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PubMed1979

Turtle-dove ovomucoid, a glycoprotein proteinase inhibitor with P1-blood-group antigen activity.

Ovomucoid from the egg white of turtle-dove (Streptopelia risoria) was purified and shown to be a glycoprotein of mol. wt. 29 400, with valine as N-terminal residue. It is an inhibitor of both trypsin and chymotrypsin, but has a lower affinity for trypsin than has hen ovomucoid. Turtle-dove ovomucoid contains antigenic activity cross-reacting with the blood-group-P1 antigen of human erythrocytes. Hen ovomucoid has no detectable blood group-P1 activity. The carbohydrate composition of turtle-dove ovomucoid differs from hen ovomucoid in having substantially higher galactose content. The possible relationship between carbohydrate composition and antigenic activity is discussed.

Amino Acids↗
PubMed1994

An Outbreak of Aeromonas hydrophila Infection in Turtles (Pseudemis scripta).

An outbreak of Aeromonas hydrophila infection with a high rate of mortality (95%) in turtles (Pseudemis scripta) in Italy is reported. Pure cultures of the pathogen were isolated from liver, lung, kidney, and heart specimens of the turtles. The A. hydrophila isolate was resistant to amoxicillin, ampicillin, cephalothin, and trimethoprim-sulfamethoxazole but was sensitive to a number of other antibiotics tested. The study indicates that pet turtles can act as reservoirs of this pathogen and may play an important role in the etiology of Aeromonas-associated human infections.

Journal Article↗
PubMed1990

31P-NMR measurements of pHi and high-energy phosphates in isolated turtle hearts during anoxia and acidosis.

We used 31P-nuclear magnetic resonance (NMR) spectroscopy to measure intracellular pH (pHi) and high-energy phosphate levels in hearts of turtles (Chrysemys picta bellii) during either 4 h of anoxia [extracellular pH (pHo) 7.8, 97% N2-3% CO2], 4 h of lactic acidosis (pHo 7.0, 97% O2-3% CO2), or 1.5 h of combined anoxia + lactic acidosis (pHo 7.0, 97% N2-3% CO2) followed by 2 h of oxygenated recovery (pHo 7.8) at 20 degrees C. We also measured heart rate, maximum ventricular-developed pressure, and rate of pressure development (dP/dtmax). 31P-NMR spectra were characterized by the seven peaks typical of mammalian hearts, although turtle spectra were dominated by a large phosphodiester peak. Anoxia caused an increase in Pi to 165% and a decrease in creatine phosphate (CP) to 42% of control, whereas ATP levels remained unchanged. pHi declined from 7.37 +/- 0.01 to 7.22 +/- 0.03 at 1 h of anoxia and remained unchanged through hour 4. Lactic acidosis caused a 59% decrease in Pi, whereas CP and ATP levels remained unchanged. pHi fell to 6.88 +/- 0.04 by hour 1 and then climbed steadily to 7.14 +/- 0.05 at hour 4. During recovery from acidosis, pHi exceeded control values and returned to control by 2 h. Combined anoxia + acidosis caused profound decreases in CP to 14% and pHi to 6.56 +/- 0.03. In anoxic hearts, cardiodynamic variables remained at control levels through hour 3, after which cardiac output, heart rate, and dP/dtmax declined. Cardiodynamic variables were essentially unchanged from control throughout 4 h of acidosis except for dP/dtmax, which declined rapidly. In the combined protocol, all measures of cardiac function decreased. Recovery in all three cases was complete by approximately 2 h. We conclude that turtle hearts were relatively resistant to the stresses imposed in all three protocols compared with mammalian hearts, although anoxia + acidosis depressed the measured cardiac variables more profoundly than predicted from responses to the conditions imposed separately. Our results from the anoxia protocol suggest no direct causal relationship between myocardial CP (or ATP) levels and cardiac function.

Acidosis↗
PubMed1994

OXYGEN TRANSPORT IN MARINE GREEN TURTLE (CHELONIA MYDAS) HATCHLINGS: BLOOD VISCOSITY AND CONTROL OF HEMOGLOBIN OXYGEN-AFFINITY

Erythrocytes from green turtle hatchlings contain a single embryonic component, unlike those from other cleidoic eggs, in which adult hemoglobin (Hb) constitutes a significant fraction of total Hb at hatching. The functional properties of the isolated and purified green turtle hatchling Hb that distinguish it from adult Hb are a high affinity for oxygen and marked sensitivity to organic phosphate modulators. Hatchling erythrocytes also contain higher concentrations of ATP and 2,3-diphosphoglyceric acid, but their oxygen affinity is indistinguishable from that of adult erythrocytes. Hatchling erythrocyte mean cell volume is approximately half of the adult value, but hematocrit, blood hemoglobin concentration and blood viscosity of hatchlings and adults are similar. Oxygen-carrying capacity in green turtles, unlike that of other diving vertebrates, corresponds with a theoretically derived optimum. The possibility of allosteric control of Hb oxygen-binding in hatchlings may relate not to the challenge of exercise during the dispersal phase but to conditions in the late embryo in the nest.

Journal Article↗
PubMed1994

A MICROCALORIMETRIC STUDY OF TURTLE CORTICAL SLICES: INSIGHTS INTO BRAIN METABOLIC DEPRESSION

In previous papers, we have examined turtle cortical neurons in vitro for mechanisms of anoxic metabolic depression ('channel arrest' and changes in electrical parameters). Negative results prompted the current study with the aim of examining more closely the energy profile and metabolism of turtle cortical slices. Calorimetry is used to measure heat dissipation during normoxia and nitrogen perfusion (120 min) and the results are converted into an ATP utilization rate. These indicate that the control rate of ATP utilization (1.72 µmol ATP g-1 min-1) agrees closely with in vivo whole-brain metabolic measurements. Both nitrogen perfusion and pharmacologically induced anoxic (cyanide+N2) groups depressed heat dissipation considerably compared with the control value (nitrogen 37 %; pharmacological anoxia 49 %). The resulting ATP utilization estimates indicate metabolic depressions of 30 % (nitrogen) and 42 % (pharmacological anoxia). The slice preparation did not exhibit a change in any measured adenylate parameter for up to 120 min of anoxia or pharmacological anoxia. Significant changes did occur in [ADP], ATP/ADP ratio and energy charge after 240 min of exposure to anoxic conditions. These results support the idea that the turtle cortical slice preparation has a profound resistance to anoxia, with both nitrogen perfusion and pharmacological anoxia causing a rapid decline in heat dissipation and metabolism.

Journal Article↗
PubMed1994

MECHANICAL PERFORMANCE OF AN IN SITU PERFUSED HEART FROM THE TURTLE CHRYSEMYS SCRIPTA DURING NORMOXIA AND ANOXIA AT 5 C AND 15 C

We developed an in situ perfused turtle (Chrysemys scripta) heart preparation to study its intrinsic mechanical properties at 5°C and 15°C using normoxic and anoxic perfusion conditions. The in situ preparation proved durable and stable. At 15°C and a spontaneous heart rate of 23.4 beats min-1, maximum stroke volume was 2.54 ml kg-1 body mass, maximum cardiac output was 62.5 ml min-1 kg-1 and maximum cardiac myocardial power output was 1.50 mW g-1 ventricular mass. There was good agreement between these values and those previously obtained in vivo. Furthermore, since the maximum stroke volume observed here was numerically equivalent to that observed in ventilating C. scripta in vivo, it seems likely that C. scripta has little scope to increase stroke volume to a level much beyond that observed in the resting animal through intrinsic mechanisms alone. The ability of the perfused turtle heart to maintain stroke volume when diastolic afterload was raised (homeometric regulation) was relatively poor. At 5°C, the spontaneous heart rate (8.1 beats min-1) was threefold lower and homeometric regulation was impaired, but maximum stroke volume (2.25 ml kg-1) was not significantly reduced compared with the value at 15°C. The significantly lower maximum values for cardiac output (18.9 ml min-1 kg-1) and power output (0.39 mW g-1 ventricular mass) at 5°C were largely related to pronounced negative chronotropy with only a relatively small negative inotropy. Anoxia had weak negative chronotropic effects and marked negative inotropic effects at both temperatures. Negative inotropy affected pressure development to a greater degree than maximum flow and this difference was more pronounced at 5°C than at 15°C. The maximum anoxic cardiac power output value at 15°C (0.77 mW g-1 ventricular mass) was not that different from values previously obtained for the performance of anoxic rainbow trout and hagfish hearts. In view of this, we conclude that the ability of turtles to overwinter under anoxic conditions depends more on their ability to reduce cardiac work to a level that can be supported through glycolysis than on their cardiac glycolytic potential being exceptional.

Journal Article↗
PubMed1999

Cardiorespiratory response to progressive hypoxia and hypercapnia in the turtle trachemys scripta

The ventilatory responses of chelonian reptiles to hypoxic and hypercapnic stress have been fairly well described. As turtles are capable of large cardiac shunts, changes in pulmonary perfusion may be an equally viable and potent response to these stressors. To test this hypothesis, conscious unrestrained turtles were unidirectionally ventilated while blood flow in the left pulmonary artery ( q_dot (LPA)) and left aortic arch ( q_dot (LAo)) was monitored. Turtles were exposed to step changes (2.5 h step(-)(1)) in O(2) tension (30, 15, 5, 2.5 or 0 % O(2); CO(2) inflow maintained constant) on day 1 followed by step changes in CO(2) tension (0, 2, 4, 8 % CO(2); O(2) inflow maintained constant) on day 2. Steady-state cardiorespiratory variables were recorded for the last 30 min of each step change in gas tension.Progressive hypoxia resulted in progressive increases in ventilation, q_dot (LPA) and q_dot (LAo) and a small, but non-significant, increase in heart rate. Progressive hypercapnia resulted in a progressive increase in ventilation, while q_dot (LPA) and q_dot (LAo) did not change at any level of CO(2). These results suggest that information from the O(2)-sensitive chemoreceptors appears to be stimulatory to both the cardiovascular and ventilatory control systems, while CO(2) chemoreception appears to affect primarily the ventilatory control system. These results also suggest that, in animals capable of intracardiac shunting, increasing pulmonary perfusion may be an integral component of the reflex response to hypoxia.

Journal Article↗
PubMed2004

Cutting the longline to extinction: new sea turtle campaign takes aim at industrial longline fishing and mercury-poisoned seafood.

Chanting "Get on the right track . . . stop killing the leatherback!," a festive protest of people of many ages dressed in colorful turtle costumes wound its way along the busy streets of San Francisco's Fishermen's Wharf. The action last October marked the launching of the Bay Area-based Sea Turtle Restoration Project's Save the Leatherback (www.savetheleatherback.com) campaign for a moratorium on longline fishing in the Pacific Ocean. Longline fishing in the Pacific kills tens of thousands of sea turtles annually to serve up swordfish, shark, and tuna poisoned with high levels of methylmercury for lucrative seafood markets in Japan, the United States, and Europe.

Journal Article↗
PubMed1999

Stimulation with N-methyl-D-aspartate or kainic acid increases cyclic guanosine monophosphate-like immunoreactivity in turtle retina: involvement of nitric oxide synthase.

In brain and retina, stimulation with excitatory amino acids (EAA) can generate nitric oxide (NO) and increase levels of cyclic guanosine monophosphate (cGMP). Because nitric oxide synthase (NOS) has been found in retinas of all species examined to date, an NO signal-transduction pathway is likely to be present in all retinas. We tested the hypothesis that stimulation of ionotropic glutamate receptors in turtle retina would result in increases in cGMP through an NOS/NO/cGMP pathway. Following in vitro incubations of turtle eye cups with the glutamate receptor agonists, N-methyl-D-aspartate (NMDA) or kainic acid (KA), we quantified the increases in cGMP-like immunoreactivity (cGMP-LI) by using enzyme-linked immunosorbant assay (ELISA) and localized the increased cGMP-LI by using an antibody against cGMP. Stimulation with NMDA or KA increased cGMP-LI in bipolar and amacrine cells as well as in some somata in the ganglion cell layer. Either KA or NMDA produced statistically significant increases in total retinal cGMP-LI by ELISA. To test the involvement of NO, we used the NOS inhibitors 7-nitroindazole and L-nitroarginine. Both inhibitors blocked virtually all of the KA- or NMDA-stimulated increases in cGMP-LI. These results indicate that activation of ionotropic glutamate receptors can increase cGMP in select retinal neurons. Differences between the agonist-evoked increases of retinal cGMP-LI suggest that there can be specificity in the activation of the NOS/NO/cGMP signal-transduction pathway by glutamate. This suggests that, in addition to short-term electrical changes, activation of ionotropic glutamate receptors also may produce longer term modulatory or metabolic effects involving NO/cGMP.

Animals↗
PubMed1999

Reelin mRNA expression during embryonic brain development in the turtle Emys orbicularis.

The expression of reelin messenger ribonucleic acid (mRNA) was studied during embryonic brain development in the turtle Emys orbicularis, by using radioactive in situ hybridization. A high expression was consistently found in the olfactory bulb and in a few neurons in the marginal zone and, to a lesser extent, in the subplate of the dorsal and medial cortical sectors. In the diencephalon, the ventral division of lateral geniculate nuclei and the prospective reticular thalamic nuclei were strongly positive. High reelin signal was also associated with some layers of the tectum and with the external granule cell layer of the cerebellum. A more moderate signal was detected in the septal nuclei, striatum, dorsal ventricular ridge, retina, habenular nuclei, and hypothalamus, and in some reticular nuclei of the midbrain and hindbrain and in ventral spinal cord. The cortical plate, basal forebrain, amygdala, and tegmentum were weakly labeled. When they are compared to reelin expression during mammalian brain development, our data reveal an evolutionarily conserved canvas of reelin expression and significant differences, particularly in developing cortical fields. Most significantly, the developing turtle cortex does not display the heavy reelin expression in subpial Cajal-Retzius cells that is so typical of its mammalian counterpart. Given the key role of reelin in laminar cortical development, our data suggest that the increase in the number of reelin-producing cells and/or the amplification of reelin expression in the cortical marginal zone might have been a driving factor during the evolution of the laminated cerebral cortex from stem reptiles to mammals, as indicated in previous comparative analyses.

Animals↗
PubMed1997

Protein synthesis under conditions of anoxia and changing workload in ventricle strips from turtle heart.

An earlier study determined that protein synthesis in isolated perfused turtle (Trachemys [= Pseudemys] scripta elegans) hearts was three-fold lower under conditions of anoxia than under conditions of normoxia. However, the earlier study did not attempt to define the role of work in the isolated perfused preparation. In this study, the effects of varying workload, as defined by changing frequency of contraction, and anoxia on protein synthesis were examined. The ventricle strip preparation allows for comparison of multiple strips from a single heart, which aids in eliminating the variability found between individuals chosen from wild populations. Ventricle strips forced to contract at 24 contractions.min-1 under anoxic conditions failed more rapidly than strips forced to contract at 24 contractions.min-1 under normoxic conditions. Protein synthesis decreased by 32% when compared to normoxic controls. When stimulation was terminated after 2 hr of contraction, the rate of protein synthesis in strips under anoxic conditions was similar to that in strips under normoxic conditions. Also, returning strips to normoxic conditions after 2 hr of anoxia restored protein synthesis to the level of the normoxic controls. A significant correlation between pacing rate and protein synthesis was found under normoxic conditions but not under anoxic conditions when strips were paced at 12, 18 and 24 contractions.min-1. Protein synthesis increased by 30% at the 18 contractions.min-1 frequency and 45% at the 24 contractions.min-1 frequency over the rate at 12 contractions.min-1 frequency. Force-frequency studies revealed that under normoxic conditions force generation did not change until above 24 contractions.min-1, but under anoxic conditions there was a significant negative inotropic effect (20% decrease in force) at 24 contractions.min-1 and fell to 50% of initial at 36 contractions.min-1. These studies indicate that, in the turtle heart, anoxia per se is not the only determinant of protein synthesis but rather that work plays an important role in protein synthesis, as in the mammalian heart.

Animals↗
PubMed1999

Basal and stress-induced corticosterone levels in olive ridley sea turtles (Lepidochelys olivacea) in relation to their mass nesting behavior.

Adrenocortical responsiveness to turning stress was examined in wild, reproductively-active olive ridley sea turtles (Lepidochelys olivacea) in relation to their mass nesting (arribada) behavior. We hypothesized that the high sensitivity threshold (HST) observed in ovipositing sea turtles is associated with a diminished sensitivity of the hypothalamo-pituitary-adrenal (HPA) axis to stressful stimuli in arribada females. We tested this hypothesis by determining whether arribada females exhibited an increased activation threshold of the HPA axis to an imposed stressor (turning stress). Mean basal corticosterone (B) and glucose levels were below 1.0 ng/ml and 60 mg/dl, respectively. Basal B remained unchanged throughout a 24-hr period in basking females. Most animals responded to turning stress with elevated mean B levels (up to 6.5 ng/ml after 6 hr) and no increase in circulating glucose. Nearly 50% of females (and none of the males) were refractory to the stimulation. Males exhibited the most rapid response, with B levels significantly elevated by 20 min over basal levels. Among females, arribada and solitary nesters exhibited a slower rate of response than basking, non-nesting animals. These results demonstrate that olive ridleys exhibit stress-induced changes in circulating B which are slower than those observed in most reptilian and in mammalian, avian, and piscine species. Furthermore, the presence of refractory females and the relatively slower increase in B in arribada and solitary nesters indicate a hyporesponsiveness of the HPA axis to turning stress in nesting olive ridleys. The hyporesponsiveness may be part of a mechanism to facilitate arribada nesting. J. Exp. Zool. 284:652-662, 1999.

Animals↗
PubMed1996

Motor pool organization of the external gastrocnemius muscle in the turtle, Pseudemys (Trachemys) scripta elegans.

The spinal cord of the adult turtle, Pseudemys (Trachemys) scripta elegans, is now considered a promising model for the study of the segmental motor system in the generalized tetrapod. To facilitate such studies we have examined the location, soma geometry, soma size, and number of motoneurons innervating the external gastrocnemius (EG) muscle in this species, as this muscle is ideally suited to the study of interrelations between the neuronal and muscular components of the segmental motor system. Motoneurons were retrogradely labeled following application of horseradish peroxidase to the EG muscle nerve. In both horizontal and transverse planes, labeled motoneurons innervating the EG muscle were concentrated in the S1 lumbosacral segment, and extended rostrally and caudally as far as the exists of the D10 and S2 spinal nerves, respectively. In the transverse plane, motoneurons were arranged in a longitudinal column which occupied the dorsolateral quadrant of the ventral horn. EG motoneurons are fusiform in shape and present their largest dimension in the transverse plane with their long axis oriented in the ventromedial to dorsolateral plane. The soma diameters of EG motoneurons were normally distributed, reflecting the absence of separate fusimotor innervation in reptilian species. In individual turtles, there was a two- to threefold range in soma diameter while soma surface area extended over a seven- to tenfold range. Based on cell counts from five animals, the EG motor pool was composed of approximately 75 motoneurons. Taken together, the results of this study provide valuable information for interpreting the results of future studies on the segmental motor system of this species under both normal and pathophysiological conditions.

Animals↗
PubMed2000

Mitogen-activated protein kinases and anoxia tolerance in turtles.

The response of two vertebrate mitogen-activated protein kinase (MAPK) family members, the extracellular signal-regulated kinases (ERKs) and c-Jun NH2-terminal kinases (JNKs), to anoxia exposure in vivo was examined in organs (liver, heart, kidney, brain, spleen) of the anoxia-tolerant adult turtle, Trachemys scripta elegans. ERKs activities rose during anoxia only in spleen (a 2.8-fold increase). JNK activity showed a significant increase only in liver (4-fold increase) after 5 hr of anoxic submergence but declined thereafter. Levels of the transcription factor c-Fos were strongly suppressed in liver, heart, and kidney of anoxia-exposed turtles, whereas levels increased 2-fold in anoxic brain. The effect of anoxia on c-Myc was organ-specific and variable with 2- and 1.5-fold increases in protein expression in kidney and brain, respectively, and a 60% decrease in anoxic spleen. These results for an anoxia-tolerant animal suggest the potential importance of the MAPKs and of the immediate-early genes (c-fos, c-myc) in mediating adaptive responses to oxygen deprivation.

Adaptation, Biological↗
PubMed2001

Influence of diminished respiratory surface area on survival of sea turtle embryos.

It has been suggested that fungal presence on sea turtle eggs may impede gas exchange. To investigate the influence of diminished gas exchange surface upon embryo survivorship, flatback (Natator depressus) and green (Chelonia mydas) eggs were painted with petroleum jelly. Variable proportions of the egg surface were covered, including both respiratory and nonrespiratory domains. Embryo survival varied with site inhibited, proportion of eggshell affected, and species of turtle. If fungi on the exterior of the eggshell are able to impede respiratory gas exchange, their presence on the upper hemisphere (primary gas exchange area in early incubation) will result in the highest embryo mortality. Large eggs are likely to demonstrate a higher survivorship than small eggs, due to their larger available respiratory area and/or to variation in weight or stage-specific embryonic metabolic demands. Interspecific differences in egg size may therefore be a contributory factor to observed mortality rate differences in the natural presence of fungi.

Animals↗
PubMed1991

Ultrastructure of the turtle spermatozoon.

The turtle spermatozoon is vermiform in shape with a narrow pointed head that is curved. In general, the turtle sperm contains a typical head, midpiece and tail, similar in morphology to that of birds, amphibians and other reptiles. However, several structures are unique. These unusual features include (1) a perforatorial cap over the proximal end of the nucleus, which contains 2-3 rods that are contiguous with intranuclear tubules; (2) a connecting collar of dense material that surrounds the base of the nucleus; (3) a distal centriole containing central microtubules that extend its entire length and having outer triplicate microtubules that open toward the central cavity of the centriole; and (4) unusual spherical mitochondria containing 7-8 outer laminated membranes.

Animals↗
PubMed2001

Turtles as hopeful monsters.

A recently published study on the development of the turtle shell highlights the important role that development plays in the origin of evolutionary novelties. The evolution of the highly derived adult anatomy of turtles is a prime example of a macroevolutionary event triggered by changes in early embryonic development. Early ontogenetic deviation may cause patterns of morphological change that are not compatible with scenarios of gradualistic, stepwise transformation.

Animals↗
PubMed2002

Associations between the morphology and physiology of ventral-horn neurons in the adult turtle.

This study compared some morphologic and physiological properties of adult turtle spinal motoneurons (MNs) vs. interneurons (INs). Reconstructions were made of 20 biocytin-stained cells, which had been previously studied physiologically in 2-mm-thick slices of lumbosacral spinal cord. The intracellularly measured physiological properties included resting potential, input resistance (R(N)), threshold (rheobase, I(Rh)), and slope of the stimulus current (I) -spike frequency (f) relation. The seven morphologic properties that were quantified for each cell included three indices of somal size (diameter, area, volume), and four of dendritic size: the number of first- and last-order branches, rostrocaudal extent, and sigma individual lengths. Significant differences were shown between all seven morphologic parameters for MNs vs. INs. Despite the small sample size, significant differences were also shown for five of seven parameters for high-threshold vs. low-threshold MNs, and three of seven for low-threshold MNs vs. INs. These latter three parameters were the number of terminal dendritic branches, their rostrocaudal extent, and the sigma dendritic lengths. Linear associations for the MN + IN and the MN samples were stronger between the four dendritic parameters than between soma-dendritic ones. Exponential associations between morphologic and physiological properties were mostly significant (28 of 30), and their strength was in the order I(Rh) < R(N) < f/I slope for the MN +IN sample and I(Rh) < R(N) = f/I slope for the MN sample. There is discussion of the relevance of the above findings to the provisional classification of turtle ventral-horn neurons on the basis of electrophysiology alone.

Action Potentials↗