Evidence for the neural crest origin of turtle plastron bones

Summary: The migrating cranial neural crest cells of birds, fish, and mammals have been shown to form the membranous bones of the cranium and face. These findings have been extrapolated to suggest that all the dermal bones of the vertebrate exoskeleton are derived from the neural crest ectomesenchyme. However, only one group of extant animals, the Chelonians, has an extensive bony exoskeleton in the trunk. We have previously shown that the autapomorphic carapacial and plastron bones of the turtle shell arise from dermal intramembranous ossification. Here, we show that the bones of the plastron stain positively for HNK‐1 and PDGFRα and are therefore most likely of neural crest origin. This extends the hypothesis of the neural crest origin of the exoskeleton to include the turtle plastron. genesis 31:111–117, 2001. © 2001 Wiley‐Liss, Inc.     

The fine structure of the pupal plastron of simuliid flies

The outer surface of the plastron of the pupal spiracular gills of the Simuliidae was previously thought to be the network formed by the apical branches of the vertical struts. It is now shown that this horizontal network supports a thin laminate outer epicuticle and also a fine meshwork, both of which have been previously overlooked. A simple method is described for quickly producing three-dimensional micrographs of great depth with the transmission electron microscope.     

Structural characterization of the micropatterned egg plastron in the mosquito,Aedes albopictus

Eggs of the Asian tiger mosquito Aedes albopictus are fully covered with an air‐covering plastron network that enables them to stay below the surface of the water. Scanning electron microscopy revealed that the chorionic surface was covered with clusters of globular tubercles of different sizes. Histologically, the eggshell provides a typical water‐repellent microarchitecture consisting of an outer exochorion, a membranous endochorion and an intermediate pillar layer. The eggshell has a distinct chorionic tubercle that increases the surface area for gas exchange to enhance respiration capacity. In particular, the chorionic wall gives rise to a hexagonal pattern with smooth and elevated boundaries. In A. albopictus, a set of micropatterns for each hexagon was specified, and each central tubercle was surrounded by 20 small peripheral tubercles. Our fine structural analysis revealed that the partitioning of the surface into numerous hexagonal chorionic sculptures is associated with the Voronoi partition (tessellation), based on the distance to a point in a specific area of the plane. Therefore, the micropatterned surface of the mosquito eggshell appears to not only resist wetting by hydrostatic pressure but also provide resistance to lysate deposition by the biofouling process.     

Relating annual increments of the endangered Blanding's turtle plastron growth to climate

This research is the first published study to report a relationship between climate variables and plastron growth increments of turtles, in this case the endangered Nova Scotia Blanding's turtle (Emydoidea blandingii). We used techniques and software common to the discipline of dendrochronology to successfully cross‐date our growth increment data series, to detrend and average our series of 80 immature Blanding's turtles into one common chronology, and to seek correlations between the chronology and environmental temperature and precipitation variables. Our cross‐dated chronology had a series intercorrelation of 0.441 (above 99% confidence interval), an average mean sensitivity of 0.293, and an average unfiltered autocorrelation of 0.377. Our master chronology represented increments from 1975 to 2007 (33 years), with index values ranging from a low of 0.688 in 2006 to a high of 1.303 in 1977. Univariate climate response function analysis on mean monthly air temperature and precipitation values revealed a positive correlation with the previous year's May temperature and current year's August temperature; a negative correlation with the previous year's October temperature; and no significant correlation with precipitation. These techniques for determining growth increment response to environmental variables should be applicable to other turtle species and merit further exploration.     

Surviving the flood: plastron respiration in the non-tracheate arthropod Phrynus marginemaculatus (Amblypygi: Arachnida)

Specimens of Phrynus marginemaculatus can remain responsive when submerged in water for more than 24 hours. Behavioral data indicate that P. marginemaculatus utilizes dissolved oxygen from the surrounding water. Scanning electron miscroscopy and light microscope sections show cuticular modifications for plastron respiration. All previous examples of plastron respiration have involved animals with tracheal systems, but amblypygids respire through the use of two pairs of book lungs. This study provides the first example of plastron respiration not only in the order Amblypygi, but also, in any non-tracheate arthropod.     

The contribution of neural crest cells to the nuchal bone and plastron of the turtle shell

The origin of the turtle plastron is not well understood, andthese nine bones have been homologized to the exoskeletal componentsof the clavicles, the interclavicular bone, and gastralia. Earlierdata from our laboratory showed that the plastral bone-formingcells stained positively for HNK-1 and PDGFR, two markers ofskeletogenic neural crest cells. We have now shown that theHNK-1⁺ cells are also positive for p75 and FoxD3, affirmingtheir neural crest identity. These cells originate from thedorsal neural tube of stage-17 turtle embryos, several daysafter the original wave of neural crest cells have migratedand differentiated. Moreover, we have demonstrated the existenceof a staging area, above the neural tube and vertebrae, wherethese late-emigrating neural crest cells collect. After residingin the carapacial staging area, these cells migrate to formthe plastral bones. We also demonstrate that one bone of thecarapace, the nuchal bone, also stains with HNK-1 and with antibodiesto PDGFR. The nuchal bone shares several other properties withthe plastral bones, suggesting that it, too, is derived fromneural crest cells. Alligator gastralia stain for HNK-1, whiletheir ribs do not, thus suggesting that the gastralial precursormay also be derived from neural crest cells.     

Are diminutive turtles miniaturized? The ontogeny of plastron shape in emydine turtles

Miniaturization, or the evolution of a dramatically reduced body size compared to related lineages, is an extraordinarily widespread phenomenon among metazoans. Evolutionary biologists have been fascinated by miniaturization because this transition has occurred numerous times, often among close relatives, providing a model system for studying convergent evolution and its underlying mechanisms. Much of the developmental work describing the ontogeny of miniature species suggests that paedomorphosis is the predominant avenue of miniaturization. Nevertheless, specific alterations to ontogeny appear highly variable, so that even related lineages with similar miniaturized traits produce those similarities via distinct ontogenetic paths. One major vertebrate group that has been overlooked in research on miniaturization is turtles. In the present study, we examined patterns of shape change in the plastron (the ventral part of the shell) over the course of ontogeny in a small clade of turtles (Emydinae) aiming to investigate whether two independently evolved diminutive members of the clade (Glyptemys muhlenbergii and Clemmys guttata) should be considered as miniaturized. We employ geometric morphometric methods to quantify the patterns of shape change these potentially miniaturized species and their relatives undergo during ontogeny, and use molecular phylogenetic trees to reconstruct ancestral conditions and provide information on the polarity of shape changes. We find that differing changes in ontogenetic parameters relative to ancestral conditions accompany the evolution of small size in emydines: G. muhlenbergii changes the duration of ontogeny and rate of shape change, whereas C. guttata changes growth rate. The observed ontogenetic repatterning of these species is reminiscent of changes in ontogeny and life history often found in miniaturized taxa. However, we conclude that C. guttata and G. muhlenbergii are not truly miniaturized because they still produce typical adult shell morphologies, and larger emydines display comparable ontogenetic flexibility. Because no emydines carry juvenile shell features forward into adulthood, we speculate that few, if any turtles, will show paedomorphic shell traits without corresponding changes in defensive strategy because such shells may offer insufficient protection. © 2013 The Linnean Society of London     

A test of an antipredatory function of conspicuous plastron coloration in hatchling turtles

Bright colorations in animals are sometimes an antipredatory signal meant to startle, warn, or deter a predator from consuming a prey organism. Freshwater turtle hatchlings of many species have bright ventral coloration with high internal contrast that may have an antipredator function. We used visual modeling and field experiments to test whether the plastron coloration of Chrysemys picta hatchlings deters predators. We found that bird predators can easily distinguish hatchling turtles from their backgrounds and can easily see color contrast within the plastron. Raccoons cannot easily discriminate within-plastron color contrast but can see hatchlings against common backgrounds. Despite this, we found that brightly-colored, high contrast, replica turtles were not attacked less than low contrast replica turtles, suggesting that the bright coloration is not likely to serve an antipredatory function in this context. We discuss the apparent lack of innate avoidance of orange coloration in freshwater turtles by predators and suggest that preference and avoidance of colors are context-dependent. Since the bright colors are likely not a signal, we hypothesize that the colors may be caused by pigments deposited in tissue from maternal reserves during development. In most species, these pigments fade ontogenetically but they may have important physiological functions in species that maintain the bright coloration throughout adulthood.     

Evidence that a late-emerging population of trunk neural crest cells forms the plastron bones in the turtle Trachemys scripta

The origin of the turtle plastron is not known, but these nine bones have been homologized to the exoskeletal components of the clavicles, the interclavicular bone, and gastralia. Earlier evidence from our laboratory showed that the bone-forming cells of the plastron were positive for HNK-1 and PDGFRalpha, two markers of the skeletogenic neural crest. This study looks at the embryonic origin of these plastron-forming cells. We show that the HNK-1+ cells are also positive for p75 and FoxD3, confirming their neural crest identity, and that they originate from the dorsal neural tube of stage 17 turtle embryos, several days after the original wave of neural crest cells have migrated and differentiated. DiI studies show that these are migratory cells, and they can be observed in the lateral regions of the embryo and can be seen forming intramembranous bone in the ventral (plastron) regions. Before migrating ventrally, these late-emerging neural crest cells reside for over a week in a carapacial staging area above the neural tube and vertebrae. It is speculated that this staging area is where they lose the inability to form skeletal cells.     

A review of the North American genus Epimartyria (Lepidoptera, Micropterigidae) with a discussion of the larval plastron

The indigenous North American micropterigid genus Epimartyria Walsingham,1898 is revised. Three species are recognized, including Epimartyria auricrinella Walsingham, 1898 which occurs widely over much of the northeastern United States and Canada, a new species, Epimartyria bimaculella Davis & Landry from northwestern United States and Canada, and Epimartyria pardella (Walsingham, 1880) from northern California to northern Oregon. The larva of Epimartyria auricrinella is described in detail, supplemented with illustrations of the external structure of the larval integument. The larval plastron is described and illustrated for Epimartyria, and this is compared with the plastrons of Neomicropteryx Issiki, 1931 and Micropterix Hübner, 1825. COI barcode sequences show that the three species are genetically distinct, congruent with morphological differences. Marked haplotype divergence within some Epimartyria auricrinella populations appears to be unrelated to morphology, geography or phenology.     

Carapace and plastron sensitivity to touch and vibration in the tortoise (Testudo hermanni and T. graeca)

Neural impulses in response to tactile stimulation of the shell were recorded from afferent nerve fibres in tortoises ( T. graeca and T. hermanni). It was found that there is a mechanoreceptive innervation in the superficial layers of the shell which is sensitive to transient stimuli, particularly to vibration at frequencies up to 100 Hz. Receptive fields pertaining to single and small groups of individual afferent fibres were mapped: the fields were sharply circumscribed and distributed in relation to the scutes of the shell. The tactile innervation that was found would be consistent with a capacity for recognition and accurate localization of innocuous stimuli and may play a central role in courtship and mating behaviour.     

Relationships among turtles of the genus Clemmys (Reptilia, Testudines, Emydidae) as suggested by plastron scute morphology

The Conservative morphology of hardshelled turtles has fostered the use of size relationships between epidermal scutes (scales) on the shell to differentiate between species and subspecies of many taxa. The size relationship of the six major pairs of plastral scutes were used to compare the four currently recognized species of the genus Clemmys with each other. as well as with the distantly related Graptemys barbouri using Jaccard Coefficients. Shannon-Weiner diversity indices, and multivariate analysis. Results were concordant among the three techniques used and confirm our prediction that plastral morphology varies little among closely related species and widely among distantly related taxa. Clemmys muhlenbergii appears to he more different from Clemmys guttata than previously suggested. Analysis of plastral morphology shows promise as a taxonomic tool for turtle systematists.     

Physical gills in Elmidae (Coleoptera: Byrrhoidea): Structure and evolutionary pattern of plastron in Stenelmis and related genera

Many species within Elmidae (Coleoptera: Byrrhoidea) have plastrons composed of flattened setae. However, some genera display fine plastrons on the epicuticle, called plastron hairs. In Japanese elmids, members of the genera Stenelmis, Ordobrevia, Nomuraelmis and Leptelmis bear ventral plastron hairs. Based on a maximum likelihood tree including most Japanese genera within Elmidae, we found that these genera are monophyletic and that plastron hairs are a derived character in Elmidae. We also found that the genus Graphelmis bears jigsaw puzzle‐like plastron scales with plastron hair‐like projections, and is sister to the group with plastron hairs.