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PubMed1992

Downregulation of sodium channels during anoxia: a putative survival strategy of turtle brain.

In contrast to mammalian brain, which exhibits rapid degeneration during anoxia, the brains of certain species of turtles show an extraordinary capacity to survive prolonged anoxia. The decrease in energy expenditure shown by the anoxic turtle brain is likely to be a key factor for anoxic survival. The "channel arrest" hypothesis proposes that ion channels, which regulate brain electrical activity in normoxia, may be altered during anoxia in the turtle brain as a mechanism to spare energy. Goals of present research were to test this hypothesis and to determine whether down-regulation of sodium channels is a possible explanation for spike threshold shifts seen during anoxia in isolated turtle cerebellum. We report here that anoxia induced a significant (42%) decline in voltage-gated sodium channel density as determined by studies of the binding of a sodium channel ligand, [3H]brevetoxin. This study demonstrates that sodium channel densities in brain may be regulated by tissue oxygenation or by physiological events associated with anoxia. Moreover, it also suggests that downregulation of sodium channels may be a basis for changes in action potential thresholds, the electrical depression and energy conservation that provide the unique anoxic tolerance of turtle brain.

Animals↗
PubMed1995

Upregulation of the GABAA/benzodiazepine receptor during anoxia in the freshwater turtle brain.

The freshwater turtle brain survives anoxia by decreasing its energy expenditure. During this anoxic period there is a sustained release of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). This study investigated whether there was a corresponding change in the binding properties of the GABAA/benzodiazepine (GABA/BDZ) receptor. Turtles (Trachemys scripta) were subjected to a 100% N2 atmosphere for up to 24 h. After exposure, the cerebral cortex was dissected out, and saturation binding assays for GABA/BDZ receptors were performed using the radioligand [3H]flunitrazepam. Control turtles had a dissociation constant (Kd) of 1.97 +/- 0.54 nM and a receptor density (Bmax) of 2,404 +/- 221 fmol/mg protein. The Kd showed no significant change over 24 h of anoxia. However, significant increases were seen in Bmax after 12 h (21%, P < 0.05) and 24 h (29%, P < 0.01) of anoxia. We suggest that a long-term upregulation of GABAA receptors occurs in the anoxic turtle brain that acts to increase the inhibitory effectiveness of the released GABA and thereby contributes to anoxia survival of the turtle.

Animals↗
PubMed1985

Three forms of the scratch reflex in the spinal turtle: movement analyses.

The scratch reflex is a motor task exhibited by the hindlimb of a turtle after complete transection of the spinal cord just posterior to the forelimb enlargement. The scratch is elicited by applying gentle tactile stimulation to a site innervated by spinal segments caudal to the level of the transection. During the scratch response, the ipsilateral hindlimb reaches toward and rubs against the stimulated site. Each scratch response consists of either a single rub or a rhythmic series of rubs. There are three types or "forms" of the scratch reflex in the spinal turtle: rostral scratch (previously termed the scratch reflex in Refs. 5, 43, 45), pocket scratch, and caudal scratch. We have characterized each scratch form according to which portion of the hindlimb is used to rub against the stimulated site. During a rostral scratch, the dorsum of the foot and/or toes is used to rub against the stimulated site. During a pocket scratch, the side of the thigh, knee, and/or calf is used to rub. During a caudal scratch, the heel or side of the foot is used to rub. Each scratch form has a distinct receptive field on the turtle shell and skin. A rhythmic scratch response to maintained tactile stimulation consists of as many as 10-25 cycles. We have divided each cycle of movement into a prerub, rub, and postrub phase. Extension of the knee joint occurs during the rub phase of each scratch cycle for all scratch forms. The timing of the extension phase of the knee joint movement within the protraction-retraction (flexion-extension) cycle of the hip joint is different for each form of the scratch. During a rostral scratch, extension of the knee joint occurs when the hip is protracted. During a pocket scratch, knee extension occurs when the hip is retracting. During a caudal scratch, knee extension occurs when the hip is retracted. The spinal turtle therefore shifts the timing of movements of a distal joint (knee) with respect to the timing of movements of a proximal joint (hip) to obtain appropriate scratching movements to different regions of the body. A narrow transition zone exists between the receptive field for one form of the scratch and the nearby receptive field for another form of the scratch. There is a rostral-pocket transition zone and caudal-pocket transition zone in the spinal turtle.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1999

Immunohistochemical localization of NMDA- and AMPA-type glutamate receptor subunits in the basal ganglia of red-eared turtles.

Corticostriatal and thalamostriatal projection systems have been shown to utilize glutamate as a neurotransmitter in mammals and birds. Although corticostriatal and thalamostriatal projection systems have been demonstrated in turtles, it is uncertain whether they too use glutamate as their neurotransmitter. Immunohistochemical localization of glutamate and of NMDA- and AMPA-type ionotropic glutamate receptor subunits (NMDAR2A/B, GluR1, GluR2/3, and GluR4) were used to address this issue. Numerous medium-sized neurons that were rich in NMDAR2A/B and GluR2/3 were observed in the striatal part of the basal ganglia of red-eared turtles. Smaller numbers of medium-sized neurons and some large neurons rich in the GluR1 and GluR4 subunits were also observed in the striatum. The striatal neuropil was notably rich in GluR1, GluR2/3 and NMDAR2A/B subunits. The pallidal region was specifically rich in large neurons possessing GluR4 subunits. Consistent with the glutamate receptors on striatal and pallidal neurons, sources of input to the striatum and pallidum in turtle such as the dorsomedial and dorsolateral thalamic nuclei (which appear to correspond to intralaminar thalamic nuclei), telencephalic pallial cell groups, and the apparent subthalamic nucleus homologue were rich in glutamatergic neurons. The results show that the thalamostriatal, corticostriatal and subthalamo-pallidal projection systems of turtles are glutamatergic and that similar basal ganglia cell types in turtles and mammals have largely similar glutamate receptor characteristics.

Animals↗
PubMed2001

Conservation of chromosome 1 in turtles over 66 million years.

Fluorescence in situ hybridization of a whole chromosome 1-specific probe from the yellow-bellied slider turtle (Trachemys scripta) to cells from four other species of turtle ranging from a desert tortoise to a loggerhead sea turtle resulted in specific and exclusive hybridization to chromosome 1 in all five species. Previous observations of conservation in the giemsa banding pattern and chromosome morphology and number among turtles are thus extended to the DNA sequence level, revealing a cytogenetic stability of chromosome 1 in these turtles during the past 66-144 million years. This contrasts with the situation for various hominoid species where, in many instances, extensive chromosomal rearrangements have been reported in one third of that time period. Our probe, which was prepared by microdissecting whole chromosomes from embryonic T. scripta fibroblasts and amplifying using DOP-PCR, is the first report of a whole-chromosome FISH probe for any reptile.

Animals↗
PubMed1977

Individual differences in the sea-finding mechanism of hatchling leatherback turtles.

When hatchling leatherback turtles, Dermochelys coriacea, are unilarerally blindfolded, some circle toward their open eyes and some toward their covered eyes. If tests were performed immediately after one eye had been blindfolded, in cases where the turtle circled toward the open eye there was a transient potentiation of circling at the start of the test. In contrast, in cases where the turtle circled toward the covered eye, circling was decreased at the start of the test. Similar transient phenomena occurred when the position of a blindfold was reversed from one eye to another. Following removal of a blindfold, circling toward the previously covered eye was increased. The transient effects in all these tests waned by about 2 min. There was no evidence of transient circling tendencies in turtles that were tested after a unilateral blindfold had been in position for 4 min. The results suggest that ipsilateral and contralateral retinotectal turning systems differ in strength among individual turtles and individual eyes, and that following unilateral blindfolding there is a potentiation only of the ipsilateral turning system associated with stimulation of the still open eye.

Animals↗
PubMed2006

Phenology of marine turtle nesting revealed by statistical model of the nesting season.

BACKGROUND: Marine turtles deposit their eggs on tropical or subtropical beaches during discrete nesting seasons that span several months. The number and distribution of nests laid during a nesting season provide vital information on various aspects of marine turtle ecology and conservation. RESULTS: In the case of leatherback sea turtles nesting in French Guiana, we developed a mathematical model to explore the phenology of their nesting season, derived from an incomplete nest count dataset. We detected 3 primary components in the nest distribution of leatherbacks: an overall shape that corresponds to the arrival and departure of leatherback females in the Guianas region, a sinusoidal pattern with a period of approximately 10 days that is related to physiological constraints of nesting female leatherbacks, and a sinusoidal pattern with a period of approximately 15 days that likely reflects the influence of spring high tides on nesting female turtles. CONCLUSION: The model proposed here offers a variety of uses for both marine turtles and also other taxa when individuals are observed in a particular location for only part of the year.

Animals↗
PubMed1979

Estrogen-binding proteins in the oviduct of the turtle, Chrysemys picta: evidence for a receptor species.

Estradiol-binding proteins in the reproductive tract of the turtle, Chrysemys picta, were characterized. Cytosol was prepared from the oviducts of mature female turtles, and estradiol binding was measured using charcoal adsorption and glycerol density gradient centrifugation. A sex steroid-binding protein (SBP) similar to that found in turtle plasma was demonstrated in oviduct cytosol. The characteristics of this SBP-like binding were as follows: Ka = 10(8) M-1; capacity, 10(-12) mol/mg protein; and sedimentation coefficient, 6--7S in low salt gradients. The SBP-like protein binds testosterone and progesterone as well as 17 beta-estradiol but does not bind diethylstilbestrol. No receptor-like binding activity could be demonstrated using these techniques. Explant culture and DNA cellulose affinity chromatography were used to remove the SBP-like material before assay of [3H]estradiol binding. Using these techniques, a high affinity (Ka = 10(9) M-1), low capacity (n = 10(-14) mol/mg cytosol protein) estradiol receptor was demonstrated. The putative turtle receptor exhibits steroid specificity and sedimentation profiles (6S and 8S in low salt, 4S and 5S in high salt) comparable to those of estrogen receptors in mammalian species. These results suggest a certain degree of physiochemical similarity between putative estrogen receptors in mammalian and turtle reproductive tracts.

Animals↗
PubMed1989

The hormonogenic sites of turtle thyroglobulin and their homology with those of mammals.

Thyroglobulin (Tg) from turtles previously injected with 125I was reduced, alkylated, and digested with trypsin. We purified the resultant peptides on HPLC columns, determined their amino acid sequences and the locations of [125I]T4 and [125I]T3 residues, and compared them with established sequences from humans, cows, rabbits, rats, and guinea pigs. We found five major T4 peptides, three of which were homologous with the major hormonogenic sites A, B, and D of mammalian Tg. Site A, the highly conserved major T4 site in mammals, had substitutions in three residues near the T4 residue and had much less of Tg's newly synthesized T4 than is found in mammalian Tg (25% in turtle vs. 44% in rabbit). Site B contained correspondingly more of Tg's new T4 (42% vs. 24% in rabbit). Turtle Tg contained little [125I]T3, and we did not find site C (Ser-T3/T4-Ser, the major T3 site in guinea pig and rabbit) in turtles, but did find Val-T4, a possible homolog. Site D was quantitatively less important than in mammals. The fifth turtle hormonogenic site, containing 12% of Tg's newly formed T4, had a tyrosyl residue substituted for the phenylalanine at residue 632 in the human sequence. We conclude that Tg's major hormonogenic sites are generally conserved across a considerable evolutionary distance, but that differences in primary structure occur and may contribute to changes in priority of hormone synthesis among these sites.

Alkylation↗
PubMed2003

Magnet-induced disorientation in hatchling loggerhead sea turtles.

Laboratory experiments have indicated that hatchling loggerhead sea turtles (Caretta caretta) are able to establish and maintain courses using information from the Earth's magnetic field. In previous experiments, turtles were tested in relatively uniform magnetic fields generated by large coil systems surrounding an orientation arena. In this study, we investigated the orientation behavior of hatchlings with either magnets or magnetically inert brass bars attached to their carapaces. Control turtles (with brass bars) oriented significantly towards the east whereas turtles bearing magnets were not significantly oriented as a group. The two distributions were statistically different. These results indicate that magnetic orientation behavior of hatchling sea turtles can be disrupted by attaching a small magnet to the carapace. This finding may prove useful both in field experiments and in efforts to localize magnetoreceptors.

Animals↗
PubMed2004

To freeze or not to freeze: adaptations for overwintering by hatchlings of the North American painted turtle.

Many physiologists believe that hatchling painted turtles (Chrysemys picta) provide a remarkable, and possibly unique, example of 'natural freeze-tolerance' in an amniotic vertebrate. However, the concept of natural freeze-tolerance in neonatal painted turtles is based on results from laboratory studies that were not placed in an appropriate ecological context, so the concept is suspect. Indeed, the weight of current evidence indicates that hatchlings overwintering in the field typically withstand exposure to ice and cold by avoiding freezing altogether and that they do so without benefit of an antifreeze to depress the equilibrium freezing point for bodily fluids. As autumn turns to winter, turtles remove active nucleating agents from bodily fluids (including bladder and gut), and their integument becomes a highly efficient barrier to the penetration of ice into body compartments from frozen soil. In the absence of a nucleating agent or a crystal of ice to 'catalyze' the transformation of water from liquid to solid, the bodily fluids remain in a supercooled, liquid state. The supercooled animals nonetheless face physiological challenges, most notably an increased reliance on anaerobic metabolism as the circulatory system first is inhibited and then caused to shut down by declining temperature. Alterations in acid/base status resulting from the accumulation of lactic acid may limit survival by supercooled turtles, and sublethal accumulations of lactate may affect behavior of turtles after the ground thaws in the spring. The interactions among temperature, circulatory function, metabolism (both aerobic and anaerobic), acid/base balance and behavior are fertile areas for future research on hatchlings of this model species.

Adaptation, Physiological↗
PubMed2005

Bioenergetics and diving activity of internesting leatherback turtles Dermochelys coriacea at Parque Nacional Marino Las Baulas, Costa Rica.

Physiology, environment and life history demands interact to influence marine turtle bioenergetics and activity. However, metabolism and diving behavior of free-swimming marine turtles have not been measured simultaneously. Using doubly labeled water, we obtained the first field metabolic rates (FMRs; 0.20-0.74 W kg(-1)) and water fluxes (16-30% TBW day(-1), where TBW=total body water) for free-ranging marine turtles and combined these data with dive information from electronic archival tags to investigate the bioenergetics and diving activity of reproductive adult female leatherback turtles Dermochelys coriacea. Mean dive durations (7.8+/-2.4 min (+/-1 s.d.), bottom times (2.7+/-0.8 min), and percentage of time spent in water temperatures (Tw) < or =24 degrees C (9.5+/-5.7%) increased with increasing mean maximum dive depths (22.6+/-7.1 m; all P< or =0.001). The FMRs increased with longer mean dive durations, bottom times and surface intervals and increased time spent in Tw< or =24 degrees C (all r2> or =0.99). This suggests that low FMRs and activity levels, combined with shuttling between different water temperatures, could allow leatherbacks to avoid overheating while in warm tropical waters. Additionally, internesting leatherback dive durations were consistently shorter than aerobic dive limits calculated from our FMRs (11.7-44.3 min). Our results indicate that internesting female leatherbacks maintained low FMRs and activity levels, thereby spending relatively little energy while active at sea. Future studies should incorporate data on metabolic rate, dive patterns, water temperatures, and body temperatures to develop further the relationship between physiological and life history demands and marine turtle bioenergetics and activity.

Animals↗
PubMed1999

Comparative pathology and pathogenesis of spontaneous and experimentally induced fibropapillomas of green turtles (Chelonia mydas).

Tumor biopsy samples from 25 Floridian and 15 Hawaiian green turtles (Chelonia mydas) with spontaneous green turtle fibropapillomatosis (GTFP) and from 27 captive-reared green turtles with experimentally induced GTFP were examined microscopically to differentiate the histologic features that result from GTFP pathogenesis and those that result from incidental factors that may vary according to geographic region. Common histologic features for spontaneous and experimentally induced tumors included fibroblast proliferation in the superficial dermis, epidermal acanthosis and hyperkeratosis, epidermal basal cell degeneration with dermal-epidermal cleft formation, spinous layer degeneration with intraepidermal vesicle and pustule formation, and ulceration. Visceral tumors, found in eight of 10 (80%) free-ranging turtles with cutaneous disease that were examined after death, had extensive interstitial fibrous proliferation. The presence of spirorchid trematode eggs and associated foreign body granulomas, common secondary findings within spontaneous tumors, varied by geographic location, and these findings were not observed in experimentally induced tumors. Eosinophilic intranuclear inclusions and intranuclear herpesvirus-associated antigen immunoreactivity were found in 18 of 38 (47%) experimentally induced cutaneous tumors and nine of 119 (7.5%) spontaneous tumors from Floridian but not Hawaiian turtles. The possible involvement of GTFP-associated herpesvirus in the pathogenesis of epidermal degenerative changes and GTFP pathogenesis is discussed.

Animals↗
PubMed2005

Renal myxozoanosis in crowned river turtles Hardella thurjii: description of the putative agent Myxidium hardella n. sp. by histopathology, electron microscopy, and DNA sequencing.

Chelonian myxozoanosis is rarely reported and has previously not been documented to cause disease. This report describes myxozoanosis associated with significant renal disease in two Crowned River turtles (Hardella thurjii). One turtle presented with emaciation and died. The cage mate presented with emaciation and was euthanized. Histologically, renal intratubular myxozoan spores were associated with renal tubular necrosis, tubular mineralization, and chronic interstitial nephritis, with membranoproliferative and mes-angioproliferative glomerulopathy. Both turtles also had disseminated metastatic mineralization. On the basis of these findings, chronic renal insufficiency from myxozoanosis and subsequent metastatic mineralization were considered the primary problems. By light and electron microscopy, the myxozoan spores had features of the genus Myxidium. Maximum parsimony analysis of small-subunit rDNA sequences placed the turtle myxozoan basal to a clade containing Myxidium truttae and a Myxidium sp. with strong bootstrap support. This myxozoan agent appears to be a significant pathogen in H. thurjii on the basis of morphologic changes in the kidneys of in the infected turtles.

Animals↗
PubMed2002

Pharmacokinetics of fluconazole in loggerhead sea turtles (Caretta caretta) after single intravenous and subcutaneous injections, and multiple subcutaneous injections.

Superficial and systemic mycotic infections are common among clinically ill sea turtles, which places growing importance on the establishment of pharmacokinetic-based dosage regimens for antifungal drugs. The pharmacokinetic properties of the antifungal drug fluconazole, after intravenous (i.v.) and subcutaneous (s.c.) injections, were studied in juvenile loggerhead sea turtles (Caretta caretta) housed at 23.0-26.5 degrees C. Fluconazole pharmacokinetic properties were further assessed in a multiple-dose s.c. regimen derived from the pharmacokinetic parameters determined in the single-dose study. Pharmacokinetic parameters were calculated, using a two-compartment model, from plasma concentration-time data obtained after single i.v. and s.c. administrations of fluconazole at a dosage of 2.5 mg/ kg body weight in six juvenile sea turtles. Blood samples were collected at intervals through 120 hr after each dose, and the concentration of fluconazole in plasma was measured by reverse-phase high-performance liquid chromatography. The i.v. and s.c. elimination half-lives were 139.5 +/- 36.0 and 132.6 +/- 48.7 hr (mean +/- SD), respectively. Systemic clearance of fluconazole was 8.2 +/- 4.3 ml/kg x hr, and the apparent volume of distribution at steady state was 1.38 +/- 0.29 L/kg. A multiple-dose regimen was derived, which consisted of a loading dose of 21 mg/kg body weight and subsequent doses of 10 mg/kg administered through s.c. injection every 120 hr (5 days). This regimen was administered to four juvenile sea turtles for 10 days, and blood samples were taken to determine peak and trough plasma concentrations of fluconazole. The mean concentrations for the two peak concentrations were 16.9 +/- 1.1 and 19.1 +/- 2.8 microg/ml 4 hr after dosing, and the mean concentrations for the three trough concentrations were 7.2 +/- 2.2, 10.4 +/- 2.7, and 10.7 +/- 2.9 microg/ml 120 hr after dosing. The terminal half-life after the last dose was calculated at 143 hr. Throughout the multiple dosing, fluconazole concentrations remained above approximately 8 microg/ml, a concentration targeted when treating mycotic infections in humans. The results of this study suggest that fluconazole can be effectively administered to sea turtles at a dosage of 10 mg/kg every 5 days after a loading dose of 21 mg/kg.

Animals↗
PubMed2003

Steady-state plasma concentrations of itraconazole after oral administration in Kemp's ridley sea turtles, Lepidochelys kempi.

Pharmacokinetic studies of antifungal agents in reptiles are uncommon. Itraconazole, which has been used prophylactically in juvenile sea turtles suffering from hypothermia (cold stunning) on a regular basis, was evaluated for steady-state plasma concentrations. Five Kemp's ridley sea turtles (Lepidochelys kempi) receiving itraconazole at several dosages in a rehabilitation program had blood collected within 24 hr to estimate dosing frequency. Subsequently, serial blood samples of Kemp's ridley sea turtles that were given itraconazole at several dosages for 30 days to treat cold stunning were collected at various intervals to evaluate itraconazole plasma concentrations. Tissue samples were collected from one Kemp's ridley that died during rehabilitation. Plasma concentrations of itraconazole (and of hydroxyitraconazole [OH-ITRA], one of its major bioactive metabolites) were determined using a modified, validated reverse-phase high-performance liquid chromatography technique. Itraconazole concentrations in tissues were determined by bioassay to be far greater than the plasma concentrations measured in any of the turtles. At a 15-mg/kg dosage, the half-life (t1/2) was 75 hr for itraconazole and 55 hr for OH-ITRA. All dosages produced adequate concentrations in some turtles, but consistent therapeutic concentrations were produced only at 15 mg/kg q72hr and 5 mg/kg s.i.d., with the latter producing the highest plasma concentrations.

Administration, Oral↗
PubMed2006

Vitellogenin in black turtle (Chelonia mydas agassizii): purification, partial characterization, and validation of an enzyme-linked immunosorbent assay for its detection.

Black turtle plasmatic vitellogenin (VTG) was purified from 17beta-estradiol-induced males using ion-exchange chromatography. The isolated protein was identified as VTG by its glycolipoprotein nature and amino acid sequence homology with other vertebrate VTG. It was characterized as a 500-kDa dimer composed of two identical, 200- to 240-kDa monomers. Polyclonal antibodies raised against black turtle VTG showed high titer and specificity, as demonstrated by enzyme-linked immunosorbent assay and Western blot analysis, respectively. The range of the assay was estimated to be between 15 ng/ml and 2 microg/ml, and the inter- and intra-assay coefficients of variation were 9.4 and 7.3%, respectively. Black turtle antibody cross-reacted with VTG of two other sea turtle species, Caretta caretta (loggerhead) and Eretmochelys imbricata (hawksbill), extending the applicability of the assay as part of a sea turtle health assessment program.

Animals↗
PubMed2003

Use of rocuronium for endotracheal intubation of North American Gulf Coast box turtles.

OBJECTIVE: To determine whether rocuronium, a reversible neuromuscular blocking agent, would provide safe, short-term immobilization to facilitate endotracheal intubation in turtles. DESIGN: Prospective study. ANIMALS: 30 healthy adult Gulf Coast box turtles. PROCEDURE: Turtles were given rocuronium, and responses were recorded every 3 minutes. Times to onset of effects, intubation, and recovery were recorded and analyzed for associations with dose and patient characteristics to determine an optimal dose range. Neostigmine and glycopyrrolate were given to augment recovery from neuromuscular blockade. RESULTS: Rocuronium administered at a dose of 0.25 to 0.5 mg/kg (0.11 to 0.23 mg/lb), IM, permitted intubation; lower doses were not effective. Mean +/- SD time to loss of the palpebral reflex was 6.4 +/- 4.0 minutes, and mean time to intubation was 9.2 +/- 6.4 minutes. Mean time to return of the palpebral reflex was 44 +/- 13.2 minutes, and mean time to walking was 55 +/- 16.6 minutes. Time to onset of effects was not associated with dose, but recovery times were prolonged with higher doses of rocuronium. Cardiac arrhythmias were observed in 13 (43%) turtles. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of rocuronium at a dose of 0.25 to 0.5 mg/kg is a safe and effective adjunct to general anesthesia in Gulf Coast box turtles. Because rocuronium does not provide any analgesic or sedative effects, the duration of neuromuscular blockade without anesthesia should be minimized to avoid undue distress.

Androstanols↗