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1.
J Evol Biol ; 20(5): 2056-66, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17714322

ABSTRACT

The present study extends evidence that Drosophila heat-shock genes are distinctively evolvable because of insertion of transposable elements by examining the genotypic diversity and phenotypic consequences of naturally occurring P element insertions in the proximal promoter regions of two small heat-shock genes. Detailed scrutiny of two populations revealed 16 distinctive P transposable elements collectively segregating in proximal promoters of two small heat-shock genes, Hsp26 and Hsp27. These elements vary in size, orientation and insertion site. Frequencies of P element-containing alleles varied from 5% to 100% in these populations. Two Hsp26 elements chosen for detailed study, R(s)P(26) and D(2)P(m), reduced or abolished Hsp26 expression respectively. The R(s)P(26) element increased or did not affect inducible tolerance of high temperature, increased fecundity, but decreased developmental rate. On the other hand, the D(2)P(m) element decreased thermotolerance and fecundity. In lines subjected to experimental evolution, the allelic frequency of the R(s)P(26)P element varied considerably, and was at lower frequencies in lines selected for increased longevity and for accelerated development than in controls. Transposable element insertions into small Hsp genes in Drosophila populations can have dramatic fitness consequences, and therefore create variation on which selection can act.


Subject(s)
DNA Transposable Elements/physiology , Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Heat-Shock Proteins/genetics , Promoter Regions, Genetic , Animals , Gene Frequency , Phenotype , Polymorphism, Genetic
2.
Heredity (Edinb) ; 96(5): 353-9, 2006 May.
Article in English | MEDLINE | ID: mdl-16552433

ABSTRACT

We examined whether a remarkable occurrence - the physiological evolution of two Drosophila melanogaster populations, despite a spatial separation of only 100-400 m, was idiosyncratic and temporary, or persisted over multiple years. We ascertained the high-temperature tolerance of Drosophila descended from populations on the north-facing slope (NFS) and south-facing slope (SFS) of 'Evolution Canyon' (Lower Nahal Oren, Mt Carmel, Israel), which were collected in 1997, 1999, and 2000. Results for these Drosophila uniformly resembled other studies in many respects: an inverse relationship between survival and heat-shock temperature, male-female differences in thermotolerance, and inducible thermotolerance. Importantly, for all years of collection, SFS flies consistently exceeded NFS flies in basal and inducible thermotolerance after diverse heat shocks, with and without thermal pretreatment, and whether isofemale lines, synthetic populations, or inbred lines were compared. Inbred lines, however, had lower thermotolerance than outbred lines. Several nonexclusive processes may explain the evolution of such physiological differentiation.


Subject(s)
Acclimatization/genetics , Biological Evolution , Drosophila melanogaster/genetics , Animals , Female , Heat-Shock Response/genetics , Male , Microclimate , Microsatellite Repeats/genetics , Phenotype , Selection, Genetic
3.
J Evol Biol ; 18(4): 901-10, 2005 Jul.
Article in English | MEDLINE | ID: mdl-16033562

ABSTRACT

Global analysis of mRNA abundance via genomic arrays (i.e. transcriptomics or transcriptional profiling) is one approach to finding the genes that matter to organisms undergoing environmental stress. In evolutionary analyses of stress, mRNA abundance is often invoked as a proxy for the protein activity that may underlie variation in fitness. To provoke discussion of the utility and sensible application of this valuable approach, this manuscript examines the adequacy of mRNA abundance as a proxy for protein activity, fitness and stress. Published work to date suggests that mRNA abundance typically provides little information on protein activity and fitness and cannot substitute for detailed functional and ecological analyses of candidate genes. While the transcriptional profile can be an exquisitely sensitive indicator of stress, simpler indicators will often suffice. In view of this outcome, transcriptomics should undergo careful cost-benefit analysis before investigators deploy it in studies of stress responses and their evolution.


Subject(s)
Adaptation, Biological/genetics , Biological Evolution , Environment , Gene Expression Profiling/methods , Proteins/metabolism , RNA, Messenger/metabolism , Stress, Physiological , Cost-Benefit Analysis
4.
Proc Natl Acad Sci U S A ; 98(23): 13195-200, 2001 Nov 06.
Article in English | MEDLINE | ID: mdl-11687637

ABSTRACT

Substantial genetic differentiation, as great as among species, exists between populations of Drosophila melanogaster inhabiting opposite slopes of a small canyon. Previous work has shown that prezygotic sexual isolation and numerous differences in stress-related phenotypes have evolved between D. melanogaster populations in "Evolution Canyon," Israel, in which slopes 100-400 m apart differ dramatically in aridity, solar radiation, and associated vegetation. Because the canyon's width is well within flies' dispersal capabilities, we examined genetic changes associated with local adaptation and incipient speciation in the absence of geographical isolation. Here we report remarkable genetic differentiation of microsatellites and divergence in the regulatory region of hsp70Ba which encodes the major inducible heat shock protein of Drosophila, in the two populations. Additionally, an analysis of microsatellites suggests a limited exchange of migrants and lack of recent population bottlenecks. We hypothesize that adaptation to the contrasting microclimates overwhelms gene flow and is responsible for the genetic and phenotypic divergence between the populations.


Subject(s)
Adaptation, Physiological/genetics , Drosophila melanogaster/genetics , Evolution, Molecular , Genetic Variation , Microclimate , Animals , Drosophila melanogaster/physiology , HSP70 Heat-Shock Proteins/genetics , Israel , Loss of Heterozygosity , Microsatellite Repeats/genetics , Species Specificity
5.
J Exp Biol ; 204(Pt 11): 1869-81, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11441029

ABSTRACT

Drosophila melanogaster collected in sub-equatorial Africa in the 1970s are remarkably tolerant of sustained laboratory culture above 30 degrees C and of acute exposure to much warmer temperatures. Inducible thermotolerance of high temperatures, which in Drosophila melanogaster is due in part to the inducible molecular chaperone Hsp70, is only modest in this strain. Expression of Hsp70 protein and hsp70 mRNA is likewise reduced and has slower kinetics in this strain (T) than in a standard wild-type strain (Oregon R). These strains also differed in constitutive and heat-inducible levels of other molecular chaperones. The lower Hsp70 expression in the T strain apparently has no basis in the activation of the heat-shock transcription factor HSF, which is similar in T and Oregon R flies. Rather, the reduced expression may stem from insertion of two transposable elements, H.M.S. Beagle in the intergenic region of the 87A7 hsp70 gene cluster and Jockey in the hsp70Ba gene promoter. We hypothesize that the reduced Hsp70 expression in a Drosophila melanogaster strain living chronically at intermediate temperatures may represent an evolved suppression of the deleterious phenotypes of Hsp70.


Subject(s)
Drosophila melanogaster/genetics , Drosophila melanogaster/physiology , HSP70 Heat-Shock Proteins/genetics , HSP70 Heat-Shock Proteins/physiology , Acclimatization , Africa South of the Sahara , Animals , Base Sequence , DNA Primers/genetics , Genes, Insect , Hot Temperature , Kinetics , Multigene Family , Phenotype , Polymorphism, Genetic , RNA, Messenger/genetics , RNA, Messenger/metabolism
6.
Mol Biol Evol ; 18(7): 1272-82, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11420366

ABSTRACT

To determine how the modern copy number (5) of hsp70 genes in Drosophila melanogaster evolved, we localized the duplication events that created the genes in the phylogeny of the melanogaster group, examined D. melanogaster genomic sequence to investigate the mechanisms of duplication, and analyzed the hsp70 gene sequences of Drosophila orena and Drosophila mauritiana. The initial two-to-four hsp70 duplication occurred 10--15 MYA, according to fixed in situ hybridization to polytene chromosomes, before the origin and divergence of the melanogaster and five other species subgroups of the melanogaster group. Analysis of more than 30 kb of flanking sequence surrounding the hsp70 gene clusters suggested that this duplication was likely a retrotransposition. For the melanogaster subgroup, Southern hybridization and an hsp70 restriction map confirmed the conserved number (4) and arrangement of hsp70 genes in the seven species other than D. melanogaster. Drosophila melanogaster is unique; tandem duplication and gene conversion at the derived cluster yielded a fifth hsp70 gene. The four D. orena hsp70 genes are highly similar and concertedly evolving. In contrast, the D. mauritiana hsp70 genes are divergent, and many alleles are nonfunctional. The proliferation, concerted evolution, and maintenance of functionality in the D. melanogaster hsp70 genes is consistent with the action of natural selection in this species.


Subject(s)
Drosophila melanogaster/genetics , Gene Duplication , Genes, Insect , HSP70 Heat-Shock Proteins/genetics , Animals , Base Sequence , DNA Primers/genetics , Drosophila/genetics , Evolution, Molecular , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Multigene Family , Phylogeny , Sequence Homology, Nucleic Acid , Species Specificity , Time Factors
7.
J Exp Biol ; 204(Pt 2): 315-23, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11136617

ABSTRACT

The magnitude and time course of activation of the heat-shock transcription factor (HSF) differ among Drosophila melanogaster lines evolving at 18 degrees C, 25 degrees C or 28 degrees C for more than 20 years. At lower heat-shock temperatures (27-35 degrees C), flies from the 18 degrees C population had higher levels of activated HSF (as detected by an electrophoretic mobility shift assay) than those reared at 25 degrees C and 28 degrees C. At higher temperatures (36 and 37 degrees C), however, the 28 degrees C flies had the highest levels of HSF. These differences persisted after one generation of acclimation at 25 degrees C, suggesting that phenotypic plasticity was limited. In addition, larvae from the 28 degrees C lines activated HSF less rapidly after a 35 degrees C heat shock than those from the 18 degrees C and 25 degrees C populations. These results are similar but not identical to previously reported differences in expression of Hsp70 (the major heat-inducible stress protein in Drosophila melanogaster) among the experimental lines. We conclude that HSF activation evolves rapidly during laboratory culture at diverse temperatures and could play an important role in the evolution of the heat-shock response.


Subject(s)
DNA-Binding Proteins/metabolism , Drosophila melanogaster/metabolism , Insect Proteins/metabolism , Transcription Factors/metabolism , Animals , Base Sequence , Binding Sites/genetics , DNA/genetics , DNA/metabolism , Drosophila Proteins , Drosophila melanogaster/genetics , Heat Shock Transcription Factors , Larva/metabolism , Oligonucleotide Probes/genetics , Temperature
8.
J Evol Biol ; 14(1): 75-82, 2001 Jan 08.
Article in English | MEDLINE | ID: mdl-29280581

ABSTRACT

To examine how the duration of laboratory domestication may affect Drosophila stocks used in studies of thermotolerance, we measured expression of the inducible heat-shock protein Hsp70 and survival after heat shock in D. melanogaster strains recently collected from nature and maintained in laboratory culture for up to 50 or more generations. After an initial increase in both Hsp70 expression and thermotolerance immediately after transfer to laboratory medium, both traits remained fairly constant over time and variation among strains persisted through laboratory domestication. Furthermore, variation in heat tolerance and Hsp70 expression did not correlate with the length of time populations evolved in the laboratory. Therefore, while environmental variation likely contributed most to early shifts in strain tolerance and Hsp70 expression, other population parameters, for example genetic drift, inbreeding, and selection likely affected these traits little. As long as populations are maintained with large numbers of individuals, the culture of insects in the laboratory may have little effect on the tolerance of different strains to thermal stress.

9.
Oecologia ; 123(4): 460-465, 2000 Jun.
Article in English | MEDLINE | ID: mdl-28308753

ABSTRACT

The expression of two temperature-sensitive reporter genes, hsp70 and an hsp70-LacZ fusion, in free-ranging adult Drosophila melanogaster indicates that natural thermal stress experienced by such small and mobile insects may be either infrequent or not severe. Levels of the heat-shock protein Hsp70, the major inducible Hsp of Drosophila, were similar in most wild Droso- phila captured after warm days to levels previously reported for unstressed flies in the laboratory. In a transgenic strain transformed with an hsp70-LacZ fusion (i.e., the structural gene encoding bacterial ß-galactosidase under control of a heat shock promoter), exposure to temperatures ≥32°C in the laboratory typically resulted in ß-galactosidase activities exceeding 140 mOD450 h-1µg-1 soluble protein. Flies caged in sun frequently had ß-galactosidase activities in excess of this level, whereas flies caged in shade and flies released and recaptured on cool days did not. Most flies (>80%) released on warm, sunny days had low ß-galactosidase activities upon recapture. Although the balance of recaptured flies had elevated ß-galactosidase activities on these days, their ß-galactosidase activities were <50% of levels for flies caged in direct sunlight or exposed to laboratory heat shock. These data suggest that even on warm days most flies may avoid thermal stress, presumably through microhabitat selection, but that a minority of adult D. melanogaster undergo mild thermal stress in nature. Both temperature-sensitive reporter genes, however, are limited in their ability to infer thermal stress and demonstrate its absence.

10.
Annu Rev Physiol ; 61: 243-82, 1999.
Article in English | MEDLINE | ID: mdl-10099689

ABSTRACT

Molecular chaperones, including the heat-shock proteins (Hsps), are a ubiquitous feature of cells in which these proteins cope with stress-induced denaturation of other proteins. Hsps have received the most attention in model organisms undergoing experimental stress in the laboratory, and the function of Hsps at the molecular and cellular level is becoming well understood in this context. A complementary focus is now emerging on the Hsps of both model and nonmodel organisms undergoing stress in nature, on the roles of Hsps in the stress physiology of whole multicellular eukaryotes and the tissues and organs they comprise, and on the ecological and evolutionary correlates of variation in Hsps and the genes that encode them. This focus discloses that (a) expression of Hsps can occur in nature, (b) all species have hsp genes but they vary in the patterns of their expression, (c) Hsp expression can be correlated with resistance to stress, and (d) species' thresholds for Hsp expression are correlated with levels of stress that they naturally undergo. These conclusions are now well established and may require little additional confirmation; many significant questions remain unanswered concerning both the mechanisms of Hsp-mediated stress tolerance at the organismal level and the evolutionary mechanisms that have diversified the hsp genes.


Subject(s)
Heat-Shock Proteins/physiology , Molecular Chaperones/physiology , Stress, Physiological/physiopathology , Animals , Biological Evolution , Ecology , Heat-Shock Proteins/genetics , Heat-Shock Response/physiology , Humans
11.
Oecologia ; 121(3): 323-329, 1999 Nov.
Article in English | MEDLINE | ID: mdl-28308320

ABSTRACT

We demonstrate that natural heat stress on wild larval Drosophila melanogaster results in severe developmental defects in >10% of eclosing adults, and that increased copy number of the gene encoding the major inducible heat shock protein of D. melanogaster, Hsp70, is sufficient to reduce the incidence of such abnormalities. Specifically, non-adult D. melanogaster inhabiting necrotic fruit experienced severe, often lethal heat stress in natural settings. Adult flies eclosing from wild larvae that had survived natural heat stress exhibited severe developmental anomalies of wing and abdominal morphology, which should dramatically affect fitness. The frequency of developmental abnormalities varied along two independent natural thermal gradients, exceeding 10% in adults eclosing from larvae developing in warm, sunlit fruit. When exposed to natural heat stress, D. melanogaster larvae with the wild-type number of hsp70 genes (n=10) developed abnormal wings significantly more frequently than a transgenic sister strain with 22 copies of the hsp70 gene.

12.
Proc Biol Sci ; 265(1394): 391-5, 1998 Mar 07.
Article in English | MEDLINE | ID: mdl-9523438

ABSTRACT

The endocellular microbe Wolbachia pipientis infects a wide variety of invertebrate species, in which its presence is closely linked to a form of reproductive failure termed cytoplasmic incompatibility (CI). CI renders infected males unable to father offspring when mated to uninfected females. Because CI can dramatically affect fitness in natural populations, mechanisms that abate CI can have equally large impacts on fitness. We have discovered that repeated copulation by Wolbachia-infected male Drosophila simulans significantly diminishes CI. Repeated copulation does not prevent Wolbachia from populating developing spermatids, but may reduce the time during spermatogenesis when Wolbachia can express CI. This restoration of fertility in premated infected males could have important implications for Wolbachia transmission and persistence in nature and for its exploitation as an agent of biological pest control.


Subject(s)
Drosophila/microbiology , Drosophila/physiology , Reproduction/physiology , Rickettsiaceae Infections/physiopathology , Rickettsiaceae/physiology , Spermatozoa/physiology , Animals , Female , Male , Spermatozoa/microbiology
13.
J Insect Physiol ; 44(11): 1091-1101, 1998 Nov.
Article in English | MEDLINE | ID: mdl-12770408

ABSTRACT

Heat shock proteins (Hsps) and other molecular chaperones perform diverse cellular roles (e.g., inducible thermotolerance) whose functional consequences are concentration dependent. We manipulated Hsp70 concentration quantitatively in intact larvae of Drosophila melanogaster to examine its effect on survival, developmental time and tissue damage after heat shock. Larvae of an extra-copy strain, which has 22 hsp70 copies, produced Hsp70 more rapidly and to higher concentrations than larvae of a control strain, which has the wild-type 10 copies of the gene. Increasing the magnitude and duration of pretreatment increased Hsp70 concentrations, improved tolerance of more severe stress, and reduced delays in development. Pretreatment, however, did not protect against acute tissue damage. For larvae provided a brief or mild intensity pretreatment, faster expression of Hsp70 in the extra-copy strain improved survival to adult and reduced tissue damage 21h after heat shock. Negative effects on survival ensued in extra-copy larvae pretreated most intensely, but their overexpression of Hsp70 did not increase tissue damage. Because rapid expression to yield a low Hsp70 concentration benefits larvae but overexpression harms them, natural selection may balance benefits and costs of high and low expression levels in natural populations.

14.
J Exp Biol ; 200(Pt 14): 2007-15, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9246784

ABSTRACT

All tissues of larval Drosophila melanogaster express Hsp70, the major heat-shock protein of this species, after both mild (36 degrees C) and severe (38.5 degrees C) heat shock. We used Hsp70-specific immunofluorescence to compare the rate and intensity of Hsp70 expression in various tissues after these two heat-shock treatments, and to compare this with related differences in the intensity of Trypan Blue staining shown by the tissues. Trypan Blue is a marker of tissue damage. Hsp70 was rarely detectable before heat shock. Brain, salivary glands, imaginal disks and hindgut expressed Hsp70 within the first hour of heat shock, whereas gut tissues, fat body and Malpighian tubules did not express Hsp70 until 4-21 h after heat shock. Differences in Hsp70 expression between tissues were more pronounced at the higher heat-shock temperature. Tissues that expressed Hsp70 slowly stained most intensely with Trypan Blue. Gut stained especially intensely, which suggests that its sensitivity to heat shock may limit larval thermotolerance. These patterns further suggest that some cells respond primarily to damage caused by heat shock rather than to elevated temperature per se and/or that Hsp70 expression is itself damaged by heat and requires time for recovery in some tissues.


Subject(s)
Drosophila melanogaster/metabolism , HSP70 Heat-Shock Proteins/biosynthesis , Heat Stress Disorders/metabolism , Animals , Drosophila melanogaster/embryology , Organ Specificity
15.
Cell Stress Chaperones ; 2(1): 60-71, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9250396

ABSTRACT

We compared transgenic Drosophila larvae varying in hsp70 copy number to assess the consequences of Hsp70 overexpression for growth and development after heat shock. Exposure to a mildly elevated temperature (36 degrees C) induced expression of Hsp70 (and presumably other heat shock proteins) and improved tolerance of more severe heat stress, 38.5-39.5 degrees C. We examined this pattern in two independently derived pairs of extra-copy and excision strains that differed primarily in hsp70 copy number (with 22 and 10 copies, respectively). Extra-copy larvae produced more Hsp70 in response to high temperature than did excision larvae, but surpassed the excision strain in survival only immediately after thermal stress. Excision larvae survived to adulthood at higher proportions than did extra-copy larvae and grew more rapidly after thermal stress. Furthermore, multiple pretreatment reduced survival of 1st-instar extra-copy larvae, but did not affect the corresponding excision strain. While extra Hsp70 provides additional protection against the immediate damage from heat stress, abnormally high concentrations can decrease growth, development and survival to adulthood.


Subject(s)
Drosophila melanogaster/genetics , HSP70 Heat-Shock Proteins/genetics , Analysis of Variance , Animals , Animals, Genetically Modified , Chromosomes/physiology , Drosophila melanogaster/chemistry , Drosophila melanogaster/growth & development , Female , Gene Expression Regulation, Developmental/physiology , Genes, Insect/physiology , Hot Temperature , Larva/chemistry , Larva/genetics , Larva/growth & development , Male , Stress, Physiological/metabolism
16.
EXS ; 83: 155-73, 1997.
Article in English | MEDLINE | ID: mdl-9342848

ABSTRACT

Classical adaptational and genetic engineering approaches offer complementary insights to understanding biological variation: the former elucidates the origins, magnitude and ecological context of natural variation, while the latter establishes which genes can underlie natural variation. Studies of the stress or heat shock response in Drosophila illustrate this point. At the cellular level, heat shock proteins (Hsps) function as molecular chaperones, minimizing aggregation of peptides in non-native conformations. To understand the adaptive significance of Hsps, we have characterized thermal stress that Drosophila experience in nature, which can be substantial. We used these findings to design ecologically relevant experiments with engineered Drosophila strains generated by unequal site-specific homologous recombination; these strains differ in hsp70 copy number but share sites of transgene integration. hsp70 copy number markedly affects Hsp70 levels in intact Drosophila, and strains with extra hsp70 copies exhibit corresponding differences in inducible thermotolerance and reactivation of a key enzyme after thermal stress. Elevated Hsp70 levels, however, are not without penalty; these levels retard growth and increase mortality. Transgenic variation in hsp70 copy number has counterparts in nature: isofemale lines from nature vary significantly in Hsp70 expression, and this variation is also correlated with both inducible thermotolerance and mortality in the absence of stress.


Subject(s)
Biological Evolution , Drosophila/physiology , Genetic Variation , Heat-Shock Proteins/biosynthesis , Animals , Animals, Genetically Modified , Drosophila/genetics , Genetic Engineering , HSP70 Heat-Shock Proteins/biosynthesis , HSP70 Heat-Shock Proteins/genetics , Heat-Shock Proteins/genetics , Hot Temperature , Models, Genetic , Reproduction , Selection, Genetic , Stress, Physiological
17.
J Exp Biol ; 199(Pt 8): 1837-44, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8708583

ABSTRACT

To determine how the accumulation of the major Drosophila melanogaster heat-shock protein, Hsp70, affects inducible thermotolerance in larvae and pupae, we have compared two sister strains generated by site-specific homologus recombination. One strain carried 12 extra copies of the Hsp70 gene at a single insertion site (extra-copy strain) and the other carried remnants of the transgene construct but lacked the extra copies of Hsp70 (excision strain). Hsp70 levels in whole-body lysates of larvae and pupae were measured by ELISA with an Hsp70-specific antibody. In both extra-copy and excision strains, Hsp70 was undetectable prior to heat shock. Hsp70 concentrations were higher in the extra-copy strain than in the excision strain at most time points during and after heat shock. Pretreatment (i.e. exposure to 36 degrees C before heat shock) significantly improved thermotolerance, and this improvement was greater and more rapid in larvae and pupae of the extra-copy strain than in those of the excision strain. The experimental conditions resemble thermal regimes actually experienced by Drosophila in the field. Thus, these findings represent the best evidence to date that the amount of a heat-shock protein affects the fitness of a complex animal in the wild.


Subject(s)
Drosophila melanogaster/physiology , Gene Expression , HSP70 Heat-Shock Proteins/genetics , HSP70 Heat-Shock Proteins/physiology , Hot Temperature , Protein Engineering , Animals , Drosophila melanogaster/genetics , Kinetics , Larva/physiology , Pupa/physiology , Recombination, Genetic
18.
J Exp Biol ; 174: 235-46, 1993 Jan.
Article in English | MEDLINE | ID: mdl-8440967

ABSTRACT

To examine the potential impact of fluid dynamic boundary layers on cutaneous ion exchange, we investigated how bulk flow of dilute Na+ solutions (< or = 1.0 mmol l-1) over the skin of intact frogs (Rana catesbeiana and Rana pipiens) affects cutaneous Na+ uptake (JNa(in)) and transepithelial potential (TEP). Cessation of stirring resulted in a 14-35% decrease in TEP and a 14-65% decrease in JNa(in). Two weeks' acclimation to an unstirred bath increased JNa(in) to levels 70% greater than in frogs acclimated to a continuously stirred bath and to levels comparable to those of frogs acclimated to deionized water. These effects are consistent with depletion of Na+ in the boundary layer, but are also consistent with depletion of O2 in the boundary layer, which might limit generation of ATP consumed by ATPases responsible for cutaneous Na+ uptake. To investigate this latter possibility, we measured TEP and JNa(in) while manipulating the PO2 of well-stirred external media at constant [Na+]. Hyperoxia (PO2 > or = 97 kPa) increased JNa(in) by 28% and had little or no effect on TEP. Hypoxia (PO2 < or = 1.5 kPa) reduced JNa(in) by 48% and decreased TEP by 22%. These results suggest that ionic and gaseous boundary layers may interact to affect cutaneous ion transport.


Subject(s)
Membranes/metabolism , Oxygen/metabolism , Skin/metabolism , Sodium/metabolism , Animals , Membrane Potentials , Models, Biological , Rana catesbeiana , Rana pipiens
19.
J Comp Physiol B ; 162(7): 646-50, 1992.
Article in English | MEDLINE | ID: mdl-1469156

ABSTRACT

Rates of O2 uptake across isolated perfused skin of bullfrogs (Rana catesbeiana) were measured in relation to blood flow at three levels of ambient O2 tension: normoxia (O2 tension = 152 torr), hypoxia (12% O2, 87 torr) and hyperoxia (42% O2, 306 torr). At bulk perfusion rates ranging from 3.4 to 10.1 microliters.cm-2 x min-1, O2 uptake was positively correlated with hemoglobin delivery rate in both normoxia and hyperoxia, but was independent of delivery rate in hypoxia. Mean O2 uptake in normoxia was 3.8 nmol O2 x cm-2 x min-1 at a delivery rate of 9.8 nmol.cm-2 x min-1 and 6.5 nmol O2 x cm-2 x min-1 at a delivery rate of 28.3 nmol.cm-2 x min-1. At any given bulk perfusion rate, oxygen uptake averaged about 49% lower in hypoxia than in normoxia, decreasing in proportion to the reduction of O2 tension difference between medium and blood. In hyperoxia, O2 uptake did not increase proportionally with the difference in O2 tension between blood and medium, averaging only 50% higher at a 2.4-fold greater O2 tension difference. Cutaneous diffusing capacity for O2 averaged 0.041 nmol O2 x cm-2 x torr-1 x min-1 during the first hour of perfusion in normoxia, and was not affected by reduction of ambient O2 tension. The results indicate that cutaneous O2 uptake in hypoxia is highly diffusion limited, and consequently, increases in cutaneous perfusion can not effectively compensate for reduction of ambient O2 tension. In hyperoxia, O2 uptake may be substantially perfusion limited because of reduced blood O2 capacitance at high O2 saturations.


Subject(s)
Hypoxia/blood , Oxygen/physiology , Rana catesbeiana/physiology , Skin Physiological Phenomena , Animals , Blood Gas Monitoring, Transcutaneous , Partial Pressure , Perfusion , Regional Blood Flow , Skin/blood supply
20.
Respir Physiol ; 85(1): 1-14, 1991 Jul.
Article in English | MEDLINE | ID: mdl-1947447

ABSTRACT

Patches of skin were removed from bullfrogs and perfused with a red cell suspension through the cutaneous artery to define the gas exchange characteristics of frog skin without complications caused by in vivo regulatory mechanisms. Oxygen uptake was primarily perfusion limited at low perfusion rates but primarily diffusion limited at perfusion rates that were similar to cutaneous blood flows previously reported in vivo. Diffusing capacity (DO2) increased only slightly as perfusion rate increased. Because the DO2 in isolated skin, in which DO2 should be maximal, was not significantly higher than estimates of DO2 in vivo, there seems to be little opportunity for increasing cutaneous DO2 or oxygen uptake in vivo.


Subject(s)
Oxygen/metabolism , Skin/metabolism , Animals , Biological Transport, Active , In Vitro Techniques , Oxygen/blood , Perfusion , Rana catesbeiana , Skin/blood supply
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