The origin and evolution of segmentation

Arthropods, annelids and chordates all possess segments. It remains unclear, however, whether the segments of these animals evolved independently or instead were derived from a common ancestor. Considering this question involves examining not only the similarities and differences in the process of segmentation between these phyla, but also how this process varies within phyla, where the homology of segments is generally accepted. This article reviews what is known about the segmentation process and considers various proposals to explain its evolution.     

The origin and evolution of segmentation

Arthropods, annelids and chordates all possess segments. It remains unclear, however, whether the segments of these animals evolved independently or instead were derived from a common ancestor. Considering this question involves examining not only the similarities and differences in the process of segmentation between these phyla, but also how this process varies within phyla, where the homology of segments is generally accepted. This article reviews what is known about the segmentation process and considers various proposals to explain its evolution.     

Genetic regulation of engrailed and wingless in Tribolium segmentation and the evolution of pair-rule segmentation

In Drosophila, primary pair-rule genes establish the parasegmental boundaries and indirectly control the periodic expression of the segment polarity genes engrailed (en) and wingless (wg) via regulation of secondary pair-rule genes. Although orthologs of some Drosophila pair-rule genes are not required for proper segmentation in Tribolium, segmental expression of Tc-en and Tc-wg is conserved. To understand how these segment polarity genes are regulated, we examined the results of expressing one or two pair-rule genes in the absence of the other known pair-rule genes. Expression of one or both of the secondary pair-rule genes, Tc-sloppy-paired (Tc-slp) and Tc-paired (Tc-prd), activated Tc-wg in the absence of the primary pair-rule genes, Tc-even-skipped (Tc-eve), Tc-runt (Tc-run) and Tc-odd-skipped (Tc-odd). Tc-eve alone failed to activate Tc-wg or Tc-en, but in combination with Tc-run or Tc-prd activated Tc-en. These results, interpreted within the pair-rule gene expression patterns, suggest separate models for the genetic regulation of the juxtaposed expression of Tc-wg and Tc-en at odd- and even-numbered parasegmental boundaries, respectively. Conserved interactions between eve and prd at the anterior boundary of odd-numbered parasegments may reflect an ancestral segmentation mechanism that functioned in every segment prior to the evolution of pair-rule segmentation.     

From variable to constant cell numbers: cellular characteristics of the arthropod nervous system argue against a sister-group relationship of Chelicerata and “Myriapoda” but favour the Mandibulata concept

In the new debate on arthropod phylogeny, structure and development of the nervous system provide important arguments. The architecture of the brain of Hexapoda, Crustacea and Chelicerata in recent years has been thoroughly compared against an evolutionary background. However, comparative aspects of the nervous systems in these taxa at the cellular level have been examined in only a few studies. This review sets out to summarize these aspects and to analyse the existing data with respect to the concept of individually identifiable neurons. In particular, mechanisms of neurogenesis, the morphology of serotonergic interneurons, the number of motoneurons, and cellular features and development of the lateral eyes are discussed. We conclude that in comparison to the Mandibulata, in Chelicerata the numbers of neurons in the different classes examined are much higher and in many cases are not fixed but variable. The cell numbers in Mandibulata are lower and the majority of neurons are individually identifiable. The characters explored in this review are mapped onto an existing phylogram, as derived from brain architecture in which the Hexapoda are an in-group of the Crustacea, and there is not any conflict of the current data with such a phylogenetic position of the Hexapoda. Nevertheless, these characters argue against a sister-group relationship of Myriapoda and Chelicerata as has been recently suggested in several molecular studies, but instead provide strong evidence in favour of the Mandibulata concept.Edited by D. Tautz     

Pax3/7 genes reveal conservation and divergence in the arthropod segmentation hierarchy

Several features of Pax3/7 gene expression are shared among distantly related insects, including pair-rule, segment polarity, and neural patterns. Recent data from arachnids imply that roles in segmentation and neurogenesis are likely to be played by Pax3/7 genes in all arthropods. To further investigate Pax3/7 genes in non-insect arthropods, we isolated two monoclonal antibodies that recognize the products of Pax3/7 genes in a wide range of taxa, allowing us to quickly survey Pax3/7 expression in all four major arthropod groups. Epitope analysis reveals that these antibodies react to a small subset of Paired-class homeodomains, which includes the products of all known Pax3/7 genes. Using these antibodies, we find that Pax3/7 genes in crustaceans are expressed in an early broad and, in one case, dynamic domain followed by segmental stripes, while myriapods and chelicerates exhibit segmental stripes that form early in the posterior-most part of the germ band. This suggests that Pax3/7 genes acquired their role in segmentation deep within, or perhaps prior to, the arthropod lineage. However, we do not detect evidence of pair-rule patterning in either myriapods or chelicerates, suggesting that the early pair-rule expression pattern of Pax3/7 genes in insects may have been acquired within the crustacean-hexapod lineage.     

Myriapod metamerism and arthropod segmentation

Outstanding progress in understanding segmentation of tracheate arthropods (Atelocerata), i.e. Chilopoda, Diplopoda, Pauropoda, Symphyla and Insecta, has been gained through experimental studies carried out on a single, very derivative organism, i.e. Drosophila. We stress the need for a broader comparative approach. We have studied the segmental structure of the trunk in geophilomorph centipedes, where we can identify morphogenetic units of two, four, eight or 16 segments. Accordingly, we sketch an improved model for arthropod segmentation, with the following initial steps: (a) biochemical marking of a very few repetitive units (eosegments); (b) iterative duplications of this first periodicity, until the embryo acquires an array of biochemical markings matching the whole number of segments of the future larva or juvenile specimen; (c) transpatterning, stabilization and interpretation of this 'segmental' arrangement; (d) possible repatterning, to give a final repetitive pattern we define as metasegmental. Finally, we express some doubt about the homology between annelid and arthropod segmentation.     

The urochordate larva and archichordate organization: chordate origin and anagenesis revisited†

A critical analysis of relevant characters in Epineuralia (Tentaculata + Deuterostomia) in order to trace the emergence of chordate organization demonstrates that a reorganization of already highly adapted planktotrophic larvae is inconceivable. In connection with the clearly regressive state of urochordate organization, an alternative concept is presented. It proposes the chordate origin via adaptation of simply ciliated, short-ranged lecithotrophic *hemichordate* larvae which retained a dorsoposterior primary anus (see Pterobranchia, Enteropneusta-Harrimaniidae, lower Chordata). The extension of larval life to exploit the food-rich layers of the sea enabled the posterior, pre-anal body (rather than a ‘tail’) to develop into a powerful propulsive complex. This was prepared by a mesoblastema heterochrony and was co-evolutively accompanied by (1) the precocious differentiation of the middorsal nerve cord of *Hemichordata* to become a neural plate extending onto the metasoma (meeting the dorsoposterior anus: neurenteric canal) (2) a cerebral sensory centre, and (3) the precociousness of the filter-feeding branchial gut. Such larval, primitive Archichordata were coupled in their life-cycle most probably with semi-vagile *hemichordate* adults (lophophore already reduced; with new endostyle). This biphasic organization subsequently radiated into the two clades of (1) Tunicata (elaboration of the branchial gut with sedentary life, peribranchial cavity, tunica/cuticle, and regressive larvae; secondarily neotene and or pelagic) and (2) Holochordata (prematurity of larvae with extension of the somatic chordate organization forwards over the branchial gut). The latter level gave rise to (a) the Acrania (Cephalochordata) turning to an epibenthic life (fairly stationary habit with asymmetry, regression of the cerebral sensory equipment) and finally becoming infaunal: (b) the Craniata (Vertebrata) elaborating the pelagic existence and changing to a macrophagous biology (selective particle-swallowing rather than mucociliary filter-feeding), correlated with a highly adapted sensory control system (cephalization) and neural crest organization.     

Nervous and muscle system development in <Emphasis Type="Italic">Phascolion strombus</Emphasis> (Sipuncula)

Recent interpretations of developmental gene expression patterns propose that the last common metazoan ancestor was segmented, although most animal phyla show no obvious signs of segmentation. Developmental studies of non-model system trochozoan taxa may shed light on this hypothesis by assessing possible cryptic segmentation patterns. In this paper, we present the first immunocytochemical data on the ontogeny of the nervous system and the musculature in the sipunculan Phascolion strombus. Myogenesis of the first anlagen of the body wall ring muscles occurs synchronously and not subsequently from anterior to posterior as in segmented spiralian taxa (i.e. annelids). The number of ring muscles remains constant during the initial stages of body axis elongation. In the anterior-posteriorly elongated larva, newly formed ring muscles originate along the entire body axis between existing myocytes, indicating that repeated muscle bands do not form from a posterior growth zone. During neurogenesis, the Phascolion larva expresses a non-metameric, paired, ventral nerve cord that fuses in the mid-body region in the late-stage elongated larva. Contrary to other trochozoans, Phascolion lacks any larval serotonergic structures. However, two to three FMRFamide-positive cells are found in the apical organ. In addition, late larvae show commissure-like neurones interconnecting the two ventral nerve cords, while early juveniles exhibit a third, medially placed FMRFamidergic ventral nerve. Although we did not find any indications for cryptic segmentation, certain neuro-developmental traits in Phascolion resemble the conditions found in polychaetes (including echiurans) and myzostomids and support a close relationship of Sipuncula and Annelida.     

Mechanisms of eye development and evolution of the arthropod visual system: The lateral eyes of myriapoda are not modified insect ommatidia

The lateral eyes of Crustacea and Insecta consist of many single optical units, the ommatidia, that are composed of a small, strictly determined and evolutionarily conserved set of cells. In contrast, the eyes of Myriapoda (millipedes and centipedes) are fields of optical units, the lateral ocelli, each of which is composed of up to several hundreds of cells. For many years these striking differences between the lateral eyes of Crustacea/Insecta versus Myriapoda have puzzled evolutionary biologists, as the Myriapoda are traditionally considered to be closely related to the Insecta. The prevailing hypothesis to explain this paradox has been that the myriapod fields of lateral ocelli derive from insect compound eyes by disintegration of the latter into single ommatidia and subsequent fusion of several ommatidia to form multicellular ocelli. To provide a fresh view on this problem, we counted and mapped the arrangement of ocelli during postembryonic development of a diplopod. Furthermore, the arrangement of proliferating cells in the eyes of another diplopod and two chilopods was monitored by labelling with the mitosis marker bromodeoxyuridine. Our results confirm that during eye growth in Myriapoda new elements are added to the side of the eye field, which extend the rows of earlier-generated optical units. This pattern closely resembles that in horseshoe crabs (Chelicerata) and Trilobita. We conclude that the trilobite, xiphosuran, diplopod and chilopod mechanism of eye growth represents the ancestral euarthropod mode of visual-system formation, which raises the possibility that the eyes of Diplopoda and Chilopoda may not be secondarily reconstructed insect eyes.     

The origin of prokaryotic life and molecular evolution

The origin of prokaryotic life is discussed with an emphasis on the self-assembly of early life in a microscale environment where ordered cellular structures and integrated functions evolved from disorder. Early molecular evolution may have been due to both molecular chaos and evolving molecular order.     

Evidence for Wg-independent tergite boundary formation in the millipede Glomeris marginata

The correlation between dorsal and ventral segmental units in diplopod myriapods is complex and disputed. Recent results with engrailed (en), hedgehog (hh), wingless (wg), and cubitus-interruptus (ci) have shown that the dorsal segments are patterned differently from the ventral segments. Ventrally, gene expression is compatible with the classical autoregulatory loop known from Drosophila to specify the parasegment boundary. In the dorsal segments, however, this Wg/Hh autoregulatory loop cannot be present because the observed gene expression patterns argue against the involvement of Wg signalling. In this paper, we present further evidence against an involvement of Wg signalling in dorsal segmentation and propose a hypothesis about how dorsal segmental boundaries may be controlled in a wg-independent way. We find that (1) the Notum gene, a modulator of the Wg gradient in Drosophila, is not expressed in the dorsal segments. (2) The H15/midline gene, a repressor of Wg action in Drosophila, is not expressed in the dorsal segments, except for future heart tissue. (3) The patched (ptc) gene, which encodes a Hh receptor, is strongly expressed in the dorsal segments, which is incompatible with Wg-Hh autoregulation. The available data suggest that anterior-posterior (AP) boundary formation in dorsal segments could instead rely on Dpp signalling rather than Wg signalling. We present a hypothesis that relies on Hh-mediated activation of Dpp signalling and optomotor-blind (omb) expression to establish the dorsal AP boundary (the future tergite boundary). The proposed mechanism is similar to the mechanism used to establish the AP boundary in Drosophila wings and ventral pleura.     

The segmentation cascade in the centipede Strigamia maritima: involvement of the Notch pathway and pair-rule gene homologues

The centipede Strigamia maritima forms all of its segments during embryogenesis. Trunk segments form sequentially from an apparently undifferentiated disk of cells at the posterior of the germ band. We have previously described periodic patterns of gene expression in this posterior disc that precede overt differentiation of segments, and suggested that a segmentation oscillator may be operating in the posterior disc. We now show that genes of the Notch signalling pathway, including the ligand Delta, and homologues of the Drosophila pair-rule genes even-skipped and hairy, show periodic expression in the posterior disc, consistent with their involvement in, or regulation by, such an oscillator. These genes are expressed in a pattern of apparently expanding concentric rings around the proctodeum, which become stripes at the base of the germ band where segments are emerging. In this transition zone, these primary stripes define a double segment periodicity: segmental stripes of engrailed expression, which mark the posterior of each segment, arise at two different phases of the primary pattern. Delta and even-skipped are also activated in secondary stripes that intercalate between primary stripes in this region, further defining the single segment repeat. These data, together with observations that Notch mediated signalling is required for segment pattern formation in other arthropods, suggest that the ancestral arthropod segmentation cascade may have involved a segmentation oscillator that utilised Notch signalling.     

The role of the segmentation gene <Emphasis Type="Italic">hairy</Emphasis> in <Emphasis Type="Italic">Tribolium</Emphasis>

Hairy stripes in Tribolium are generated during blastoderm and germ band extension, but a direct role for Tc-h in trunk segmentation was not found. We have studied here several aspects of hairy function and expression in Tribolium, to further elucidate its role. First, we show that there is no functional redundancy with other hairy paralogues in Tribolium. Second, we cloned the hairy orthologue from Tribolium confusum and show that its expression mimics that of Tribolium castaneum, implying that stripe expression should be functional in some way. Third, we show that the dynamics of stripe formation in the growth zone is not compatible with an oscillatory mechanism comparable to the one driving the expression of hairy homologues in vertebrates. Fourth, we use parental RNAi experiments to study Tc-h function and we find that mandible and labium are particularly sensitive to loss of Tc-h, reminiscent of a pair-rule function in the head region. In addition, lack of Tc-h leads to cell death in the gnathal region at later embryonic stages, resulting in a detachment of the head. Cell death patterns are also altered in the midline. Finally, we have analysed the effect of Tc-h knockdown on two of the target genes of hairy in Drosophila, namely fushi tarazu and paired. We find that the trunk expression of Tc-h is required to regulate Tc-ftz, although Tc-ftz is itself also not required for trunk segmentation in Tribolium. Our results imply that there is considerable divergence in hairy function between Tribolium and Drosophila.     

Blood markers and genetic evolution in Cercopithecinae

Serum proteins and RBC enzymes were surveyed in 16 species (183 animals) of African guenons (tribe Cercopithecini) in order to determine their genetic polymorphism and to establish dendrograms on the basis of their allele frequencies. The molecular data obtained were compared with those of mangabeys (16 animals tested) and discussed in the light of our results inPapio andMacaca. The species surveyed wereCercopithecus neglectus, C. hamlyni, C. l'hoesti (C. l'h. l'hoesti, C. l'h. preussi, andC. l'h. solatus), C. nictitans, C. mitis (C. m. kolbi, C. m. albotorquatus, C. m. stuhlmanni, andC. m. albogularis), C. cephus, C. ascanius, C. erythrotis, C. petaurista, C. mona, C. pogonias, C. wolfi, andC. aethiops, Miopithecus talapoin, Allenopithecus nigroviridis andErythrocebus patas, Lophocebus albigena, andCercocebus torquatus. Eleven loci (ten systems) were studied in red blood cell enzymes and the Gc, Gm, Km, and Bm systems in DBP and immunoglobulin serum proteins. Most of the loci were polymorphic. Similar and different polymorphisms occur in closely related species or subspecies, particularly inCercopithecus. Guenons have phenotypes clearly distinct from mangabeys.     

The origin of annelids

Annelids are a phylum of segmented bilaterian animals that have become important components of ecosystems spanning terrestrial realms to the deep sea. Annelids are remarkably diverse, possessing high taxonomic diversity and exceptional morphological disparity, and have evolved numerous feeding strategies and ecologies. Their interrelationships and evolution have been the source of much controversy over the past century with the composition of the annelid crown group, the relationship of major groups and the body plan of the ancestral annelid having undergone major recent revisions. There is a convincing body of molecular evidence that polychaetes form a paraphyletic grade and that clitellates are derived polychaetes. The earliest stem group annelids from Cambrian Lagerstätten are errant, epibenthic polychaetes, confirming that biramous parapodia, head appendages and diverse, simple chaetae are primitive for annelids. Current evidence from molecular clocks and the fossil record suggest that crown group annelids are a Late Cambrian – Ordovician radiation, with clitellates radiating in the Late Palaeozoic. Their body fossil record is largely confined to deposits showing exceptional preservation and is punctuated by the acquisition of hard parts in major groups. The discovery of an Ordovician fossil with soft tissues has shown that machaeridians are in fact a clade of crown polychaetes. They were in existence for more than 200 million years and possess unique calcitic dorsal armour, allowing their mode of life and phylogeny to be interpreted in the context of the annelid body plan. We identify a novel clade of machaeridians, the Cuniculepadida, which exhibit a series of adaptations for burrowing.     

Phospholipids of sea worms,mollusks, and arthropods

The phospholipid composition of organs and tissues was investigated in representatives of five phyla of marine invertebrates: Annelida (Chaetopterus variopedatus, Serpula vermicularis), Echiuroidea (Urechis unicinctus), Sipunculoidea (Phascolosoma japonicum), Mollusca (Gastropoda: Tectonatica janthostoma, Neptunea polycostata; Bivalvia: Mactra sulcataria, Peronidia venuloza, Patinopecten yessoensis, Crenomytilus grayanus; Cephalopoda: Octopus conispadiceus, Todarodes pacificus), and Arthropoda (Paralythodes camtschatica, Erimacrus isenbekii, Hemigrapsus sanguineus, Pinixa rathbunii). The specificity of phospholipid distribution was shown to be related to the taxonomic position of marine invertebrates and functional properties of their organs and tissues. In Echiuroidea, Sipunculoidea, and Arthropoda, ceramide aminoethylphosphonate was found only in the digestive organs. This suggests an exogenous origin of this phospholipid.     

Origin,evolution, phylogeny and taxonomy of Pulex irritans

The human flea Pulex irritans Linnaeus, 1758 (Siphonaptera: Pulicidae) is one of the most studied species together with the cat flea Ctenocephalides felis Bouché, 1835, because they have a cosmopolitan distribution and are closely related to humans. The present study aimed to carry out a comparative morphometric and molecular study of two different populations of P. irritans (Spain and Argentina). Accordingly, internal transcribed spacer (ITS)1 and ITS2 of rDNA and the partial cytochrome c oxidase subunit 1 (cox1) and cytochrome b (cytb) mtDNA genes of these taxa were sequenced. Furthermore, the taxonomy, origin, evolution and phylogeny of P. irritans was assessed. The morphometric data obtained did not show significant differences between P. irritans specimens from Spain and Argentina, even when these two populations were collected from different hosts; however, there was a considerable degree of molecular divergence between both populations based on nuclear and mitochondrial markers. Thus, it is proposed that P. irritans, in contrast with other generalist fleas, maintains a certain degree of morphological similarity, at least between Western Palearctic and Neotropical areas. Furthermore, two well defined geographical genetic lineages within the P. irritans species are indicated, suggesting the existence of two cryptic species that could be discriminated by a polymerase chain reaction‐linked restriction fragment length polymorphism.