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Serum gonadotropins and gonadal steroids associated with ovulation and egg production in sea turtles.

Changes in serum concentrations of gonadotropins and gonadal steroids during the periovulatory period were monitored in green, Chelonia mydas, and loggerhead, Caretta caretta, sea turtles. Turtles were from natural populations that nest on a coral island on the Great Barrier Reef. After nesting, each turtle was transferred to a holding tank and held for a maximum of 8 days. A time series of blood samples was obtained from each of five sea turtles (three C. mydas and two C. caretta) starting immediately after nesting and then at approximately 12-hr intervals until the time of release. Prior to release back into the ocean, each turtle was examined by laparoscopy to verify that ovulation had occurred. Serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (PRO), and testosterone (T) in both species exhibited significant changes during this period. Surges of FSH, LH, and PRO were evident within approximately 20 to 50 hr after each turtle had nested. The significant change in FSH concentration during the periovulatory period is the first such report for a reptile. Coincident with maximal concentrations of FSH, LH, and PRO was a decline in T concentrations in both species. Estradiol-17 beta concentrations were near or below assay sensitivity in the C. mydas, whereas those in the C. caretta were detectable but exhibited no significant changes. The dynamic changes in FSH, LH, PRO, and T concentrations are consistent with the hypothesis that these hormones facilitate specific physiological events during ovulation and egg production.

Animals↗

Ventilation, gas exchange and metabolic scaling of a sea turtle.

Ventilation of green turtles (Chelonia mydas) was affected by the position in which the animal was placed: supine animals breathed slowly 0.07 breaths/min) and deeply (8.0 L/breath); prone animals breathed more rapidly (0.43 breaths/min) and more shallowly (3.5 L/breath). From the respiratory exchange ratio and other indicators it appears that green turtles hyperventilate during exercise and hypoventilate during recovery. O2 consumption of the resting sea turtle (0.024 L-kg-1-h-1) is similar to that of other large turtles. Maximal O2 consumption (0.25L-kg-1-h-1) is greater than that of other large turtles. Minimal O2 consumption scaled in proportion to the -0.17 power of the body mass of green turtles over the range of 0.030 to 141.5 kg. The maximal O2 consumption scaled in proportion to the -0.06 power of body mass for the same range of body masses.

Animals↗

Extracellular and intracellular acid-base effects of submergence anoxia and nitrogen breathing in turtles.

We compared extracellular and intracellular acid-base state in turtles (Chrysemys picta bellii) subjected to anoxic submergence to turtles made anoxic by N2-breathing. Measurements made on control animals and on animals after 1, 2, 4, or 6 h of anoxia included blood pH, PO2, PCO2, and lactate as well as liver, heart, skeletal muscle, and brain pHi (using DMO equilibration), lactate, and glycogen concentrations. We hypothesized that the anaerobic metabolic rate of submerged turtles would be depressed by the more severe extra- and intracellular acidosis, and that this would be indicated by reduced lactate accumulation and glycogen depletion. Submerged turtles became extremely acidemic due to a combined metabolic and respiratory acidosis and had significantly lower arterial pH than N2-breathing animals (6.98 and 7.34, respectively, after 6 h). In spite of this disparity in pHa, 6 h pHi values for liver, heart, and brain were similar. Likewise, our data on glycogen depletion and lactate accumulation at h 6 in these tissues suggest no dramatic differences in anaerobic metabolic rate. While skeletal muscle pHi was somewhat lower at h 6 in the submerged group (6.73 vs 6.91 for N2-breathers), we observed no differences in either glycogen depletion or lactate accumulation in this tissue between our two treatments. Thus, at h 6, in spite of a 0.37 pH unit difference in pHa and a nearly 70 mm Hg difference in arterial and presumably cytosolic PCO2, pHi and tissue lactate and glycogen concentrations were similar. These results can be explained if the in vivo intracellular buffer values (beta) of turtle tissues are very high. We conclude that extracellular acid-base state is not necessarily reflected intracellularly in vivo in turtles and care must be taken in extrapolating from one compartment to another when attempting to make inferences about metabolic depression or acid-base regulation in this species.

Acid-Base Equilibrium↗

Surface immunoglobulin on granular and agranular leukocytes in the thymus and spleen of the snapping turtle, Chelydra serpentina.

Spleen and thymus suspensions from the turtle, Chelydra serpentina were examined by indirect immunofluorescence and found to contain both agranulocytes and granulocytes positive for surface Ig. Among the splenic agranulocytes , 50% of the lymphocyte and monocyte population was positive for surface Ig. In the thymus only 7% of the thymocytes were positive. Ninety-two percent of the granulocytes, composed primarily of basophils and eosinophils, were positive for surface Ig in both the thymus and spleen. The presence of surface immunoglobulins on turtle splenic leukocytes was confirmed by sheep red blood cell (SRBC) immunization studies. Basophils, lymphocytes and monocytes from immunized turtles formed SRBC rosettes, while eosinophils from immunized turtles were found to specifically phagocytose SRBC. Splenic leukocytes from control turtles did not phagocytose or rosette with SRBC. This study demonstrates that indirect immunofluorescent techniques can be used to identify surface immunoglobulin on turtle granulocytes as well as agranulocytes .

Animals↗

Neurotransmitter organization and connections of turtle cortex: implications for the evolution of mammalian isocortex.

Telencephalic cortex in turtles is a simple three layered-structure. The dorsal most part of this structure is thought to resemble the reptilian forerunner of at least parts of mammalian isocortex. This dorsal part of turtle cortex contains several functionally distinct regions that show similarity in their connections and function to specific areas in mammalian isocortex. The types of neurons found in turtle dorsal cortex (as defined by their morphology and neurotransmitter content) also show great similarity to those observed in mammals, with the major exception that turtle cortex appears to lack the types of neurons found in granular and supragranular layers of mammalian isocortex. Similar results have also been observed in other living reptiles. Thus, one major step in the evolution of reptilian cortex into mammalian cortex must have been the addition of the types of neurons found in the granular and supragranular layers of mammalian isocortex. These observations for turtles also suggest that turtle cortex in particular and reptilian telencephalic cortex in general must differ functionally from mammalian isocortex with respect to those features associated with the laminar and columnar organization of isocortex. These issues are discussed in more detail below and in Reiner (1991).

Animals↗

Ganglion cell densities in normal and dark-reared turtle retinas.

In dark-reared, neonatal turtle retinas, ganglion cell receptive fields and dendritic trees grow faster than normal. As a result, their areas may become, on average, up to twice as large as in control retinas. This raises the question of whether the coverage factor of dark-reared ganglion cells is larger than normal. Alternatively, dark rearing may lead to smaller-than-normal cell densities by accelerating apoptosis. To test these alternatives, we investigated the effect of light deprivation on densities and soma sizes of turtle retinal ganglion cells. For this purpose, we marked these cells using retrograde labeling of fixed turtle retinas with DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). Control turtles were maintained in a regular 12-h light/dark cycle from hatching until 4 weeks of age, whereas dark-reared turtles were maintained in total darkness for the same period. Ganglion cells in the control and dark-reared retinas were found to be similar in density and soma sizes. These results show that the mean coverage factor of turtle dark-reared ganglion cells is larger than normal.

Animals↗

Comparison of sea turtle thrombocyte aggregation to human platelet aggregation in whole blood.

The endangered sea turtles are living "fossils" that afford us an opportunity to study the hemostatic process as it likely existed millions of years ago. There are essentially no data about turtle thrombocyte aggregation prior to our studies. Thrombocytes are nucleated cells that serve the same hemostatic functions as the anucleated mammalian platelet. Sea turtle thrombocytes aggregate in response to collagen and beta-thrombin. Ristocetin induces an agglutination/aggregation response indicating the presence of a von Willebrand-like receptor, GPIb, found in all mammalian platelets. Samples treated with alpha-thrombin plus gamma-thrombin followed by ristocetin results in a rapid, stronger response than ristocetin alone. These responses are inhibited by the RGDS peptide that blocks fibrinogen cross-linking of mammalian platelets via the fibrinogen receptor, GPIIb/IIIa. Three platelet-like proteins, GPIb, GPIIb/IIIa and P-selection are detected in sea turtle thrombocytes by fluorescence activated cell sorting. Turtle thrombocytes do not respond to ADP, epinephrine, serotonin, thromboxane A2 mimetic, U46619, trypsin, or alpha-thrombin and gamma-thrombin added alone. Comparison of hemostasis in sea turtles to other vertebrates could provide a framework for understanding the structure/function and evolution of these pathways and their individual components.

Animals↗

Depot fatty acid composition in immature green turtles (Chelonia mydas) residing at two near-shore foraging areas in the Hawaiian Islands.

The lipid content and fatty acid composition of depot fat were determined for 58 immature green turtles (Chelonia mydas) residing at two near-shore foraging areas, Ahu-O-Laka, located in Kaneohe Bay on Oahu, and Kiholo Bay located on the island of Hawaii. Benthic flora at Kiholo was limited to a single algal species but included algae and seagrass at Ahu-O-Laka. Turtle straight carapace length ranged from 38.6 to 59.2 cm, suggesting that the sample set included new recruits to up to 12-year residents. Fatty acid data were analyzed using principal components analysis (PCA). PC1 accounted for over 50% of the variance. Turtles were generally delineated along PC1 by the length of time on benthic foraging grounds, with high (>0.75) negative loadings for the fatty acids 22:6n-3, 7M7H, t16:1n-10, 15:0, and 17:0 associated with relatively new recruits (suggesting a pelagic dietary source for these fatty acids) and high positive loadings for 12:0 and 14:0 associated with long-term residents. PC2 separated turtles primarily by capture location, with high positive loadings for 18:2n-6 and 18:3n-3 [the primary seagrass polyunsaturated fatty acids] associated with the Ahu-O-Laka turtles. Fatty acid profiles of turtles from both locations differed substantially from those of their benthic diets, suggesting considerable modification of dietary fatty acids and de novo biosynthesis.

Adaptation, Physiological↗

Partial development of the steroidogenic ultrastructural features in degenerative corpora lutea after a single injection of pituitary extract in the Western painted turtle (Chrysemys picta).

Pituitary glands were removed from sexually mature female turtles (Chrysemys picta) and they were injected intraperitoneally (i.p.) into other mature females of the same species (experimental). In addition mature females of the same species received saline injection only (controls). Initially all the turtles used in this study were steroidogenically inactive with corpora lutea already undergoing luteolysis (degeneration) as these turtles had ovioposited their eggs approximately 2 weeks earlier. Forty-eight hour post injection the corpora lutea were removed from the control and experimental turtles. In the experimental turtles, the lutein granulosa cells developed ultrastructural features such as tubular and cisternal smooth endoplasmic reticulum (SER) and mitochondria with tubular cristae associated with lipid droplets. However, the controls maintained degenerative corpora lutea without steroidogenic ultrastructural features. The circulating progesterone (Pro) levels in the experimental turtles were significantly higher than the controls (P<0.049). Although the 48h development of steroidogenic ultrastructural features in the lutein granulosa cells was only partial in development, the effect of the pituitary taken from the inactive donor triggered an activating process within a short period, clear evidence of gonadotropic effect on the inactive corpora lutea. The present data offer interesting information on the short-term effect of gonadotropins during the non-reproductive period. This information may have useful implication under natural conditions particularly during the onset of a new reproductive cycle where the ovary is still inactive.

Animals↗

Molecular cloning of the cDNA encoding follicle-stimulating hormone beta subunit of the Chinese soft-shell turtle Pelodiscus sinensis, and its gene expression.

Follicle-stimulating hormone (FSH) is a member of the pituitary glycoprotein hormone family. These hormones are composed of two dissimilar subunits, alpha and beta. Very little information is available regarding the nucleotide and amino acid sequence of FSHbeta in reptilian species. For better understanding of the phylogenetic diversity and evolution of FSH molecule, we have isolated and sequenced the complementary DNA (cDNA) encoding the Chinese soft-shell turtle (Pelodiscus sinensis, Family of Trionychidae) FSHbeta precursor molecule by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) methods. The cloned Chinese soft-shell turtle FSHbeta cDNA consists of 602-bp nucleotides, including 34-bp nucleotides of the 5'-untranslated region (UTR), 396-bp of the open reading frame, and 3'-UTR of 206-bp nucleotides. It encodes a 131-amino acid precursor molecule of FSHbeta subunit with a signal peptide of 20 amino acids followed by a mature protein of 111 amino acids. Twelve cysteine residues, forming six disulfide bonds within beta-subunit and two putative asparagine-linked glycosylation sites, are also conserved in the Chinese soft-shell turtle FSHbeta subunit. The deduced amino acid sequence of the Chinese soft-shell turtle FSHbeta shares identities of 97% with Reeves's turtle (Family of Bataguridae), 83-89% with birds, 61-70% with mammals, 63-66% with amphibians and 40-58% with fish. By contrast, when comparing the FSHbeta with the beta-subunits of the Chinese soft-shell turtle luteinizing hormone and thyroid stimulating hormone, the homologies are as low as 38 and 39%, respectively. A phylogenetic tree including reptilian species of FSHbeta subunits, is presented for the first time. Out of various tissues examined, FSHbeta mRNA was only expressed in the pituitary gland and can be up-regulated by gonadotropin-releasing hormone in pituitary tissue culture as estimated by fluorescence real-time PCR analysis.

Amino Acid Sequence↗

Molecular cloning and sequence analysis of a cDNA encoding pituitary thyroid stimulating hormone beta-subunit of the Chinese soft-shell turtle Pelodiscus sinensis and regulation of its gene expression.

A cDNA encoding thyroid stimulating hormone beta-subunit (TSHbeta) was cloned from pituitary of the Chinese soft-shell turtle, Pelodiscus sinensis, and its regulation of mRNA expression was investigated for the first time in reptile. The Chinese soft-shell turtle TSHbeta cDNA was cloned from pituitary RNA by reverse transcription and polymerase chain reaction (RT-PCR), and rapid amplification cDNA end (RACE) methods. The Chinese soft-shell turtle TSHbeta cDNA consists of 580-bp nucleotides, including 67-bp nucleotides of 5'-untranslated region (UTR), 402-bp of the open reading frame, and 97-bp of 3'-UTR followed by a 14 poly (A) trait. It encodes a precursor protein molecule of 133 amino acids with a putative signal peptide of 19 amino acids and a putative mature protein of 114 amino acids. The number and position of 12 cysteine residues, presumably forming six disulfide bonds, one putative asparagine-linked glycosylation site, and six proline residues that are found at positions for changing the backbone direction of the protein have been conserved in the turtle as in other vertebrate groups. The deduced amino acid sequence of the Chinese soft-shell turtle TSHbeta mature protein shares identities of 82-83% with birds, 71-72% with mammals, 49-57% with amphibians, and 44-61% with fish. The Chinese soft-shell turtle pituitaries were incubated in vitro with synthetic TRH (TSH-releasing hormone), thyroxine and triiodothyronine at doses of 10(-10) and 10(-8)M. TRH stimulated, while thyroid hormones suppressed, TSHbeta mRNA levels in dose-related manner. The sequences of cDNA and its deduced peptide of TSHbeta as well as the regulation of its mRNA level were reported for the first time in reptile.

Amino Acid Sequence↗

Turtle phylogeny: insights from a novel nuclear intron.

Introns have gained considerable popularity as markers for molecular phylogenetics. However, no primers exist for a nuclear intron that amplifies across all turtles. Available data from morphology and mitochondrial DNA have not unambiguously resolved relationships within the superfamily Trionychoidea and the family Chelidae, which together form a large portion of extant turtle diversity. We tested the phylogenetic utility of a novel intron from the RNA fingerprint protein 35 (R35) as applied to these two areas of turtle systematics. We found the intron to be a single-copy locus that provides excellent resolving power for lineages among turtles, though problems with alignment made it impossible to infer deeper amniote relationships. Maximum parsimony and maximum likelihood both demonstrated the polyphyly of Trionychoidea and the reciprocal monophyly of Australian/New Guinea and South American chelid turtles. This is the first study to resolve such relationships with strong statistical support, and we suggest that R35 holds great promise for resolving additional persistent problems in the phylogeny of living turtles.

Animals↗

Seasonal changes in hepatic progesterone receptor mRNA, estrogen receptor mRNA, and vitellogenin mRNA in the painted turtle, Chrysemys picta.

Previous studies using the fresh water turtle Chrysemys picta have demonstrated that progesterone (P) inhibits estradiol (E)-induced vitellogenin (vtg) secretion in this species. Further, there is evidence for the differential expression of the two P receptor isoforms (PRA and PRB) in the liver during the turtle seasonal cycle, correlating with hepatic vitellogenesis. In this study we report changes in the hepatic PR mPNA, ER mRNA, and vitellogenin (vtg) mRNA transcripts during the reproductive cycle of the turtle. Fragments of the turtle hepatic PR and ER cDNAs were cloned and sequenced and a previously cloned turtle vtg cDNA were used as probes in Northern blotting. No 3.7-kb PR mRNA, corresponding to the smaller PR transcript, PRA of other species was found, although, a smaller 1.8-kb transcript (putative PRC mRNA) was present. These observations suggest that the turtle as in the chicken and human, the 4.5-kb PR mRNA transcript encodes both PRA and PRB proteins. Only the larger PR mRNA transcript (4.5-kb), was found to vary significantly during the annual cycle, being highest when vitellogenesis was inhibited in winter and summer. Vtg mRNA could not be detected during the summer or winter, was highest during vitellogenesis in the spring, and reappeared during the fall period of vitellogenesis and ovarian recrudescence. ER mRNA followed a similar pattern, being highest during spring and early fall, when vtg synthesis is high. The data suggest that P/PR, as well as E/ER, may be involved in the seasonal regulation of hepatic vitellogenesis in this species.

Amino Acid Sequence↗

Subcellular distribution of trace elements in the liver of sea turtles.

Subcellular distribution of Cu, Zn, Se, Rb, Mo, Ag, Cd and Pb was determined in the liver of green turtles (Chelonia mydas) and hawksbill turtles (Eretmochelys imbricata) from Yaeyama Islands, Japan. Also, hepatic cytosol from sea turtles was applied on a Sephadex G-75 column and elution profiles of trace elements were examined. Copper, Zn, Se, Rb, Ag and Cd were largely present in cytosol in the liver of both species, indicating that cytosol was the significant site for the accumulation of these elements in sea turtles. In contrast, Mo and Pb were accumulated specifically in nuclear and mitochondrial fraction and microsomal fraction, respectively. Gel filtration analysis showed that Cu, Zn, Ag and Cd were bound to metallothionein (MT) in the cytosol of sea turtles. To our knowledge, this is the first report on the association of trace elements with MT in sea turtles.

Animals↗

Identification and expression of gastrin and cholecystokinin mRNAs from the turtle, Pseudemys scripta: evidence of tissue-specific tyrosyl sulfation(1).

Gastrin and cholecystokinin (CCK) are related peptide hormones expressed in the brain and gut of vertebrates. In this study, complementary DNAs have been characterised from the red-eared slider turtle, Pseudemys scripta. The encoded preproCCK contains mono and dibasic endoproteolytic processing sites for formation of the previously identified CCK-70, CCK-40 and CCK-8 products, whereas preprogastrin contains two dibasic processing sites for the generation of gastrin-52. Alignment of the predicted preprohormone structures with those of other species, showed that preproCCK has been well conserved among all vertebrates, whereas progastrin is less conserved. Both gastrin and CCK mRNA display expression patterns similar to their mammalian counterparts, with CCK being expressed in the brain, duodenum and small intestine, and gastrin in the antrum. Heterologous expression of turtle preprogastrin in a mammalian endocrine cell line led to production of carboxyamidated gastrin-52 as observed in turtle antrum. However, in contrast to the non-sulfated endogenous peptide, the heterologously expressed gastrin was completely Tyr sulfated. Consequently, it appears that either gastrin producing cells in the turtle gut do not express tyrosylprotein sulfotransferases or the enzyme(s) present in turtle antrum is unable to sulfate turtle gastrin.

Amino Acid Sequence↗

Cytochromes P450 in liver of the turtle Chrysemys picta picta and the induction and partial purification of CYP1A-like proteins.

Cytochromes P450 (CYP) in hepatic microsomes from the turtle Chrysemys picta picta and their response to inducers were examined. Freshly caught turtles had one protein (59 kDa) detected in western blot with monoclonal antibody 1-12-3 to scup CYP1A. That same band and a second band were detected with polyclonal anti-mouse Cyp1a1. Polyclonal anti-scup P450B (putative CYP2B) recognized three bands and anti-scup P450A (putative CYP3A), one band. TCB (3,3',4,4'-tetrachlorobiphenyl) at 5 mg kg-1 injected once induced EROD activity 3-fold. Repeated high-dose injections of TCB, 2,3,3',4,4'-pentachlorobiphenyl, Aroclor 1254 or beta-naphthoflavone induced CYP1A 20-fold and P450B-related proteins 2-3-fold. Rates of ethoxy- (EROD) methoxy- (MROD) and pentoxyresorufin O-dealkylases and benzo[a]pyrene (B[a]P) hydroxylase (AHH) were induced by these treatments, and were correlated with putative CYP1A content. Phenobarbital slightly elevated only MROD activity. Ethoxycoumarin (EC) O-deethylase rates were high, 1.6-2.2 nmol min-1 mg-1 in control and treated turtles, suggesting that EC is not a turtle CYP1A substrate. Highly induced EROD rates were 0.06 nmol min-1 mg-1, while AHH rates exceeded 4 nmol min-1 mg-1, suggesting that C. picta picta CYP1A may prefer PAH substrates. Induction of AHH was reflected in the formation of metabolites 3-OH-, 9-OH- and 7-OH-BP and BP-7,8-dihydrodiol (DHD). BP-4,5-DHD was not detected. Chromatographic procedures resolved the 59 kDa putative CYP1A from the second protein recognized by anti-Cyp1a1. The 59-kDa protein was also specifically and highly immunopurified by Mab 1-12-3. Thus, several CYP including two CYP1A-related proteins are expressed in turtle liver. Multiple CYP1A genes in reptiles may provide an insight into the origin of divergence in the CYP1A subfamily. Induction of a CYP1A may be a useful indicator of exposure to Ah receptor agonists in turtles.

Animals↗

Phylogenetic position of turtles among amniotes: evidence from mitochondrial and nuclear genes.

Maximum likelihood analysis, accounting for site-heterogeneity in evolutionary rate with the Gamma-distribution model, was carried out with amino acid sequences of 12 mitochondrial proteins and nucleotide sequences of mitochondrial 12S and 16S rRNAs from three turtles, one squamate, one crocodile, and eight birds. The analysis strongly suggests that turtles are closely related to archosaurs (birds+crocodilians), and it supports both Tree-2: (((birds, crocodilians), turtles), squamates) and Tree-3: ((birds, (crocodilians, turtles)), squamates). A more traditional Tree-1: (((birds, crocodilians), squamates), turtles) and a tree in which turtles are basal to other amniotes were rejected with high statistical significance. Tree-3 has recently been proposed by Hedges and Poling [Science 283 (1999) 998-1001] based mainly on nuclear genes. Therefore, we re-analyzed their data using the maximum likelihood method, and evaluated the total evidence of the analyses of mitochondrial and nuclear data sets. Tree-1 was again rejected strongly. The most likely hypothesis was Tree-3, though Tree-2 remained a plausible candidate.

Animals↗

Ultrastructural study of the dorsal lingual epithelium of the Asian snail-eating turtle, Malayemys subtrijuga.

The Asian snail-eating turtle, Malayemys subtrijuga, is classified phylogenetically as a member of the family Emydinae. Members of this family usually live in small rivers or ponds. However, this species is relatively well-adapted to terrestrial life. We describe here the light, scanning electron and transmission electron microscopic appearance of the dorsal lingual epithelium of the snail-eating turtle and we compare the results to those obtained from other freshwater turtles in an attempt to clarify the relationship between the histological and ultrastructural differences in the lingual epithelium and the living circumstances of the turtles. The tongue is triangular with a rounded apex when viewed dorsally but it appears flattened when viewed laterally. Under the scanning electron microscope, no lingual papillae were visible on the dorsal surface of the tongue. Instead, plicae were seen all over the dorsal surface. On the surface of the epithelium of the outermost side, dome-shaped bulges, each of which was coincident with an individual cell, were compactly distributed. At higher magnification, scanning electron microscopy revealed numerous microvilli and microridges on the surface of these cells, and the thickening of cell-margins was clearly seen. Light microscopy revealed that the mucosal epithelium of the tongue was of the non-keratinized, stratified squamous type. Under the transmission electron microscope, the cells of the basal and deep intermediate layers of the epithelium appeared irregularly elliptical in shape. The nucleus was large and also irregularly elliptical, lying in the central region of each epithelial cell. The cytoplasm of these cells contained mitochondria, free ribosomes, rough endoplasmic reticulum and bundles of tonofibrils. Cell membranes formed processes around individual cells. Desmosomes were intercalated between the processes of adjacent cells. In the shallow intermediate layer, the cells were also elliptical, and the elliptical nucleus was located in the central area of each cell. A large part of the cytoplasm was occupied by electron-dense, discoid granules. Filamentous structures filled the spaces between these granules. Small numbers of free ribosomes, mitochondria and rough endoplasmic reticulum were scattered in the cytoplasm. Cell membranes still formed processes around cells. Desmosomes were intercalated between the processes of adjacent cells. The cells of the surface layer were still elliptical, as were their nuclei. Most of the cytoplasm was filled with electron-dense, discoid granules. Fine filamentous structures were dispersed between these granules. Cell membranes formed processes around cells which were coincident with microvilli and microridges. Intercalated desmosomes were also seen. In some cells, many of the electron-dense, discoid granules were secreted into the oral cavity. In conclusion, the histology of the lingual epithelium of the snail-eating turtle is very similar to that of the freshwater turtle, reflecting similarities in the gross morphology of the tongues of these species, in spite of the differences in their life styles.

Animals↗