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PubMed1986

Circadian rhythms in serotonin, norepinephrine, and epinephrine contents of the pineal-paraphyseal complex of the soft-shelled turtle (Lissemys punctata punctata).

Diurnal variations in pineal serotonin, norepinephrine, and epinephrine levels were measured at four different time intervals (06:00, 12:00, 18:00, and 24:00 hr) in adult male soft-shelled turtles. The serotonin level increased during day and decreased during night. But the norepinephrine and epinephrine levels showed reverse changes. The findings reveal that there is a distinct circadian rhythm in serotonin, norepinephrine, and epinephrine contents of the pineal-paraphyseal complex in adult male soft-shelled turtles.

Animals↗
PubMed1987

Effects of reserpine and chlorpromazine on the serotonin, norepinephrine, and epinephrine levels in the pineal-paraphyseal complex of soft-shelled turtles (Lissemys punctata punctata).

Turtles were injected intramuscularly with reserpine (0.5 mg/100 g body wt) and chlorpromazine (10 mg/100 g body wt) at 2400, 1800, and 0600 hr and were sacrificed at 6, 12, and 24 hr, respectively after the treatments, i.e., at 0600 hr. The spectrofluorometric findings revealed that the serotonin, norepinephrine, and epinephrine contents of the pineal-paraphyseal complex decreased at 6 hr, increased at 12 hr, and declined again at 24 hr after the treatments. It is suggested that reserpine and chlorpromazine act largely by favoring release with transient resynthesis of these amines in the pineal-paraphyseal complex of soft-shelled turtles.

Animals↗
PubMed1987

Localization of a salmon calcitonin-like molecule in one type of ultimobranchial cells in the freshwater turtle Pseudemys scripta.

Immunohistochemical methods using affinity adsorbed antibodies raised against the three families of calcitonins (CT) were applied to ultimobranchial (UB) cells in situ to investigate the nature of the Chelonian calcitonin molecule and its distribution in the ultimobranchial bodies of the freshwater turtle, Pseudemys scripta. In this species, the UB glands were present on both sides and consisted of scattered cell clumps between epithelial vesicular structures. The neighboring parathyroid tissue also contained two components, the majority being composed of similar vesicles, with occasional solid cell cords evenly distributed. Calcitonin immunoreactivity was found in the cell clumps of the UB gland and in the cell cords of the associated parathyroid, but not in the epithelial component lining the vesicles or in the amorphous material which sometimes filled the lumen. Turtle calcitonin was exclusively of the salmon type, as determined by the negative results obtained in situ after the use of antibodies raised against human and porcine molecules. The salmon-like calcitonin content of the ultimobranchial area was estimated as 15.2 ng; however, the molecule was undetectable in the circulation. In this work we localize the quantitate a salmon-like CT molecule in one type of ultimobranchial and parathyroid cell of a reptile for the first time.

Animals↗
PubMed1989

Increased levels of arginine vasotocin and neurophysin during nesting in sea turtles.

Arginine vasotocin (AVT) and neurophysin (NP) levels were measured by radioimmunoassay in two species of sea turtle, the olive ridley, Lepidochelys olivacea, and the loggerhead, Caretta caretta, during the brief period of nesting and oviposition. In both species, AVT was low in animals which were not reproductively active. AVT was also low at the time animals emerged from the surf to nest, but increased significantly during oviposition and then declined as the animals returned to the water. NP increased in concert with AVT, also reaching highest levels during oviposition. In both species, however, NP levels remained elevated over prenesting levels at the time of return to the water. These findings are consistent with the hypothesis that an AVT-neurophysin complex is released from the neurohypophysis during nesting, and that AVT is a physiological regulator of oviducal contractions in sea turtles.

Animals↗
PubMed1989

Effects of hypophysectomy and ovariectomy on hepatic estrogen receptor content in the turtle, Chrysemys picta.

Estrogen receptor (ER) content in the cytosol and nuclear extract of the liver of adult female Chrysemys picta (control females) was measured in February, May, early June, and late June. Both cytosolic (C) and nuclear (N) hepatic ER content rose from low levels (C: 189 +/- 19; N: 52 +/- 23 fmol/g tissue) in February to significantly higher levels in May (C: 437 +/- 15; N: 124 +/- 25 fmol/g tissue) before declining to lower values in early June (C: 298 +/- 19; N: 118 +/- 20 fmol/g tissue) and late June (C: 274 +/- 22; N: 89 +/- 13 fmol/g tissue). This pattern of seasonal variation in hepatic ER content was also observed in gonadectomized females. Levels of hepatic ER content in ovariectomized turtles fluctuated in the same manner as those found in control females. However, elevations in hepatic ER contents were not observed in hypophysectomized females. Removal of the pituitary resulted in a drastic reduction in the May value (316 +/- 49 fmol/g tissue) of cytosolic ER content and a complete blockade in a rise of nuclear ER content in the liver of the turtle. Nuclear hepatic ER content remained at around 50 fmol/g tissue throughout the months of May and June. The two surgeries procedures caused no significant changes in the ER's affinity for estrogen. The present findings demonstrate a direct control of the pituitary over hepatic ER content which signifies important pituitary regulation of the vitellogenic process, at the liver level, during the spring reproductive season.

Animals↗
PubMed1990

Steroidal modulation of pituitary gonadotropin-releasing hormone responsiveness in young turtles, Pseudemys scripta.

Steroid-modulated pituitary secretion and glandular content of gonadotropin (Gth: LH and FSH) was studied in young slider turtles. Injection (ip) of both 17 beta-estradiol (E2) and testosterone (T) reduced pituitary content of both Gths and caused significant inhibition of basal LH secretion and gonadotropin-releasing hormone (GnRH)-stimulated LH and FSH secretion measured in vitro. However, gonadectomy did not affect pituitary Gth secretion or response in these juveniles, and anti-estrogen and anti-androgen compounds had some steroid agonistic action on the pituitary gland. Exposure to E2, T, and 5 alpha-dihydrotestosterone (DHT) in vitro for 4, 24, or 48 hr either had no effect or completely inhibited pituitary GnRH responsiveness. Progesterone (P) alone had no effect on pituitary GnRH response and in combination did not alter the typical inhibitory effect of E2. There were several indications of differential effects of steroids on secretion of the two Gths, especially in response to GnRH and tetraethyl chloride (receptor independent) stimulation. The results suggest that steroids may act directly at the pituitary level to alter Gth secretion and that steroidal modulation of pituitary secretion might play a role in differential regulation of LH and FSH in turtles.

Animals↗
PubMed1991

Isolation, purification and primary structure of insulin from the turtle Chrysemys dorbigni.

Insulin A and B chains from pancreas of the turtle Chrysemys dorbigni have been purified to homogeneity, and their primary structures have been determined. The sequence of the A chain is G-I-V-E-Q-C-C-H-N-T-C-S-L-Y-Q-L-E-N-Y-C-N, and that of the B chain is A-A-N-Q-H-L-C-G-S-H-L-V-E-A-L-Y-L-V-C-G-E-R-G-F-F-Y-S-P-K-A. The amino acid sequence of Chrysemys insulin is identical to that of another turtle (Pseudemys scripta), the chicken, and turkey. When compared with alligator insulin, it has three conservative substitutions in the B chain. However, there are seven substitutions when compared with the insulin of the rattlesnake.

Amino Acid Sequence↗
PubMed1991

In vitro effect of insulin on the uptake of glucose and alpha-aminoisobutyric acid in the thyroid gland of the turtle (Chrysemys dorbigni).

The presence of specific insulin binding sites in the thyroid gland of the turtle Chrysemys dorbigni has been previously reported. The purpose of the present work was to investigate the probable action of insulin on the uptake of [14C]deoxy-D-glucose ([14C]DG) and [14C]alpha-aminoisobutyric acid ([14C]AIB) in turtle (C. dorbigni) thyroid glands in vitro. Thyroid fragments (+/- 40 mg) were incubated at 25 degrees in Krebs-Ringer-bicarbonate buffer containing 0.2 microCi of [14C]DG or [14C]AIB without or with bovine insulin at different periods of time. The uptake of [14C]DG and [14C]AIB increased with incubation time. The presence of insulin (7 x 10(-6) M) in the incubation medium during 240 min did not modify the [14C]DG uptake. However if the thyroid fragments were previously incubated with insulin (60 min) and then incubated (240 min) with the same concentration of the hormone, the [14C]DG uptake was markedly increased. This stimulatory effect of insulin was dose-dependent. In similar experimental conditions, insulin also produced a significant increase in the uptake of [14C]AIB. Therefore, these findings strongly support the hypothesis that insulin might exert a direct action on the thyroid function.

Aminoisobutyric Acids↗
PubMed1991

Plasma estradiol-17 beta, progesterone, prostaglandin F, and prostaglandin E2 concentrations during natural oviposition in the loggerhead turtle (Caretta caretta).

Changes in plasma concentrations of steroids and prostaglandins (PGs) during natural nesting and oviposition in the loggerhead turtle were studied. Blood samples were obtained during nine distinct behavioral stages of oviposition. Emerging females had no detectable prostaglandin F (PGF) or prostaglandin E2 (PGE2) whereas plasma estradiol-17 beta averaged 255 pg/ml and mean plasma progesterone was 395 pg/ml. Plasma steroid concentrations did not vary significantly during nesting. In contrast, plasma PGF and PGE2 exhibited significant elevations during nest digging about 15 min after emergence. A further significant increase in plasma PGs was observed 10 min later during early oviposition. Plasma PGE2 peaked during mid oviposition whereas maximal plasma PGF levels occurred during nest covering although mean values were not significantly different than those observed during oviposition. Both PGs showed an abrupt decline (within 10 min) during body pit covering to concentrations similar to those observed during nest construction. Our data suggest that PGs have an active role during oviposition and nesting in the loggerhead turtle and are consistent with hypotheses that PGF2 alpha stimulates uterine contractions promoting egg expulsion while PGE2 may be more important in promoting cervical relaxation.

Animals↗
PubMed1992

Identification and purification of a high-affinity thyroxine binding protein that is distinct from albumin and prealbumin in the blood of a turtle, Trachemys scripta.

Fractionation of plasma proteins in the turtle, Trachemys scripta, confirmed the presence of a high-affinity thyroxine (T4) binding protein (TBP) that was distinct from albumin (ALB) and prealbumin (PA). The TBP was isolated by adsorption on a T4-affinity column and a high degree of purification was achieved by gel filtration and preparative electrophoresis. Analysis by reversed-phase HPLC showed a single peak of protein with T4 binding activity. The electrophoretic mobility of the TBP, based on staining and binding to [125I]T4 on nondenaturing polyacrylamide gels (PAGE), corresponded to that of the major T4 binding activity previously identified in plasma (ca. 60 kDa). PA was fractionated as a complex with retinol binding protein (PA-RBP) based on retinol associated fluorescence using ion exchange chromatography on DEAE-Sephacel and gel filtration. This complex behaved as a larger and more highly charged molecule than TBP; it was partially dissociated in low ionic strength basic solution. SDS-PAGE of the PA-RBP-enriched fraction revealed a major component of about 48 kDa (possibly free PA), with smaller components corresponding to those expected for free RBP (ca. 22 kDa) and subunits of PA (e.g., 14 and 28 kDa). ALB was purified by ion exchange chromatography on DEAE and gel filtration; it behaved as less basic than PA with MW approximately 67 kDa. TBP accounted for virtually all the T4 binding activity of whole plasma: TBP was about 100 times as active and PA and ALB were less than 1% as active as plasma. The binding affinity of purified TBP was similar to that of whole plasma from turtle and human (e.g., approx. 10(9) M-1 on Sephadex G-25).

Animals↗
PubMed1977

Properties of blood oxygen transport in the turtle Pseudemys scripta and the tortoise Testudo graeca: effects of temperature, CO2 and pH.

Properties of oxygen-haemoglobin binding have been investigated in the aquatic turtle Psuedemys scripta and the terrestrial tortoise Testudo graeca. Haematocrit (30-35%) and haemoglobin concentration (12-14 g/100 ml blood) were similar in both species. P50 at physiological levels of PCO2 (20-25 mm Hg) was 21 mm Hg in Pseudemys, compared with 23 mm Hg in Testudo. The Bohr shift of the blood of both the turtle and the tortoise was almost identical at -0.28. The heat of oxygenation, deltaH, reflecting the temperature sensitivity of O2-Hb affinity, was -10.55 in Pseudemys and -8.12 kcal/mol in Testudo. These data on whole blood do not support previous generalizations in the literature suggesting marked differences in oxygen-haemoglobin binding between aquatic and terrestrial chelonian reptiles.

Animals↗
PubMed1978

Respiratory control and CO2 conductance: temperature effects in a turtle and a frog.

The respiratory control systems of the predominantly lung-breathing turtle. Pseudemys scripta, and the bimodal breathing frog, Rana catesbeiana, are compared. At constant temperature, control is effected in both species primarily by pulmonary ventilation, and appears to be similar in basic organization to the mammalian system. The skin CO2 exchange of the frog is largely passive and apparently under little adaptive control. CO2 exchange at different temperature is analyzed by means of the conductance equations: convective conductance [MCO2 = (VA/RT).pACO2] for lung exchange, and diffusive conductance (MCO2 = GCO2.transcutaneous deltaPCO2) for skin exchange. Predictions of conductance values, based on data for PaCO2 and MO2, agree well with measured values of both the turtle's ventilation and the frog's skin CO2 conductance at different temperatures. In each case the conductances increase only moderately, if at all, with temperature, a result that can be attributed to the similar Q10 values of PaCO2 and MO2. Because the conductances exhibit this blunted temperature dependence, both lung and skin breathers are provided with the respiratory effector mechanisms necessary to regulate PaCO2 and pHa at their characteristic values over the ectothermic temperature range.

Animals↗
PubMed1980

Distribution of ventilation in the turtle Pseudemys scripta.

Spontaneously ventilating turtles, Pseudemys scripta, were allowed to rebreathe 133Xe to equilibrium. Frequent gamma camera images were obtained and processed to allow regional evaluation of ventilation. The distribution of ventilation was markedly inhomogeneous, with washin to equilibrium most rapid in anterior lung compartments and slowest in posterior lung compartments. Similarly washout from equilibrium was most rapid in anterior compartments and slowest in posterior compartments. During apnea intercompartmental gas exchange occurred rapidly during spontaneous activity of the limbs or, more slowly, during episodes ascribed to regional contraction of lung smooth muscle. Despite inhomogeneous ventilation, overall washout of 133Xe could be monoexponential. A simple series model of the turtle lung is consistent with the inhomogeneous ventilation observed.

Animals↗
PubMed1982

Oxygen affinity of sea turtle blood.

The oxygen binding properties of whole blood in adult sea turtles (green, Chelonia mydas; loggerhead, Caretta caretta) were measured as a function of pH, and carbon dioxide partial pressure using the 'Hemoscan Oxygen Dissociation Analyser' with modifications that improved the performance of the method. Under approximately physiological conditions (25.7 degrees C, PCO2 = 37 Torr; pH 7.45) there is a marked affinity difference between the two species (green, P50 = 29 Torr; loggerhead, 47 Torr). It is suggested that the large affinity difference between blood of green and loggerhead turtles may be related to differences in depths to which they typically dive. The Hill coefficient at 50% saturation is 2.4 in the green under these conditions, and 2.7 in loggerhead. Hill plots of both species curve upward and have significant regions with n approaching 1 at low saturation with the effect that the lower end of the binding curve is more hyperbolic than sigmoidal. The fixed acid and CO2 Bohr effects in both species are strongly saturation dependent, being very low at low saturation and increasing markedly at higher saturation. The fixed acid Bohr effects are similar in the two species but the green is more sensitive to CO2, at constant pH, and so has a larger CO2 Bohr effect. The hyperbolic shape of the lower end of the binding curves, and low pH sensitivity in this region, would tend to favor oxygen loading during a dive, when lung PO2 and blood pH are falling. These properties might therefore favor more complete utilization of the lung oxygen store. We suggest that diving vertebrates can usefully be divided into two groups, those that use the lung as the main oxygen stores and those that use tissue (blood and myoglobin) stores.

Animals↗
PubMed1982

Effects of pH, CO2 and organic phosphates on oxygen affinity of sea turtle hemoglobins.

The affinities of adult-type hemoglobins of green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles were determined at 25.7 degrees C as a function of pH, PCO2 and each of the organic phosphates found in erythrocytes of these species (ATP, IHP (substitute for IPP) and DPG). Each species showed a single hemoglobin band in cellulose acetate electrophoresis which isoelectric focusing resolved into one major and three minor components having isoelectric points near pH 7.2. Oxygen binding curves were recorded using a modified 'Hemoscan' (Instrument Division, Travenol Laboratories, Savage, MD). Hemolysates were kept saturated with CO during preparation to prevent methemoglobin formation. CO was driven off immediately before performing each oxygen binding determination, using the photodissociative effect of the Hemoscan light beam. In both species, phosphate-free hemoglobin has higher affinity than whole blood, and had lower pH sensitivity. CO2 (37 Torr) reduces affinity, particularly in the green sea turtle, and eliminates pH sensitivity. Each organic phosphate tested reduces affinity, with IHP being most effective at a given concentration and DPG least effective. Organic phosphates are most effective at low pH, and restore both pH sensitivity and oxygen affinity to whole-blood levels taking into account the difference between plasma and intracellular pH. Reasons are suggested why the organic phosphate effect was not detected in a previous study. Hemolysates of both species are distinctive in giving upward-curving Hill plots, with n = 1 asymptotes at low saturation that are insensitive to pH, CO2 and organic phosphates. The oxygen affinities corresponding to these asymptotes are about 100 Torr in both species. Results are compared to oxygen dissociation curves obtained from whole blood.

Animals↗
PubMed1988

Acid-base balance and the control of respiration during anoxic and anoxic-hypercapnic gas breathing in turtles.

We studied the ventilatory and blood acid-base response of turtles to 6 h of breathing either 100% N2 (anoxic) or 95% N2-5% CO2 (anoxic-hypercapnic). In both groups, minute ventilation (VE) increased promptly with anoxia, with peak ventilation occurring between 1 and 3 h. VE then decreased but was still significantly above control at 6 h. The increase in VE resulted from increases in both respiratory frequency (f) and tidal volume (VT) but after ventilation peaked, f declined to control while VT remained elevated. We observed no significant differences in VE between the two groups in spite of significantly lower arterial pH and higher arterial PCO2 in the anoxic-hypercapnic turtles. During normoxic recovery, VE quickly increased to the peak anoxic values due primarily to a greatly increased f. In both groups, plasma [lactate-] increased during anoxia. Plasma cation concentrations also increased, partially compensating for the elevated blood lactate. We conclude that the anoxic hyperventilation did not depend on arterial pH and central chemoreceptor control but rather on peripheral hypoxic chemoreceptor control. We believe that the decline in VE during prolonged anoxic breathing results from a metabolic arrest response and/or a depression in central nervous function.

Acid-Base Equilibrium↗
PubMed1989

Respiratory mechanics of the loggerhead sea turtle, Caretta caretta.

Respiratory mechanics were evaluated in excised lungs and in spontaneously breathing loggerhead sea turtles (Caretta caretta). Respiratory compliance curves reflect the pressure volume characteristics of the body wall. Compliance values are comparable to those of reptiles having simpler lung structure. Maximum flow rates in excised lungs (18-25 ml.sec-1.kg-1) were only slightly below the range reported for marine mammals. During spontaneous tidal breathing expiratory flow rates (11.8 ml.sec-1.kg-1) were lower than maximum values. In the sea turtle, respiratory adaptations including reduced airway resistance and muscular contribution to breathing are similar to marine mammals. These mechanical specializations shorten breathing time, which contributes to diving performance.

Animals↗
PubMed1986

The effects of continuous superfusion of L-aspartate and L-glutamate on horizontal cells of the turtle retina.

We have studied the effects of prolonged superfusion with L-aspartate and L-glutamate on the membrane potential and photoresponses recorded in luminosity type horizontal cells in the turtle retina using an everted eyecup preparation. These acidic amino acids produce effects which are a function of the past history of the impaled cell. Initial prolonged superfusions with 30 mM of either drug has no pronounced effect on the membrane potential and photoresponses of horizontal cells. Subsequent superfusions with either agent eventually produce depolarizations with reductions in the amplitudes of the light evoked responses. These effects, however, are transient; the horizontal cell rehyperpolarizes and the light evoked response grows with time. In a retina which had been stored at 4 degrees centrigrade for 20 hours, an initial superfusion with L-aspartate solution produced an immediate depolarization of the horizontal cell and complete suppression of the light evoked response for as long as the amino acid was present. The data are consistent with the existence of powerful amino acid uptake mechanisms operating at a number of sites within the inner and outer retina but also raise questions about the role of acidic amino acids in the outer plexiform layer of the turtle retina.

Animals↗