Biomedical subjects

Noel W Davies

Publications and source records attributed to Noel W Davies.

At least 19 recordsLinked to original sources

PubMed2006

How do soil nutrients affect within-plant patterns of herbivory in seedlings of Eucalyptus nitens?

This study assessed how the palatability of leaves of different age classes (young, intermediate and older) of Eucalyptus nitens seedlings varied with plant nutrient status, based on captive feeding trials with two mammalian herbivores, red-bellied pademelons (Thylogale billardierii), and common brushtail possums (Trichosurus vulpecula). Seedlings were grown under three nutrient treatments (low, medium and high), and we determined how palatability was related to chemical and physical characteristics of the leaves. Pademelons ate more older leaves than young and intermediate leaves for all treatments. This pattern was best explained by sideroxylonals (formylated phloroglucinol compounds known to deter herbivory by other marsupials), and/or essential oil compounds that were present in lower concentrations in older leaves. In the low-nutrient treatment, possums also ate more of the older leaves. However, in the medium- and high-nutrient treatments, possums ate more intermediate leaves than older leaves and showed a behavioural preference for young leaves (consuming younger leaves first) over intermediate and older leaves, in spite of high levels of sideroxylonals and essential oils. The young leaves did, however, have the highest nitrogen concentration of all the leaf age classes. Thus, either sideroxylonals and essential oils provided little or no deterrent to possums, or the deterrent was outweighed by other factors such as high nitrogen. This study indicates that mammalian herbivores show different levels of relative use and damage to leaf age classes at varying levels of plant nutrient status and, therefore, their impact on plant fitness may vary with environment.

Animals↗

PubMed2006

Behavioral responses of a generalist mammalian folivore to the physiological constraints of a chemically defended diet.

Mammalian herbivores, particularly browsers and folivores, encounter and consume a range of plant chemical defenses [plant secondary metabolites (PSMs)] on a regular basis. The physiological regulation of PSM ingestion and the resulting behavioral responses of mammalian herbivores directly affect their feeding decisions and the subsequent foraging strategies that they adopt. Generalist mammalian herbivores are hypothesized to consume a generalized diet because of physiological limitations of their detoxification systems. The consumption of a generalized diet is proposed to enable toxin (PSM) dilution through the use of multiple detoxification pathways. We tested the predictions of the detoxification-limitation hypothesis by offering two chemically different plant species, Eucalyptus regnans and E. globulus, to a generalist mammalian folivore, the common brushtail possum (Trichosurus vulpecula), as single- and mixed-species diets. By feeding more efficiently, brushtail possums benefited more, through increased intake, on the mixed-species diet than on either of the single-species diets. We argue that frequently switching between chemically diverse foliage reduces the physiological constraints imposed by a PSM-rich diet and enables more efficient feeding. The behavioral responses of brushtail possums were consistent with the proposed physiological constraints of a chemically defended diet, offering support for predictions of the detoxification-limitation hypothesis. We suggest that feeding behavior of herbivores may be a useful indicator of the physiological constraints imposed by a chemically defended diet.

Animals↗

PubMed2006

Regiospecificity profiles of storage and membrane lipids from the gill and muscle tissue of Atlantic salmon (Salmo salar L.) grown at elevated temperature.

Regiospecific and traditional analysis, of both storage and membrane lipids, was performed on gill, white muscle, and red muscle samples taken from Atlantic salmon (Salmo salar) to gauge the effect of elevated water temperature. The fish, fed a commercial diet, were held at an elevated water temperature of 19 degrees C. Total n-3 PUFA, total PUFA, and n-3/n-6 and unsaturated/saturated fatty acid (UFA/SFA) ratios in the FA profile of the total lipid extract in the white muscle were fairly low compared with fish grown at 15 degrees C. Adaptation of structural and storage lipids at elevated temperatures was shown by a significant (P < 0.01) reduction in PUFA especially in the percentage of EPA (6-8%). Further adaptation was indicated by the percentages of SFA, which were significantly (P< 0.05) higher in gill (56%) and white muscle (58%) polar lipid fractions and coincided with lower percentages of n-3, n-6, and total PUFA. The regiospecific profiles indicated a high affinity of DHA to the sn-2 position in both the TAG (61-68%) and polar lipid (35-60%) fractions. The combination of detailed regiospecific and lipid analyses demonstrated adaptation of cell membrane structure in Atlantic salmon grown at an elevated water temperature.

Adaptation, Physiological↗

PubMed2006

A role for ethylene in the phytochrome-mediated control of vegetative development.

Members of the phytochrome family of photoreceptors play key roles in vegetative plant development, including the regulation of stem elongation, leaf development and chlorophyll accumulation. Hormones have been implicated in the control of these processes in de-etiolating seedlings. However, the mechanisms by which the phytochromes regulate vegetative development in more mature plants are less well understood. Pea (Pisum sativum) mutant plants lacking phytochromes A and B, the two phytochromes present in this species, develop severe defects later in development, including short, thick, distorted internodes and reduced leaf expansion, chlorophyll content and CAB gene transcript level. Studies presented here indicate that many of these defects in phyA phyB mutant plants appear to be due to elevated ethylene production, and suggest that an important role of the phytochromes in pea is to restrict ethylene production to a level that does not inhibit vegetative growth. Mutant phyA phyB plants produce significantly more ethylene than WT plants, and application of an ethylene biosynthesis inhibitor rescued many aspects of the phyA phyB mutant phenotype. This deregulation of ethylene production in phy-deficient plants appears likely to be due, at least in part, to the elevated transcript levels of key ethylene-biosynthesis genes. The phytochrome A photoreceptor appears to play a prominent role in the regulation of ethylene production, as phyA, but not phyB, single-mutant plants also exhibit a phenotype consistent with elevated ethylene production. Potential interactions between ethylene and secondary plant hormones in the control of the phy-deficient mutant phenotype were explored, revealing that ethylene may inhibit stem elongation in part by reducing gibberellin levels.

Ethylenes↗

PubMed2006

Spatial scale of the patchiness of plant poisons: a critical influence on foraging efficiency.

Generalist mammalian browsers and folivores feed on a range of chemically different plant species, which may assist them in diluting toxins and diversifying nutrient consumption. The frequency and order in which their diets are mixed are important determinants of intake. As a result, the degree of plant heterogeneity in an environment, and the spatial scale at which this occurs, should directly influence herbivore foraging decisions. We tested whether altering the Spatial scale of plants, and thus plant secondary metabolites (PSMs), affected foraging efficiency of a generalist folivore, the common brushtail possum (Trichosurus vulpecula). First, we demonstrated that possums were able to consume more from a mixed diet of two chemically different species, Eucalyptus globulus and E. tenuiramis, than when either of these species was offered alone. We then tested whether altering the spatial scale between E. globulus and E. tenuiramis, as small- or large-scale plant heterogeneity "patches," affected possum foraging behavior and, ultimately, their foraging efficiency. Possums increased their foraging efficiency when the spatial scale of plant heterogeneity was small rather than large. We argue that the ability to regularly switch diets, when plant spatial distribution is at a small scale, reduces the negative effects of PSM ingestion. We predict that the heterogeneity of plant patches, in relation to PSM distribution, and the scale at which this occurs across a landscape, are critical factors that influence foraging efficiency and, ultimately, fitness of mammalian herbivores. This research provides a fundamental link between plant chemistry, foraging, and habitat heterogeneity.

Animals↗

PubMed2006

In situ lubricant degradation in Antarctic marine sediments. 1. Short-term changes.

A large-scale, in situ experiment was set up near the Bailey Peninsula area (Casey Station, East Antarctica) to monitor the natural attenuation of synthetic lubricants in marine sediments over five years. Here, we report the short-term changes after 5 and 56 weeks. The lubricants tested were an unused and used Mobil lubricant (0W/40; Exxon Mobil, Irving, TX, USA) and a biodegradable alternative (0W/20; Fuchs Lubricants, Harvey, IL, USA). Clean sediment was collected, contaminated with the lubricants, and deployed by divers onto the seabed in a randomized block design. The sampled sediments were analyzed by gas chromatography-flame-ionization detector and gas chromatography-mass spectrometry with selective ion monitoring. The base fluid of all lubricant treatments did not decrease significantly after 56 weeks in situ. Alkanoate esters of 1,1,1-tris(hydroxymethyl)propane in the biodegradable and unused lubricants were degraded extensively in situ; however, these esters constituted only a minor proportion of the lubricant volume. The additives, alkylated naphthalenes and substituted diphenylamines, were fairly resistant to degradation, which is of environmental concern because of their toxicity. The biodegradable lubricant did not break down to recognized biodegradable thresholds and, as such, should not be classified as biodegradable under Antarctic marine conditions. A separate experiment was conducted to determine the influence of sediment preparation and deployment on compound ratios within the lubricants, and we found that preparation and deployment of the contaminated sediments had only a minor effect on compound recovery. Further monitoring of this in situ experiment will provide much needed information about the long-term natural attenuation of lubricants.

Antarctic Regions↗

PubMed2005

Proteomic analysis of Myrmecia pilosula (jack jumper) ant venom.

Ant sting allergy in Australia is predominantly due to the Myrmecia pilosula species complex. Gel separation of M. pilosula venom is necessary so that the allergenic importance of each component can be defined by western blotting. However, previous PAGE methods produced suboptimal resolution and the components of each band were not precisely defined. Venom was resolved in both non-reduced and reduced form by one-dimensional acid urea PAGE, SDS-PAGE and two-dimensional acid urea-SDS PAGE. Resolved peptides were extracted and analysed by HPLC-MS. Acid urea PAGE and acid urea-SDS PAGE proved more effective than SDS-PAGE for resolution of peptides smaller than 10 kDa. All of the major peptides previously observed in M. pilosula venom were observed in gel resolved venom. Venom was found to primarily consist of peptides with molecular weight <10 kDa, most of which contain disulfide bridges. SDS-PAGE of non-reduced venom clearly defined six higher molecular weight proteins between 26 and 90 kDa. An 8546 Da dimer named pilosulin 5 was observed, but pilosulin 4, a peptide recently proposed to be present in venom was not. A variant of pilosulin 4 here named pilosulin 4.1a, existing as an 8198 Da dimer, was observed and has been characterised.

Animals↗

PubMed2005

Diet switching in a generalist mammalian folivore: fundamental to maximising intake.

Generalist mammalian herbivores exploit a diverse diet. A generalised feeding strategy utilises a mixed diet to obtain a range of nutrients and to reduce the detoxication load of similar groups of plant secondary metabolites (PSMs). There is limited research investigating how mammalian herbivores achieve this dietary mixing in their daily foraging activities. We investigated the patterns of, and behaviours associated with, dietary mixing in a generalist mammalian folivore, the common brushtail possum (Trichosurus vulpecula). Possums were offered foliage of two eucalypt species (Eucalyptus globulus and E. regnans) as either (a) Full choice: both species offered for 8 h; (b) Restricted choice: both species offered for 2x2 h blocks; (c) G-R no choice: E. globulus offered for the first 4 h, E. regnans offered for next 4 h; and (d) R-G no choice: E. regnans offered for first 4 h, E. globulus offered for next 4 h. We hypothesised that possums would maximise intake on the Full choice diet, where time availability was greatest in combination with a choice of foliage. We also hypothesised that diet switching, defined as the frequency of changing between food types while feeding, would play a fundamental role in maximising intake. Possums achieved maximum intake on the Full choice diet and minimum intake on the No choice diets. Although intake was similar between the Restricted choice and No choice diets, possums achieved this intake in half the amount of time when able to switch regularly between foliage on the Restricted choice diet. We conclude that a generalist herbivore's ability to effectively switch diets when foraging is fundamental to maximising intake. Hence, the degree of plant heterogeneity in an environment, and the spatial scale at which it occurs, may affect an herbivore's foraging decisions and, ultimately, influence its foraging efficiency.

Animals↗

PubMed2005

Effect of drying temperature and air flow on the production and retention of secondary metabolites in saffron.

Safranal is the compound most responsible for the aroma of saffron spice and is, together with the suite of crocin pigments, the major determinant of the product quality. The content of safranal and pigments in saffron is determined by the method of postharvest treatment of the Crocus stigmas. A range of drying treatments involving different temperatures, with or without air flow, was applied to stigmas from three harvest dates. Dual solvent extractions combined with quantitative measurement using GC and HPLC-UV-vis techniques were used to analyze the secondary metabolite contents of the products. It was demonstrated that these methods overcame the previously reported problems in measuring the concentration of both pigments and safranal in saffron caused by the very different polarities and thus solubilities of these compounds. The results showed that a brief (20 min) initial period at a relatively high temperature (between 80 and 92 degrees C) followed by continued drying at a lower temperature (43 degrees C) produced saffron with a safranal content up to 25 times that of saffron dried only at lower temperatures. Evidence was provided suggesting that drying with significant air flow reduced the safranal concentration. The results, moreover, indicated that high-temperature treatment had allowed greater retention of crocin pigments than in saffron dried at intermediate temperatures (46-58 degrees C). The biochemical implications of the various treatments are discussed in relation to the potential for optimizing color and fragrance quality in the product.

Air Movements↗

PubMed2005

Stems of the Arabidopsis pin1-1 mutant are not deficient in free indole-3-acetic acid.

The pin1-1 mutant of Arabidopsis thaliana has been pivotal for studies on auxin transport and on the role of auxin in plant development. It was reported previously that when whole shoots were analysed, levels of the major auxin, indole-3-acetic acid (IAA) were dramatically reduced in the mutant, compared with the WT (Okada et al. 1991). The cloning of PIN1, however, provided evidence that this gene encodes a facilitator of auxin efflux, raising the question of how the pin1-1 mutation might reduce overall IAA levels as well as IAA transport. We therefore re-examined IAA levels in individual parts of pin1-1 and WT plants, focusing on inflorescence stems. Our data show that there is in fact no systemic IAA deficiency in the mutant. The previously reported difference between mutant and WT may have been due to the inclusion of reproductive structures in the WT harvest: we show here that the inflorescence itself contains high levels of IAA. We reconcile the normal IAA levels of pin1-1 inflorescence stems with their (previously-reported) reduced ability to transport IAA by presenting evidence that the auxin in mutant stems is not imported from their apical portion. Our data also indicate that levels of another auxin, indole-3-butyric acid (IBA), are very low in stems of the genotypes used in this study.

Arabidopsis↗

PubMed2005

Patterns of peripheral steroid metabolism vary with sex, season, and tissue type in blotched blue-tongued lizards (Tiliqua nigrolutea).

We examined sexual and seasonal variation in the ability of reproductively relevant tissues (liver, skin, adrenal gland, cloaca, kidney, renal sexual segment, epididymis, oviduct, muscle, testis, and ovary) to metabolise a primary steroid [testosterone (T) or estradiol (E2)] in the scincid lizard, Tiliqua nigrolutea. We observed considerable variation between sexes and across seasons in the patterns of conjugation and derivatisation of the primary steroids by these tissues. All tissues demonstrated the ability to conjugate the relevant primary steroid. Other general trends included increased conjugation by all tissues of gestating females, reduced metabolism of E2 by female tissues during late vitellogenesis, and reduced metabolism of T by males during early spermatogenesis. 5alpha-Dihydrotestosterone was the most commonly detected derivative in males, and production varied with season and tissue type. We suggest that seasonal variation in the ability of reproductively relevant tissues may be important in the physiological regulation of reproduction in this species.

Animals↗

PubMed2005

Inheritance of resistance to mammalian herbivores and of plant defensive chemistry in an Eucalyptus species.

Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their Fl hybrids. We assessed browsing damage of 3-year-old trees in a common environment field trial on four hybrid types of known progeny. The progeny were artificial intra-race crosses and reciprocal inter-race F1 hybrids of two geographically distinct populations (races) of E. globulus north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race Fl hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the Fl hybrids varied among major groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race Fl hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing.

Animals↗

PubMed2005

Inheritance of resistance to mammalian herbivores and of plant defensive chemistry in a Eucalyptus species.

Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their F1 hybrids. We assessed browsing damage of three-year-old trees in a common environment field trial on four hybrid types of known progeny. The progency were artificial intra-race crosses and reciprocal inter-race F1 hybrids of two geographically distinct populations (races) of E. globulus; north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race F1 hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the F1 hybrids varied among groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race F1 hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing.

Animals↗

PubMed2005

Unsaturated diether lipids in the psychrotrophic archaeon Halorubrum lacusprofundi.

The major phospholipids of Halorubrum lacusprofundi grown at 25 degrees C were archaeol phosphatidylglycerol, archaeol phosphatidylglycerylsulphate and archaeol phosphatidylglycerylphosphate methyl ester. Glycolipids included a monoglycosyl archaeol and the sulphate ester of a diglycosyl archaeol. Cultures grown at 12 degrees C contained the same suite of phospho- and glycolipids, with the addition of a series of unsaturated analogues with up to six double bonds. The patterns of unsaturation were similar for all the phospholipid series, but a different pattern occurred in the glycolipids. The analytical techniques used in this study allow facile detection of unsaturated archaeal cell membrane lipids that are degraded by commonly used chemical derivatization procedures.

Fatty Acids, Unsaturated↗

PubMed2004

Effects of nutrient variability on the genetic-based resistance of Eucalyptus globulus to a mammalian herbivore and on plant defensive chemistry.

Plant resistance to herbivores can be influenced not only by the independent effects of plant genotype and environmental variation, but by interactions between the two. The main aim of this study was to assess the effects of environmental variability (nutrient treatment) on the known genetic-based expression of resistance and defensive chemistry of Eucalyptus globulus to browsing by the generalist mammalian herbivore Trichosurus vulpecula. In a captive feeding trial, we measured intake of seedlings from one relatively resistant (Blue Gum Hill) and one relatively susceptible (St Helens) population of E. globulus grown under two nutrient treatments (no fertiliser, plus fertiliser). There was a significant genotypexfertiliser interaction effect on intake of E. globulus foliage by T. vulpecula, and the predicted genetic-based resistance of the two populations was expressed only for the non-fertilised treatment. Expression of resistance largely reflected the combined and inverse effects of nitrogen and condensed tannin concentrations. The expression of plant secondary metabolite concentration differed between compounds, but in all cases the effects of plant genotype and fertiliser treatment were independent. The formylated phloroglucinol compounds differed significantly between genotypes but not between fertiliser treatments. In contrast, the effect of plant genotype on the expression of condensed tannins was weak but they were significantly reduced by fertiliser. Essential oils were influenced by both plant genotype and fertiliser treatment and were significantly higher in the fertilised seedlings than in the non-fertilised seedlings. This study highlights interactive effects of plant genotype and environment in influencing the phenotypic expression of resistance in a eucalypt species to a mammalian browser. It also demonstrates that this interactive effect is the net result of independent effects of genotype and environment on plant chemistry and finally, that different groups of compounds within a plant can respond very differently to variation in environmental conditions.

Adaptation, Physiological↗

PubMed2004

The dissipation of tebuconazole and propiconazole in boronia (Boronia megastigma Nees).

The broad spectrum, systemic fungicides tebuconazole and propiconazole are used to control rust in boronia (Boronia megastigma Nees). Gas chromatography combined with either a benchtop quadrupole mass spectrometer or a high-resolution mass spectrometer allowed for the monitoring of both pesticides in boronia leaves, flowers, and concrete. Field trials were established at two sites to determine the rate of dissipation of tebuconazole and propiconazole in boronia. At site 1, two application rates of 125 and 250 g active ingredient/hectare (ai/ha) tebuconazole were employed. Treatments were repeated 17 days later. At harvest, 286 days after the final application, tebuconazole was detected at levels of 0.06 +/- 0.05 and 0.5 +/- 0.1 [mg/kg +/- standard error, on a dry matter basis (DMB)] in the leaves collected from plots treated with 125 and 250 g ai/ha of tebuconazole, respectively. The oil produced from the flowers collected at the final harvest had residues of tebuconazole at levels of 0.06 +/- 0.03 and 0.10 +/- 0.08 mg/kg for the 125 and 250 g ai/ha application rates, respectively. Two repeat applications of 125 g ai/ha propiconazole were also used at site 1. Residues of propiconazole were detected at 0.09 +/- 0.03 mg/kg (DMB) 286 days after the final application. At site 2, treatments of 125 g ai/ha of tebuconazole were applied twice. At harvest, 279 days after the final application of tebuconazole, residues were recorded at 0.30 +/- 0.09 mg/kg in the leaves (DMB) while the oil produced had 0.20 +/- 0.07 mg/kg.

Flowers↗

PubMed2004

Proximal C-terminal domain of sulphonylurea receptor 2A interacts with pore-forming Kir6 subunits in KATP channels.

Functional KATP (ATP-sensitive potassium) channels are hetero-octamers of four Kir6 (inwardly rectifying potassium) channel subunits and four SUR (sulphonylurea receptor) subunits. Possible interactions between the C-terminal domain of SUR2A and Kir6.2 were investigated by co-immunoprecipitation of rat SUR2A C-terminal fragments with full-length Kir6.2 and by analysis of cloned KATP channel function and distribution in HEK-293 cells (human embryonic kidney 293 cells) in the presence of competing rSUR2A fragments. Three maltose-binding protein-SUR2A fusions, rSUR2A-CTA (rSUR2A residues 1254-1545), rSUR2A-CTB (residues 1254-1403) and rSUR2A-CTC (residues 1294-1403), were co-immunoprecipitated with full-length Kir6.2 using a polyclonal anti-Kir6.2 antiserum. A fourth C-terminal domain fragment, rSUR2A-CTD (residues 1358-1545) did not co-immunoprecipitate with Kir6.2 under the same conditions, indicating a direct interaction between Kir6.2 and a 65-amino-acid section of the cytoplasmic C-terminal region of rSUR2A between residues 1294 and 1358. ATP- and glibenclamide-sensitive K+ currents were decreased in HEK-293 cells expressing full-length Kir6 and SUR2 subunits that were transiently transfected with fragments rSUR2A-CTA, rSUR2A-CTC and rSUR2A-CTE (residues 1294-1359) compared with fragment rSUR2A-CTD or mock-transfected cells, suggesting either channel inhibition or a reduction in the number of functional KATP channels at the cell surface. Anti-KATP channel subunit-associated fluorescence in the cell membrane was substantially lower and intracellular fluorescence increased in rSUR2A-CTE expressing cells; thus, SUR2A fragments containing residues 1294-1358 reduce current by decreasing the number of channel subunits in the cell membrane. These results identify a site in the C-terminal domain of rSUR2A, between residues 1294 and 1358, whose direct interaction with full-length Kir6.2 is crucial for the assembly of functional KATP channels.

ATP-Binding Cassette Transporters↗

PubMed2004

Diazoxide causes early activation of cardiac sarcolemmal KATP channels during metabolic inhibition by an indirect mechanism.

OBJECTIVE: We have used isolated myocytes to investigate the effects of diazoxide on sarcolemmal KATP channel (sarcoKATP) activity and action potential failure during metabolic inhibition, and the role of these channels in protection of functional recovery on reperfusion. MATERIALS AND METHODS: Isolated adult rat ventricular myocytes were exposed to metabolic inhibition (NaCN and iodoacetate) and reperfusion. Functional recovery was assessed from the ability of cells to contract on electrical stimulation and to recover calcium homeostasis, measured with fura-2. Action potentials and KATP currents were measured using patch clamp. RESULTS: Pretreatment with diazoxide (100 microM, 5 min) increased the proportion of cells that recovered contractile function after MI and reperfusion from 16.8 +/- 2.4% to 65.0 +/- 2.2% (p<0.001) and the proportion of cells in which [Ca2+]i recovered to <250 nM. Pretreatment also accelerated action potential and contractile failure during MI. In cell-attached patches, MI activated sarcoKATP channels after 224 +/- 11 s, and diazoxide pretreatment decreased this to 145 +/- 24 s (p<0.01). However, diazoxide present in the patch pipette did not accelerate sarcoKATP channel activation. Intracellular Mg2+ rose earlier in diazoxide-pretreated cells. The sarcoKATP blocker HMR 1883 delayed action potential failure and reduced diazoxide protection. CONCLUSIONS: Diazoxide pretreatment increases recovery of function and [Ca2+]i following reperfusion. Protection is coupled with early action potential failure, due to early activation of sarcoKATP channels during metabolic inhibition (MI), which is likely to involve an indirect effect of diazoxide.

Action Potentials↗