Larval Types Research Articles

Olfactory metamorphosis in the coastal tailed frog Ascaphus truei (Amphibia, Anura, Leiopelmatidae)

The structure of the olfactory organ in larvae and adults of the basal anuran Ascaphus truei was examined using light micrography, electron micrography, and resin casts of the nasal cavity. The larval olfactory organ consists of nonsensory anterior and posterior nasal tubes connected to a large, main olfactory cavity containing olfactory epithelium; the vomeronasal organ is a ventrolateral diverticulum of this cavity. A small patch of olfactory epithelium (the "epithelial band") also is present in the preoral buccal cavity, anterolateral to the choana. The main olfactory epithelium and epithelial band have both microvillar and ciliated receptor cells, and both microvillar and ciliated supporting cells. The epithelial band also contains secretory ciliated supporting cells. The vomeronasal epithelium contains only microvillar receptor cells. After metamorphosis, the adult olfactory organ is divided into the three typical anuran olfactory chambers: the principal, middle, and inferior cavities. The anterior part of the principal cavity contains a "larval type" epithelium that has both microvillar and ciliated receptor cells and both microvillar and ciliated supporting cells, whereas the posterior part is lined with an "adult-type" epithelium that has only ciliated receptor cells and microvillar supporting cells. The middle cavity is nonsensory. The vomeronasal epithelium of the inferior cavity resembles that of larvae but is distinguished by a novel type of microvillar cell. The presence of two distinct types of olfactory epithelium in the principal cavity of adult A. truei is unique among previously described anuran olfactory organs. A comparative review suggests that the anterior olfactory epithelium is homologous with the "recessus olfactorius" of other anurans and with the accessory nasal cavity of pipids and functions to detect water-borne odorants.

The fate of the larval epidermis in the Desor-larva of Lineus viridis (Pilidiophora, Nemertea) displays a historically constrained functional shift from planktotrophy to lecithotrophy

Pilidiophora constitutes a clade of nemerteans characterized by a peculiar larval type, the pilidium. A characteristic of this larva is the transitory epidermis in which the juvenile develops from imaginal discs. The primary function of this larval envelope is assumed to be feeding and dispersal. When juvenile development is complete, the larval epidermis is ruptured and swallowed by the juvenile. According to recent cladistic and molecular analyses of the Nemertea, the intracapsular Desor-larva of the sibling species Lineus viridis and L. ruber is thought to have evolved from a pelagic pilidium. The general course of development has been demonstrated to be similar to that of the pilidium, in which the juvenile forms from imaginal discs under the larval epidermis. The two Lineus species, however, differ in their mode of larval feeding: L. ruber being ootrophic and L. viridis being lecithotrophic. In order to elucidate the transition from the planktotrophic pilidum to lecithotrophic development, I studied the early cleavage and metamorphosis from intracapsular Desor-larva to juvenile stages in L. viridis from the island of Sylt, using light microscopical, electron microscopical, and fluorescent staining methods. Due to the specific cleavage pattern with equally sized 1st quartet animal blastomeres and vegetal blastomeres in L. viridis, the larval epidermis later contains a considerable amount of the yolk reserve. During metamorphosis, the larval epidermis is ingested by the juvenile thus displaying behavior similar to that of the pilidium larva. In contrast to the pilidium, the function of the larval epidermis of the Desor-larva has shifted from feeding and dispersal to direct food supply. Thus, the development of L. viridis is a perfect example for strong historical constraints that prevent ancestral larval structures from being lost.

Patterns of early colony development (astogeny) in four genera of trepostome bryozoans from the Upper Ordovician

The early astogeny of specimens representing the bryozoan genera Dekayia, Parvohallopora, Heterotrypa, and Homotrypella follows the same pattern documented in other Ordovician trepostomes. After larval settlement and metamorphosis, typically three primary, periancestrular, autozooids bud distally from the founder zooid, the ancestrula, with secondary zooids budding from the primaries, tertiary zooids from the secondaries, and so on. Progressive lateral displacement of buds, through backbudding, eventually produces a subcircular ancestrular disk. Significant intergeneric differences in this basic pattern do not appear to exist. Intrageneric variability in budding pattern occurs during colony development. That variability might have resulted from any or all of the following sources: within-genus plasticity, the development of multiple ancestrulae, or environmental variation. The basic pattern of three primary buds developed from individual ancestrulae has been recognized in other trepostomes, some cyclostomes, and two suborders of cryptostomes. Cladistically, the trait must be considered polyphyletic until more is known about the number of primary buds in the remaining bryozoan suborders. Ancestrular dimensions are statistically similar across genera except between the smaller Dekayia and larger Parvohallopora, but ancestrular sizes in all four Dillsboro genera are consistent with the inference that they possessed a polyembryonic lecithotrophic larval type.

Synergistic effects of geographical strain, temperature and larval food on insecticide tolerance in Callosobruchus maculatus (F.)

AbstractThis study investigated the multifactorial interaction of various environmental factors including geographical strain (Brazil, Cameroon and Yemen strains), temperature, dose and larval food (cowpea and mungbean) on the response of Callosobruchus maculatus adult to insecticide. All the main factors, their two‐way interactions and the four‐way interaction had significant effects on C. maculatus response to malathion (an organophosphate insecticide). However, the three‐way interactions were not statistically significant (except strain × food × dose, P = 0.002). The Brazil strain was the most responsive to temperature irrespective of the larval food type. The impact of food type differs from one strain to the other; for instance, the food that imparts higher tolerance in a strain might reduce the tolerance in another. Likewise, the hierarchy of tolerance among the cowpea‐reared strains (Brazil > Cameroon > Yemen) was totally different from that of the mungbean‐reared strains (Cameroon > Yemen > Brazil). The reasons for these differences were discussed in the light of their impact on C. maculatus management. Both the management of C. maculatus and development of resistance could be complex; hence, the states of a variety of environmental factors need to be considered. This is necessary to maximize management success of this bruchid, especially in tropical/subtropical developing countries.

Evaluating neurophylogenetic patterns in the larval nervous systems of brachiopods and their evolutionary significance to other bilaterian phyla

Recent structural analyses of invertebrate nervous systems have supported hypotheses stating that specific developmental and cytological aspects of larval and adult brains are conserved among bilaterian animals. Opposing views argue that structural similarities in larval nervous systems may be the result of convergent evolution and that the developmental diversity of adult brains is more indicative of several independent origins. Here, I use various cytological probes, confocal microscopy, and reconstruction techniques to investigate the cellular diversity within the larval nervous systems of Glottidia pyramidata and Terebratalia transversa (Brachiopoda). Neuronal cell types are compared among the rhynchonelliform, linguliform, and craniiform brachiopods as well as the phoronids. Although the respective larval types of the previously mentioned systematic groups clearly diverge in the neuroarchitecture of their larval apical organs (and nervous systems in general), a ground plan is proposed based on shared, centrally-located, peptidergic neuronal cell types that can be compared with similar cell types in other lophotrochozoan phyla (bryozoans and spiralians). Assessing hierarchal levels of homology within and among the nervous systems of morphologically disparate phyla is challenging in that many phyla share early developmental signals that induce the specification of the neural ectoderm, clouding our ability to discern divergent larval and juvenile brain structure. Solving these problems will require a combined effort involving both traditional and more recent cytological techniques with a diversity of molecular probes that will better map the neuronal complexity of diverse invertebrate nervous systems.

Effects of Triclocarban, Triclosan, and Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems

Triclosan (TCS) and triclocarban (TCC) are widely used broad spectrum bactericides that are common pollutants of waterways and soils. Methyl triclosan (mTCS) is the predominant bacterial TCS metabolite. Previous studies have shown that TCS disrupts thyroid hormone (TH) action; however, the effects of mTCS or TCC are not known. The present study uses the cultured frog tadpole tail fin biopsy (C-fin) assay and the TH-responsive rat pituitary GH3 cell line to assess the effects of these three chemicals (1-1000 nM) on TH signaling and cellular stress within 48 h. mRNA abundance of TH receptor β, Rana larval keratin type I (TH-response), heat shock protein 30, and catalase (stress-response) was measured using quantitative real-time polymerase chain reaction in the C-fin assay. The TH-responsive gene transcripts encoding growth hormone, deiodinase I, and prolactin were measured in GH3 cells with the heat shock protein 70 transcript acting as a cellular stress indicator. We found alteration of stress indicators at a wide range of concentrations of TCS, mTCS, and TCC in both test systems. mTCS and TCC affected TH-responsive gene transcripts at the highest concentration in mammalian cells, whereas a modest effect included lower concentrations in the C-fin assay. In contrast, TCS did not affect TH-responsive transcripts. These results identify nontarget biological effects of these bacteriocides on amphibian and mammalian cells and suggest the TH-disrupting effects observed for TCS could be mediated through its metabolite.

The Hawaiian amphibious caterpillar guild: new species of Hyposmocoma (Lepidoptera: Cosmopterigidae) confirm distinct aquatic invasions and complex speciation patterns

Although aquatic caterpillars are a globally rare lifestyle, we have found them in multiple, independent lineages of the endemic moth genus Hyposmocoma across all of the Hawaiian Islands with flowing water. We formally describe 11 new species of Hyposmocoma that belong to four different larval case types: cone, bugle, medium burrito, and large burrito: Hyposmocoma kahamanoasp. nov. from Oahu Island, Hyposmocoma kamakou sp. nov. from Molokai Island, Hyposmocoma kahaiaosp. nov., Hyposmocoma waihohonusp. nov., and Hyposmocoma moopalikeasp. nov. from Maui Island, andHyposmocoma aumakuawaisp. nov.,Hyposmocoma eepawaisp. nov., Hyposmocoma ipowainuisp. nov., Hyposmocoma kawaikoisp. nov., Hyposmocoma uhauiolesp. nov., and Hyposmocoma wailuasp. nov. from Kauai Island. We also illustrate and describe in detail the aquatic case-bearing larva of Hyposmocoma kahamanoa. Despite having similar ecologies as algae and lichen grazers at and below the water line of streams, prior research indicates that species with each case type constitute an independent lineage, with terrestrial sister taxa, and therefore the different groups of species bearing unique case types each represent an independent aquatic invasion. The case-bearing larvae often occur sympatrically, and on Kauai even species with similar case-types occur together, suggesting complex patterns of speciation and either past periods of isolation or sympatric speciation. Phylogenetic analysis of 2243 base pairs from two nuclear and one mitochondrial gene for 18 species confirm that each species is endemic to a single volcano, and that morphological divergence within case-types has not been dramatic. Diversification has been complex, and superficially similar case type lineages are not all monophyletic. Kauai, the oldest but smallest of the major high islands, supports more species in the aquatic guild than any other island, thus island age, rather than size, may be important in generating diversity in this group. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162, 15–42.

Dry season ecology of Anopheles gambiae complex mosquitoes at larval habitats in two traditionally semi-arid villages in Baringo, Kenya

BackgroundPre-adult stages of malaria vectors in semi-arid areas are confronted with highly variable and challenging climatic conditions. The objective of this study was to determine which larval habitat types are most productive in terms of larval densities in the dry and wet seasons within semi-arid environments, and how vector species productivity is partitioned over time.MethodsLarval habitats were mapped and larvae sampled longitudinally using standard dipping techniques. Larvae were identified to species level morphologically using taxonomic keys and to sub-species by polymerase chain reaction (PCR) methods. Physical characteristics of larval habitats, including water depth, turbidity, and presence of floating and emergent vegetation were recorded. Water depth was measured using a metal ruler. Turbidity, pH, conductivity, dissolved oxygen, temperatures salinity and total dissolved solids (TDS) were measured in the field using the hand-held water chemistry meters.ResultsMean larval densities were higher in the dry season than during the wet season but the differences in density were not statistically significant (F = 0.04, df = 1, p = 0.8501). Significantly higher densities of larvae were collected in habitats that were shaded and holding turbid, temporary and still water. Presence of emergent or floating vegetation, habitat depth, habitat size and habitat distance to the nearest house did not significantly affect larval density in both villages. There was a weakly positive relationship between larval density and salinity (r = 0.19, p < 0.05), conductivity (r = 0.05, p = 0.45) and total dissolved solids (r = 0.17, p < 0.05). However, the relationship between water temperature and larval density was weakly negative (r = 0.15, p = 0.35). All statistical tests were significant at alpha = 0.05.ConclusionBreeding of malaria vector mosquitoes in Baringo is driven by predominantly human-made and permanent breeding sites in which Anopheles arabiensis and Anopheles funestus breed at a low level throughout the year. Permanent water sources available during the dry season serve as inocula by providing "larval seed" to freshly formed rain-fed habitats during the rainy season. The highly localized and focal nature of breeding sites in these semi-desert environments provides a good opportunity for targeted larval control since the habitats are few, well-defined and easily traceable.

SEM description of the first larval instar ofAchelia assimilis(Pycnogonida: Ammotheidae)

The present paper is the first scanning electron microscope (SEM) description of a protonymphon larva of an Ammotheidae. The morphology of the first larval instar ofAchelia assimilisis described in detail and illustrated for the first time. The morphological characters are compared to previous larval descriptions of other pycnogonid species. The larvae are integrated into Bain's classification of larval types within the pycnogonids (Bain, 2003a) as a ‘typical protonymphon’. Larvae were obtained from ovigerous males, caught in Punta Huinay, Huinay, Chile and analysed with light microscope and SEM. Descriptions of pycnogonid protonymphons at a species-specific level in the future will contribute to a deeper understanding of larval pycnogonid taxonomy at the level of differential diagnoses.

Extrinsic and intrinsic influences on the phylogeography of the Austrocochlea constricta species group

Although extrinsic factors, such as oceanic currents and isolation induced by sea level maxima during Plio-Pleistocene glacial cycles, are often suggested as principal determinants of marine phylogeography, they are not always complete explanations. The counterexamples to predominant phylogeographic patterns in southeastern Australia suggest for example, that intrinsic factors such a habitat preference or reproductive mode can have significant influence. We collected DNA sequences from mitochondrial cytochrome c oxidase I (COI) and two nuclear gene introns from the Austrocochlea constricta species group to determine whether its porcata and constricta phenotypes are genetically distinguishable, to examine the phylogeographic effects of the Bassian Isthmus landbridge formed between Tasmania and Victoria during glacial maxima, and to investigate the importance of intrinsic factors in structuring its genetic variability. No fixed genetic differences between the porcata and constricta phenotypes were identified in any gene so the number of species comprised by the group cannot yet be determined. The two major clades recovered in COI analyses were respectively found principally east or west of the Bassian Isthmus. A. constricta is the first lecithotrophic or estuarine invertebrate known to show this pattern. There were no fixed differences, for any of the three genes, between specimens from estuarine and marine populations within either the eastern or western COI clades. Other intrinsic factors such as breeding period (possibly) and larval type (probably) may play roles in modulating phylogeographic patterns.

First-instar larva of Palaestra rufipennis (Westwood, 1841) and other Australian blister beetles (Coleoptera, Meloidae, Nemognathinae).

This is the first study of Australian blister beetle larvae. The first‐instar larva of one species of the endemic genus Palaestra Laporte de Castelnau, 1840, is described as well as those of other phoretic species without adult association and consequently unplaced to genus. Two main larval types (Groups 1 and 2) are recognised, as are distinct morphotypes within each group. Although both groups are assignable to the Nemognathinae, they are characterised by traits not paralleled by subfamily representatives from other regions of the world. Group 1, which includes Palaestra, is the more distinctive, whereas Group 2 shares most characteristics with members of the cosmopolitan tribe Nemognathini. A tabular comparison of larval groups and morphotypes is included.

Mutation scanning-coupled sequencing of nuclear ribosomal DNA spacers as a tool for the specific identification of different Contracaecum (Nematoda: Anisakidae) larval types

In the present study, various morphotypes of Contracaecum (Nematoda) larvae in different developmental stages were described and then genetically characterised using sequence data obtained from the first and second internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA). The alignment of ITS-1 and ITS-2 sequence data for larvae investigated with those from well characterised adults in previous studies showed that fourth-stage Contracaecum larvae from the Australian pelican, Pelecanus conspicillatus, had the same sequence as Contracaecum microcephalum reported from the little pied cormorant, Phalacrocorax melanoleucos. In addition, four different morphotypes of third-stage larvae, including types I to IV, were found in various species of fish. Contracaecum type I larvae were composed of two distict genotypes; those collected from mullet, Mugil cephalus, had the same sequence as C. multipapillatum, and those from an unknown species of hardyhead (family Atherinidae) were C. pyripapillatum. Contracaecum type II larvae from mackerel, Scomber australasicus, had the same sequence as adult C. ogmorhini sensu stricto, found in the Australian and New Zealand fur seals ( Arctocephalus spp). Contracaecum type III larvae from flathead, Platycephalus laevigatus, had the same sequence as C. rudolphii D found in the black cormorant, Phalacrocorax carbo. The approach used herein was an effective means of matching incompletely identifiable larval nematodes with reference sequences and provides a basis for exploring the composition of populations of anisakid larvae in fish as well as their ecology, particularly their life cycles and transmission patterns.

Immature Mosquitoes Of Serra Do Mar Park, São Paulo State, Brazil

With the objective of providing knowledge about the natural habitat of mosquito larvae, we conducted a study on the culicid fauna and identified larval habitat types at 3 different locations in the Serra do Mar State Park, Brazil, over the 12 months of 1991. We collected 1,425 larvae belonging to 23 species, of which the most frequent were Culex iridescens, Culex spp., Limatus durhami, and Trichoprosopon pallidiventer. The mosquito larval habitats presenting the greatest densities of specimens were bamboo, bromeliads, rubber boots, and streams. We observed that the culicids used a variety of containers as larval habitats and bred under a diversity of ecological conditions. Most of the habitats were natural, formed by bamboo, bromeliads, streams, and depressions in the ground and in rocks, along with artificial habitats consisting of rubber boots and abandoned metal cans. Seven species occurred preferentially in the internodes of closed bamboo stems, 6 in internodes of open stems, and 4 in bromeliads.

Structure and occurrence of cyphonautes larvae (bryozoa, ectoprocta)

We have studied larvae of the freshwater ctenostome Hislopia malayensis with scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and LM of serial sections. Some additional observations on larvae of M. membranacea using SEM and CLSM are also reported. The overall configuration of muscles, nerves, and cilia of the two larvae are identical. However, the larva of H. malayensis is much smaller than that of M. membranacea, which may explain most of the differences observed. Although all major nerves and muscle strands are present in H. malayensis, they are generally composed of fewer fibers. The H. malayensis larva lacks the anterior and posterior intervalve cilia. Its pyriform organ is unciliated with only a small central depression. The adhesive epithelium is not invaginated as an adhesive sac and lacks the large muscles interpreted as adhesive sac muscles in the M. membranacea larva. The velum carries two rows of ciliated cells, though the lower "row" consists of only one or two cells. Both rows of ciliated cells are innervated by nerves, which have not been detected in the M. membranacea larva. The ciliated ridge of H. malayensis lacks the frontal cilia. The planktotrophic cyphonautes larvae in a number of ctenostome clades and in the "basal" cheilostome clade Malacostega (and probably in the earliest cheilostomes) support the idea that the cyphonautes larva is the ancestral larval type of the Eurystomata. It may even represent the ancestral larval type of the bryozoans (= ectoprocts).

Anopheles larval abundance and diversity in three rice agro-village complexes Mwea irrigation scheme, central Kenya

BackgroundThe diversity and abundance of Anopheles larvae has significant influence on the resulting adult mosquito population and hence the dynamics of malaria transmission. Studies were conducted to examine larval habitat dynamics and ecological factors affecting survivorship of aquatic stages of malaria vectors in three agro-ecological settings in Mwea, Kenya.MethodsThree villages were selected based on rice husbandry and water management practices. Aquatic habitats in the 3 villages representing planned rice cultivation (Mbui Njeru), unplanned rice cultivation (Kiamachiri) and non-irrigated (Murinduko) agro-ecosystems were sampled every 2 weeks to generate stage-specific estimates of mosquito larval densities, relative abundance and diversity. Records of distance to the nearest homestead, vegetation coverage, surface debris, turbidity, habitat stability, habitat type, rice growth stage, number of rice tillers and percent Azolla cover were taken for each habitat.ResultsCaptures of early, late instars and pupae accounted for 78.2%, 10.9% and 10.8% of the total Anopheles immatures sampled (n = 29,252), respectively. There were significant differences in larval abundance between 3 agro-ecosystems. The village with 'planned' rice cultivation had relatively lower Anopheles larval densities compared to the villages where 'unplanned' or non-irrigated. Similarly, species composition and richness was higher in the two villages with either 'unplanned' or limited rice cultivation, an indication of the importance of land use patterns on diversity of larval habitat types. Rice fields and associated canals were the most productive habitat types while water pools and puddles were important for short periods during the rainy season. Multiple logistic regression analysis showed that presence of other invertebrates, percentage Azolla cover, distance to nearest homestead, depth and water turbidity were the best predictors for Anopheles mosquito larval abundance.ConclusionThese results suggest that agricultural practices have significant influence on mosquito species diversity and abundance and that certain habitat characteristics favor production of malaria vectors. These factors should be considered when implementing larval control strategies which should be targeted based on habitat productivity and water management.

Larval behavioral syndrome does not affect emergence behavior in a damselfly (Lestes congener)

Activity is a key behavioral trait that often mediates a trade-off between finding food for growth and evading predation. We investigated how activity of the damselfly Lestes congener is affected by larval state and predator presence and if larval behavioral type (BT) can be used to predict larval emergence behavior. Activity level of individual larvae was studied without predators at two different physiological states (hungry, fed) and in two predator treatments (familiar or unfamiliar predator cues). Larvae did not adjust their activity depending on state or when subjected to unfamiliar predator cues but a general reduction in activity was seen in the familiar predator treatment. Hence, active individuals remained active compared to their conspecifics, independent of state or predator treatment illustrating the presence of a behavioral syndrome. However, we found no correlation between larval BT and emergence behavior. Active individuals did not differ from less active individuals in any emergence characteristics. The results illustrate that the larval BT occurs in many situations keeping active larvae active even in maladaptive situations. Furthermore, we show that damselfly emergence behavior can be completely decoupled from larval BT, indicating a loss of stability in individual BT during critical stages in ontogeny.

The Invention of the Pilidium Larva in an Otherwise Perfectly Good Spiralian Phylum Nemertea

One of the most remarkable larval types among spiralians, and invertebrates in general, is the planktotrophic pilidium. The pilidium is found in a single clade of nemerteans, called the Pilidiophora, and appears to be an innovation of this group. All other nemerteans have either planktotrophic or lecithotrophic juvenile-like planuliform larvae or have direct development. The invention of the pilidium larva is associated with the formation of an extensive blastocoel that supports the delicate larval frame and elaborate ciliary band. Perhaps the most striking characteristic of the pilidium is the way the juvenile worm develops inside the larva from a series of isolated rudiments, called the imaginal discs. The paired cephalic discs, cerebral organ discs, and trunk discs originate as invaginations of larval epidermis and subsequently grow and fuse around the larval gut to form the juvenile. The fully formed juvenile ruptures the larval body and, more often than not, devours the larva during catastrophic metamorphosis. This review is an attempt to examine the pilidium in the context of recent data on development of non-pilidiophoran nemerteans, and speculate about the evolution of pilidial larval development. The author emphasizes the difference between the planuliform larvae of Palaeonemerteans and Hoplonemerteans, and suggest a new name for the hoplonemertean larvae--the decidula.

Early development of medaka Oryzias latipes muscles as revealed by transgenic approaches using embryonic and larval types of myosin heavy chain genes

We cloned three full-length cDNAs encoding myosin heavy chains (MYHs) previously found to be expressed in embryos or larvae of medaka Oryzias latipes. Based on cDNA sequence information, the three medaka MYH genes, mMYH(emb1), mMYH(L1) and mMYH(L2), were localized on the chromosomes. In vivo promoter assay using the gene encoding green or red fluorescent protein and linked to the 5'-flanking region of mMYH demonstrated that the transcripts of fast-type mMYH(emb1), first expressed in embryos but belonging to the adult type in phylogenetic analysis, were located in the horizontal myoseptum. On the other hand, embryonic fast-type mMYH(L1) and mMYH(L2) were expressed in the whole myotomes. Interestingly, cells expressing mMYH(emb1) were localized together with engrailed, and cyclopamine, which blocks hedgehog signaling, inhibited mMYH(emb1) expression as well as the formation of the horizontal myoseptum, suggesting that muscle pioneer cells express mMYH(emb1) as a key protein in the formation of the horizontal myoseptum.

A microscopic and microanalytical study (Fe, Ca) of the teeth of the larval and juvenile Ambystoma mexicanum (Amphibia: Urodela: Ambystomatidae)

We studied the teeth of larvae and one juvenile of the axolotl Ambystoma mexicanum, a urodele species that undergoes partial metamorphosis, by light microscopy of ground sections, backscattered electron imaging and semi-quantitative microanalysis in the scanning electron microscope. By applying these techniques it was possible to identify enamel, enameloid and dentin in the teeth. Iron was found to be present in enamel and enameloid, the concentrations being highest in the enamel. A staining indicative of the presence of iron was observed in the inner dental epithelium of tooth germs. Dentinal tubules mostly followed a straight course, but some recurved over a short distance distally. In larval teeth and teeth of “larval type” in the juvenile individual the dentinal tubules ended in the basal portion of the enameloid. Our results show that in the axolotl, monocuspid teeth of the “larval type” that developed after partial transformation still possess an enameloid layer beneath a thin enamel cap. The findings of the present study are consistent with the view that enameloid matrix is secreted by odontoblasts, while enameloid maturation is (largely) controlled by ameloblasts.

Difference in larval type explains patterns of nonsynonymous substitutions in two ancient paralogs of the histone H3 gene in sea stars

Paralogous genes frequently show differences in patterns and rates of substitution that are typically attributed to different selection regimes, mutation rates, or local recombination rates. Here, two anciently diverged paralogous copies of the histone H3 gene in sea stars, the tandem-repetitive early-stage gene and a newly isolated gene with lower copy number that was termed the "putative late-stage histone H3 gene" were analyzed in 69 species with varying mode of larval development. The two genes showed differences in relative copy number, overall substitution rates, nucleotide composition, and codon usage, but similar patterns of relative nonsynonymous substitution rates, when analyzed by the d(N)/d(S) ratio. Sea stars with a nonpelagic and nonfeeding larval type (i.e., brooding lineages) were observed to have d(N)/d(S) ratios that were larger than for nonbrooders but equal between the two paralogs. This finding suggested that demographic differences between brooding and nonbrooding lineages were responsible for the elevated d(N)/d(S) ratios observed for brooders and refuted a suggestion from a previous analysis of the early-stage gene that the excess nonsynonymous substitutions were due to either (1) gene expression differences at the larval stage between brooders and nonbrooders or (2) the highly repetitive structure of the early-stage histone H3 gene.