Stage Of Vertebrate Evolution Research Articles

Amphibian myelopoiesis

Macrophage-lineage cells are indispensable to immunity and physiology of all vertebrates. Amongst these, amphibians represent a key stage in vertebrate evolution and are facing decimating population declines and extinctions, in large part due to emerging infectious agents. While recent studies indicate that macrophages and related innate immune cells are critically involved during these infections, much remains unknown regarding the ontogeny and functional differentiation of these cell types in amphibians. Accordingly, in this review we coalesce what has been established to date about amphibian blood cell development (hematopoiesis), the development of key amphibian innate immune cells (myelopoiesis) and the differentiation of amphibian macrophage subsets (monopoiesis). We explore the current understanding of designated sites of larval and adult hematopoiesis across distinct amphibian species and consider what mechanisms may lend to these species-specific adaptations. We discern the identified molecular mechanisms governing the functional differentiation of disparate amphibian (chiefly Xenopus laevis) macrophage subsets and describe what is known about the roles of these subsets during amphibian infections with intracellular pathogens. Macrophage lineage cells are at the heart of so many vertebrate physiological processes. Thus, garnering greater understanding of the mechanisms responsible for the ontogeny and functionality of these cells in amphibians will lend to a more comprehensive view of vertebrate evolution.

The Role of Maternal pou5f3.3/oct60 Gene in the Regulation of Initial Stages of Tissue Differentiation during Xenopus laevis Embryogenesis

The role of maternal Pou5f3.3/Oct60 factor of Xenopus laevis, a homolog of the pluripotency regulator of mammalian stem cells, the Oct4 protein, in early embryonic development was studied. It was shown that the maximum concentration of the Pou5f3.3/Oct60 protein is observed at the blastula stage, and then its degradation, stimulated by posttranslational modifications after the genome activation of the embryo, occurs. Using an approach based on Pou5f3.3/Oct60 overexpression in embryos, it was found that this protein activates expression of early pluripotency marker genes, pou5f3.2, vent2.2/2.1, klf 4/5, but inhibits expression of cytoskeletal and cell adhesion genes: actin, claudin, zyxin, ldb3. In addition, Pou5f3.3/Oct60 inhibits expression of ag1/2 and agr2 genes involved in the regulation of regenerative blastema differentiation in amputated limbs, as well as in the regulation of early brain development. Given the conservatism of early stages of development in vertebrates, the data obtained in the embryos of Xenopus laevis can be used to better understand the early development in mammals, including humans.

The origins and developments of sulfation-prone tyrosine-rich and acidic N- and C-terminal extensions of class ll and lll small leucine-rich repeat proteins shed light on connective tissue evolution in vertebrates

BackgroundSmall leucine-rich repeat protein (SLRP) family members contain conserved leucine-rich repeat motifs flanked by highly variable N- and C-terminal regions. Most class II and III SLRPs have tyrosine-rich N-terminal regions and some of these are sulfated. However, the evolutionary origin and conservation of the tyrosine-rich and acidic terminal regions remain undetermined. In this study, we present the most comprehensive multiple sequence alignment (MSA) analyses of all eight class II and III SLRPs to date. Based on the level of conservation of tyrosine residues and adjacent sequences, we predict which tyrosine residues are most likely to be sulfated in the terminal regions of human class II and III SLRPs.ResultsUsing this novel approach, we predict a total of 22 tyrosine sulfation sites in human SLRPs, of which only 8 sites had been experimentally identified in mammals. Our analyses suggest that sulfation-prone, tyrosine-rich and acidic terminal regions of the class II and III SLRPs emerged via convergent evolution at different stages of vertebrate evolution, coinciding with significant evolutionary events including the development of endochondral bones and articular cartilage, the aquatic to terrestrial transition, and the formation of an amnion.ConclusionsOur study suggests that selective pressures due to changes in life conditions led to the formation of sulfotyrosine-rich and acidic terminal regions. We believe the independent emergence and evolution of sulfotyrosine-rich and acidic N- and C-terminal regions have provided each class II and III SLRP member with novel vital functions required to develop new specialized extracellular matrices and tissues in vertebrate species.

Reconstruction of the Carbohydrate 6-O Sulfotransferase Gene Family Evolution in Vertebrates Reveals Novel Member, CHST16, Lost in Amniotes.

Glycosaminoglycans are sulfated polysaccharide molecules, essential for many biological processes. The 6-O sulfation of glycosaminoglycans is carried out by carbohydrate 6-O sulfotransferases (C6OSTs), previously named Gal/GalNAc/GlcNAc 6-O sulfotransferases. Here, for the first time, we present a detailed phylogenetic reconstruction, analysis of gene synteny conservation and propose an evolutionary scenario for the C6OST family in major vertebrate groups, including mammals, birds, nonavian reptiles, amphibians, lobe-finned fishes, ray-finned fishes, cartilaginous fishes, and jawless vertebrates.The C6OST gene expansion likely started early in the chordate lineage, giving rise to four ancestral genes after the divergence of tunicates and before the emergence of extant vertebrates. The two rounds of whole-genome duplication in early vertebrate evolution (1R/2R) only contributed two additional C6OST subtype genes, increasing the vertebrate repertoire from four genes to six, divided into two branches. The first branch includes CHST1 and CHST3 as well as a previously unrecognized subtype, CHST16 that was lost in amniotes. The second branch includes CHST2, CHST7, and CHST5. Subsequently, local duplications of CHST5 gave rise to CHST4 in the ancestor of tetrapods, and to CHST6 in the ancestor of primates.The teleost-specific gene duplicates were identified for CHST1, CHST2, and CHST3 and are result of whole-genome duplication (3R) in the teleost lineage. We could also detect multiple, more recent lineage-specific duplicates. Thus, the vertebrate repertoire of C6OST genes has been shaped by gene duplications and gene losses at several stages of vertebrate evolution, with implications for the evolution of skeleton, nervous system, and cell–cell interactions.

Descriptions of spawning of the pouched lamprey (Geotria australis) in tanks and of morphological changes leading up to and following spawning

The anadromous Geotria australis, one of only three lamprey species representing the early agnathan (jawless) stage of vertebrate evolution in Australia and New Zealand, is declining in abundance. Its adults were caught soon after they had entered rivers on their non-trophic upstream migration and maintained in laboratory tanks for 13–15 months through to spawning. As adult G. australis are susceptible to haemorrhagic septicaemia, they were treated prophylactically and maintained in 3-m3 aquaria supplied with a flow-through charcoal filtration system and UV steriliser. Air temperature and the light:dark regime were constantly adjusted to parallel those in the environment. Males developed the very large suctorial disc and gular pouch characteristic of maturity and both sexes matured at the same time as in the wild. While males frequently showed aggressive behaviour towards each other, the same male and female mated on several occasions. The male coiled around the female and, with his urogenital papilla close to the female’s cloaca, twisted and vibrated, leading to egg release. These eggs formed coagulated clusters as in the wild, with many progressing through to the eight-cell stage. Remarkably, numerous G. australis were still alive 95–392 days after the end of the short spawning period, and one male after a further 119 days. Postspawning survival would be facilitated inter alia by extensive proteolysis, reflected in a shortening of the body. The data in this paper emphasise that G. australis is a highly atypical lamprey and provides invaluable information for conserving this declining species.

Double maternal-effect: duplicated nucleoplasmin 2 genes, npm2a and npm2b, with essential but distinct functions are shared by fish and tetrapods

BackgroundNucleoplasmin 2 (npm2) is an essential maternal-effect gene that mediates early embryonic events through its function as a histone chaperone that remodels chromatin. Recently, two npm2 (npm2a and npm2b) genes have been annotated in zebrafish. Thus, we examined the evolution of npm2a and npm2b in a variety of vertebrates, their potential phylogenetic relationships, and their biological functions using knockout models via the CRISPR/cas9 system.ResultsWe demonstrated that the two npm2 duplicates exist in a wide range of vertebrates, including sharks, ray-finned fish, amphibians, and sauropsids, while npm2a was lost in coelacanth and mammals, as well as some specific teleost lineages. Using phylogeny and synteny analyses, we traced their origins to the early stages of vertebrate evolution. Our findings suggested that npm2a and npm2b resulted from an ancient local gene duplication, and their functions diverged although key protein domains were conserved. We then investigated their functions by examining their tissue distribution in a wide variety of species and found that they shared ovarian-specific expression, a key feature of maternal-effect genes. We also demonstrated that both npm2a and npm2b are maternally-inherited transcripts in vertebrates, and that they play essential, but distinct, roles in early embryogenesis using zebrafish knockout models. Both npm2a and npm2b function early during oogenesis and may play a role in cortical granule function that impact egg activation and fertilization, while npm2b is also involved in early embryogenesis.ConclusionThese novel findings will broaden our knowledge on the evolutionary history of maternal-effect genes and underlying mechanisms that contribute to vertebrate reproductive success. In addition, our results demonstrate the existence of a newly described maternal-effect gene, npm2a, that contributes to egg competence, an area that still requires further comprehension.

The splicing factor SRSF1 modulates pattern formation by inhibiting transcription of tissue specific genes during embryogenesis

Alternative splicing is a major mechanism regulating pattern of gene expression through the production of multiple mRNAs from a single gene transcript. Any misregulation can cause various human diseases and also have severe effects on embryogenesis. SRSF1 is one of the critical factors regulating alternative splicing at many stages of vertebrate development and any disturbance in SRSF1 leads to serious consequences. In current study, we investigated the effects of loss of the SRSF1 gene using antisense morpholino oligonucleotides (MO) in Xenopus embryogenesis. It is evident from the results of RT-PCR and whole-mount in situ hybridization that SRSF1 is a maternal gene having strong expression in head, eyes and central nervous system. Moreover, SRSF1 morphants exhibited malformed phenotypes, including miscoiled guts, heart and cartilage formation, edema in the head and heart, and small eyes. Especially, in SRSF1 morphants, bone cartilage formation was reduced in the brain and Nkx-2.5 expression was dramatically reduced in the heart of SRSF1 morphants. In addition, a dramatic reduction in functional chordin RNA in SRSF1 morphants was observed suggesting that chordin is one of the targets of SRSF1. Thus, we concluded that SRSF1 is an essential factor for pattern formation including heart, cartilage and germ layers through the regulation of specific genes.

Morphology, Characterization and Distribution of Retinal Photoreceptors in the South American (Lepidosiren paradoxa) and Spotted African (Protopterus dolloi) Lungfishes

Lungfishes are the closest living relatives of the ancestors to all terrestrial vertebrates and have remained relatively unchanged since the early Lochkovin period (410 mya). Lungfishes, therefore, represent a critical stage in vertebrate evolution and their sensory neurobiology is of considerable interest. This study examines the ultrastructure of the retina of two species of lungfishes: the South American lungfish, Lepidosiren paradoxa and the spotted African lungfish, Protopterus dolloi in an attempt to assess variations in photoreception in these two ancient groups of sarcopterygian (lobe-finned) fishes. In juvenile P. dolloi, the retina contains one rod and two cone photoreceptor types (one containing a red oil droplet), while only one rod and one cone photoreceptor type is present in adult L. paradoxa. Both species lack double cones. The large size and inclusion of oil droplets in both species apart from one of the cone photoreceptor types in P. dolloi suggests that L. paradoxa and P. dolloi are adapted for increasing sensitivity. However, the complement of photoreceptor types suggests that there may be a major difference in the capacity to discriminate colour (dichromatic and monochromatic photoreception in P. dolloi and L. paradoxa, respectively). This study suggests that the visual needs of these two species may differ.

Ontogenetic shifts in brain scaling reflect behavioral changes in the life cycle of the pouched lamprey Geotria australis.

Very few studies have described brain scaling in vertebrates throughout ontogeny and none in lampreys, one of the two surviving groups of the early agnathan (jawless) stage in vertebrate evolution. The life cycle of anadromous parasitic lampreys comprises two divergent trophic phases, firstly filter-feeding as larvae in freshwater and secondly parasitism as adults in the sea, with the transition marked by a radical metamorphosis. We characterized the growth of the brain during the life cycle of the pouched lamprey Geotria australis, an anadromous parasitic lamprey, focusing on the scaling between brain and body during ontogeny and testing the hypothesis that the vast transitions in behavior and environment are reflected in differences in the scaling and relative size of the major brain subdivisions throughout life. The body and brain mass and the volume of six brain structures of G. australis, representing six points of the life cycle, were recorded, ranging from the early larval stage to the final stage of spawning and death. Brain mass does not increase linearly with body mass during the ontogeny of G. australis. During metamorphosis, brain mass increases markedly, even though the body mass does not increase, reflecting an overall growth of the brain, with particularly large increases in the volume of the optic tectum and other visual areas of the brain and, to a lesser extent, the olfactory bulbs. These results are consistent with the conclusions that ammocoetes rely predominantly on non-visual and chemosensory signals, while adults rely on both visual and olfactory cues.

Age distribution patterns of human gene families: divergent for Gene Ontology categories and concordant between different subcellular localizations

The age distribution of gene duplication events within the human genome exhibits two waves of duplications along with an ancient component. However, because of functional constraint differences, genes in different functional categories might show dissimilar retention patterns after duplication. It is known that genes in some functional categories are highly duplicated in the early stage of vertebrate evolution. However, the correlations of the age distribution pattern of gene duplication between the different functional categories are still unknown. To investigate this issue, we developed a robust pipeline to date the gene duplication events in the human genome. We successfully estimated about three-quarters of the duplication events within the human genome, along with the age distribution pattern in each Gene Ontology (GO) slim category. We found that some GO slim categories show different distribution patterns when compared to the whole genome. Further hierarchical clustering of the GO slim functional categories enabled grouping into two main clusters. We found that human genes located in the duplicated copy number variant regions, whose duplicate genes have not been fixed in the human population, were mainly enriched in the groups with a high proportion of recently duplicated genes. Moreover, we used a phylogenetic tree-based method to date the age of duplications in three signaling-related gene superfamilies: transcription factors, protein kinases and G-protein coupled receptors. These superfamilies were expressed in different subcellular localizations. They showed a similar age distribution as the signaling-related GO slim categories. We also compared the differences between the age distributions of gene duplications in multiple subcellular localizations. We found that the distribution patterns of the major subcellular localizations were similar to that of the whole genome. This study revealed the whole picture of the evolution patterns of gene functional categories in the human genome.

Three-dimensional limb joint mobility in the early tetrapod Ichthyostega

The origin of tetrapods and the transition from swimming to walking was a pivotal step in the evolution and diversification of terrestrial vertebrates. During this time, modifications of the limbs—particularly the specialization of joints and the structures that guide their motions—fundamentally changed the ways in which early tetrapods could move. Nonetheless, little is known about the functional consequences of limb anatomy in early tetrapods and how that anatomy influenced locomotion capabilities at this very critical stage in vertebrate evolution. Here we present a three-dimensional reconstruction of the iconic Devonian tetrapod Ichthyostega and a quantitative and comparative analysis of limb mobility in this early tetrapod. We show that Ichthyostega could not have employed typical tetrapod locomotory behaviours, such as lateral sequence walking. In particular, it lacked the necessary rotary motions in its limbs to push the body off the ground and move the limbs in an alternating sequence. Given that long-axis rotation was present in the fins of tetrapodomorph fishes, it seems that either early tetrapods evolved through an initial stage of restricted shoulder and hip joint mobility or that Ichthyostega was unique in this respect. We conclude that early tetrapods with the skeletal morphology and limb mobility of Ichthyostega were unlikely to have made some of the recently described Middle Devonian trackways.

Molecular cloning, expression analysis of insulin-like growth factor I (IGF-I) gene and IGF-I serum concentration in female and male Tongue sole (Cynoglossus semilaevis)

Insulin-like growth factor I (IGF-I) is a polypeptide hormone that regulates growth during all stages of development in vertebrates. To examine the mechanisms of the sexual growth dimorphism in the Tongue sole (Cynoglossus semilaevis), molecular cloning, expression analysis of IGF-I gene and IGF-I serum concentration analysis were performed. As a result, the IGF-I cDNA sequence is 911 bp, which contains an open reading frame (ORF) of 564 bp encoding a protein of 187 amino acids. The sex-specific tissue expression was analyzed by using 14 tissues from females, normal males and extra-large male adults. The IGF-I mRNA was predominantly expressed in liver, and the IGF-I expression levels in females and extra-large males were 1.9 and 10.2 times as much as those in normal males, respectively. Sex differences in IGF-I mRNA expressions at early life stages were also examined by using a full-sib family of C. semilaevis, and the IGF-I mRNA was detected at all of the 27 sampling points from 10 to 410 days old. An increase in IGF-I mRNA was detected after 190 day old fish. The significantly higher levels of IGF-I mRNA in females were observed after 190 days old in comparison with males (P<0.01). The IGF-I concentrations in serum of mature individuals were detected by ELISA. The IGF-I level in the serum of females was approximately two times as much as that of males. Consequently, IGF-I may play an important role in the endocrine regulation of the sexually dimorphic growth of C. semilaevis.

Gene expression throughout a vertebrate's embryogenesis

BackgroundDescribing the patterns of gene expression during embryonic development has broadened our understanding of the processes and patterns that define morphogenesis. Yet gene expression patterns have not been described throughout vertebrate embryogenesis. This study presents statistical analyses of gene expression during all 40 developmental stages in the teleost Fundulus heteroclitus using four biological replicates per stage.ResultsPatterns of gene expression for 7,000 genes appear to be important as they recapitulate developmental timing. Among the 45% of genes with significant expression differences between pairs of temporally adjacent stages, significant differences in gene expression vary from as few as five to more than 660. Five adjacent stages have disproportionately more significant changes in gene expression (> 200 genes) relative to other stages: four to eight and eight to sixteen cell stages, onset of circulation, pre and post-hatch, and during complete yolk absorption. The fewest differences among adjacent stages occur during gastrulation. Yet, at stage 16, (pre-mid-gastrulation) the largest number of genes has peak expression. This stage has an over representation of genes in oxidative respiration and protein expression (ribosomes, translational genes and proteases). Unexpectedly, among all ribosomal genes, both strong positive and negative correlations occur. Similar correlated patterns of expression occur among all significant genes.ConclusionsThese data provide statistical support for the temporal dynamics of developmental gene expression during all stages of vertebrate development.

Proinsulin: from hormonal precursor to neuroprotective factor

In the last decade, non-canonical functions have been described for several molecules with hormone-like activities in different stages of vertebrate development. Since its purification in the 1960s, proinsulin has been one of the best described hormonal precursors, though it has been overwhelmingly studied in the context of insulin, the mature protein secreted by the pancreas. Beginning with our discovery of the presence and precise regulation of proinsulin mRNA in early neurulation and neurogenesis, we uncovered a role for proinsulin in cell survival in the developing nervous system. We subsequently demonstrated the ability of proinsulin to prevent pathological cell death and delay photoreceptor degeneration in a mouse model of retinitis pigmentosa. In this review, we focus on the evolution of proinsulin/insulin, beginning with insulin-like peptides expressed in mainly the neurosecretory cells of some invertebrates. We summarize findings related to the regulation of proinsulin expression during development and discuss the possible effects of proinsulin in neural cells or tissue, and its potential as a neuroprotective molecule.

Analysis of the Albumin/α-Fetoprotein/Afamin/Group specific component gene family in the context of zebrafish liver differentiation

Members of the Albumin/α-Fetoprotein/Afamin/Group specific component ( Alb/Afp/Afm/Gc) multi-gene family perform physiological functions essential for body homeostasis and are among the earliest genes to be expressed in the fetal liver in mammals. A comprehensive search of the zebrafish genome has led to the isolation of a single member of this gene family, exhibiting close homology to group specific component (gc; also described as vitamin D binding protein (dbp)). Our phylogenetic analyses did not uncover albumin in the genome, indicating its likely absence in zebrafish, whereas the absence of afp and afm is in agreement with previous findings that both genes arose at a later stage of vertebrate evolution. gc mRNA expression is initiated weakly around 55 hours post fertilisation (hpf) in the developing liver, and increases until it reaches a continuously high level from about 72 hpf onwards. Investigation of gc mRNA in hdac1 mutants revealed a severe delay of expression, indicating a defect in progression of hepatic differentiation. This provides further evidence for Hdac1 regulating the precise timely execution of hepatic gene expression programmes. Conversely, onset of gc expression was unaltered in cloche mutant embryos, which lack hepatic vasculature, suggesting that this particular step of hepatic differentiation occurs independently from endothelial cells. Our studies identify gc as the likely only member of the Alb/Afp/Afm/Gc gene family in zebrafish, providing important insights into the evolution of this multi-gene family in vertebrates. Furthermore, the identification of gc adds a valuable temporal marker for investigating progressive hepatic differentiation in zebrafish.

Comparison of two kinds of functional asymmetry in the roach Rutilus rutilus (Teleostei: Cyprinidae)

In the higher vertebrates, asymmetries of different functions of individual are poorly connected with each other. At present it is unknown whether this poor link is a secondary phenomenon or it is characteristic of different kinds of functional asymmetry already at early stages of vertebrate evolution. In connection with this problem, in the roach Rutilus rutilus we studied asymmetry of two behavioral reactions--movements in an annular corridor and of the C-shaped body bending at a sudden action of electrical stimulus. Different individuals prefer a certain direction of movement in the corridor (to the right or to the left). Comparison of these two kinds of asymmetry in the same individuals has shown the absence of significant association between asymmetries of different reactions. The obtained data indicate that already in the lower vertebrates, different kinds of functional asymmetry are controlled by different mechanisms and are poorly associated with each other.

Gene Structure and Functional Characterization of Growth Hormone in Dogfish, Squalus acanthias

Dogfish (Squalus acanthias) growth hormone (GH) was identified by cDNA cloning and protein purification from the pituitary gland. Dogfish GH cDNA encoded a prehormone of 210 amino acids (aa). Sequence analysis of purified GH revealed that the prehormone is composed of a signal peptide of 27 aa and a mature protein of 183 aa. Dogfish GH showed 94% sequence identity with blue shark GH, and also showed 37-66%, 26%, and 48-67% sequence identity with GH from osteichtyes, an agnathan, and tetrapods. The site of production was identified through immunocytochemistry to be cells of the proximal pars distalis of the pituitary gland. Dogfish GH stimulates both insulin-like growth factor-I and II mRNA levels in dogfish liver in vitro. The dogfish GH gene consisted of five exons and four introns, the same as in lamprey, teleosts such as cypriniforms and siluriforms, and tetrapods. The 5'-flanking region within 1082 bp of the transcription start site contained consensus sequences for the TATA box, Pit-1/GHF-1, CRE, TRE, and ERE. These results show that the endocrine mechanism for growth stimulation by the GH-IGF axis was established at an early stage of vertebrate evolution, and that the 5-exon-type gene organization might reflect the structure of the ancestral gene for the GH gene family.

Early evolution of multifocal optics for well-focused colour vision in vertebrates

Jawless fishes (Agnatha; lampreys and hagfishes) most closely resemble the earliest stage in vertebrate evolution and lamprey-like animals already existed in the Lower Cambrian [about 540 million years ago (MYA)]. Agnathans are thought to have separated from the main vertebrate lineage at least 500 MYA. Hagfishes have primitive eyes, but the eyes of adult lampreys are well-developed. The southern hemisphere lamprey, Geotria australis, possesses five types of opsin genes, three of which are clearly orthologous to the opsin genes of jawed vertebrates. This suggests that the last common ancestor of all vertebrate lineages possessed a complex colour vision system. In the eyes of many bony fishes and tetrapods, well-focused colour images are created by multifocal crystalline lenses that compensate for longitudinal chromatic aberration. To trace the evolutionary origins of multifocal lenses, we studied the optical properties of the lenses in four species of lamprey (Geotria australis, Mordacia praecox, Lampetra fluviatilis and Petromyzon marinus), with representatives from all three of the extant lamprey families. Multifocal lenses are present in all lampreys studied. This suggests that the ability to create well-focused colour images with multifocal optical systems also evolved very early.

Parathyroid hormone-related protein production in the lamprey Geotria australis: developmental and evolutionary perspectives

This study explored the distribution of parathyroid hormone-related protein (PTHrP) and its mRNA in tissues of the lamprey Geotria australis, a representative of one of the two surviving groups of an early and jawless stage in vertebrate evolution. For this purpose, antibodies to N-terminal and mid-molecule human PTHrP were used to determine the locations of the antigen. Sites of mRNA production were demonstrated by in situ hybridisation with a digoxigenin-labelled riboprobe to exon VI of the human PTHrP gene. The results revealed that antigen and its mRNA were widely distributed among similar sites of tissue localisation to those described for mammalian and avian species. However, some novel sites of localisation, such as in the gill and notochord, were also found. Some differences in PTHrP localisation were noted among individuals at different intervals of the life cycle, indicating that the distributions of PTHrP, and possibly its roles, change with the stage of development in this species. The widespread tissue distribution in G. australis implies diverse physiological roles for this protein. The presence of PTHrP in the lamprey, a representative of a group of vertebrates, which apparently evolved over 540 million years ago, strongly suggests that it is a protein of ancient origin. In addition, the successful use of antibodies and probes based on the human sequence in the lamprey also provides evidence that the PTHrP molecule may have been conserved from lampreys through to humans.

Adaptive Radiation of Lampreys

Lampreys, together with hagfishes, are the sole survivors of the jawless (agnathan) stage in vertebrate evolution. The 34 species of northern hemisphere lampreys are placed in the Petromyzontidae, whereas each of the four species of southern hemisphere lampreys are separated into either the monospecific Geotriidae or monogeneric Mordaciidae. Eighteen species feed parasitically as adults in either marine or fresh waters, whereas 20 species are nonparasitic and have apparently evolved from parasitic species. This account of adaptive radiation is based on the hypothesis that the basal stock of contemporary lampreys possessed many characters that were similar to those of contemporary Ichthyomyzon and Petromyzon species, and that the Mordacia species and Geotria australis have been separated for a long period. Most of the morphological differences among the adults of northern hemisphere lampreys (i.e., variations in dentitional arrangement and size of buccal glands and velar tentacles) are related to whether the species feeds mainly on blood, as with Ichthyomyzon and Petromyzon species, or predominantly on flesh, as with the more derived Lampetra species. Mordacia species have apparently remained blood feeders, while G. australis has become a flesh feeder. The eyes of the adults of southern hemisphere lampreys are unique amongst lampreys, with those of G. australis possessing two types of cone photoreceptor (as well as a rod photoreceptor) and an irideal flap, while those of Mordacia mordax possess only a rod photoreceptor but contain a tapetum. These differences represent adaptations to life in bright surface waters by G. australis and to diurnal burrowing and nocturnal activity by Mordacia species. The restriction of larval intestinal diverticula to Mordacia species and G. australis may represent adaptations for increasing the ability to digest the particularly tough plant material that is found in southern hemisphere rivers. The bile duct of the larvae of southern hemisphere lampreys enters the apex of the above intestinal diverticula and thus, unlike the situation in northern hemisphere lampreys, does not play a role in forming the endocrine pancreas of adult lampreys during metamorphosis. This accounts for the absence of intermediate and caudal pancreatic cords in the adults of southern hemisphere species. The amino acid sequences of intestinal molecules in different lamprey taxa support the view that the lampreys are monophyletic and that polyploidy occurred early in or just prior to their evolution.