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1.
Mol Cell Proteomics ; 19(9): 1523-1532, 2020 09.
Article in English | MEDLINE | ID: mdl-32581039

ABSTRACT

Communication between individuals via molecules, termed chemosignaling, is widespread among animal and plant species. However, we lack knowledge on the specific functions of the substances involved for most systems. The femoral gland is an organ that secretes a waxy substance involved in chemical communication in lizards. Although the lipids and volatile substances secreted by the femoral glands have been investigated in several biochemical studies, the protein composition and functions of secretions remain completely unknown. Applying a proteomic approach, we provide the first attempt to comprehensively characterize the protein composition of femoral gland secretions from the Galápagos marine iguana. Using samples from several organs, the marine iguana proteome was assembled by next-generation sequencing and MS, resulting in 7513 proteins. Of these, 4305 proteins were present in the femoral gland, including keratins, small serum proteins, and fatty acid-binding proteins. Surprisingly, no proteins with discernible roles in partner recognition or inter-species communication could be identified. However, we did find several proteins with direct associations to the innate immune system, including lysozyme C, antileukoproteinase (ALP), pulmonary surfactant protein (SFTPD), and galectin (LGALS1) suggesting that the femoral glands function as an important barrier to infection. Furthermore, we report several novel anti-microbial peptides from the femoral glands that show similar action against Escherichia coli and Bacillus subtilis such as oncocin, a peptide known for its effectiveness against Gram-negative pathogens. This proteomics data set is a valuable resource for future functional protein analysis and demonstrates that femoral gland secretions also perform functions of the innate immune system.


Subject(s)
Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Iguanas/metabolism , Immune System/metabolism , Immunity, Innate , Proteome/metabolism , Transcriptome , Animals , Apoproteins/genetics , Apoproteins/metabolism , Bacillus subtilis/drug effects , Brain/metabolism , Chemotactic Factors/genetics , Chemotactic Factors/metabolism , Ecuador , Endopeptidases/genetics , Endopeptidases/metabolism , Escherichia coli/drug effects , Galectins/genetics , Galectins/metabolism , Heart/physiology , High-Throughput Nucleotide Sequencing , Humans , Iguanas/genetics , Iguanas/immunology , Immunity, Innate/genetics , Lung/metabolism , Muramidase/genetics , Muramidase/metabolism , Muscles/metabolism , Myocardium/metabolism , Organ Specificity , Proteome/genetics , Proteome/immunology , Proteomics , Pulmonary Surfactant-Associated Proteins/genetics , Pulmonary Surfactant-Associated Proteins/metabolism , Skin/metabolism , Tandem Mass Spectrometry , Transcriptome/genetics
2.
Gen Comp Endocrinol ; 294: 113468, 2020 08 01.
Article in English | MEDLINE | ID: mdl-32201233

ABSTRACT

Ecological factors, such as habitat quality, influence the survival and reproductive success of free-living organisms. Urbanization, including roads, alters native habitat and likely influences physiology, behavior, and ultimately Darwinian fitness. Some effects of roads are clearly negative, such as increased habitat fragmentation and mortality from vehicle collision. However, roads can also have positive effects, such as decreasing predator density and increased vegetation cover, particularly in xeric habitats due to increased water run-off. Glucocorticoids are metabolic hormones that reflect baseline metabolic needs, increase in response to acute challenges, and may mediate endogenous resource trade-offs between survival and reproduction. Here we examined circulating concentrations of corticosterone (baseline and stress-induced) in desert iguanas (Dipsosaurus dorsalis) in relation to the distance from a major anthropogenic disturbance, a high-traffic road in Palm Springs, CA. Additionally, we analyzed body condition and population density as additional predictors of glucocorticoid physiology. Surprisingly, we found lower baseline CORT levels closer to the road, but no effect of distance from road on stress-induced CORT or stress responsiveness (difference between baseline and stress-induced concentrations). Both population density and body condition were negative predictors of baseline CORT, stress-induced CORT, and stress responsiveness. Given the known effect of roads to increase run-off and vegetation density, increased water availability may improve available forage and shade, which may then increase the carrying capacity of the habitat and minimize metabolic challenges for this herbivorous lizard. However, it is important to recognize that surfaces covered by asphalt are not usable habitat for iguanas, likely resulting in a net habitat loss.


Subject(s)
Glucocorticoids/metabolism , Human Activities , Iguanas/metabolism , Animals , Corticosterone/blood , Female , Iguanas/blood , Male , Population Density , Stress, Physiological
4.
J Zoo Wildl Med ; 49(1): 86-91, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29517463

ABSTRACT

The objective of this study was to establish the pharmacokinetic parameters of ceftiofur crystalline free acid (CCFA) for a single intramuscular injection in green iguanas ( Iguana iguana). Six green iguanas received an injection of 5 mg/kg CCFA into the triceps muscle. Using high-performance liquid chromatography, concentrations of ceftiofur free acid equivalents in plasma samples collected at predetermined time points were evaluated up to 21 days following drug administration. Noncompartmental pharmacokinetic analysis was applied to the data. The observed maximum plasma concentration (Cmax obs) was 2.765 ± 0.864 µg/mL, and the time of observed maximum concentration (Tmax obs) was 6.1 ± 9.2 hr. The area under the curve (0 to infinity) was 239.3 ± 121.1 µg·hr/mL. No significant adverse drug reactions were clinically observed, and no visible injection site reactions were noted. Minimum inhibitory concentrations of bacterial isolates from iguanas were used to establish a target plasma concentration of 2.0 µg/mL. Based on the results from this study, a potential dosing interval for ceftiofur crystalline free acid administered at 5 mg/kg intramuscularly for iguanas maintained at a temperature of 30°C would be 24 hr based on a target plasma concentration of 2 µg/mL; however, multidose studies still need to be performed.


Subject(s)
Cephalosporins/pharmacokinetics , Iguanas/metabolism , Animals , Area Under Curve , Bacteria/drug effects , Cephalosporins/administration & dosage , Chromatography, High Pressure Liquid/veterinary , Female , Half-Life , Iguanas/blood , Injections, Intramuscular/veterinary , Male , Microbial Sensitivity Tests
5.
Gen Comp Endocrinol ; 230-231: 76-86, 2016 05 01.
Article in English | MEDLINE | ID: mdl-27044512

ABSTRACT

Growth hormone (GH), together with thyroid hormones (TH), regulates growth and development, and has critical effects on vertebrate metabolism. In ectotherms, these physiological processes are strongly influenced by environmental temperature. In reptiles, however, little is known about the direct influences of this factor on the somatotropic and thyroid axes. Therefore, the aim of this study was to describe the effects of both acute (48h) and chronic (2weeks) exposure to sub-optimal temperatures (25 and 18°C) upon somatotropic and thyroid axis function of the green iguana, in comparison to the control temperature (30-35°C). We found a significant increase in GH release (2.0-fold at 25°C and 1.9-fold at 18°C) and GH mRNA expression (up to 3.7-fold), mainly under chronic exposure conditions. The serum concentration of insulin-like growth factor-I (IGF-I) was significantly greater after chronic exposure (18.5±2.3 at 25°C; 15.92±3.4 at 18°C; vs. 9.3±1.21ng/ml at 35°C), while hepatic IGF-I mRNA expression increased up to 6.8-fold. Somatotropic axis may be regulated, under acute conditions, by thyrotropin-releasing hormone (TRH) that significantly increased its hypothalamic concentration (1.45 times) and mRNA expression (0.9-fold above control), respectively; and somatostatin (mRNA expression increased 1.0-1.2 times above control); and under chronic treatment, by pituitary adenylate cyclase-activating peptide (PACAP mRNA expression was increased from 0.4 to 0.6 times). Also, it was shown that, under control conditions, injection of TRH stimulated a significant increase in circulating GH. On the other hand, while there was a significant rise in the hypothalamic content of TRH and its mRNA expression, this hormone did not appear to influence the thyroid axis activity, which showed a severe diminution in all conditions of cold exposure, as indicated by the decreases in thyrotropin (TSH) mRNA expression (up to one-eight of the control), serum T4 (from 11.6±1.09 to 5.3±0.58ng/ml, after 2weeks at 18°C) and T3 (from 0.87±0.09 to 0.05±0.01ng/ml, under chronic conditions at 25°C), and Type-2 deiodinase (D2) activity (from 992.5±224 to 213.6±26.4fmolI(125)T4/mgh). The reduction in thyroid activity correlates with the down-regulation of metabolism as suggested by the decrease in the serum glucose and free fatty acid levels. These changes apparently were independent of a possible stress response, at least under acute exposure to both temperatures and in chronic treatment to 25°C, since serum corticosterone had no significant changes in these conditions, while at chronic 18°C exposure, a slight increase (0.38 times above control) was found. Thus, these data suggest that the reptilian somatotropic and thyroid axes have differential responses to cold exposure, and that GH and TRH may play important roles associated to adaptation mechanisms that support temperature acclimation in the green iguana.


Subject(s)
Growth Hormone/metabolism , Iguanas/metabolism , Temperature , Thyroid Gland/metabolism , Thyrotropin-Releasing Hormone/metabolism , Animals , Blood Glucose/analysis , Corticosterone/blood , Growth Hormone/genetics , Hypothalamus/drug effects , Hypothalamus/metabolism , Iguanas/blood , Iguanas/genetics , Insulin-Like Growth Factor I/genetics , Iodide Peroxidase/metabolism , Liver/drug effects , Liver/metabolism , Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , RNA, Messenger/metabolism , Receptors, Cell Surface/blood , Somatostatin/genetics , Thyroid Gland/drug effects , Thyroid Hormones/blood , Thyroid Hormones/genetics , Thyroid Hormones/metabolism , Thyrotropin/genetics , Thyrotropin-Releasing Hormone/administration & dosage , Thyrotropin-Releasing Hormone/genetics , Thyrotropin-Releasing Hormone/pharmacology
6.
PLoS One ; 10(9): e0136770, 2015.
Article in English | MEDLINE | ID: mdl-26367118

ABSTRACT

Electron microscopy analyses of Iguana iguana blood preparations revealed the presence of mitochondria within erythrocytes with well-structured cristae. Fluorescence microscopy analyses upon incubation with phalloidin-FITC, Hoechst 33342 and mitochondrial transmembrane potential (Δψm)-sensitive probe MitoTracker Red indicated that mitochondria i) widely occur in erythrocytes, ii) are polarized, and iii) seem to be preferentially confined at a "perinuclear" region, as confirmed by electron microscopy. The analysis of NADH-dependent oxygen consumption showed that red blood cells retain the capability to consume oxygen, thereby providing compelling evidence that mitochondria of Iguana erythrocytes are functional and capable to perform oxidative phosphorylation.


Subject(s)
Erythrocytes/metabolism , Iguanas/metabolism , Mitochondria/metabolism , Animals , Erythrocytes/ultrastructure , Membrane Potential, Mitochondrial , Mitochondria/ultrastructure , NAD/metabolism , Oxidative Phosphorylation
7.
Gen Comp Endocrinol ; 203: 281-95, 2014 Jul 01.
Article in English | MEDLINE | ID: mdl-24769041

ABSTRACT

Pituitary growth hormone (GH) has been studied in most vertebrate groups; however, only a few studies have been carried out in reptiles. Little is known about pituitary hormones in the order Squamata, to which the green iguana (gi) belongs. In this work, we characterized the hypophysis of Iguana iguana morphologically. The somatotrophs (round cells of 7.6-10 µm containing 250- to 300-nm secretory granules where the giGH is stored) were found, by immunohistochemistry and in situ hybridization, exclusively in the caudal lobe of the pars distalis, whereas the lactotrophs were distributed only in the rostral lobe. A pituitary giGH-like protein was obtained by immuno-affinity chromatography employing a heterologous antibody against chicken GH. giGH showed molecular heterogeneity (22, 44, and 88 kDa by SDS-PAGE/Western blot under non-reducing conditions and at least four charge variants (pIs 6.2, 6.5, 6.9, 7.4) by isoelectric focusing. The pituitary giGH cDNA (1016 bp), amplified by PCR and RACE, encodes a pre-hormone of 218 aa, of which 190 aa correspond to the mature protein and 28 aa to the signal peptide. The giGH receptor cDNA was also partially sequenced. Phylogenetic analyses of the amino acid sequences of giGH and giGHR homologs in vertebrates suggest a parallel evolution and functional relationship between the GH and its receptor.


Subject(s)
Growth Hormone/genetics , Growth Hormone/metabolism , Iguanas/genetics , Iguanas/metabolism , Receptors, Somatotropin/genetics , Receptors, Somatotropin/metabolism , Amino Acid Sequence , Animals , Base Sequence , Electrophoresis, Polyacrylamide Gel , Evolution, Molecular , Immunohistochemistry , In Situ Hybridization , Molecular Sequence Data , Phylogeny , Pituitary Gland/metabolism , RNA, Messenger/metabolism , Sequence Homology, Amino Acid , Somatotrophs/metabolism
8.
Am J Vet Res ; 74(12): 1536-41, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24274892

ABSTRACT

OBJECTIVE: To determine an efficient method for the collection of semen samples by means of electroejaculation, characterize spermatozoa quality and quantity, and determine the effect of refrigerated storage on motility of spermatozoa obtained from green iguanas (Iguana iguana). ANIMALS: 18 adult green iguanas. PROCEDURES: Green iguanas were anesthetized, and semen samples were obtained by means of electroejaculation. Up to 3 series of electrostimulations were performed; the procedure was stopped after a semen sample was obtained. Various semen sample variables were evaluated. RESULTS: Semen samples were obtained from 16 iguanas; most (n = 10) iguanas produced a semen sample after the second series of electrostimulations. Median semen sample volume was 0.05 mL. Mean spermatozoa concentration was 2 69.0 × 10(6) spermatozoa/mL. Median percentage of motile spermatozoa was 78%. The only morphological abnormality of spermatozoa was bent tails (mean percentage in a semen sample, 5.7%). Spermatozoa motility decreased significantly during refrigeration (4°C); median percentage motility after 24, 48, and 72 hours of refrigeration was 60%, 33%, and 0%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study suggested electroejaculation can be performed to collect semen samples from green iguanas, characteristics of iguana semen samples are similar to those for semen samples obtained from other reptiles, and motility of iguana spermatozoa decreases during refrigeration within 48 to 72 hours.


Subject(s)
Iguanas/metabolism , Semen/chemistry , Specimen Handling/veterinary , Sperm Retrieval/veterinary , Spermatozoa/cytology , Animals , Electric Stimulation/methods , Male , Refrigeration/methods , Refrigeration/veterinary , Specimen Handling/methods , Sperm Motility/physiology , Statistics, Nonparametric
9.
PLoS One ; 7(9): e45871, 2012.
Article in English | MEDLINE | ID: mdl-23029286

ABSTRACT

Amelogenin, the major enamel matrix protein in tooth development, has been demonstrated to play a significant role in tooth enamel formation. Previous studies have identified the alternative splicing of amelogenin in many mammalian vertebrates as one mechanism for amelogenin heterogeneous expression in teeth. While amelogenin and its splicing forms in mammalian vertebrates have been cloned and sequenced, the amelogenin gene, especially its splicing forms in non-mammalian species, remains largely unknown. To better understand the mechanism underlying amelogenin evolution, we previously cloned and characterized an amelogenin gene sequence from a squamate, the green iguana. In this study, we employed RT-PCR to amplify the amelogenin gene from the black spiny-tailed iguana Ctenosaura similis teeth, and discovered a novel splicing form of the amelogenin gene. The transcript of the newly identified iguana amelogenin gene (named C. Similis-T2L) is 873 nucleotides long encoding an expected polypeptide of 206 amino acids. The C. Similis-T2L contains a unique exon denominated exon X, which is located between exon 5 and exon 6. The C. Similis-T2L contains 7 exons including exon 1, 2, 3, 5, X, 6, and 7. Analysis of the secondary and tertiary structures of T2L amelogenin protein demonstrated that exon X has a dramatic effect on the amelogenin structures. This is the first report to provide definitive evidence for the amelogenin alternative splicing in non-mammalian vertebrates, revealing a unique exon X and the splicing form of the amelogenin gene transcript in Ctenosaura similis.


Subject(s)
Amelogenin/genetics , Iguanas/genetics , Reptilian Proteins/genetics , Amelogenin/metabolism , Amino Acid Sequence , Animals , Base Sequence , DNA Primers/genetics , Exons , Gene Expression , Iguanas/metabolism , Models, Molecular , Molecular Sequence Data , Organ Specificity , Protein Isoforms/genetics , Protein Isoforms/metabolism , Protein Structure, Secondary , Reptilian Proteins/metabolism , Sequence Alignment , Sequence Analysis, DNA , Tooth/metabolism
10.
Article in English | MEDLINE | ID: mdl-22710252

ABSTRACT

Development within the cleidoic egg of birds and reptiles presents the embryo with the problem of accumulation of wastes from nitrogen metabolism. Ammonia derived from protein catabolism is converted into the less toxic product urea or relatively insoluble uric acid. The pattern of nitrogen excretion of the green iguana, Iguana iguana, was determined during embryonic development using samples from allantoic fluid and from the whole homogenized egg, and in hatchlings and adults using samples of blood plasma. Urea was the major excretory product over the course of embryonic development. It was found in higher concentrations in the allantoic sac, suggesting that there is a mechanism present on the allantoic membrane enabling the concentration of urea. The newly hatched iguana still produced urea while adults produced uric acid. The time course of this shift in the type of nitrogen waste was not determined but the change is likely to be related to the water relations associated with the terrestrial habit of the adult. The green iguana produces parchment-shelled eggs that double in mass during incubation due to water absorption; the eggs also accumulate 0.02 mM of urea, representing 82% of the total measured nitrogenous residues that accumulate inside the allantois. The increase in egg mass and urea concentration became significant after 55 days of incubation then were unchanged until hatching.


Subject(s)
Iguanas/embryology , Iguanas/metabolism , Nitrogen/metabolism , Ammonia/blood , Ammonia/metabolism , Analysis of Variance , Animals , Ovum/metabolism , Urea/blood , Urea/metabolism , Uric Acid/blood , Uric Acid/metabolism , Yolk Sac/metabolism
11.
Gen Comp Endocrinol ; 177(1): 93-7, 2012 May 15.
Article in English | MEDLINE | ID: mdl-22414390

ABSTRACT

Quantification of glucocorticoid metabolites in feces has been shown to be a powerful tool in evaluating well-being in vertebrates. Little is known however about the hypothalamic-pituitary-adrenal axis response to stressors, and consequent glucocorticoid excretion, in reptiles. In a longitudinal study, fecal corticosterone metabolite (FCM) levels in green iguanas (Iguana iguana) were quantified during periods of rest and exposure to hypothesized stressors. FCM quantification was combined with behavioral analysis to further contextualize the measured increases. It was shown that both daily 5-minute handling/restraint, as well as housing devoid of climbing opportunity, resulted in increased FCM excretion. Behavioral analysis suggested that the iguanas were chronically stressed by the lack of climbing opportunity, whereas handling may have induced only a transient stress response. The experimental design, using repeated periods of stressor-exposure, also revealed a facilitating effect, where the two stressors potentiated one another. Furthermore, the order of the two stressors was found to be important. The study provides insight into the functioning of the hormonal stress response in green iguanas, and to the refining of their housing and handling.


Subject(s)
Corticosterone/metabolism , Feces/chemistry , Iguanas/metabolism , Stress, Physiological/physiology , Animals , Female , Iguanas/physiology , Male
12.
Comp Biochem Physiol B Biochem Mol Biol ; 161(2): 117-23, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22036614

ABSTRACT

Indicator models of sexual selection suggest that signal honesty is maintained via costs of ornament expression. Carotenoid-based visual signals are a well-studied example, as carotenoids may be environmentally limited and impact signaler health. However, not all bright yellow, orange and red ornaments found in vertebrates are carotenoid-based; pteridine pigments may also produce these colors. We examine the contribution of carotenoid and pteridine pigments to the orange reproductive color of female striped plateau lizards (Sceloporus virgatus). This color ornament reliably indicates female mate quality, yet costs maintaining signal honesty are currently unknown. Dietary carotenoid manipulations did not affect orange color, and orange skin differed from surrounding white skin in drosopterin, not carotenoid, content. Further, orange color positively correlated with drosopterin, not carotenoid, concentration. Drosopterin-based female ornaments avoid the direct trade-offs of using carotenoids for ornament production vs egg production, thus may relax counter-selection against color ornament exaggeration in females. Direct experimentation is needed to determine the actual costs of pteridine-based ornaments. Like carotenoids, pteridines influence important biological processes, including immune and antioxidant function; predation and social costs may also be relevant.


Subject(s)
Iguanas/metabolism , Pigments, Biological/metabolism , Pteridines/metabolism , Sex Characteristics , Skin Pigmentation , Animals , Carotenoids/metabolism , Epidermis/physiology , Female , Male , Pharynx/physiology
13.
Horm Behav ; 58(5): 792-9, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20708010

ABSTRACT

Anthropogenic disturbance is a relevant and widespread facilitator of environmental change and there is clear evidence that it impacts natural populations. While population-level responses to major anthropogenic changes have been well studied, individual physiological responses to mild disturbance can be equally critical to the long-term survival of a species, yet they remain largely unexamined. The current study investigated the impact of seemingly low-level anthropogenic disturbance (ecotourism) on stress responsiveness and specific fitness-related immune measures in different breeding stages of the marine iguana (Amblyrhynchus cristatus). Specifically, we found stress-induced elevations in plasma corticosterone among tourist-exposed populations relative to undisturbed populations. We also found changes in multiple immunological responses associated with stress-related effects of human disturbance, including bacterial killing ability, cutaneous wound healing, and hemolytic complement activity, and the responses varied according to reproductive state. By identifying health-related consequences of human disturbance, this study provides critical insight into the conservation of a well-known species that has a very distinct ecology. The study also broadens the foundation of knowledge needed to understand the global significance of various levels of human disturbance.


Subject(s)
Endocrine System/physiology , Human Activities , Iguanas/physiology , Immune System/physiology , Animals , Aquatic Organisms/physiology , Blood Bactericidal Activity/physiology , Complement System Proteins/metabolism , Corticosterone/blood , Corticosterone/metabolism , Ecuador , Endocrine System/metabolism , Female , Humans , Iguanas/blood , Iguanas/metabolism , Immune System/metabolism , Male , Stress, Psychological/blood , Stress, Psychological/physiopathology , Testosterone/blood , Wound Healing/physiology
14.
J Zoo Wildl Med ; 40(3): 495-500, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19746865

ABSTRACT

Heavy metal toxicosis is a well-known phenomenon in wild, captive-animal, and domestic animal medicine. However, the occurrence among reptiles is not well documented. One reason for this is the lack of information regarding reference blood and tissue levels of heavy metals in reptiles. To determine normal blood lead, plasma zinc, and liver lead and zinc concentrations, blood and liver samples were collected from 4 adult and 16 juvenile, healthy green iguanas (Iguana iguana). Lead and zinc levels were measured using atomic absorption spectroscopy. Using the mean +/- two SD as the normal reference range, the present study suggests the following for captive common green iguana: 1) whole blood lead level: 0.06 +/- 0.06 microg/ml; 2) plasma zinc level: 2.68 +/- 1.66 microg/ml; 3) liver lead level (wet-weight basis): <1.0 +/- 0.0 microg/g; 4) liver lead level (dry-weight basis): <3.0 +/- 0.0 microg/g; 5) liver zinc level (wet-weight basis): 24.9 +/- 11.6 microg/g; and 6) liver zinc level (dry-weight basis): 83.4 +/- 44.6 microg/g. These values are fairly consistent with published reference levels in other mammalian and avian species.


Subject(s)
Iguanas/metabolism , Lead/metabolism , Liver/metabolism , Zinc/metabolism , Animals , Animals, Zoo/blood , Animals, Zoo/metabolism , Iguanas/blood , Lead/blood , Reference Values , Spectrophotometry, Atomic/veterinary , Zinc/blood
15.
Physiol Biochem Zool ; 82(5): 430-7, 2009.
Article in English | MEDLINE | ID: mdl-19659444

ABSTRACT

Abstract Physiological responses to organismal stress can have direct impacts on individual fitness. While responses to stressors mediated by glucocorticoid hormones are well studied, the regulation of the redox system via pro-oxidant and antioxidant balance as well as the natural causes of oxidative stress in nature remain poorly known, especially for reptiles. In this study, we investigate the interpopulation and intersex variation in oxidative damage and plasma antioxidant capacity in the Galápagos land iguana, Conolophus subcristatus, over a 3-yr study to evaluate what factors (e.g., season, food availability, reproductive activity) can explain levels and patterns of oxidative damage and of plasma antioxidant capacity. Our results indicate that (1) males showed lower levels of oxidative damage, higher levels of plasma antioxidant capacity, and better body condition than females and (2) significant interactions exist among patterns of oxidative damage across sexes, sampling localities, body condition, and season. These results suggest that reproductive activity and food abundance might act as determinants shaping levels and patterns of oxidative stress of land iguanas.


Subject(s)
Iguanas/metabolism , Animals , Antioxidants/metabolism , Ecuador , Female , Iguanas/physiology , Male , Oxidative Stress , Population Dynamics , Reproduction , Seasons , Sex Characteristics , Time Factors
16.
Article in English | MEDLINE | ID: mdl-18313960

ABSTRACT

The Galápagos marine iguana, Amblyrhynchus cristatus, is unique among lizards in foraging subtidally, leading to activity across a broad range of ambient temperatures ( approximately 14-40 degrees C). To determine whether the marine iguana shows any biochemical changes consistent with maintaining enzyme function at both warm and cold body temperatures, we examined the function of the aerobic enzyme citrate synthase (CS) and the muscle isoform of the anaerobic enzyme lactate dehydrogenase (A(4)-LDH) in A. cristatus and a confamilial species, Iguana iguana, from 14 to 46 degrees C. We also deduced amino acid sequences from cDNA of each enzyme. In CS, despite two amino acid substitutions, we found no difference in the apparent Michaelis-Menten constant K(m) of oxaloacetate at any temperature, indicating that the substrate affinity of CS in A. cristatus has not adapted to changes in thermal environment. In A(4)-LDH, we used site-directed mutagenesis to show that the substitutions T9A and I283V (A. cristatus --> I. iguana) individually have no effect on kinetics, but together significantly decrease the K(m) of pyruvate and catalytic rate constant (k(cat)) of the A. cristatus ortholog. Thus, our data show that A. cristatus A(4)-LDH has not become cold adapted in response to this species' aquatic foraging behavior, and instead may be consistent with moderate warm adaptation with respect to the I. iguana ortholog.


Subject(s)
Citrate (si)-Synthase/metabolism , Iguanas/metabolism , L-Lactate Dehydrogenase/metabolism , Muscles/enzymology , Temperature , Amino Acid Sequence , Animals , Catalysis , Citrate (si)-Synthase/chemistry , Ecuador , Kinetics , L-Lactate Dehydrogenase/chemistry , Molecular Sequence Data , Mutation/genetics , Protein Structure, Secondary , Pyruvic Acid/metabolism , Sequence Alignment , Substrate Specificity
17.
J Vet Pharmacol Ther ; 31(1): 9-17, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18177313

ABSTRACT

When body size varies greatly, drug disposition can best be described as an allometric function of body weight. Therefore, the allometry of standard metabolic rate (SMR; 3/4 power) and body surface area (BSA; 2/3 power) have been advocated as surrogate markers for the prediction of key pharmacokinetic parameters. The goal of the present study was to examine the allometric basis of pharmacokinetic scaling within a species, green iguanas. Enrofloxacin was administered intravenously to 20 green iguanas (322-3824 g), and noncompartmental analysis was used to calculate standard pharmacokinetic parameters, which were log(10) transformed and regressed against log(10) body weight. The slopes of significant regressions were compared with the values of unity, 3/4, and 2/3. The slope of enrofloxacin total body clearance (Cl) was also compared with the slopes relating SMR and renal Cl of (99m)Tc-diethylenetriamine penta-acetic acid ((99m)DTPA) to body weight in iguanas. Enrofloxacin Cl depended allometrically on body weight with the power of 0.32. The slope of enrofloxacin Cl was significantly less than those of SMR, Cl of (99m)DTPA, and the 2/3 value. Therefore, the relationship between enrofloxacin Cl and body weight does not directly depend on the allometry of BSA, SMR, or renal Cl of (99m)DTPA in iguanas.


Subject(s)
Anti-Infective Agents/pharmacokinetics , Fluoroquinolones/pharmacokinetics , Iguanas/metabolism , Kidney/metabolism , Animals , Anti-Infective Agents/administration & dosage , Anti-Infective Agents/blood , Area Under Curve , Enrofloxacin , Female , Fluoroquinolones/administration & dosage , Fluoroquinolones/blood , Infusions, Intravenous/veterinary , Male , Metabolic Clearance Rate , Radiopharmaceuticals , Technetium Tc 99m Pentetate
18.
Article in English | MEDLINE | ID: mdl-17658283

ABSTRACT

One of the proposed ecological functions of sleep is to conserve energy. The majority of studies that support this theory have been done on endothermic animals whose body temperatures drop during sleep due to the reduced neurological control of thermoregulation. In the present study, we examined typical temperatures to which the Desert Iguana, Dipsosaurus dorsalis, is exposed to in the field and found that mean high temperatures ranged from 24-58 degrees C throughout the active portion of the year. We also examined the ecological savings that sleep could provide for this ectothermic iguana using a closed system respirometer. We found that laboratory-acclimated iguanas are able to save significantly more (27.6%) energy by sleeping than by being awake and that field iguanas also had significant savings of energy (69.1%) while asleep. However, iguanas could save more energy by remaining awake at cooler temperatures than by sleeping at warmer temperatures. In addition, we found no correlation for time of night with metabolic rate. Our study supports the hypothesis that one potential function of sleep is to conserve energy.


Subject(s)
Acclimatization , Body Temperature Regulation , Desert Climate , Energy Metabolism , Iguanas/physiology , Sleep , Temperature , Wakefulness , Animals , Basal Metabolism , California , Iguanas/metabolism , Male , Seasons
19.
Gene ; 398(1-2): 94-102, 2007 Aug 15.
Article in English | MEDLINE | ID: mdl-17560742

ABSTRACT

Cytoglobin (Cygb), a recently discovered vertebrate cytoplasmic heme-binding globin, is considered to be in a clade with vertebrate myoglobin (Mb), which is exclusively distributed in the cytoplasm of cardiac and skeletal muscles as an oxygen storage protein. GenBank databases (NCBI and JGI) and gene synteny analyses showed the absence of the Mb gene (mb) in two anuran amphibians, Xenopus laevis and X. tropicalis. Here we conducted comparative studies on the gene expression and tissue distribution of Cygb and Mb in anuran and reptilian tissues. Cygb and Mb genes were cloned from a reptile, iguana (Iguana iguana). Two types of cygb (cygb-1 and -2) were cloned, with lengths of 1066 and 1034 bp, and 196 and 193 amino acid residues, respectively. Their nucleotide and amino acid sequence identities were 90 and 87%, respectively. The Mb gene covered 1416 bp with an open reading frame of 465 bp, giving rise to a 154 amino acid protein. The distal ligand-binding histidine at E7, the proximal heme-binding histidine at F8, and the phenylalanine residue at CD1 were conserved in Mb and Cygb. The nucleotide and amino acid sequence identity of I. iguana cygb-1 and cygb-2 against X. laevis cygb were approximately 67% and 65%, respectively. RT-PCR demonstrated that X. laevis cygb was uniquely expressed in the heart and skeletal muscles, and faintly in the liver and spleen, which was quite contrasted with Iguana and the other vertebrates, where mb is exclusively expressed in the heart and skeletal muscles. Immunohistochemical analyses showed the distribution of Cygb in the cytoplasm of skeletal muscle cells. Interestingly, Cygb in the heart was localized in the nuclei. Considering the absence of mb in the Anura, we hypothesize that Cygb in muscle cells of anurans compensates for the lack of Mb for the storage and intracellular transportation of oxygen.


Subject(s)
Gene Expression Profiling , Globins/genetics , Iguanas/genetics , Myoglobin/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Cloning, Molecular , DNA, Complementary/chemistry , DNA, Complementary/genetics , Globins/metabolism , Iguanas/metabolism , Immunohistochemistry , Male , Molecular Sequence Data , Muscle, Skeletal/metabolism , Myoglobin/metabolism , Phylogeny , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Salamandridae/genetics , Salamandridae/metabolism , Sequence Analysis, DNA , Sequence Homology , Synteny , Xenopus/genetics , Xenopus/metabolism , Xenopus laevis/genetics , Xenopus laevis/metabolism
20.
Am J Vet Res ; 67(10): 1670-4, 2006 Oct.
Article in English | MEDLINE | ID: mdl-17014314

ABSTRACT

OBJECTIVE: To test the hypothesis that differences in anesthetic uptake and elimination in iguanas would counter the pharmacokinetic effects of blood:gas solubility and thus serve to minimize kinetic differences among inhaled agents. ANIMALS: 6 green iguanas (Iguana iguana). PROCEDURES: Iguanas were anesthetized with isoflurane, sevoflurane, or desflurane in a Latin-square design. Intervals from initial administration of an anesthetic agent to specific induction events and from cessation of administration of an anesthetic agent to specific recovery events were recorded. End-expired gas concentrations were measured during anesthetic washout. RESULTS: Significant differences were not detected for any induction or recovery events for any inhalation agent in iguanas. Washout curves best fit a 2-compartment model, but slopes for both compartments did not differ significantly among the 3 anesthetics. CONCLUSIONS AND CLINICAL RELEVANCE: Differences in blood:gas solubility for isoflurane, sevoflurane, and desflurane did not significantly influence differences in pharmacokinetics for the inhalation agents in iguanas.


Subject(s)
Anesthesia, Inhalation/veterinary , Iguanas/metabolism , Isoflurane/analogs & derivatives , Isoflurane/pharmacokinetics , Methyl Ethers/pharmacokinetics , Anesthetics, Inhalation/pharmacokinetics , Animals , Desflurane , Female , Half-Life , Male , Sevoflurane
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