was not lowered. The inhibitory effect of OTC is specific for bacterial MDH and LDH. The same enzymes of mammalian origin are not inhibitedin vitro even by high OTC concentrations (100 μg/ml).     

Biochemical polymorphism of starry flounder, Platichthys stellatus, from the northwestern and northeastern Pacific Ocean

Three enzyme systems, MDH, PGM and AGPDH, were investigated in starry flounder collected from off northwestern Hokkaido Island, Japan; western Kamchatka; USSR; Puget Sound, Washington, USA; and Yaquina Bay, Oregon, USA. Significant differences in MDH allele distribution were found between the Asian-Puget Sound and Yaquina Bay-Puget Sound samples suggesting that this system may be useful to separate different population units.
The allele frequencies of MDH and PGM did not show significant variation with body length and handedness in starry flounder collected from Puget Sound, Washington, and did not support the hypothesis of differential viability between dextral and sinistrial forms of starry flounder.     

Allozyme polymorphism of Mdh and α-Gpdh in Ixodes ricinus populations: comparison of borreliae-infected and uninfected ticks

Ixodes ricinus Linnaeus (Acari: Ixodidae) ticks are vectors of numerous infectious diseases in humans and animals. The allozyme variability of MDH and α-Gpdh was detected by native polyacrylamide gel electrophoresis in I. ricinus natural populations in three localities in Serbia. Four alleles of Mdh locus (MDH 1, MDH 2, MDH 3 and MDH X) and four alleles of α-Gpdh locus (VS, S, F and VF) were detected. Interpopulation differences in Mdh and α-Gpdh allele frequencies were statistically insignificant. Significant difference in α-Gpdh allele frequencies between males and females was recorded in the largest sample only. Differences in allele frequencies, detected between borreliae-infected and uninfected I. ricinus ticks, were close to the level of statistical significance, especially for α-Gpdh locus. Clear significant difference appeared in females when sexes were tested separatelly (P = 0.037). It is interesting that genotypes containing rarer alleles (MDH 1 and S) were infected in higher proportion in comparison to other genotypes. Our results point towards a possible role of Mdh and α-Gpdh loci in I. ricinus ticks in the determination of energy requirements for host seeking. Sex differences in α-Gpdh allele frequencies suggest that selective pressure, concerning efficiency of reserve materials utilisation, points to α-Gpdh rather than to Mdh locus.     

家蝇幼虫同工酶研究

本通过聚丙烯酰凝胶电泳法对家蝇幼虫体内的过氧化物酶、乳酸脱氢酶、苹果酸脱氢酶、酯酶和超氧化物歧化酶,以及对超氧化物歧化酶的特性进行了研究。实验结果为家蝇幼虫资源开发利用提供了理论依据。