Some electrophysiological consequences of electrogenic sodium and potassium transport in cardiac muscle: a theoretical study

A previous paper described a kinetic model for electrogenic sodium-potassium transport in cardiac muscle, combining a thermodynamically-constrained transport model with simple passive permeabilities for sodium and potassium to generate a cardiac action potential (Chapman, Kootsey & Johnson, 1979). The present paper explores the extent to which this simplest of active-passive transport models can account (without further modification) for the electrophysiological behavior of cardiac muscle. The long term (several minutes) changes in the duration of the action potential observed following a change in stimulation rate are predicted by the model through a shift in the steady-state current-voltage relationship caused by small changes in inside ion concentrations. The diastolic hyperpolarization observed following an increase in rate is also predicted, including the linear relationship between the maximum diastolic depolarization and the rate of stimulation. Varying the outside potassium concentration in the model produces changes in the rest potential and current-voltage relationship similar to published data. Deviations from ideal potassium electrode behavior occur at both high and low concentrations because of effects on the pump. The model not only predicts the observed shift of the current-voltage curve in the depolarizing direction with increasing [K⁺]₀, but also the crossing of the curve in normal [K ⁺]₀ without having to assume a variation in g_K. Anoxia was introduced into the model by changing the concentrations of ATP and ADP, thereby enabling the model to account for the rapid diastolic depolarization observed in myocardial ischemia.     

Virion nucleic acid of Ebola virus.

The virion nucleic acid of Ebola virus consists of a single-stranded RNA with a molecular weight of approximately 4.0 x 10(6). The virion RNA did not bind to oligodeoxythymidylic acid-cellulose under conditions known to bind RNAs rich in polyadenylic acid and was not infectious under conditions which yielded infectious RNA from Sindbis virus, suggesting that Ebola virus virion nucleic acid is a negative-stranded RNA.     

Mutations within an intron and its flanking sites: Patterns of novel polypeptides generated by mutants in one segment of the cob-box region of yeast mitochondrial DNA

Summary We have undertaken a systematic examination of the polypeptides accumulating in thirteen (out of 23) mutants in the intron cluster box7 and its flanking clusters box2 and box9 of the cob-box (cytochrome b) region of the mitochondrial genome of Saccharomyces cerevisiae. We have subjected these polypeptides to fingerprint analysis, both sequential and in parallel, with two proteases in order to disclose sequence homologies and differences between the different novel polypeptides themselves, and between them and the wild type product of the gene, i.e. apocytochrome b. One of our aims has been to establish the existence of possible correlations between the nature of the novel polypeptides and the fine structure genetic map of that segment of the mitochondrial genome.Our results show that all box7 mutants accumulate the following set of polypeptides not seen in wild type cells: a) a characteristic set of large polypeptides consisting of three species: p56, p42 and p35 or p34.5; b) a polypeptide p23; c) a much shorter fragment (of which the apparent molecular weight varies from 12.5 to 13, according to the mutation) with the exception of two mutants; d) in addition, the majority accumulate in varying amounts a polypeptide p30 closely related to but not identical with apocytochrome b.Moreover only two box7 mutants accumulate a polypeptide in the range of mobilities corresponding to 25–27 Kd (referred to as class p26) while such a polypeptide is seen in all box9 and box2 mutants examined with one exception in box2.Only one mutant in box2 resembles box7 mutants with respect to criterion a), and no box2 or box9 mutants resemble box7 mutants with respect to criterion c); criteria b) and d) appear to apply equally well to mutants in all three clusters.Fingerprint analysis, carried out with polypeptides p56, p42, p35, p34.5, p30, p26, p23, discloses that a) The polypeptides belonging to the same class of mobility exhibit very similar if not identical sequences in various cases. b) These polypeptide classes, except p56, p42 and p26, share considerable sequence homologies with wild type apocytochrome b, perhaps encompassing 50% or more of the wild type sequences. b) Polypeptides belonging to the classes p42 and p26 exhibit less extensive but nevertheless significant homologies with the wild type sequence. c) Sequences in polypeptides belonging to class p56 are virtually indistinguishable from ones in cytochrome oxidase subunit I.The inferences from these findings are 1) one gene can produce a multiplicity of polypeptide products that share a common sequence at the promoter-proximal (N-terminal) portion, but diverge beyond these regions of homology. 2) Both the multiplicity of products in single mutants, and the protein structure found, argue against the divergent segments to be due to frameshift terminations, and instead suggest that the novel products are consequences of mRNA processing defects (excision and/or ligation) at and near intron regions. 3) Mutations at edges and the center of an intron can generate similar polypeptide patterns, i.e. produce analoguous mRNA processing defects. 4) Mutations in exons, at their boundary with introns, can produce polypeptide patterns indistinguishable from those at the neighbouring intron; they diverge and eventually become typically exonlike in mutants localized at increasing distances from the boundary. 5) Taken together these findings argue that pre-mRNA processing extends beyond the boundaries of the intron proper and that certain exonsequences participate in excision and ligation. 6) Accumulated pre-mRNAs, resulting from defects in splicing can be translated. 7) Product p56 is formally analogous to p23, as a faulty but highly conserved partial product of the wild type protein, the former of Cox I (oxi3 gene), the latter of the cob-box gene proper. Therefore both genes may utilize identical RNA processing elements which are affected by box7 mutations. 8) A small amount of product similar to p56 may accumulate even in some wild types but not in others. This observations suggests that in certain nuclear backgrounds RNA processing may be more error-prone than in others.Publication No. XXXX from the Department of Chemistry, Indiana UniversitySupported by Research Grant GM 12228 from the National Institute of General Medical Sciences, National Institutes of Health; recipient of Research Career Award K06 05060 from the same Institute.Supported by Délégation Générale à la Recherche Scientifique et Technique grant n⁰ 78-70341     

Embryo-uterine interaction in implantation

Pregnant donor (day 3) and non-pregnant recipient rats were hypophysectomized and injected daily for 6 days with 2 mg of progesterone. A single dose of 20 ng of estradiol-17β in saline was administered via a tail vein to either the donor, the recipient, or to both animals; blastocysts were transferred 60 to 90 minutes after the latter injection. Twenty-four hours later uterine implantation sites were delineated by injection of Chicago Blue-B dye. The results indicate that both the blastocyst and the uterus must be exposed to estrogen to obtain normal implantation rates. While 43.2% of the embryos implanted when both the donor and the recipient received estrogen, only 6.3% implanted when only the recipient was injected with estrogen. No implantations were found in animals in which only the embryos had been exposed to estrogen, suggesting that if this steroid was synthesized by the embryo it was insufficient to induce implantation in the rat.     

Affinity labeling of eukaryotic elongation factors using N epsilon-bromoacetyl-Lys-tRNA.

eEF-T and eEF-Tu from rabbit reticulocyte and from Artemia were affinity labeled using N epsilon-bromoacetyl-Lys-tRNA prepared with either yeast or E. coli tRNA. Only the eEF-Tu polypeptide was crosslinked when eEF-T was incubated with the reactive aminoacyl-tRNA analogue, which indicates that at least part of the aminoacyl-tRNA binding site is the same in both eEF-Tu and the multisubunit eEF-T. Complex formation (eEF-Tu x aa-tRNA x GTP) was required for crosslinking, since no covalent reaction with eEF-Tu occurred in the absence of GTP. The yield of crosslinked product was greatly reduced by adding either unmodified rabbit liver aminoacyl-tRNA or unmodified E. coli Lys-tRNA to the incubation to compete for the aminoacyl-tRNA binding site on eEF-T or eEF-Tu, indicating that the covalent reaction occurs while the N epsilon-bromoacetyl-Lys-tRNA is bound in this site. The affinity labeling of a prokaryotic and two different eukaryotic elongation factors by the same reagent suggests that there may be conservation of structure in the region of the proteins which binds the aminoacyl end of the aminoacyl-tRNA.     

Effects of α-methylphenylalanine plus phenylalanine treatment during development on myelin in rat brain

The effects of hyperphenylalaninemia induced by treatment with -methylphenylalanine (MPA) plus phenylalanine (PHE) on body and brain weight, on myelin and synaptosome formation, and on the lipids and fatty acids of myelin were studied in rats. The administration of MPA (2.4 mol/g body wt) plus PHE (2.6 mol/g body wt) for 25 and 35 days beginning on the fifth postnatal day did not affect brain development. On doubling the dosage of PHE, body and brain weights and myelin yields were significantly lowered. The lipid composition of myelin from the brains of treated animals was largely unaffected; however, the concentration of sulfatides was significantly reduced. Unsaturated fatty acid levels in myelin from hyperphenylalaninemic rat brains were reduced while long-chain fatty acids were unaffected. We conclude that as in hyperphenylalaninemia induced by other methods, MPA+PHE treatment impairs body and brain growth, reduces myelin formation, and causes inhibition of fatty acid desaturation in the brain.     

The site of VX2 tumor transplantation affects the development of hypercalcemia in rabbits

The relationship between plasma levels of 15-keto-13, 14-dihydro-prostaglandin E₂ (15K-H₂-PGE₂) and serum calcium levels was studied in nontumor-bearing rabbits and in rabbits bearing the VX₂ carcinoma intramuscularly and intra-abdominally. The plasma levels of 15K-H₂-PGE₂ in the two groups of tumor-bearing animals did not vary significantly but was several fold greater than in nontumor-bearing rabbits. Rabbits bearing the VX₂ carcinoma intramuscularly developed hypercalcemia between the second and third week after implantation of neoplastic tissue and remained hypercalcemic until they expired. The serum calcium levels in rabbits bearing the VX₂ carcinoma intra-abdominally did not vary significantly from those of nontumor-bearing rabbits. The differences in the serum calcium levels in rabbits bearing the VX₂ carcinoma at intramuscular and intra-abdominal implant sites may be related to different extents of metabolism by the lung and by the liver of prostaglandin E₂ or other cyclooxygense products of polyenoic fatty acids produced by the tumor.