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Effects of kainic acid and piperidine dicarboxylic acid on displaced bipolar cells in the turtle retina.

An immunoreaction against glutamate was used to visualize photoreceptors, bipolar, and ganglion cells in the turtle retina. Incubation of the retina prior to fixation in kainic acid (9 microM) led to selective loss of glutamate-like immunoreactivity in OFF-centre bipolar cells, as judged by the loss of staining in the distal half of the inner plexiform layer. In addition, displaced bipolar cells and ganglion cells lost their immunoreactivity. Incubation of the retina in 2,3-cis piperidine dicarboxylate (1 mM) did not result in noticeable glutamate depletion in any cell but enhanced labelling in displaced bipolar cells. These findings suggest that all displaced bipolar cells in the turtle retina are depolarized by kainic acid and hyperpolarized by 2,3-cis piperidine dicarboxylate.

Animals↗

Molecular evolution and population genetics of Greater Caribbean green turtles (Chelonia mydas) as inferred from mitochondrial DNA control region sequences.

The molecular evolution and population genetics of migratory green turtles (Chelonia mydas) in the Greater Caribbean were examined with mitochondrial DNA (mtDNA) control region I sequences. A total of 488 base positions (bp) per individual were aligned for 44 individuals from four nesting populations in Florida, Costa Rica, Aves Island (Venezuela), and Surinam. Twelve sequence polymorphisms were detected, representing ten transitions, one transversion, and one 10-bp repeat. Sequence analyses of within- and between-population diversity revealed a deep divergence between western and eastern Caribbean nesting colonies and an inverse relationship between reproductive female population size and mtDNA diversity. In small populations, genetic admixture was important to maintaining high diversity, whereas larger populations appear to have experienced historical bottlenecks or resulted from founder effects. Mitochondrial DNA sequences of the control region offer an order of magnitude greater resolution than restriction site data for addressing questions about mtDNA variation, both within and between populations of green turtles.

Animals↗

Distribution and morphology of dopaminergic amacrine cells in the retina of the turtle (Pseudemys scripta elegans).

A light microscopical study of the cell types that stain by immunohistochemistry for the synthesizing enzyme for dopamine, tyrosine hydroxylase, has been performed on the retina of the turtle Pseudemys scripta elegans. The immunostain can be localized to a single morphological type of amacrine cell. The cells are like A28 cells of a Golgi classification. They have medium sized dendritic fields that range in diameter from 200 to 700 micron with eccentricity from the visual streak. The amacrines have a tri-stratified dendritic tree with tiers of fine, curved dendrites ramifying in strata S1, lower S2 and the S4/5 border of the inner plexiform layer. We, like others, can find no good evidence that these cells are interplexiform cells. The dopaminergic amacrine cells have a low frequency (approximately 1300-1500 total cells in 130 mm2 retina), with their highest density occurring in the visual streak (60 cells per mm2). The density profiles fall in elliptical isodensity rings from the visual streak towards the peripheral retina. At all points on the retina the dendritic fields maintain a constant coverage factor independent of eccentricity. A comparison of the dopaminergic amacrine cells in the turtle and other vertebrate retinae is made.

Animals↗

Serosal Na/Ca exchange and H+ and Na+ transport by the turtle and toad bladders.

A Na/Ca exchange system has been described in the plasma membrane of several tissues and seems to regulate the concentration of calcium in cytosol. Replacement of extracellular Na by sucrose increases calcium uptake into and decreases calcium efflux from the cell, leading to an increase in cytosolic calcium. The effect of an increase in cytosolic calcium mediated by the Na/Ca exchange system on H+ and Na transport in the turtle and toad bladder was investigated by replacing serosal Na isosmotically by sucrose or choline. Replacement of serosal by sucrose was associated with a significant inhibition of H+ secretion or Na transport which was reversible by addition of NaCl. Replacement of mucosal Na by sucrose failed to alter H+ secretion. Removal of serosal Na was associated with a significant increase in 45Ca uptake which could be blocked by pretreatment with lanthanum chloride. Pretreatment with lanthanum chloride blunted the inhibitory effect of replacement of serosal Na by sucrose on H+ and Na transport, thus suggesting that the increase in calcium uptake and the inhibition of transport are causally related. Under anaerobic conditions the rate of H+ or Na transport are linked to the rate of lactate production. The inhibition of Na or H+ transport by removal of serosal Na was accompanied by a proportional decrease in lactate production, thus suggesting that an increase in cytosolic calcium does not inhibit transport by uncoupling glycolysis from transport. Replacement of serosal Na by sucrose did not alter the force of the H+ or Na pump but led to an increase in resistance of the active pathway of H+ and Na transport. The inhibition of Na transport by replacement of serosal Na with sucrose could be reversed by addition of amphotericin B, an agent which increases luminal permeability to Na, thus suggesting that decreased Na entry across the apical membrane is the mechanism responsible for the inhibition of Na transport. The results of the present studies strongly suggest that an increase in cytosolic calcium through the serosal Na/Ca exchange system inhibits H+ and Na transport in the turtle and toad bladder probably by increasing the resistance of the luminal membrane.

Amphotericin B↗

Metabolic pathways coupled to H+ transport in turtle urinary bladder.

Active H+ transport in the turtle urinary bladder is mediated by an ATPase. Although the source of ATP is usually mitochondrial oxidative phosphorylation, it is possible because of intracellular compartmentalization or cellular heterogeneity that one metabolic pathway exclusively provides ATP to the pump. To examine this we performed several types of experiments. In one, the coupling between the rate of transport and the rate of oxidation of 14C-labeled substrates was studied. We found that there was coupling between H+ transport and glucose, butyrate oleate, and beta-OH-butyrate oxidation. In another set of experiments we depleted turtle bladders of their endogenous substrates and tested the effect of a number of substrates on the rate of transport. We found that glucose, pyruvate, lactate, actetate, butyrate and beta-OH butyrate all stimulated H+ transport. In a third set of experiments we found no coupling between H+ transport and lactate production. Finally, we found that reduction of H+ transport by mucosal acidification resulted in an increase in epithelial cell ATP concentrations and a decrease in ADP levels. These results suggest that the H+ pump receives its ATP from carbohydrate and fatty acid oxidation. The changes in ATP and ADP levels provide an initial explanation for the coupling of H+ transport to the rate of cellular oxidative metabolism.

Adenine Nucleotides↗

Sodium uptake across the apical border of the isolated turtle colon: confirmation of the two-barrier model.

The initial rate of Na uptake by the turtle colon from the mucosal bathing solution consists of two operationally distinct components. One component is a linear function of mucosal Na concentration, is unaffected by amiloride, and appears to represent Na uptake into the paracellular shunt path. The major component of Na uptake is abolished by amiloride and is virtually equal to the short-circuit current over a wide range of mucosal Na concentrations, suggesting that this portion of Na uptake represents Na movement into Na-transporting cells of the colon. The amiloride-sensitive component of Na uptake, at low mucosal Na concentrations, was unaffected if net Na transport was abolished by ouabain. Similarly, at low mucosal Na concentrations the amiloride-sensitive conductance of the colon was identical in the presence and in the absence of net Na transport. These results show that the isolated turtle colon behaves as two distinct barriers to transmural Na transport, an apical barrier blocked by amiloride and a more basal-lying barrier where active, transmural Na transport is blocked by ouabain. In addition, these experiments appear to provide the first unambiguous demonstration that the initial-rate isotope uptake technique can provide a direct measure of the properties of the amiloride-sensitive barrier to transmural Na movement, presumably the apical membranes of the Na-transporting cells. The results are consistent with the notion that the rate of transmural active Na transport and the conductance of the active Na-transport path are determined by the properties of the apical membrane.

Amiloride↗

Coupling between H+ transport and anaerobic glycolysis in turtle urinary bladder: effect of inhibitors of H+ ATPase.

The coupling between H+ transport (JH) and anaerobic glycolysis was examined in vitro in an anaerobic preparation of turtle urinary bladder. JH was measured as the short-circuit current after Na+ transport was abolished with ouabain and by pH stat titration. The media were gassed with N2 and 1% CO2 (PO2 less than 0.5 mm Hg) and contained 10 mM glucose. Under these conditions, JH was not inhibited by 3 mM serosal (S) cyanide or by 0.1 mM mucosal (M) dinitrophenol. Control anaerobic lactate production (Jlac) of 47 bladders was plotted as a function of simultaneously measured JH. The slope of Jlac on JH was 0.58 0.12 with an intercept for Jlac at JH = 0 of 0.55 micromol/hr. Values for delta Jlac/delta JH were determined in groups of individual bladders when JH was inhibited by an opposing pH gradient (0.55 0.16), by acetazolamide (0.58 0.19) and by dicyclohexylcarbodiimide, DCCD (0.58 0.14). The constancy of delto Jlac/ delta JH indicates a high degree of coupling between JH and Jlac. Since the anaerobic metabolism of glucose produces one ATP for each lactate formed, the delta Jlac/ delta JH values can be used to estimate the stoichiometry of H+ translocation. The movement of slightly less than 2H+ ions is coupled to the hydrolysis of one ATP. During anaerobiosis (absence of mitochondrial ATPase function) the acidification pump was not inhibited by M addition of oligomycin but was inhibited by M addition of DCCD and Dio-9, inhibitors of H+ flow in the proteolipid portion of H+-translocating ATPases. DCCD inhibited anaerobic JH without change in delta Jlac/delta JH or basal Jlac and, therefore, acted primarily on the H+ pump. S addition of vanadate also inhibited JH, but the inhibition was associated with an increase in Jlac. The site of this apparent uncoupling remains to be defined. The acidification pump of the luminal cell membrane of the turtle bladder has H+-ATPase characteristics that differ from mitochondrial ATPase in that H+ transport is oligomycin-resistant and vanadate-sensitive. As judged from the flows of H+ and lactate, the H+/ATP stoichiometry of the pump is about 2.

Adenosine Triphosphatases↗

Radioimmunoassay of plasma progesterone, testosterone, total estrogen and immunoreactive gonadotropin in the nesting and non-nesting green sea turtle, Chelonia mydas (L.).

Plasma progesterone, testosterone, total estrogens and immunoreactive gonadotropin were measured in nesting and non-nesting sea turtles, Chelonia mydas. Progesterone and gonadotropin concentrations were significantly higher in nesting than in non-nesting turtles, testosterone was not significantly different in either group and total estrogens appeared to be slightly in the nesting group.

Animals↗

A study about the possible characteristics of cardiovascular histamine receptors of the turtle.

Histamine produced pressor, depressor or biphasic responses in the systemic blood pressure of the turtle. The pressor response or component reversed to depressor effect after the injection of mepyramine, an H1 receptor blocker. It was thought that this response is mediated through the H1 receptors. The depressor response or component was abolished by the treatment of metiamide, a new histamine H2 receptor blocker. Also, the effect reversed after metiamide. These results indicated the presence of H2 receptors in the turtle.

Animals↗

Odor discrimination of "IP3-" and "cAMP-increasing" odorants in the turtle olfactory bulb.

The ability of the turtle olfactory system to discriminate between various odorants that increase levels of adenosine 3',5'-cyclic monophosphate (cAMP) and inositol trisphosphate (IP3) in the olfactory bulb was examined by the cross-adaptation technique and analyzed by multidimensional scaling. The mean values of the degree of discrimination among the IP3-increasing odorants were higher than those among the cAMP-increasing odorants, and were similar to those between cAMP- and IP3-increasing odorants, suggesting that the features of the receptors of cAMP-increasing odorants are different from those which respond to IP3-increasing odorants. Analysis by multidimensional scaling suggested that differences in second messenger pathways are not related to detecting odor quality in the turtle olfactory system.

Acyclic Monoterpenes↗

Morphological changes induced in turtle retinal neurons by exposure to 6-hydroxydopamine and 5,6-dihydroxytryptamine.

Following intraocular injection of the dopamine neurotoxin 6-hydroxydopamine (10-50 micrograms on two successive days in a Ringer vehicle containing ascorbate and pargyline) and an incubation period of 1 to 18 days, degeneration was noted in presumptive amacrine cells in the retina of the turtle, Pseudemys scripta elegans. Injection of vehicle alone produced no effect. Affected perikarya initially showed swollen mitochondria, lysosomes and distended cisternae. At later stages the cells took on a darkened appearance. In contrast, affected amacrine processes in the inner plexiform layer became markedly distended and lost their cytoplasmic contents, resulting in empty, very swollen profiles. No degeneration was noted distal to the affected cell bodies, i.e. the affected cells were not interplexiform neurons. Cells lesioned by 6-hydroxydopamine were shown to accumulate [3H]dopamine. Intraocular administration of 5,6-dihydroxytryptamine (a single dose of 10-40 micrograms in the same vehicle) followed by 4-6 days incubation resulted in a marked darkening of certain bipolar cell axon terminals, cell bodies and Landolt's clubs. The toxic effects of 5,6-dihydroxytryptamine were blocked by zimelidine, a serotonin uptake blocker. Thus, these two neurotoxins have different targets in the turtle retina. At the highest dose tested, however, 6-hydroxydopamine did produce degenerative changes in the presumed serotonergic bipolar cell.

5,6-Dihydroxytryptamine↗

Initiation and ultrastructure of a reptilian fibroblast cell line obtained from cutaneous fibropapillomas of the green turtle, Chelonia mydas.

Two fibroblastic cell lines were established from explants of fibropapillomas of each of two different green turtles (Chelonia mydas). These cells, designated GTFP (Green Turtle Fibropapilloma), were subcultured approximately 30 times at 30 degrees C in Eagle's minimal essential media supplemented with 2 to 10% fetal bovine serum. The ultrastructural morphology of the cultured fibroblasts is described. The cells contained abundant rough endoplasmic reticulum, polyribosomes, and mitochondria; collagen fibrils were visible in the extracellular space. No viruslike particles or evidence of other pathogenic agents could be demonstrated by electron microscopy in any of the cultured cells examined.

Animals↗

Human salmonellosis transmitted by a domestic turtle.

Salmonella typhimurium was isolated in the culture test of a small child admitted to hospital suffering from febrile gastroenteritis with stools containing traces of mucus and blood. Her mother also resulted positive for this microorganism. The family had recently bought a small turtle, imported from Florida, at the city fair. Further tests revealed Salmonella typhimurium in both the turtle's feces and the water in its tank.

Animals↗

Localization of NADPH-diaphorase containing neurons in the spinal dorsal horn and spinal sensory ganglia of the turtle Chrysemys d'orbigny.

NADPH-diaphorase positive (NDP) neurons and nerve fibers were found in the spinal dorsal horn (DH) and sensory ganglia of the turtle Chrysemys d'orbigny. Three well-defined types of NDP neurons were found in the DH: (a) elongated nerve cells with two radially arranged dendritic branches, (b) neurons with rostrocaudal dendritic branches, (c) bitufted neurons with two, practically symmetric branches that project to the ipsilateral and contralateral dorsal horns. A combination of the techniques that reveal NADPH-diaphorase activity with the horseradish peroxidase transganglionic labeling of the dorsal root collaterals, suggested that NDP neurons of the DH are second-order cells of the spinal sensory pathway. NDP neurons were also found in the spinal sensory ganglia at all metameric levels. Our findings indicate that the DH of turtles, like that of mammals, contains both the enzymatic machinery and the neural connections required to postulate the participation of nitric oxide in "plastic phenomena" such as hyperalgesia and central sensitization. Two other alternatives or complementary hypotheses are discussed: (a) NDP neurons in the DH and sensory ganglia may represent specific cell populations involved in the processing of sensory visceral information; (b) NADPH-diaphorase reactivity may indicate sustained levels of neuronal activity.

Afferent Pathways↗

The sea-finding behavior of hatchling olive ridley sea turtles, Lepidochelys olivacea, at the beach of San Miguel (Costa Rica).

Newly hatched olive ridley sea turtles (Lepidochelys olivacea) were tested for their directional preferences in a sand-filled circular arena in total darkness. Hatchlings that had crawled about 5 m on the beach, toward the sea preferred the southwesterly direction that would have brought them to the water line, whereas hatchlings that had been denied this experience headed eastward, a direction of unclear origin. These data suggest that a short crawl across the natural beach can set the direction in which the young turtles subsequently move. The crawling experience was sufficient to acquire the compass course that they later follow, probably with the help of a magnetic compass, not only in the water, but already while still on land.

Animals↗

Synaptic pharmacology in the turtle accessory optic system.

The accessory optic system of the turtle (the basal optic nucleus, BON) receives both excitatory and inhibitory inputs that are direction-sensitive. When the dorsal midbrain is ablated, only the monosynaptic direction-sensitive input from the retina to the BON remains. To better understand the central visual processing performed by the accessory optic system, this study identifies the neurotransmitters and their receptors that mediate the synaptic excitation and inhibition of BON cells. We used a reduced in vitro turtle brainstem preparation in which the two eyes and brain were isolated pharmacologically. Patch recordings were made on BON neurons while drugs were applied to the brain, with the eyes bathed in control media and either exposed to visual pattern motion or subjected to electrical stimulation. An antagonist of the AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) subtype of glutamate receptor applied within the brain chamber blocked the visual responses. In response to electrical stimulation both excitatory and inhibitory synaptic events were blocked in BON cells, presumably by blocking direct excitation by retinal ganglion cell axons in the BON and indirect excitation of inhibitory interneurons elsewhere in the brainstem. An NMDA receptor antagonist was ineffective, even when the response was measured in a BON cell depolarized in Mg(2+)-free media. A GABA(A) receptor on the BON cell mediates the inhibitory responses to retinal stimulation. Injection of lidocaine into the contralateral eye caused an increase in spontaneous inhibitory post-synaptic potentials (IPSPs), suggesting that a tonic retinal output exists that reduces brainstem inhibition of BON cells. Also, there may be tonic inhibition of an excitatory path to BON neurons from within the brainstem, because bicuculline increased spontaneous excitatory post-synaptic potentials (EPSPs) observed in a BON cell without retinal input. These results indicate that the BON is a site of complex visual processing of competing visual signals and provide insight into how an interaction of excitation and inhibition creates a retinal slip signal in the accessory optic system.

2-Amino-5-phosphonovalerate↗

Evidence for a photosensitive region in the caudal mesencephalon of the turtle brain.

Using an in vitro brainstem-cerebellum preparation from the turtle Chrysemys picta, burst discharge was recorded from the abducens nerve when light was directed on the brainstem. This burst discharge likely represents a neural correlate of the eye-blink reflex. Increasing the intensity of the light stimulus reduced the response latency from a duration of many seconds to approximately 1-2 s. No response was recorded when the light source was covered. The response was present when infrared light was blocked, and it could only be produced when the light spectra contained wavelengths below approximately 550 nm. Lesion experiments reveal that the photosensitive area is located caudal to the trochlear nerve and rostral to the trigeminal nerve. Single-unit microelectrode recordings demonstrate that this region is tonically active in the dark and that activity is suppressed by light. Pharmacological results show that the light response is blocked by application of compounds that act as serotonergic antagonists, and that antagonists of noradrenergic receptors (alpha or beta) either have no effect or their actions are variable. Taken together, these data suggest that an isthmo-optic-like area in the turtle brain is photosensitive to direct light and activates neural circuits that control eye movements.

Adrenergic Antagonists↗

Accumulation and selective maternal transfer of contaminants in the turtle Trachemys scripta associated with coal ash deposition.

Coal combustion wastes are enriched in a number of potentially toxic compounds and may pose risks to biota exposed to the wastes. Slider turtles (Trachemys scripta) are common inhabitants of coal ash settling basins in South Carolina, USA, where they feed on contaminated prey items and accumulate high levels of potentially toxic compounds in their tissues. Furthermore, female sliders sometimes nest in contaminated spill piles and thus may expose embryos to contaminated soils. We examined two potential pathways by which female T. scripta may influence the survivorship and quality of their offspring in a contaminated habitat: (1) nesting in contaminated soil and (2) maternal transfer of pollutants. Eggs were collected from turtles captured in coal ash-polluted or unpolluted sites; individual clutches were incubated in both ash-contaminated and uncontaminated soil in outdoor, artificial nests. Incubation in contaminated soil was associated with reduced embryo survivorship. Adult females from the polluted site accumulated high levels of As, Cd, Cr, and Se in their tissues, yet Se was the only element transferred maternally to hatchlings at relatively high levels. Hatchlings from polluted-site females exhibited reduced O2 consumption rates compared to hatchlings from reference sites. Relatively high levels of Se transferred to hatchlings by females at the ash-polluted site might contribute to the observed differences in hatchling physiology.

Animals↗