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PubMed1994

Complexity and scaling properties of amacrine, ganglion, horizontal, and bipolar cells in the turtle retina.

In the present study we have evaluated the complexity and scaling properties of the morphology of retinal neurons using fractal dimension as a quantitative parameter. We examined a large number of cells from Pseudemys scripta and Mauremys caspica turtles that had been labeled using Golgi-impregnation techniques, intracellular injection of Lucifer Yellow followed by photooxidation, intracellular injection of rhodamine conjugated horseradish peroxidase, or intracellular injection of Lucifer Yellow or horseradish peroxidase alone. The fractal dimensions of two-dimensional projections of the cells were calculated using a box counting method. Discriminant analysis revealed fractal dimension to be a significant classification parameter among several other parameters typically used for placing turtle retinal neurons in different cell classes. The fractal dimension of amacrine cells was significantly correlated with dendritic field diameters, while the fractal dimensions of ganglion cells did not vary with dendritic field span. There were no significant differences between the same cell types in two different turtle species, or between the same types of neurons in the same species after labeling with different techniques. The application of fractal dimension, as a quantitative measure of complexity and scaling properties and as a classification criterion of neuronal types, appears to be useful and may have wide applicability to other parts of the central nervous system.

Animals↗
PubMed2004

Sox8 is expressed at similar levels in gonads of both sexes during the sex determining period in turtles.

A critical gene involved in mammalian sex determination and differentiation is the Sry-related gene Sox9. In reptiles, Sox9 resembles that of mammals in both structure and expression pattern in the developing gonad, but a causal role in male sex determination has not been established. A closely related gene, Sox8, is conserved in human, mouse, and trout and is expressed in developing testes and not developing ovaries in mouse. In this study, we tested the possibility of Sox8 being important for sex determination or sex differentiation in the red-eared slider turtle Trachemys scripta, in which sex is determined by egg incubation temperature between stages 15 and 20. We cloned partial turtle Sox8 and anti-Müllerian hormone (Amh) cDNAs, and analyzed the expression patterns of these genes in developing gonads by reverse transcriptase-polymerase chain reaction and whole-mount in situ hybridization. While Amh is expressed more strongly in males than in females at stage 17, Sox8 is expressed at similar levels in males and females throughout the sex-determining period. These observations suggest that differential transcription of Sox8 is not responsible for regulation of Amh, nor responsible for sex determination in turtle.

Amino Acid Sequence↗
PubMed2001

Sexual differentiation in the spiny softshell turtle (Apalone spinifera), a species with genetic sex determination.

It is hypothesized on the basis of sex determination theory that species exhibiting genetic sex determination (GSD) may undergo sexual differentiation earlier in development than species with environmental sex determination (ESD). Most turtle species exhibit a form of ESD known as temperature-dependent sex determination (TSD), and in such species the chronology of sex differentiation is well studied. Apalone spinifera is a species of softshell turtle (Trionychidae) that exhibits GSD. We studied sexual differentiation in this species in order to facilitate comparison to TSD species. Eggs were incubated at two different temperatures and embryos were harvested at various stages of mid to late development. Gonad length was measured with image analysis software, then prepared histologically. Indifferent gonads have differentiated in stage 19 embryos. Histological details of gonadogenesis follow the same pattern as described for other reptiles. Regression of the male paramesonephric duct closely follows testicular differentiation. Gonad lengths are longer at the warmer incubation temperature, and ovaries are generally longer than testes at each stage and for each temperature. Although sexual differentiation takes place at about the same stage as in other turtles with TSD (18-20), in A. spinifera this differentiation is irreversible at this stage, while in some of the TSD species sex is reversible until about stage 22. This immutable, definitive sexual differentiation may support the hypothesis of an accelerated chronology of sex differentiation for this species. We also note that sexual dichromatism at hatching is known in this species and may provide additional evidence of early differentiation. J. Exp. Zool. 290:190-200, 2001.

Animals↗
PubMed1986

Photothermal effects on the testicular cycle in the musk turtle, Sternotherus odoratus.

A series of photothermal treatments was initiated at different times of year to examine the relative contributions of exogenous and endogenous factors affecting the timing of the testicular cycle of the kinosternid turtle, Sternotherus odoratus. Elevated temperature appeared to be the major factor stimulating testicular recrudescence (spermatocytogenesis, spermiogenesis, and, to some extent, androgen secretion). However, depending on the time of year, short or long daylengths, in conjunction with elevated temperatures, enhanced testicular responses (recrudescence or regression). Also, high temperatures are not requisite for recrudescence. Spermatogenic activity and testis growth differ in their temperature sensitivity and may become dissociated, and an endogenous mechanism may have affected peak androgen secretion. Testicular regression exhibited a dependence on an interaction among temperature, photoperiod, and endogenous regulation. Turtles with testes in advanced stages of spermiogenesis underwent accelerated regression when exposed to even slightly reduced temperatures, especially at short daylengths. However, regression was delayed by a combination of high temperature and short daylength in fall. Results are compared to other experimental studies of photothermal effects on turtles and discussed in terms of the described latitudinal variations in the timing of the testicular cycle of S. odoratus.

Acclimatization↗
PubMed2003

Oviductal sperm storage structure and their changes during the seasonal (dissociated) reproductive cycle in the soft-shelled turtle Lissemys punctata punctata.

The oviduct of the Indian fresh water soft-shelled turtle Lissemys punctata punctata was examined throughout the year under light and scanning electron microscopes to determine the location, histomorphological characteristics, and function of sperm storage structure, as well as their changes at different phases of the seasonal reproductive cycle. Sperm storage structures in the form of tubules were observed in the wall of isthmus throughout the year. These tubules developed either by folding or fusion of the oviductal mucosal folds and were lined by both ciliated and nonciliated epithelial cells. The height and secretory activities of the epithelia were markedly high during the breeding phase (August to September) but low in the nonbreeding phase (October to June). A few short tubules lined by cuboidal epithelium appear in the wall of infundibulum only during the breeding phase. Following mating (May), inseminated sperm were stored within the tubules of isthmus up to the pre-ovulatory stage (August). Thereafter, sperm associated with PAS-positive materials secreted from the epithelium (referred to as a carrier matrix) moved forward to the infundibulum and were stored within the storage tubules of the infundibulum for a short time. Subsequently, sperm evacuated the storage tubules and entered the oviductal lumen to fertilize the subsequently ovulated eggs during or prior to ovulation. The isthmus-tubules become shorter and narrower in the regressive phase (October to November) and remained so until the early preparatory phase (April). Sperm release might have been stimulated by estrogen secreted from the ovarian follicles of pre-ovulatory turtles. Stored sperm not utilized for fertilization remained viable not less than six months in the present turtle species.

Animals↗
PubMed2004

Effect of gonadal steroids on progesterone receptor, estrogen receptor, and vitellogenin expression in male turtles (Chrysemys picta).

Hepatic vitellogenin (vtg) is a yolk precursor protein sequestered in follicular oocytes as nutrient supply for developing embryos in nonmammalian vertebrates. In prior research studies we have demonstrated that both progesterone (P) and testosterone (T) inhibit estrogen (E)-induced vitellogenesis in the male fresh water turtle (Chrysemys picta), and have suggested that these hormones may be involved in multihormonal regulation of vitellogenesis in the female turtle. However, the modes of action of progesterone and testosterone on estrogen-induced vitellogenesis are not known. We have proposed that progesterone inhibits vitellogenesis by modulation of progesterone receptor A (PRA) or B (PRB) isoforms and/or estrogen receptor (ER) gene transcription. In this study, we compare the vitellogenic responses of reproductively inactive male turtles to estradiol 17beta in the presence of exogenous testosterone or progesterone. Northern blot analysis was used to monitor the changes in vtg mRNA, ER mRNA, and PR mRNA expression; Western blotting to determine changes in PR isoform expression and a homologous ELISA for measurement of plasma vtg. Progesterone and testosterone reduced estrogen-induced vtg mRNA expression, but plasma vtg was not significantly reduced by these steroids. PRA and PRB were transcribed even though ER mRNA could not be detected, suggesting constitutive PR expression. However, in the presence of estradiol 17beta, both PR isoforms and mRNA transcripts were increased as a correlate of ER mRNA transcription, suggesting both transcriptional and translational effects; these effects were inhibited by testosterone and progesterone treatments. Since ER mRNA was sharply reduced by both testosterone and progesterone, and estradiol 17 beta increased PR mRNA transcription and translation, it is likely that the action of progesterone in reducing vtg mRNA is indirect via down regulation of ER mRNA, thus ER. This study provides further information on the role of progesterone and testosterone in the regulation of hepatic vitellogenesis, suggesting regulation of vitellogenesis mainly via modulation of hepatic ER mRNA.

Animals↗
PubMed2004

Lymphoid aggregates in gonads of embryos, hatchlings, and young of turtles with temperature-dependent sex determination.

Cellular infiltrations forming lymphoid-like aggregates were previously observed in gonads of two turtle species exhibiting temperature-dependent sex determination (TSD): at hatching in Chelydra serpentina; at and after hatching in Emys orbicularis. We show here that such aggregates are also present in gonads of Testudo graeca by the end of embryonic development, suggesting that their occurrence is general in turtles. Since in C. serpentina, infiltrations were observed mainly in testes exhibiting remnants of the germinal epithelium, it was assumed that their occurrence was an expression of maleness leading to rejection of this epithelium. The generality of this hypothesis was tested in E. orbicularis by looking for lymphoid-like aggregates in three types of gonads (testes, ovotestes, and ovaries) and for the stages at which they occur. Gonads were from embryos, hatchlings, and young incubated at various temperatures. Ovotestes obtained by treatment with an aromatase inhibitor of eggs incubated at female-producing temperature were also examined. In these gonads, the differentiation of Sertoli cells in testicular cords/tubes was ascertained by expression of SOX9. Moreover, the cell composition of aggregates was determined on electron micrographs. Aggregates appear in ovaries and ovotestes by the end of embryonic development and are present in the majority of these gonads at hatching, and at least up to one year after hatching. They are composed mainly of lymphocytes and fibroblasts. Aggregates are not present in typical testes. Since they occur in most ovaries, they cannot be seen as an expression of maleness. Rather, lymphocytic infiltration and formation of lymphoid aggregates in turtle gonads can be seen as components of the immune system, and can be under the control of gonadal endogenous sex steroids.

Animals↗
PubMed2005

Slow-tonic muscle fibers and their potential innervation in the turtle, Pseudemys (Trachemys) scripta elegans.

A description is provided of the ratio of slow-tonic vs. slow- and fast-twitch fibers for five muscles in the adult turtle, Pseudemys (Trachemys) scripta elegans. The cross-sectional area of each fiber type and an estimation of the relative (weighted) cross-sectional area occupied by the different fiber types are also provided. Two hindlimb muscles (flexor digitorum longus, FDL; external gastrocnemius, EG) were selected on the basis of their suitability for future motor-unit studies. Three neck muscles (the fourth head of testo-cervicis, TeC4; the fourth head of retrahens capitus collique, RCCQ4; transversalis cervicis, TrC) were chosen for their progressively decreasing oxidative capacity. Serial sections were stained for myosin adenosine triphosphatase (ATPase), NADH-diaphorase, and alpha-glycerophosphate dehydrogenase (alpha-GPDH). Conventional fiber-type classification was then performed using indirect markers for contraction speed and oxidative (aerobic) vs. glycolytic (anaerobic) metabolism: i.e., slow oxidative (SO, including slow-twitch and possibly slow-tonic fibers), fast-twitch, oxidative-glycolytic (FOG), and fast-twitch glycolytic (Fg) fibers. Slow-tonic fibers in the SO class were then revealed by directing the monoclonal antibody, ALD-58 (raised against the slow-tonic fiber myosin heavy chain of chicken anterior latissimus dorsi), to additional muscle cross sections. All five of the tested muscles contained the four fiber types, with the ATPase-stained fibers including both slow-tonic and slow-twitch fibers. The extreme distributions of SO fibers were in the predominately glycolytic TrC vs. the predominately oxidative TeC4 muscle (TrC-SO, 9%; FOG, 20%; Fg, 71% vs. TeC4-SO, 58%: FOG, 16%; Fg, 25%). Across the five muscles, the relative prevalence of slow-tonic fibers (4-47%) paralleled that of the SO fibers (9-58%). TeC4 had the highest prevalence of slow-tonic fibers (47%). The test muscles exhibited varying degrees of regional concentration of each fiber type, with the distribution of slow-tonic fibers paralleling that of the SO fibers. In the five test muscles, fiber cross-sectional area was usually ranked Fg > FOG > SO, and slow-twitch always > slow-tonic. In terms of weighted cross-sectional area, which provides a coarse-grain measure of each fiber type's potential contribution to whole muscle force, all five muscles exhibited a higher Fg and lower SO contribution to cross-sectional area than suggested by their corresponding fiber-type prevalence. This was also the case for the slow-twitch vs. slow-tonic fibers. We conclude that slow-tonic fibers are widespread in turtle muscle. The weighted cross-sectional area evidence suggested, however, that their contribution to force generation is minor except in highly oxidative muscles, with a special functional role, like TeC4. There is discussion of: 1) the relationship between the present results and previous work on homologous neck and hindlimb muscles in other nonmammalian species, and 2) the potential motoneuronal innervation of slow-tonic fibers in turtle hindlimb muscles.

Animals↗
PubMed1978

The pineal-paraphyseal complex of sea turtles. I. Light microscopic description.

The pineal-paraphyseal complex of sea turtles is an impressively large structure which projects dorsally and anteriorly above the prosencephalon. The complex was examined by light microscopy in several age classes of green sea turtles (Chelonia mydas) and from juvenile loggerhead turtles (Caretta caretta). The paraphysis is extensively fused to the distal portion of the pineal body, suggesting an interrelated function for these two tissues. No duct or canal was observed connecting the pineal lumen to the third ventricle. Two pineal cell types are described which appear to correspond to the neuroglial supportive cells and the secretory rudimentary photoreceptor cells of other amniotic vertebrates. A possible luminal secretion in the form of apical protrusions is produced by the latter cell type. No typical photoreceptive outer segments were observed.

Animals↗
PubMed1995

A commentary on the segmental motor system of the turtle: implications for the study of its cellular mechanisms and interactions.

A commentary is provided on the segmental motor system of the turtle Pseudemys (Trachemys) scripta elegans with an emphasis on neuronal, neuromuscular, and muscular mechanisms that control the development of force under normal, fatiguing, and pathophysiological conditions. For the central neuronal component of the segmental motor system, it has recently been shown that intracellular analysis of the firing properties of motoneurons and interneurons can be undertaken for relatively long periods of time in in vitro slices of the lumbosacral spinal cord of the adult turtle. In other less reduced in vitro preparations, analyses are available on complex motor behaviors generated by the isolated spinal cord. These behaviors of spinal neuronal networks are analogous in key aspects to those generated by the isolated in vivo cord, and by the cord in intact preparations. These results suggest that the neuronal components of the segmental motor system can not be studied from the cellular/molecular level of analysis in in vitro slice preparations to the systems level in conscious, freely moving animals. The in vitro approach can also be used for the analysis of cellular mechanisms in suprasegmental brain structures, which contribute to the control of voluntary movement. For the peripheral neuromuscular component of the segmental motor system, information is now available on muscle fiber types and selected aspects of sensory innervation, and it is feasible to study the mechanical and biochemical properties of motor units. As such, the turtle presents a valuable model for exploring interrelations between the neuronal and mechanical components of the segmental motor system of the generalized tetrapod. A prominent feature of these recent developments is the extent to which they have been driven by findings that have emphasized an evolutionary conservation of motor-control mechanisms extending from ion channels, at the cellular level, to the control of multijointed movements at the systems level of analysis.

Animals↗
PubMed1997

Synergistic responses of steroidal estrogens in vitro (yeast) and in vivo (turtles).

Many environmental agents exert estrogenic activity. Previous studies from our laboratories demonstrated that certain combinations of environmental estrogens (i) reverse the sex of male turtle embryos in a synergistic manner (Bergeron et al., (1994) Environ. Hlth Perspect. 102, 780-782), and (ii) synergistically transactivate the human estrogen receptor (hER) in yeast and mammalian cells (Arnold et al., (1996) Science 272, 1489-1492). Because our findings with synthetic estrogens suggested that combinations of naturally-occurring steroidal estrogens might also produce synergistic activity of the ER, we used the same model systems to measure the activity of combinations of steroidal estrogens. The activity of combinations of estrone, estradiol-17beta or estradiol-17alpha in yeast strains expressing hER was synergistic at submaximal concentrations of both estrogenic compounds. However, synergy was not observed with mixtures of estrogens when the concentration of one of the estrogens alone was maximally active in yeast. Ligand-binding assays in yeast performed with various radiolabeled estrogens suggested that multiple estrogens may interact with the receptor. The estrogen-dependent process of sex-reversal of turtle embryos incubated at a male-producing temperature was used to determine whether steroidal estrogens also had synergistic activity in vivo. In this instance, a combination of estriol and estradiol-17beta was effective in reversing the gonadal sex of turtle embryos from males to females in a synergistic manner. Our results suggest that the synergy of some combinations of estrogens, synthetic or steroidal, may play a role in the estrogen-dependent process of sexual development in certain species.

Animals↗
PubMed1993

Sensitive stages for the effects of temperature on gonadal aromatase activity in embryos of the marine turtle Dermochelys coriacea.

Many data suggest an involvement of estrogens in gonadal differentiation in reptiles with temperature-dependent sex determination (TSD). However, the site of estrogen synthesis in two species of freshwater turtles is unclear. In Emys orbicularis, estrogens were shown to be produced by the gonads, whereas in Trachemys scripta, gonadal steroids were not detected. The marine turtle Dermochelys coriacea exhibits TSD but in gonadal development, ovarian differentiation is delayed. Gonadal aromatase activity and estrogen content in this species were measured in embryos incubated at 27 degrees and in embryos incubated at 30.5 degrees, respectively, masculinizing and feminizing temperatures within the range of temperatures found in natural nests. At all stages studied, aromatase activity was present and found to be higher at 30.5 degrees than at 27 degrees. Estrogens were only found at 30.5 degrees. The effects of temperature shifts on gonadal aromatase activity were then examined. Eggs were shifted from 27 to 35 degrees (feminizing temperature) at different embryonic stages and exposed to 35 degrees for 6 days. An increase in gonadal aromatase activity, although with significant individual variations, was seen only when eggs were shifted between stages 23 and 27. These stages are in the range of the thermosensitive stages for sexual differentiation of the gonads determined in other turtles. These results are similar to those previously obtained in E. orbicularis and agree with a key role for endogenous estrogens in gonadal differentiation of reptiles with TSD.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Purification and characterization of plasma vitamin D binding protein from the snapping turtle, Chelydra serpentina.

Vitamin D binding protein (DBP) was isolated from the plasma of the snapping turtle, Chelydra serpentina (mixed sex adult), and compared with the previously characterized dual function binding plasma protein that binds both thyroxine (T4) and vitamin D (tsTDBP) in an emydid turtle, Trachemys scripta. Purification of Chelydra serpentina DBP (csDBP) was accomplished by ion exchange chromatography, preparative polyacrylamide gel electrophoresis, and reverse-phase high-performance liquid chromatography. Estimates of increased purification (ca. 71-fold), recovery (ca. 1.5%), and corresponding plasma concentration (ca. 0.6 mg/ml) are confounded by interference with other proteins. A protein was identified that showed the same high affinity for 25-OH-cholecalciferol (vitamin D3) as the tsTDBP, and the two exhibited similar heat resistances in binding. The csDBP and tsTDBP had similar molecular weights by SDS-PAGE (ca. 58 kDa), showed immunological cross reactivity, and differed by only three residues, representing conservative substitutions, in the 30 NH2-terminal amino acid sequence. A slightly lesser homology (up to 89% similarity based on conservative substitutions) was seen with three mammalian DBPs. However, unlike tsTDBP, the DBP from the snapper did not bind T4. These data support the view that the DBP of emydid turtles secondarily evolved a second T4 binding site, and this is probably independent of the D3 binding domain.

Amino Acid Sequence↗
PubMed1997

Plasma steroid profiles in nesting loggerhead turtles (Caretta caretta) in Queensland, Australia: relationship to nesting episode and season.

Plasma levels of four hormones-progesterone (P), testosterone (T), estradiol 17-beta (E2), and corticosterone (B)-were measured in samples taken from nesting female loggerhead turtles (Caretta caretta) by using specific radioimmunoassays. Samples were taken in an early, middle, or late period during the summer nesting season from females at first, second, third, or > fourth nesting episodes, defined as successive within-season nesting events, at Mon Repos Beach, Queensland, Australia. Data on individual patterns of nesting, collected over the past 20 years by the Queensland Turtle Research Project, and the seasonal nesting data, were analyzed with respect to influences on hormonal profiles. Circulating levels of E2 were mostly undetectable, suggesting either that this estrogen is not produced at this time of nesting, or that, perhaps, another estrogen may be present that is not detected by the specific radioimmunoassay. P, T, and B profiles in the nesting females were associated with the individual turtles' progression through successive nesting episodes, with a marked decline in all three hormones by the last (> 4) nesting episode of the season. Nesting episode accounted for significant changes that were related to season, in that nesting episode and season were significantly correlated. These patterns were observed in the population, when singly sampled at each time period or nesting episode, and in individual females sampled repeatedly over time. Moreover, T and B were highly and significantly correlated in females at all nesting episodes and time periods, and in the singly and repeatedly sampled females. The magnitude of change in T and B over time was also highly and significantly correlated in repeatedly sampled females. Together these results suggest the hypothesis that T and B interact over the period of successive nesting and may be involved in reproductive functions such as the mobilization of reserves for egg production in C. caretta.

Animals↗
PubMed1997

Overlapping distribution of receptors for atrial natriuretic peptide and angiotensin II in the kidney and the adrenal gland of the freshwater turtle, Amyda japonica.

The distribution of receptors for atrial natriuretic peptide (ANP) and angiotensin II (ANG II) in the kidney and adrenal gland of the freshwater turtle, Amyda japonica, was examined by quantitative in vitro autoradiography using 125I-labeled rat (r)ANP1-28 and 125I-labeled human ANG II as labeled ligands. Receptor subtypes were characterized by competition with des[Gln18,Ser19,Gly20, Leu21, Gly22] ANP(4-23) (C-ANP) as a selective ligand of the clearance receptors to specific 125I-rANP(1-28) binding, and with DuP 753 and PD 123319 as nonpeptide antagonists for ANG II receptors type 1 (AT1) and type 2 (AT2) subtypes, respectively, to 125I-ANG II binding. Specific 125I-rANP(1-28) binding with a single binding site was found overlying glomeruli of the kidney and the outer zone of the adrenal gland with dissociation constants (Kd) of 4.39 +/- 0.33 and 6.07 +/- 1.36 nM, respectively. C-ANP (10 microM) inhibited about 90% of the glomerular and adrenal 125I-rANP(1-28) binding. Specific 125I-ANG II binding was also localized in the glomeruli of the kidney and the outer zone of the adrenal gland with Kd of 1.02 +/- 0.22 and 0.37 +/- 0.04 nM, respectively. DuP 753 (10 microM) potently inhibited about 80% of glomerular and 90% of adrenal 125I-ANG II binding, whereas PD 123319 (10 microM) was very weak in competing for specific 125I-ANG II binding in both tissues. Therefore, these results indicate that specific ANP and ANG II receptors with high affinities have an overlapping distribution in glomeruli of the kidney and the outer zone of the adrenal gland of the freshwater turtle. They also suggest that biological and clearance ANP receptor-like subtypes coexist in both tissues, and the predominant ANG II receptor subtype in these tissues is the AT1-like receptor. The overlapping distribution of specific receptors for both peptides in these tissues provides the basis for possible receptor interactions to exert a functional antagonism between ANP and ANG II in the freshwater turtle.

Adrenal Glands↗
PubMed2000

Localization of receptors for natriuretic peptide and endothelin in the duct of the epididymis of the freshwater turtle.

The presence of receptor subtypes for natriuretic peptides (NPs) and endothelin (ET) in the epididymis of the freshwater turtle, Amyda japonica, was examined by quantitative in vitro autoradiography using iodinated mammalian-type atrial NP ((125)I-ANP((1-28))), phylogenically conserved C-type NP ((125)I-[Tyr(0)]-CNP((1-22))), and ET-1 ((125)I-ET-1) as radiolabeled ligands. To characterize NP receptor (NPR) subtypes, we also performed an activation of particulate guanylyl cyclase (GC) in membranes of the epididymis by NPs. Specific (125)I-ANP((1-28)) and (125)I-[Tyr(0)]-CNP((1-22)) bindings were localized in surrounding smooth muscle cell layer of the duct of the epididymis with an apparent dissociation constant (K(d)) of 0.84+/-0.15 and 1.74+/-0.39 nM and a maximal binding capacity (B(max)) of 0.47+/-0.11 and 0.08+/-0.01 fmol/mm(2), respectively. Bindings of (125)I-ANP((1-28)) and (125)I-[Tyr(0)]-CNP((1-22)) to these sites were also displaced by des[Gln(18),Ser(19),Gly(20), Leu(21),Gly(22)]ANF((4-23)), a specific ligand of the NP clearance receptor. Production of 3',5'-cyclic guanosine monophosphate by particulate GC in membranes of the epididymis was stimulated by ANP((1-28)), BNP((1-26)), and CNP((1-22)). Receptor subtypes for ET in the epididymis were characterized by competition with BQ 123 and BQ 788 as specific antagonists for ET receptors, type A (ET(A)) and type B (ET(B)) subtypes, respectively. Specific (125)I-ET-1 bindings were localized in the smooth muscle cell layer of the duct of the epididymis with K(d) and B(max) of 0.21+/-0.03 nM and 0.52+/-0.05 fmol/mm(2), respectively. These specific bindings were potently inhibited in a dose-dependent manner by BQ 123, whereas BQ 788 (10 microM) was not in competing for specific (125)I-ET-1 bindings in this structure. Therefore, these results indicate that specific NP and ET receptors are localized in surrounding smooth muscle cells of the duct of the epididymis of the freshwater turtle. It is also suggested that biological and clearance NPR-like subtypes coexist in these cells, and the predominant ET receptor subtype in this tissue is the ET(A)-like receptor. The localization of specific receptors for NPs and ET in the epididymis may be involved in the control of the transport of sperm in the freshwater turtle.

Animals↗
PubMed2002

Dynamic endocrine responses to stress: evidence for energetic constraints and status dependence in breeding male green turtles.

During reproduction, male vertebrates may exhibit a continuum of interactions between sex and adrenal steroids during stressful events, the outcome of which may be important in either reducing or promoting male reproductive success. We studied adult male green turtles (Chelonia mydas) to examine if they altered plasma corticosterone (CORT) and androgen levels in response to a standardized capture/restraint stressor as potential mechanisms to maintain reproductive activity during stressful events. At the population level, we found that migrant breeding males had a significantly smaller CORT response to the capture/restraint stressor compared to nonbreeding males and that this decreased response coincided with the generally poorer body condition of migrant breeders. In contrast, plasma androgen levels decreased significantly in response to the capture/restraint stressor in migrant breeding males, but not in nonbreeding and pre-migrant breeding males. For individual migrant breeding males, the magnitude of their CORT and androgen responses to the capture/restraint stressor was highly correlated with their body condition and body length, respectively. Our results demonstrate that male green turtles exhibit complex interactions in their endocrine responses to a capture/restraint stressor and that variation in these interactions is associated with differences in males' reproductive, energetic, and physical state. We hypothesize that interplay between physical status and plasma hormone responses to stressors could have important consequences for male green turtle reproduction.

Androgens↗
PubMed1995

Reptile heme protein structure: X-ray crystallographic study of the aquo-met and cyano-met derivatives of the loggerhead sea turtle (Caretta caretta) myoglobin at 2.0 A resolution.

The X-ray crystal structures of the aquo-met and cyano-met derivatives of the loggerhead sea turtle (Caretta caretta) myoglobin have been determined at 2.0 A resolution (R = 0.182, and 0.178, respectively). The results here reported, representing the first reptile globin solved by X-ray crystallography, have been analyzed in parallel with data for related monomeric hemoproteins, and indicate a strong overall structural similarity between the loggerhead sea turtle and mammalian myoglobins, reflected by the 63% amino acid identity of their primary structures. The root-mean-square deviation between the entire polypeptide backbones of loggerhead sea turtle and sperm whale myoglobins, after structure superposition, is 0.83 A. Upon cyanide binding to the protein distal site, the iron-bound water molecule present in the aquo-met form is displaced by the incoming ligand. Cyanide is oriented towards the inner part of the heme distal site forming a Fe-C-N angle of 133 degrees.

Animals↗