RFLP mapping in barely of a dominant gene conferring resistance to scald (Rhynchosporium secalis)

A progeny consisting of 52 anther-derived doubled haploid barley lines from a F₁ between the winter cultivars Igri (susceptible) and Triton (resistant) was tested for resistance to Rhynchosporium secalis. A dominant gene was detected and tagged by a series of cosegregating RFLP markers located in the proximal portion of the long arm of chromosome 3, close to the centromere. One of the cosegregating RFLP markers, cMWG680, was converted into a codominant sequence tagged site marker. Polymerase chain reaction analysis with this marker of a series of accessions carrying known resistance genes provided evidence that scald resistance in cv Triton is due to the presence of the Rh gene.     

New sources of dwarfing genes in pearl millet (Pennisetum americanum)

Summary Thirteen naturally occurring dwarf lines of pearl millet [Pennisetum americanum (L.) Leeke], identified from the world collection, varied for several morphological and agronomic characters. Extreme dwarfs were characterized by a tufted growth habit which could be distinguished from the time of germination, while the other dwarf lines could be distinguished only after anthesis. The F₁ hybrids between the tall and dwarf genotypes were tall, indicating that dwarfness is a recessive trait. In 10 out of the 13 crosses, the F₂ segregation ratio was three tall to one dwarf (31) suggesting that the dwarfness is controlled by a single recessive gene, while the height differences in 3 of the dwarfs (IP 8056, IP 8210 and IP 8214) were controlled by more than one gene as they showed continuous variation for plant height in F₂. When the remaining 10 single gene dwarfs were crossed to either d ₁ (Tift 238) or d ₂ (Tift 23 DB) dwarfs, only 2 crosses produced tall F₂ hybrids and they segregated for height in F₂ indicating that these 2 dwarfs are non-allelic to d ₁ and d ₂. Reciprocal crosses of these 2 dwarfs produced tall F₁ hybrids and showed a dihybrid segregation of 934 in F₂ indicating that the dwarfing genes of these 2 parents are non-allelic to each other. These non-allelic dwarfs were assigned the gene symbols d ₃ (IP 10401), and d ₄ (IP 10402).Submitted as J.A. No. 429 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Genetic control of Endosperm Balance Number (EBN): three additive loci in a threshold-like system

Summary The genetic control of Endosperm Balance Number (EBN) was investigated by a complete diallel of four exceptional diploid Solanum commersonii-S. chacoense hybrids (1 1/2 EBN) and backcrosses to their species parents, S. commersonii (1 EBN) and S. chacoense (2 EBN). Crosses in which the female parent had a higher EBN value than the male, S. chacoense (2 EBN)XF₁ (11/2 EBN) and F₁ (11/2 EBN)X S. commersonii (1 EBN), produced viable seed to aborted seed ratios of 11.1 and 11.3, respectively, and had average to small sized viable seed. Crosses in which the female parent had a lower EBN value than the male, S. commersonii (1 EBN)XF₁ (11/2 EBN) and F₁ (11/2 EBN)XS. chacoense (2 EBN), produced viable seed to aborted seed ratios of 1 7.9 and 1 6.7, respectively, and had average to large sized viable seeds. The results of these crosses appear to be consistent with the relative EBN values of the male and female parent. A model is proposed for the system regulating endosperm development. The assumptions of this model are: (1) three unlinked loci control the system; (2) the loci are homozygous within a species; (3) the genes have additive effects and are of equal strength within a species; (4) the genes within S. chacoense have twice the effect with respect to endosperm regulation as those within S. commersonii; and (5) a slight excess maternal dosage will produce the qualitative effect of small but viable seed. This model, in which quantitative genes operate in a dosage dependent system bears many similarities to classical, threshold-type genetic models.     

Polymorphism and stability in the histone gene cluster of Drosophila melanogaster

Histone genes in Drosophila melanogaster are organized into repeats of 4.8 and 5.0 kb (Lifton et al., 1978). We find these repeat sizes in every one of the more than 20 Drosophila strains we have examined. Strains differ in the relative amounts of the two repeat types, with ratios varying from 11 to 14, the 5.0 kb repeat always present in equal or greater amounts than the 4.8 kb repeat. Restriction enzyme digestion and blotting analysis reveals that the strains also differ in a number of far less abundant fragments containing histone DNA sequences. In the Amherst and Samarkand strains, there are, in addition, many copies of 4.0 and 5.5 kb repeat-like fragments respectively. A series of stocks were made isogenic for single second chromosomes from the Amherst strain. The hybridization patterns of the histone DNA from these stocks containing different Amherst chromosomes are very similar but a number of differences in the minor fragments were seen. The stability of the histone locus restriction pattern was tested by following the DNA derived from a single second chromosome of the b Adhn² pr cn strain over a two year period. The restriction pattern of major and minor bands remained identical. Finally, histone loci distinguishable by their restriction pattern on blots were recombined with visible markers. These chromosomes will be useful in tracing the fate of specific histone loci during genetic manipulations.     

Ribosomal gene variation in soybean (Glycine) and its relatives

Summary The genes encoding the 18S25S ribosomal RNA gene repeat in soybean (Glycine max) and its relatives in the genus Glycine are surveyed for variation in repeat length and restriction enzyme site locations. Within the wild species of subgenus Glycine, considerable differences in repeat size occur, with a maximum observed in G. falcata. Repeat length and site polymorphisms occur in several species, but within individual plants only single repeat types are observed. The rDNA of the cultivated soybean and its wild progenitor, G. soja are identical at the level of this study, and no variation is found in over 40 accessions of the two species. Data from rDNA mapping studies are congruent with those of previous biosystematic studies, and in some instances give evidence of divergences not seen with other approaches.     

The glucose kinase gene of Streptomyces coelicolor and its use in selecting spontaneous deletions for desired regions of the genome

Summary A number of deletions in the glucose kinase (glk) region of the Streptomyces coelicolor chromosome were found among spontaneous glk mutants. The deletions were identified by probing Southern blots of chromosomal DNA from glk mutants with cloned glk DNA. The deletions ranged in size from 0.3 kb to greater than 2.9 kb. When cloned glk DNA was introduced on a C31 phage vector into a glk mutant that contained a deletion of the entire homolgous chromosomal glk region, glucose kinase activity was detected in extracts of these cells. The entire coding information for at least a subunit of glucose kinase is there-fore present on the cloned glk DNA. The 0.3 kb glk chromosomal deletion was used to demonstrate that transfer of chromosomal glk mutations on the the C31::glk phage could occur by recombination in vivo. Since glk mutations frequently arise from deletion events, a method was devised for inserting the cloned glk DNA at sites in the chromosome for which cloned DNA is available, and thus facilitating the isolation of deletions in those DNA regions. C31::glk vectors containing a deletion of the phage att site cannot lysogenize S. coelicolor recipients containing a deletion of the glk chromosomal gene unless these phages contain S. coelicolor chromosomal DNA. In such lysogens, the glk gene becomes integrated into the chromosome by homologous recombination directed by the chromosomal insert on the phage DNA. In appropriate selective conditions, mutants which contain deletions of the glk gene that extend into the adjacent host DNA can be easily isolated. This method was used to insert glk into the methylenomycin biosynthetic genes, and isolate derivatives with deletions of host DNA from within the prophage into the adjacent host DNA. Phenotypic and Southern blot analysis of the deletions showed that there are no genes essential for methylenomycin biosynthesis for at least 13 kb to the left of a region concerned with negative regulation of methylenomycin biosynthesis. Many of the deletions also removed part of the C31 prophage.     

Genetic studies of self-fertility in rye (Secale cereale L.). 1. The identification of genotypes of self-fertile lines for the Sf alleles of self-incompatibility genes

Segregation for self-fertility has been studied in progenies from the crosses of self-sterile (SS) plants with interline hybrids obtained by a diallel scheme of pollinations between seven self-fertile (SF) lines (nos. 2–8) and with F₁ (SS plant x SF line) hybrids. All the offspring families from the SS plant x F₁ (SS plant x SF line) crosses demonstrated a 1SF1SS segregation. The crosses of SS plants with some interline hybrids gave only self-fertile plants, whereas the crosses with other interline hybrids gave a segregation of 3SF:1SS expected in the case of digenic segregation. The data obtained permitted us to identify three different S loci (S1, S2, S5) and to estimate the genotypes of self-fertile lines for their Sf alleles: lines 5, 6, 7 and 8 are S1f/S1f S2n/S2n S5m/S5m, line 4 is S1n/S1n S2f/S2f S5m/S5m, and lines 2 and 3 are S1n/S1n S2m/S2m S5f/S5f(Sn, Sm designate active alleles of the incompatibility genes). The identification of the particular S gene which is presented by the Sf allele in each line has been made on the basis of our data concerning the linkage of the Sf mutation with isozyme markers of particular rye chromosomes, which is reported in an accompanying paper.     

Inheritance of high oleic acid content in the seed oil of soybean mutant M23

A mutant line, M23, of soybean [Glycine max (L.) Merr.] was found to have two fold increases in oleic acid content in the seed oil compared with the original variety, Bay. Our objective was to determine the inheritance of the high oleic acid content in this mutant. Reciprocal crosses were made between M23 and Bay. There were no maternal and cytoplasmic effects for oleic acid content. The F₁ seeds and F₁ plants were significantly different from either parents or the midparent value, indicating partial dominance of oleic acid content in these crosses. The oleic acid content segregated in the F₂ seeds and F₂ plants in a trimodal pattern with normal, intermediate and high classes, satisfactorily fitting a 121 ratio. The seeds of a backcross between M23 and F₁ segregated into intermediate and high classes in a ratio of 11. These results indicated that oleic acid content was controlled by two alleles at a single locus with a partial dominant effect. Thus, the allele in M23 was designated ol and the genotypes of M23 and Bay were determined to be olol and 0l0l, respectively. The oleic acid contents of the F₂ seeds and F₂ plants were inversely related with the linoleic acid content which segregated in a trimodal pattern with normal, intermediate and low classes in a 121 ratio. Thus, it was assumed that the low linoleic acid content in M23 was also controlled by the ol alleles. Because a diet with high oleic acid content reduces the content of low density lipoprotein cholesterol in blood plasma, the mutant allele, ol, would be useful in improving soybean cultivars for high oleic acid content.     

Inheritance of heat-stable resistance to Meloidogyne incognita in Lycopersicon peruvianum and its relationship to the Mi gene

Summary The inheritance of heat-stable resistance to the root-knot nematode, Meloidogyne incognita (Kofoid and White) Chitwood, was studied in crosses between different accessions and clones of Lycopersicon peruvianum L. F₁, F₂ and BC₁ generations were evaluated for their index of resistance based on numbers of eggs and infective second-stage juveniles (J₂) per gram of root, and the segregation ratios were determined in experiments carried out at constant soil temperatures of 25 °C and 30 °C. L. peruvianum P.I. 270435 clones 3 MH and 2R2 and P.I. 126443 clone 1 MH, all heatstable resistant, were crossed with L. peruvianum P.I. 126440 clone 9 MH, which is susceptible at both 25 °C and 30 °C. All F₁ progeny were resistant at 25 °C and 30 °C; F₂ and BC₁ generations at 25 °C gave resistant: susceptible (RS) ratios of 151 and 31, respectively, which suggests that resistance is conditioned by two independently assorting genes. However, at 30 °C, RS ratios of 31 and 11 were observed for the F₂ and BC₁ generations, respectively. These results indicate that heat-stable resistance is conferred by a single dominant gene expressed at 30 °C, while the second resistance gene is heat unstable and not expressed at 30 °C. P.I. 270435 clones 2R2 and 3 MH and P.I. 126443 clone 1 MH were crossed with P.I. 128657 clone 3 R4 (source of gene Mi), which is resistant at 25 °C but susceptible at 30 °C. All of the F₁ progeny were resistant at 25 °C and 30 °C.TC₁ progeny of 270435-2 R2 x 128657-3 R4, 270435-3 MH x 128657-3 R4 and 126443-1 MH x 128657-3 R4 crossed with susceptible 126440-9 MH were all resistant at 25 °C and segregated in a 11 ratio at 30 °C. These results also suggest that the heat-stable resistance is monogenic and that it is non-allelic to gene Mi. The non-segregation of TC₁ progenies at 25 °C, suggests that the heat-unstable resistance factor in L. peruvianum P.I. 270435 clones 2 R2 and 3 MH and in P.I. 126443 clone 1 MH is allelic to or the same as gene Mi. We propose the symbol Mi-2 for the gene in P.I. 270435 that confers heat-stable resistance to M. incognita.     

Evolution of actin gene families of sea urchins

The actin gene family of the sea urchin Lytechinus pictus includes a single muscle actin gene, LpM, and four cytoskeletal actin genes: LpC1, LpC2, LpC3, and LpC4. The origin and relationship of these actin genes to members of the actin gene family of the sea urchin Strongylocentrotus purpuratus were considered. Comparison of deduced amino acid sequences suggested a close relationship between LpC1 and the CyI–CyII subfamily of S. purpuratus actin genes, and between LpC2 and the CyIII subfamily of S. purpuratus actin genes; the muscle actin genes were orthologous. It is proposed that two divergent cytoskeletal actin genes of the common ancestral sea urchin gave rise by duplication to the extant cytoskeletal actin genes of these species, some of which have changed 3 noncoding sequences while others have maintained a terminus highly conserved among sea urchin actin genes.Correspondence to: B.P. Brandhorst     

Effect of X-irradiation on the size of the mutated sector in diploid and tetraploid rice

Summary An investigation was carried out of the effect of X-irradiation on the size of the mutated sector in three diploids (A.S.D.8, T.N. 1 and G.E.B. 24) and the autotetraploid of G.E.B 24 of Oryza sativa L., by analyzing the M₂ segregation ratios of chlorophyll mutants. A total of 612 segregating M₁ panicle progenies in the diploids and 284 panicle progenies in the tetraploid was studied in the M₂ generation for the ratio of normal to chlorophyll mutant seedlings. Of the 573 segregating M₁ panicles analysed by chi square test in the diploids, 206 showed good fit with a 31 ratio, while the rest of the progenies (367) deviated significantly in both directions from the expected 31 segregation. This was interpreted as the result of M₁ panicles developing from both single cell and multicellular initials following X-irradiation, the former segregating for 31, the latter significantly deviating from 31. An assessment of fertility (as judged from the number of M₂ seedlings per mutated M₁ panicle), related to the distribution of chi square deviations from the 31 ratio in the diploids, indicated that the M₁ panicle progenies deriving from single cell initials had low fertility but those originating from multicellular initials ranged from low to high fertility. A comparison of the distribution pattern of the M₂ segregation ratios of chlorophyll mutant phenotypes in the diploid and autotetraploid G.E.B. 24 indicated that, in the tetraploid, a larger sector and only one, or fewer cell initials than in the diploid species, are involved.     

Electrogenic 2 Na+/1 H+ exchange in crustanceans

Summary Hepatopancreatic brush border membrane vesicles of the freshwater prawn,Macrobrachium rosenbergii and the marine lobster,Homarus americanus exhibited²²Na uptake which was Cl-independent, amiloride sensitive, and stimulated by a transmembrane H gradient (H _i >H _o ). Sodium influx by vesicles of both species were sigmoidal functions of [Na] _o , yielding Hill coefficients that were not significantly different (P>0.5) than 2.0. Estimations of half-saturation constants (K _Na) were 82.2mm (prawn) and 280.1mm (lobster), suggesting a possible adaptation of this transporter to environmental salinity.Trans-stimulation andcis-inhibition experiments involving variable [H] suggested that the exchangers in both species possessed single internal cation binding sites (pK 6.5–6.7) and two external cation binding sites (prawn, pK 4.0 and 5.7; lobster pK 3.5 and 6.1). Similarcis inhibition studies using amiloride as a competitive inhibitor of Na uptake supported the occurrence of dual external sites (prawn,K _i 50 and 1520 m; lobsterK _i 9 and 340 m). Electrogenic Na/H exchange by vesicles from both crustaceans was demonstrated using equilibrium shift experiments where a transmembrane potential was used as the only driving force for the transport event. Transport stoichiometries of the antiporters were determined using Static Head analysis where driving forces for cation transfer were balanced using a 101 Na gradient, a 1001 H gradient, and a stoichiometry of 2.0. These electrogenic 2 Na/1 H exchangers appear thermodynamically capable of generating sufficient gastric acidification for organismic digestive activities.     

Molecular-marker analysis of seed-weight: genomic locations,gene action,and evidence for orthologous evolution among three legume species

The objectives of this study were to use molecular markers to: (1) identify quantitative trait loci (QTL) controlling seed-weight in soybean, (2) characterize the genetic basis of seed-weight expression, and (3) determine whether soybean shares orthologous seed-weight genes with cowpea and/or mung bean. An F₂ population was developed between a large-seeded Glycine max breeding line and a small-seeded G. soja plant introduction. DNA samples from 150 F₂ individuals were analyzed with 91 polymorphic genetic markers, including RFLPs, RAPDs and SSRs. Seed-weight was analyzed by randomly sampling 100 seeds from each of 150 greenhouse-grown F₂ individuals, and their 150 F₂₃ lines, from a replicated field trial. Markers associated with seed-weight were identified using the computer program MapMaker-QTL and a one-way analysis of variance. Three and five markers were significantly associated with seed-weight variation (P<0.01) in the F₂ and F₂₃ generations, respectively. Tests for digenic epistasis revealed three significant interactions in both generations. In a combined analysis, these markers and interactions explained 50 and 60% of the phenotypic variation for seed-weight in the F₂ and F₂₃ generations, respectively. Comparison of our results in soybean (Glycine) with those previously reported in cowpea and mung bean (Vigna) indicated that soybean and cowpea share an orthologous seed-weight gene. In both species, a genomic region significantly associated with seed-weight spanned the same RFLP markers in the same linkage order. A significant digenic interaction involving this genomic region was conserved in all three species. These results suggest that the exploitation of comparative QTL mapping is an invaluable tool for quantitative geneticists working with poorly characterized plant systems.     

Increase in incidence of chromosome instability and non-conservative recombination between repeats in Saccharomyces cerevisiae hpr1Δ strains

Null hprl strains show a large increase (up to 2000-fold) over wild type in the frequency of occurrence of deletions between direct repeats on three different chromosomes. However, we show that hprl mutations have little or no effect on reciprocal exchange, gene conversion or unequal sister chromatid exchange, as determined using intrachromosomal, interchromosomal and plasmid-chromosome assay systems. A novel intrachromosomal recombination system has allowed us to determine that over 95% of deletions in hpr1 strains do not occur by reciprocal exchange. On the other hand, hpr1 strains show chromosome loss frequencies of up to 100 times the wild-type level. Our results suggest that yeast cells have a very efficient non-conservative recombination mechanism, dependent on RADI and RAD52, that causes deletions between direct DNA repeats, and this mechanism is strongly stimulated in hpr1 strains. The results indicate that the Hpr1 protein is required for stability of DNA repeats and chromosomes. We propose that in the absence of the Hprl protein the cell destabilizes the genome by allowing the initiation of events that lead to deletions of sequences between repeats, and to chromosome instability. We discuss the roles that proteins such as Hprl have in maintaining direct repeats and in preventing non-conservative recombination and consider the importance of these functions for chromosome stability.     

A New Family of “H3L-Like” Histone Genes

The H3L histone variant gene in Paracentrotus lividus (sea urchin) shows almost all typical features of the replication-dependent histone genes, but it codes for the H3.3 histone protein with the S.//. A.IG amino acid motif, which is typical of the variants synthesized in a replication-independent manner. H3L-like histone genes have been found in several unrelated organisms. These genes are intronless and encode for the typical H3.3 histone proteins. The newly described family of H3L-like variants, nearly ubiquitous within the animal kingdom, could represent the common ancestor of H3 and H3.3 histone genes.     

Characterization of a Na∶K∶2Cl cotransport system in the apical membrane of a renal epithelial cell line (LLC-PK1)

Summary ⁸⁶Rb uptake into LLC-PK₁ cells (an established renal epithelial cell line) was found to be comprised of an active ouabain-sensitive component, a loop diuretic-sensitive component which was passive and strictly dependent upon the presence of extracellular Na⁺ and Cl^– for activity, and a leak component. The diuretic-sensitive component of influx was investigated further in apical membrane vesicles derived from these cells. A large fraction of⁸⁶Rb,²²Na and³⁶Cl flux into these vesicles was sensitive to inhibition by furosemide and dependent upon the presence of the other two co-ions, in keeping with the presence of a loop diuretic-sensitive Na⁺K⁺Cl^– cotransport system. The kinetic parameters for Na⁺ and K⁺ interaction have been analyzed under initial linear zerotrans conditions. The following values were obtained:K _mNa+=0.42±0.05 mmol/liter,V _max=303±24 pmol/mg/6 sec;K _mK+=11.9±1.0 mmol/liter,V _maxK+=307±27 pmol/mg/6 sec. For Cl^– interaction evidence for two cooperative binding sites with different affinities and different specificities were obtained. Thus, a stoichiometry of 1Na⁺1K⁺2Cl^– can be calculated. It is concluded that the apical membrane of LLC-PK₁ cells contains a Na⁺K⁺2Cl^– cotransport system with properties similar to those described for the thick ascending limb of the loop of Henle.     

Sid: a Mendelian locus controlling thylakoid membrane disassembly in senescing leaves of Festuca pratensis

Summary A spontaneous mutation arising in Festuca pratensis has the effect of stabilizing the pigmentproteolipid complexes of thylakoid membranes so that leaf tissue does not turn yellow during senescence. Inheritance of the non-yellowing character was analysed in crosses between the wild-type cultivar Rossa and a mutant line Bf 993. Electrophoretic variants of cytoplasmic phosphoglucoisomerase coded by alleles of the nuclear gene Pgi-2 were used to identify hybrids during intercrossing. About 96% of the F₁ progeny were heterozygous and all were phenotypically yellowing. In the F₂ generation yellow green segregated in a ratio of 2.141, not significantly different from 31. In the backcross between F₁ and Bf 993 the ratio was 11 yellow green. There was no indication of linkage to Pgi-2. Senescence of detached Bf 993 and Rossa leaves was compared with that of the F₁ hybrid. The hybrid behaved in an essentially identical fashion to the wildtype parent, and in marked contrast to the mutant, in all aspects of the senescence syndrome investigated, including loss of chlorophyll, carotenoids and the light-harvesting chlorophyll-protein of thylakoid membranes, and elevation of the particulate protein chlorophyll ratio in the terminal stages. It is concluded that there exists in Festuca pratensis a nuclear gene, designated Sid (senescence-induced degradation) which regulates turnover of hydrophobic components of photosynthetic membranes in ageing leaf tissue and which occurs in at least two allelic forms, y (yellow) dominant over g (green).Abbreviations PGI phosphoglucoisomerase - Pgi nuclear gene coding for PGI - TSH tris buffer containing 2-mercaptoethanol - SDS sodium dodecyl sulphate - Chl chlorophyll - LHCP light-harvesting chlorophyll a/b binding protein - ELISA enzyme-linked immunosorbent assay     

Cation permeation at the amphibian motor end-plate

Summary Measurements of acetylcholine-induced single-channel conductance and null potentials at the amphibian motor end-plate in solutions containing Na, K, Li and Cs ions (Gage & Van Helden, 1979;J. Physiol. (London) (in press) were analyzed in terms of three models. Two of these models, the neutral site channel model and the charged site channel model were developed to cater for three cations. Both were shown to be able to explain the dependence of single-channel conductance on membrane potential and gave the following sequences of equilibrium constants and mobilities.K _Li/K _Na/K _K/K _Cs=71.710.9 andu _Cs/u _K/u _Na/u _Li=1.410.580.13 at 8 °C. Similar sequences were obtained at 20 °C. Although the neutral model fitted the data for relative conductances in Li-, Cs-and Na-solutions slightly better than the charged model, experiments done in normal [NaCl] and [NaCl]/2 solutions could only be fitted by the neutral model. In contrast, the third model, the Constant Field Equation, was unable to fit the conductance data in any of the above situations. The data available suggests that permeation is through long neutral channels, lined with high field-strength negative polar groups and including one or possibly more high resistance barriers for anions.     

Kinetic mechanism of Na+, K+, Cl−-cotransport as studied by Rb+ influx into HeLa cells: Effects of extracellular monovalent ions

Summary Ouabain-insensitive, furosemide-sensitive Rb⁺ influx (J _Rb) into HeLa cells was examined as functions of the extracellular Rb⁺, Na⁺ and Cl^– concentrations. Rate equations and kinetic parameters, including the apparent maximumJ _Rb, the apparent values ofK _m for the three ions and the apparentK _i for K⁺, were derived. Results suggested that one unit molecule of this transport system has one Na⁺, one K⁺ and two Cl^– sites with different affinities, one of the Cl^– sites related with binding of Na⁺, and the other with binding of K⁺(Rb⁺). A 11 stoichiometry was demonstrated between ouabain-insensitive, furosemidesensitive influxes of²²Na⁺ and Rb⁺, and a 12 stoichiometry between those of Rb⁺ and³⁶Cl^–. The influx of either one of these ions was inhibited in the absence of any one of the other two ions. Monovalent anions such as nitrate, acetate, thiocyanate and lactate as substitutes for Cl^– inhibited ouabain-insensitive Rb⁺ influx, whereas sulfamate and probably also gluconate did not inhibitJ _Rb. From the present results, a general model and a specialized cotransport model were proposed: 1) In HeLa cells, one Na⁺ and one Cl^– bind concurrently to their sites and then one K⁺ (Rb⁺) and another Cl^– bind concurrently. 2) After completion of ion bindings Na⁺, K⁺(Rb^–) and Cl^– in a ratio of 112 show synchronous transmembrane movements.     

Modification by hypoxia and drug treatment of cerebral ATPase plasticity

The plasticity of synaptosomal non-mitochondrial ATPases was evaluated in cerebral cortex from 3-month-old normoxic rats and rats subjected to either mild or severe intermittent normobaric hypoxia [12 hr daily exposure to N₂O₂ (9010 or 91.58.5) for four weeks]. The activities of Na⁺, K⁺-ATPase, low- and high-affinity Ca²⁺-ATPase, Mg²⁺-ATPase, and Ca²⁺, Mg²⁺-ATPase were assayed in synaptosomes and synaptosomal subfractions, namely synaptosomal plasma membranes and synaptic vesicles. The evaluations were performed after a 4-week treatment with saline (controls) or -adrenergic agents (-yohimbine, clonidine), a vasodilator compound (papaverine), and an oxygen-partial pressure increasing agent (almitrine). These treatments differently changed the adaptation to chronic intermittent hypoxia characterized by a decrease in the activity of Na⁺, K⁺-ATPase, Ca²⁺,Mg²⁺-ATPase, and high-affinity Ca²⁺-ATPase, concomitant with a modification in the activity of Mg²⁺-ATPase supported in a different way by the enzymatic forms located into the synaptosomal plasma membranes and synaptic vesicles.