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The segmentation clock is defined as the combination of various oscillating signals, which lead to the generation of somites at regular intervals from the presomitic mesoderm. The numerous researches conducted on the subject reveals that the segmentation clock is actually much more complex than it is thought to be and also includes oscillating expression of genes from three different signaling pathways in various coordinated networks.

Evolutionary dynamics of vertebrate body mass range were studied by regression equations, which are used to predict the origin age, initial body mass and body mass range. The results suggested that dynamics of phenotypic characteristic can help molecular clock and fossil models in dating evolutionary years.

The skeletal muscles and axial skeleton of vertebrates derive from the embryonic paraxial mesoderm. In amniotes, paraxial mesoderm is formed bilaterally to the nerve cord as a result of primitive streak and tail-bud regression during body axis formation. In chick and mouse embryos, paraxial mesoderm was proposed to derive from a population of resident cells located in the regressing primitive streak and tail bud. In contrast, in lower vertebrates, paraxial mesoderm is formed as a result of the continuation of ingression movements of gastrulation. Here, we reinvestigate paraxial mesoderm formation in the chicken embryo and demonstrate that these two modes are concomitantly at work to set up the paraxial mesoderm. Although the medial part of somites derives from stem cells resident in the primitive streak/tail bud, the lateral part derives from continuous ingression of epiblastic material. Our fate mapping further shows that the paraxial mesoderm territory in the epiblast is regionalized along the anteroposterior axis as in lower vertebrates. These observations suggest that the mechanisms responsible for paraxial mesoderm formation are largely conserved across vertebrates. vertebrate | epiblast | primitive streak | tail bud | body axis formation

Otx2 is a paired type homeobox gene that plays essential roles in each step and site of head development in vertebrates. In the mouse, Otx2 expression in the anterior neuroectoderm is regulated primarily by two distinct enhancers: anterior neuroectoderm (AN) and forebrain/midbrain (FM) enhancers at 92 kb and 75 kb 5'of the Otx2 locus, respectively. The AN enhancer has activity in the entire anterior neuroectoderm at headfold and early somite stages, whereas the FM enhancer is subsequently active in the future caudal forebrain and midbrain ectoderm. In tetrapods, both AN and FM enhancers are conserved, whereas the AN region is missing in teleosts, despite overt Otx2 expression in the anterior neuroectoderm. Here, we show that zebrafish and fugu FM regions drive expression not only in the forebrain and midbrain but also in the anterior neuroectoderm at headfold stage. The analysis of coelacanth and skate genomic Otx2 orthologues suggests that the utilization of the two enhancers, AN and FM, is an ancestral condition. In contrast, the AN enhancer has been specifically lost in the teleost lineage with a compensatory establishment of AN activity within the FM enhancer. Furthermore, the AN activity in the fish FM enhancer was established by recruiting upstream factors different from those that direct the tetrapod AN enhancer, yet zebrafish FM enhancer is active in both mouse and zebrafish anterior neuroectoderm at the headfold stage. anterior neuroectoderm | coelacanth | enhancer | tetrapod | chondrichthyes

Inhibition of Notch signaling by Numb is critical for many cell fate decisions. In this study, we demonstrate a more complex relationship between Notch and the two vertebrate Numb homologues Numb and Numblike. Although Numb and Numblike at low levels of Notch signaling negatively regulated Notch, high levels of Notch signaling conversely led to a reduction of Numb and Numblike protein levels in cultured cells and in the developing chick central nervous system. The Notch intracellular domain but not the canonical Notch downstream proteins Hes 1 and Hey 1 caused a reduction of Numb and Numblike. The Notch-mediated reduction of Numblike required the PEST domain in the Numblike protein and was blocked by the proteasome inhibitor MG132. Collectively, these observations reveal a reciprocal negative regulation between Notch and Numb/ Numblike, which may be of relevance for stabilizing asymmetric cell fate switches and for tumor development.

How does the way an organism functions affect its subsequent development? Bryozoans are colonial animals that capture suspended food particles from water currents they generate using crowns of ciliated tentacles (lophophores). In many encrusting bryozoans the water passes through and then under the lophophores until it exits the colony at 'chimneys' where the lophophores spread apart to form and opening. To determine whether these water currents can induce the formation of new chimneys, I augmented the excurrent flow by injecting seawater into the colony. New chimneys began to develop near the site of seawater injection within as little as one to two days. New chimneys rarely began to develop within this time interval at control sites where I did not inject seawater. This shows that fluid flow controls development in an external fluid transport system lacking pipe-like conduits, as has been found in the vertebrate circulatory system, an internal fluid transport system with pipe-like conduits. These fluid transport systems show feedback between the way they function and their own development. This kind of 'function-dependent development' should be differentiated from phenotypic plasticity, since the developing system, not the environment, produces the signals that induce morphological change., Introduction In many complex biological systems--including the vertebrate circulatory (Langille, 1995), skeletal (Joo et al., 2003), and nervous systems (Adams and Horton, 2002)--the way the system functions modulates subsequent morphogenesis [...]

Hemoglobin (Hb) released from erythrocytes may cause oxidation of lipids and proteins. Haptoglobin (Hp), which occurs in the plasma of all mammals, binds free Hb and inhibits its oxidative activity. It is not known whether this protective protein also exists in lower vertebrates. By analyzing available genomic sequences, we have found that bony fish, but not more primitive animals, have a gene coding for a protein homologous to mammalian Hp. Furthermore, we show that this protein is present in the plasma of Japanese pufferfish (Takifugu rubripes) and that it binds Hb. These results, together with a phylogenetic analysis, suggest that Hp evolved from a complement-associated protein (mannose-binding lectin-associated serine proteinase, MASP), with the emergence of fish. Surprisingly, we found that both chicken (Gallus gallus) and the Western clawed frog (Xenopus tropicalis) lack the Hp gene. In chicken plasma, however, we identified a different type of Hb-binding protein, PIT54, which has been reported to be a potent antioxidant. PIT54 is a soluble member of the family of scavenger receptor cysteine-rich proteins, and we found that its gene exists only in birds. We also show that the plasma of ostrich (Strutio camelus), a primitive bird, contains both PIT54 and Hp. Collectively, our data suggest that PIT54 has successively taken over the function of Hp during the evolution of the avian lineage and has completely replaced the latter protein in chicken. evolution | PIT54 | complement proteinase

During vertebrate brain development, axons are enwrapped by myelin, an insulating membrane produced by oligodendrocytes. Neuron-derived signaling molecules are temporally and spatially required to coordinate oligodendrocyte differentiation. In this study, we show that neurons regulate myelin membrane trafficking in oligodendrocytes. In the absence of neurons, the major myelin membrane protein, the proteolipid protein (PLP), is internalized and stored in late endosomes/lysosomes (LEs/Ls) by a cholesterol-dependent and clathrin independent endocytosis pathway that requires actin and the RhoA guanosine triphosphatase. Upon maturation, the rate of endocytosis is reduced, and a cAMP-dependent neuronal signal triggers the transport of PLP from LEs/Ls to the plasma membrane. These findings reveal a fundamental and novel role of LEs/Ls in oligodendrocytes: to store and release PLP in a regulated fashion. The release of myelin membrane from LEs/Ls by neuronal signals may represent a mechanism to control myelin membrane growth.

Vertebrate mineralized tissues are vital to the adaptive evolution of various traits. Among these traits is the tooth, which consists of two characteristic mineralized tissues, a highly mineralized surface layer (enamel in tetrapods and enameloid in fish) and a softer body (dentin), both supported by basal bone. However, enamel and enameloid are significantly different in development, and dentin shows many histological variations; hence their evolution has been intensively studied. Nevertheless, their genetic basis has been revealed only in tetrapods. We previously reported that many genes involved in tetrapod tissue mineralization arose from a common ancestor and constitute the secretory calcium-binding phosphoprotein (SCPP) gene family. Now we show that teleost fish also use many SCPPs for enameloid and dentin mineralization, but none of these directly corresponds to tetrapod SCPPs. This finding suggests that teleost and tetrapod SCPP genes have experienced independent parallel duplication histories. Thus, through phenogenetic drift, the tooth has remained a stable trait in jawed vertebrates, while evolving distinct genetic bases in teleosts and tetrapods. The characteristics of teleost SCPP genes and their expression domains in tooth development suggest the possibility that enameloid arose from dentin and enamel from enameloid more than once in vertebrate evolution. In fugu (puffer fish), expression of SCPP genes is also detected in an unusual beak-like structure that shelters numerous teeth. Their expression pattern suggests that the jaw consists of the dentin beak and supportive bone. These findings illustrate the complexity of the homology concept in understanding evolution, particularly the evolution of mineralized tissues. dentin | enamel | enameloid | homology | gene duplication

Ultraviolet (UV) and violet vision in vertebrates is mediated by UV and violet visual pigments that absorb light maximally ([[lambda].sub.max]) at ~360 and 390-440 nm, respectively. So far, a total of 11 amino acid sites only in transmembrane (TM) helices I-III are known to be involved in the functional differentiation of these short wavelength-sensitive type 1 (SWS1) pigments. Here, we have constructed chimeric pigments between the violet pigment of African clawed frog (Xenopus laevis) and its ancestral UV pigment. The results show that not only are the absorption spectra of these pigments modulated strongly by amino acids in TM I-VII, but also, for unknown reasons, the overall effect of amino acid changes in TM W-VII on the [[lambda].sub.max]-shift is abolished. The spectral tuning of the contemporary frog pigment is explained by amino acid replacements F86M, V91I, T93P, V109A, E113D, L116V, and S118T, in which V91I and V109A are previously unknown, increasing the total number of critical amino acid sites that are involved in the spectral tuning of SWS1 pigments in vertebrates to 13.

Vertebrates and arthropods share the common problem of controlling a rigid, articulated skeleton using neurally-controlled, striated muscle. Since this condition has arisen independently in the two groups, there is no reason to assume, a priori, that the control mechanisms used by the two groups will be the same. Indeed, there appear to be fundamental differences in the tactics used by the two groups. Insects and crustaceans use small numbers of heterogeneous motoneurons, while vertebrates (mammals especially) use many, more homogeneous, motor axons. In particular, arthropods make extensive use of peripheral neuromodulation to alter the properties of both neuromuscular junctions and muscle fibers. There has been little consideration of the functional consequences of these differences. I suggest that, faced with a size constraint on the number of motor units available, arthropuds use peripheral modulation of muscle properties to achieve the flexibility and dynamic range that vertebrates achieve through recruitment of motor units.

The vertebrate endocrine system is well-characterized, with many reports of disruption by environmental chemicals. In contrast, cnidarians are less compartmentalized, physiological regulation is poorly understood, and the potential for disruption is unknown. Endocrine-like activity has not been systematically studied in cnidarians, but several classical vertebrate hormones (e.g., steroids, iodinated organic compounds, neuropeptides, and indoleamines) have been identified in cnidarian tissues. Investigators have made progress in identifying putative bioregulatory molecules in cnidarians, and testing the effects of these individual compounds. Less progress has been made in elucidating signaling pathways. For example, putative gonadotropin-releasing hormone and sex steroids have been identified in cnidarian tissues, but it is unknown whether these compounds are components of a larger signal cascade comparable to the vertebrate hypothalamic-pituitary-gonadal axis. Further, while sex steroids and iodinated organic compounds may help to regulate cnidarian physiology, the mechanisms of action are unknown. Homologs to the vertebrate steroid and thyroid receptors have not been identified in cnidarians, so more research is needed to understand the mechanisms of endocrine-like signaling in cnidarians. Elucidation of cnidarian regulatory pathways will provide insight into evolution of hormonal signaling. These studies will also improve understanding of how cnidarians respond to environmental cues and will provide a basis to investigate disruption of physiological processes by physical and chemical stressors.

The last three decades have brought clear recognition that many populations of animals are experiencing severe declines or local and global extinctions. Many examples have become common knowledge to the general public, such as worldwide declines in amphibian populations and extensive loss of coral reefs. The mechanisms underlying these and other changes are poorly understood. However, a growing literature indicates that a wide array of chemical contaminants have the potential to disrupt normal cell-to-cell signaling mechanisms. A global pollutant of most aquatic systems, nitrate has the potential to be an endocrine disrupting contaminant. This paper reviews studies performed on vertebrates demonstrating that nitrate and/or nitrite have the potential to alter endocrine function. Further, a retrospective study of our work on alligators from various lakes in Florida suggests that nitrate could contribute to some of the altered endocrine parameters previously reported in juvenile animals. We propose hypotheses suggesting that nitrate could alter steroidogenesis by 1) conversion to nitrite and nitric oxide in the mitochondria, the site of initial steroid synthesis, 2) altering C[l.sup.-] ion concentrations in the cell by substituting for C[l.sup.-] in the membrane transport pump or 3) binding to the heine region of various P450 enzymes associated with steroidogenesis and altering enzymatic action. Future studies are needed to examine the endocrine disruptive action of this ubiquitous pollutant. A growing literature indicates that all biologists studying natural systems, whether they choose to or not, must now consider contaminant exposure as a direct influence on their studies. That is, ubiquitous global contamination has the potential to alter the endocrine, nervous and immune systems of all organisms with resulting changes in gene expression and phenotypes.

The zebrasystem system is used for the investigating vertebrate development, physiology, disease mechanisms and also the genetic control of embryonic axis formation. Nodal, FGF, Bmp, Wnt and retinoic signals provide positional information and activate transcription factors that specify cell fates during gastrulation.

Recent studies have shown that most tetrapod groups (mammals, birds, chelonians, amphibians) show general intraspecific tendencies for increasing body size with latitude, whereas squamates (lizards and snakes) show an intraspecific tendency towards decreasing body size with latitude. Here I evaluate whether these size trends are general by using independent contrasts analysis to investigate the dependence of intraspecific size-latitude relationships (r), and the magnitude alone of size-latitude relationships ([r]), for tetrapod vertebrates, on sample size, range of latitudes sampled, average latitude sampled, and body size. Range of latitudes sampled, average latitude sampled, and body size did not influence body size-latitude relationships (r) or the magnitude alone of body size-latitude relationship ([r]). Sample size did not influence size-latitude relationships (r), but did influence the magnitude alone of size-latitude relationships ([r]), possibly indicating increased precision of estimating size-latitude relationships with increased sampling. In short, intraspecific size-latitude relationships are similar for species of different sizes, occurring at different latitudes, sampled over different latitudinal ranges, and differing in number of populations sampled (though magnitude alone is influenced by sample size). These results suggest that intraspecific size-latitude trends are general, and biologically significant (i.e., are not artifacts of sampling), thus deserving explanation.

Across vertebrates, there are three principal skeletal tissues: bone, persistent cartilage, and replacement cartilage. Although each tissue has a different evolutionary history and functional morphology, they also share many features. For example, they function as structural supports, they are comprised of cells embedded in collagen-rich extracellular matrix, and they derive from a common embryonic stem cell, the osteochondroprogenitor. Occasionally, homologous skeletal elements can change tissue type through phylogeny. Together, these observations raise the possibility that skeletal tissue identity is determined by a shared set of genes. Here, we show that misexpression of either Sox9 or Runx2 can substitute bone with replacement cartilage or can convert persistent cartilage into replacement cartilage and vice versa. Our data also suggest that these transcription factors function in a molecular hierarchy in which chondrogenic factors dominate. We propose a binary molecular code that determines whether skeletal tissues form as bone, persistent cartilage, or replacement cartilage. Finally, these data provide insights into the roles that master regulatory genes play during evolutionary change of the vertebrate skeleton. Keywords: Sox9; Runx2; Persistent cartilage; Replacement cartilage; Bone; Transcription factors; Differentiation; Secondary cartilage; Dominance; Skeletal evolution

We provide genetic evidence defining a role for noncanonical Wnt function in vertebrate axis formation. In zebrafish, misexpression of Wnt-4, -5, and -11 stimulates calcium ([Ca.sup.2+]) release, defining the Wnt/[Ca.sup.2+] class. We describe genetic interaction between two Wnt/[Ca.sup.2+] members, Wnt-5 (pipetail) and Wnt-11 (silberblick), and a reduction of [Ca.sup.2+] release in Wnt-5/pipetail. Embryos genetically depleted of both maternal and zygotic Wnt-5 product exhibit cell movement defects as well as hyperdorsalization and axis-duplication phenotypes. The dorsalized phenotypes result from increased [beta]-catenin accumulation and activation of downstream genes. The Wnt-5 loss-off function defect is consistent with [Ca.sup.2+] modulation having an antagonistic interaction with Wnt/[beta]-catenin signaling.

The BMP signaling pathway plays a key role during dorsoventral pattern formation of vertebrate embryos. In zebrafish, all cloned mutants affecting this process are deficient in members of the BMP pathway. In a search for factors differentially expressed in swirl/bmp2b mutants compared with wild type, we isolated zebrafish Sizzled, a member of the secreted Frizzled-related protein family and putative Wnt inhibitor. The knockdown of sizzled using antisense morpholino phenocopied the ventralized mutant ogon (formerly also known as mercedes and short tail). By sequencing and rescue experiments, we demonstrate that ogon encodes sizzled. Overexpression of sizzled, resulting in strongly dorsalized phenotypes, and the expression domains of sizzled in wild type embryos, localized in the ventral side during gastrulation and restricted to the posterior end during segmentation stages, correlate with its role in dorsoventral patterning. The expanded expression domain of sizzled in ogon and chordino together with its downregulation in swirl suggests a BMP2b-dependent negative autoregulation of sizzled. Indicating a novel role for a secreted Frizzled-related protein, we show that, in addition to the BMP pathway, a component of the Wnt signaling pathway is required for dorsoventral pattern formation in zebrafish. Keywords: ogon; sizzled; Zebrafish; Dorsoventral patterning; Secreted Frizzled-related protein

Many fish, amphibians, reptiles, birds, and some mammals use UV vision for such basic activities as foraging, mate selection, and communication. UV vision is mediated by UV pigments in the short wavelength-sensitive type 1 (SWS1) group that absorb light maximally ([[lambda].sub.max]) at [approximately equal to] 360 nm. Reconstructed SWS1 pigments of most vertebrate ancestors have [[lambda].sub.max] values of [approximately equal to] 360 nm, whereas the ancestral avian pigment has a [[lambda].sub.max] value of 393 nm. In the nonavian lineage, UV vision in many modern species is inherited directly from the vertebrate ancestor, whereas violet vision in others has evolved by different amino acid replacements at [approximately equal to] 10 specific sites. In the avian lineage, the origin of the violet pigment and the subsequent restoration of UV pigments in some species are caused by amino acid replacements F49V/F86S/L116V/S118A and S90C, respectively. The use of UV vision is associated strongly with UV-dependent behaviors of organisms. When UV light is not available or is unimportant to organisms, the SWS1 gene can become nonfunctional, as exemplified by coelacanth and dolphin.

Tetraodon nigroviridis is among the smallest known vertebrate genomes and as such represents an interesting model for studying genome architecture and evolution. Previous studies have shown that Tetraodon contains several types of tandem and dispersed repeats, but that their overall contribution is >10% of the genome. Using genomic library hybridization, fluorescent in situ hybridization, and whole genome shotgun and directed sequencing, we have investigated the global and local organization of repeat sequences in Tetraodon. We show that both tandem and dispersed repeat elements are compartmentalized in specific regions that correspond to the short arms of small subtelocentric chromosomes. The concentration of repeats in these heterochromatic regions is in sharp contrast to their paucity in euchromatin. In addition, we have identified a number of pseudogenes that have arisen through either duplication of genes or the retro-transcription of mRNAs. These pseudogenes are amplified to high numbers, some with more than 200 copies, and remain almost exclusively located in the same heterochromatic regions as transposable elements. The sequencing of one such heterochromatic region reveals a complex pattern of duplications and inversions, reminiscent of active and frequent rearrangements that can result in the truncation and hence inactivation of transposable elements. This tight compartmentalization of repeats and pseudogenes is absent in large vertebrate genomes such as mammals and is reminiscent of genomes that remain compact during evolution such as Drosophila and Arabidopsis.

The centrosome usually replicates in a semiconservative fashion, i.e., new centrioles form in association with preexisting 'maternal' centrioles. De novo formation of centrioles has been reported for a few highly specialized cell types but it has not been seen in vertebrate somatic cells. We find that when centrosomes are completely destroyed by laser microsurgery in CHO cells arrested in S phase by hydroxyurea, new centrosomes form by de novo assembly. Formation of new centrosomes occurs in two steps: ~5-8 h after ablation, clouds of pericentriolar material (PCM) containing [gamma]-tubulin and pericentrin appear in the cell. By 24 h, centrioles have formed inside of already well-developed PCM clouds. This de novo pathway leads to the formation of a random number of centrioles (2-14 per cell). Although clouds of PCM consistently form even when microtubules are completely disassembled by nocodazole, the centrioles are not assembled under these conditions.

Four major questions can be asked about vertebrate brain evolution: 1) What major changes have occurred in neural organization and function? 2) When did these changes occur? 3) By what mechanisms did these changes occur? 4) Why did these changes occur? Comparative neurobiologists have been very successful in recognizing major changes in brain structure. They have also made progress in understanding the functional significance of these changes, although this understanding is primarily limited to sensory centers, rather than integrative or motor centers, because of the relative ease of manipulating the relevant stimuli. Although neuropaleontology continues to provide important insights into when changes occurred, this approach is generally limited to recognizing variation in overall brain size, and sometimes brain regions, as interpreted from the surface of an endocranial cast. In recent years, most new information regarding when neural changes occurred has been based on cladistical analysis of neural features in extant taxa. Historically, neurobiologists have made little progress in understanding how and why brains evolve. The emerging field of evolutionary developmental biology appears to be the most promising approach for revealing how changes in development and its processes produce neural changes, including the emergence of novel features. Why neural changes have occurred is the most difficult question and one that has been the most ignored, in large part because its investigation requires a broad interdisciplinary approach involving both behavior and ecology.

Stress inhibits feeding behavior in all vertebrates. Data from mammals suggest an important role for hypothalamic neuropeptides, in particular the melanocortins and corticotropin-releasing hormone (CRH)-like peptides, in mediating stress-induced inhibition of feeding. The effects of CRH on food intake are evolutionarily ancient, as this peptide inhibits feeding in fishes, birds, and mammals. The effects of melanocortins on food intake have not been as extensively studied, but available evidence suggests that the anorexic role of neuronal melanocortins has been conserved. Although there is evidence that CRH and the melanocortins influence hypothalamic circuitry controlling food intake, these peptides may have a more primitive role in modulating visuomotor pathways involved in the recognition and acquisition of food. Stress rapidly reduces visually guided prey-catching behavior in toads, an effect that can be mimicked by administration of CRH, while corticosterone and isoproterenol are without effect. Melanocortins also reduce prey-oriented turning movements and, in addition, facilitate the acquisition of habituation to a moving prey item. The effects of these neuropeptides are rapid, occurring within 30 min after administration. Thus, changes in neuroendocrine status during stress may dramatically influence the efficacy with which visual stimuli release feeding behavior. By modulating visuomotor processing these neuropeptides may help animals make appropriate behavioral decisions during stress.

Studies over the last two decades have shown that mammalian nonmacrophagic liver endothelial cells clear the blood from numerous physiological and foreign waste macromolecules, such as polysaccharides and proteins released during extracellular matrix turnover, intracellular macromolecules, modified serum proteins, and bacterial and fungal proteins [Smedsrod, B., Pertoft, H., Gustafson, S. & Laurent, T. C. (1990) Biochem. J. 266, 313-327]. These macromolecules are released daily in gram-amounts in a normal human body and are effectively taken up and degraded by the liver endothelial cells. Recent studies show that bony fishes harbor a similar system of specialized nonmacrophagic scavenger endothelial cells in either kidney [Smedsrod, B., Gjoen, T., Sveinbjornsson, B. & Berg, T. (1993) J. Fish Biol. 42, 279-291] or heart [Sorensen, K. K., Melkko, J. & Smedsrod, B. (1998) J. Exp. Biol. 201, 1707-1718], but not in liver. Using specific and extremely effective endocytosis, these fish scavenger endothelial cells function as their mammalian counterpart to eliminate soluble waste macromolecules from the circulation. We show here that species from all seven vertebrate classes carry a population of nonmacrophagic scavenger endothelial cells that efficiently eliminate an array of circulating waste macromolecules. Thus representing an important part of the vertebrate innate immune system, these scavenger endothelial cells display the following distribution in the different vertebrate classes: Gills in Agnatha and Chondrichtyes; heart or kidney in Osteichtyes; and liver in Amphibia, Reptilia, Aves, and Mammalia.

Segmentation of the vertebrate hindbrain into rhombomeres is a key step in the development of a complex pattern of differentiated neurons from a homogeneous neuroepithelium. Many of the transcription factors important for establishing the segmental plan and assigning rhombomere identity are now known. However, the downstream effectors that bring about the formation of rhombomere boundaries are only just being characterized. Here we discuss molecules that could be responsible for segregating populations of cells from different rhombomeres. We focus on recent work demonstrating that the Eph family of receptor tyrosine kinases and their ligands, the ephrins, function in rhombomere-specific cell sorting and initiation of a structural boundary. We discuss the contributions of two mechanisms -- cell sorting and plasticity -- to the formation of rhombomere boundaries.

Radial glia are specialized cells in the developing nervous system of all vertebrates, and are characterized by long radial processes. These processes facilitate the best known function of radial glia: guiding the radial migration of newborn neurons from the ventricular zone to the mantle regions. Recent data indicate further important roles for these cells as ubiquitous precursors that generate neurons and glia, and as key elements in patterning and region-specific differentiation of the CNS. Thus, from being regarded mainly as support cells, radial glia have emerged as multi-purpose cells involved in most aspects of brain development.

Comparative phylogeography has proved useful for investigating biological responses to past climate change and is strongest when combined with extrinsic hypotheses derived from the fossil record or geology. However, the rarity of species with sufficient, spatially explicit fossil evidence restricts the application of this method. Here, we develop an alternative approach in which spatial models of predicted species distributions under serial paleoclimates are compared with a molecular phylogeography, in this case for a snail endemic to the rainforests of North Queensland, Australia. We also compare the phylogeography of the snail to those from several endemic vertebrates and use consilience across all of these approaches to enhance biogeographical inference for this rainforest fauna. The snail mtDNA phylogeography is consistent with predictions from paleoclimate modeling in relation to the location and size of climatic refugia through the late Pleistocene-Holocene and broad patterns of extinction and recolonization. There is general agreement between quantitative estimates of population expansion from sequence data (using likelihood and coalescent methods) vs. distributional modeling. The snail phylogeography represents a composite of both common and idiosyncratic patterns seen among vertebrates, reflecting the geographically finer scale of persistence and subdivision in the snail. In general, this multifaceted approach, combining spatially explicit paleoclimatological models and comparative phylogeography, provides a powerful approach to locating historical refugia and understanding species' responses to them.

The discipline of functional morphology grew out of a comparative anatomical tradition, its transformation into a modern experimental science facilitated largely by technological advances. Early morphologists, such as Cuvier, felt that function was predictable from organismal form, to the extent that animals and plants represented perfect adaptations to their habits. However, anatomy alone could not reveal how organisms actually performed their activities. Recording techniques capable of capturing fast motion were first required to begin to understand animal movement. Muybridge is most famous for his pioneering work in fast photography in the late 19th century, enabling him to 'freeze' images of even the fastest horse at a full gallop. In fact, contemporary kinematic analysis grew directly out of the techniques Muybridge developed. Marey made perhaps an even greater contribution to experimental science through his invention of automatic apparati for recording events of animal motion. Over the first half of the 20th century, scientists developed practical methods to record activity patterns from muscles of a living, behaving human or animal. The technique of electromyography, initially used in clinical applications, was co-opted as a tool of organismal biologists in the late 1960s. Comparative anatomy, kinematic analysis and electromyography have for many years been the mainstay of vertebrate functional morphology; however, those interested in animal form and function have recently begun branching out to incorporate approaches from experimental biomechanics and other disciplines (see accompanying symposium papers), and functional morphology now stands at the threshold of becoming a truly integrative, central field in organismal biology.

Competition among the sperm of rival males is an important evolutionary phenomenon in many organisms. Yet, despite extensive research on sperm competition in some vertebrate taxa, very little progress has been made on this topic in amphibians. Urodele amphibians (newts and salamanders) are of particular interest to theories of sperm competition because most urodele females--in contrast to other vertebrate females--control the transfer of sperm from the male. Here we present a molecular study of sperm precedence and storage patterns in the rough-skinned newt (Taricha granulosa). First, we used microsatellite markers to show that female newts typically use sperm from 1-3 males under natural and seminatural conditions. Second, we mated experimental females sequentially to two males and collected fertilized eggs in a temporal series. Patterns of paternity were consistent with first-male sperm precedence and complete mixing of sperm within the female. This simple pattern of sperm usage, best described as 'topping off,' is consistent with the expectation from sexual conflict theory that free female choice before insemination eliminates selective pressures for the evolution of complex patterns of paternity manipulation involving cryptic female choice.

Vertebrate rod photoreceptors adjust their sensitivity as they adapt during exposure to steady light. Light adaptation prevents the rod from saturating and significantly extends its dynamic range. We examined the time course of the onset of light adaptation in bullfrog rods and compared it with the projected onset of feedback reactions thought to underlie light adaptation on the molecular level. We found that adaptation developed in two distinct temporal phases: (1) a fast phase that operated within seconds after the onset of illumination, which is consistent with most previous reports of a 1-2-s time constant for the onset of adaptation; and (2) a slow phase that engaged over tens of seconds of continuous illumination. The fast phase desensitized the rods as much as 80-fold, and was observed at every light intensity tested. The slow phase was observed only at light intensities that suppressed more than half of the dark current. It provided an additional sensitivity loss of up to 40-fold before the rod saturated. Thus, rods achieved a total degree of adaptation of ~3,000-fold. Although the fast adaptation is likely to originate from the well characterized [Ca.sup.2+]-dependent feedback mechanisms regulating the activities of several phototransduction cascade components, the molecular mechanism underlying slow adaptation is unclear. We tested the hypothesis that the slow adaptation phase is mediated by cGMP dissociation from noncatalytic binding sites on the cGMP phosphodiesterase, which has been shown to reduce the lifetime of activated phosphodiesterase in vitro. Although cGMP dissociated from the noncatalytic binding sites in intact rods with kinetics approximating that for the slow adaptation phase, this hypothesis was ruled out because the intensity of light required for cGMP dissociation far exceeded that required to evoke the slow phase. Other possible mechanisms are discussed. KEYWORDS: photoreceptors * phototransduction * adaptation * calcium * cGMP

The retinoic acid-inactivating enzyme CYP26 has been found to be necessary for setting up uneven distribution of retinoic acid along the anterio-posterior axis in mouse embryos. Mutant mice lacking CYP26 were generated to study the role of the enzyme. Retinoic acid is a derivative of vitamin A and plays a vital role in vertebrate development.

Neurotrophins, key regulators of cell shapes and cell fates in the nervous system of vertebrates, are discussed in this review article. A two-receptor system is used for neurotrophin signaling, and neuroprophin receptor expression is highly regulated. The neurotrophins control survival of neurons in that they prevent and cause programmed cell death. They also regulate growth of neuronal processes. Topics include signal transduction through the Trk receptors, the splice variants of these receptors, signal transduction through the neurotrophin receptor p75, variants of the p75 receptor, nuclear targets of neurotrophin signaling, and neurotrophins and synaptic transmission.

Recently published work that establishes a specific role for the nuclear receptor FXR in cholesterol catabolism pathways and in repression of the CYP7A1 gene and regulation by nuclear receptors, is discussed in this review article. The rate-limiting step of the neutral bile biosynthesis pathway is catalyzed by the cytochrome P450 cholesterol 7alpha-hydroxylase or CYP7A1. Lipid homeostasis in vertebrates, that is, the acquisition, synthesis and metabolism of cholesterol, is important. As mediators of organ physiology, nuclear hormone receptors, and other proteins are thought to play a vital role in the processes by controlling the transcription of genes encoding the transporters, the apoplipoproteins, as well as catabolic and synthetic enzymes. Implications for future drug development in treatment of hypercholesterolemia and cholestasis are important.

The strucutral linkage between the opposing globular domains in vertebrate calmodulin (CaM) has been studied by putting together a CaM mutant deficient in the last four amino acids between Met(super.145) and Lys(super.148) at the carboxyl terminal. Proximity between opposing domains of the CaM was found to be greater after the deletion. The structural collapse of CaM helps the rapid, cooperative binding of the opposing globular domains with target proteins. That is important to high-affinity binding and rapid enzyme activation.

I. INTRODUCTION This review explores the mechanisms of central control and coordination of the respiratory and cardiovascular systems in vertebrates. Animals have evolved sophisticated control mechanisms enabling them to match [...]

Vertebrate olfactory transduction is discussed with attention to controversial issues and a review of slow-gating kinetics of the cyclic nucleotide-gated channel and detection of single-odorant molecules. It cannot be said that any odorant-evoked biochemical responses seen in a ciliary preparation originate in olfactory cilia. Evidence indicates cyclic AMP is the only pathway mediating olfactory transduction and that cyclic AMP is the only second messenger. Studies have involved two knockout mice.

Timing in the vertebrate brain stem auditory nuclei is discussed. In all higher vertebrates the timing of electrical signaling carries acoustic information. Precision is in tens of microseconds in pathways from cochlea to midbrain, and different facets of timing information are used and pulled out along parallel pathways. Strong evidence that mammals and birds detect interaural phase differences for localization of low-frequency sound in the medial superior olive (MSO) and in the horizontal plane in the nucleus laminaris, respectively.

Although regional differences in mesenchymal cell affinity in the limb bud represent positional identity, the molecular basis for cell affinity is poorly understood. We found that treatment of the cell surface with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) could change cell affinity in culture. When PI-PLC was added to the culture medium, segregation of the progress zone (PZ) cells from different stage limb buds was inhibited. Similarly, sorting out of the cells from different positions along the proximodistal (PD) axis of the same stage limb buds was disturbed. Since PI-PLC can remove glycosylphosphatidylinositol (GPI)-anchored membrane bound proteins from the cell surface, the GPI-anchored cell surface proteins may be involved in sorting out. To define the GPI-anchored molecules that determine the segregation of limb mesenchymal cells, we examined the effect of neutralizing antibody on the EphA4 receptor that binds to GPI-anchored cell surface ligands, called ephrin-A. Sorting out of the PZ cells at different stages could be inhibited by the neutralizing antibody to EphA4. These results suggest that EphA4 and its GPI-anchored ligands are, at least in part, involved in sorting out of limb mesenchymal cells with different proximal-distal positional values, and that GPI-anchored cell surface proteins play important roles in determining cell affinity in the limb bud. Key Words: limb pattern formation; cell sorting out; positional identity; GPI-anchored molecule; EphA4; ephrin-A.

In vertebrates, the segmented somites, which are the medial-most component in the paraxial mesoderm, are the entity giving rise to the axial bones and skeletal muscles. We previously demonstrated that the mechanism that distinguishes the somite from the more lateral mesoderm (lateral plate) involves different levels of BMP-4 activity which is highest in the lateral plate. We report that Noggin, an antagonist of BMP-4, is expressed in the presumptive somite and appears to control effective levels of BMP-4 to differentiate somitic mesoderm from the lateral plate. When Noggin-producing cells were implanted into the presumptive lateral plate, they produced ectopic somites that were respecified from the lateral plate precursors. These somites exhibited no mediolateral (M-L) polarity, but acquired it when implanted Noggin was eliminated. Thus, in normal embryogenesis no or low BMP-4 activity realized by Noggin specifies the somites in the medial-most portion of the paraxial mesoderm, and then BMP-4 emanating from the lateral plate subsequently establishes the M-L polarity in the somites. Key Words: mediolateral axis; mesoderm subdivision; BMP-4; morphogen.