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The eta-globin gene. Its long evolutionary history in the beta-globin gene family of mammals.

In phylogenetic reconstructions by the parsimony method, utilizing 62 sequenced globin genes and pseudogenes (including 34 of the beta-globin gene family from eutherian orders Primates, Lagomorpha, Artiodactyla and Rodentia), the branch of primate psi beta pseudogenes and the goat embryonically expressed epsilon II gene group monophyletically together as orthologues of a common ancestral gene (labelled eta) distinct from orthologues of epsilon, gamma, delta and beta. This primate psi eta-goat eta branch is cladistically closer to epsilon and gamma than to delta and beta branches. In each eutherian order gene conversions replaced portions of delta by beta sequences, whereas in descent of Primates epsilon, gamma and eta mostly retained their separate ancient identities predating the radiation of Eutheria in all their exons and non-coding regions. The loci of the ancestral beta-globin gene cluster in basal eutherians and proto-primates, as deduced from beta-clusters representing the four eutherian orders, were linked 5'-epsilon-gamma-eta-delta-beta-3' with epsilon, gamma and eta being embryonically expressed genes, and delta and beta ontogenetically later expressed genes. Through deletions gamma was lost in artiodactyl evolution, eta in lagomorph and rodent evolution, and all DNA between exon 2 3' boundaries of eta and delta in prosimian lemuriform evolution (lemur having the hybrid pseudogene psi eta delta). Simian primates retained intact the five loci of the ancestral cluster. Not only did eta, after it became a pseudogene in the basal primates, persist intact in descent to present-day simians but in the line to hominoids it evolved during the last 40 million years at the decelerated rate of 1 X 10(-9) substitutions/site per year which is one-fifth the expected neutral rate. The possibility is suggested that the psi eta locus situated between fetal and adult chromosomal domains of the simian beta-globin gene cluster might play some role in a mechanism for ontogenetic switches of globin gene expression. However, not enough sequence data on genes and intergenic regions in DNA of species of primates and other mammals as yet exist to know if the slow rate of 1 X 10(-9) reflects the rate of a conserved functional gene or primarily reflects a decelerated neutral rate of hominoid DNA evolution, conceivably from enhanced DNA repair and longer generation times in hominoids. The further possibility is raised that gene correction (repair of damaged DNA that prevents emergence of new alleles) and gene conversion both more often involve strand copying of conserved than of rapidly evolving DNA.

Animals↗

Species specific sensitivity towards the hemorrhagin of Ophiophagus hannah (Elapidae).

The filtration pattern of Ophiophagus hannah venom on Sephadex G-75 shows several peaks. The first peak, S1, includes high molecular weight proteins and contains the hemorrhagic and proteolytic activities. The proteolytic fractions overlap the hemorrhagic fractions, but are not identical with them. The crude venom and the high molecular weight peak have caseinase, benzoyl-L-arginine ethyl ester hydrolase and kallikrein-like activities, but not collagenase, gelatinase, thrombin, plasmin or urokinase-like activities. The hemorrhagin of Ophiophagus hannah shows species specific differences in its hemorrhagic effects: it causes hemorrhages in rabbits and hares (Lagomorpha), but not in rats, mice or guinea-pigs (Rodentia).

Animals↗

Amino acid sequences of P1 protamines and the phylogeny of eutherian mammals: a cladistic study.

Amino acid and cDNA sequences of eutherian P1 protamines, known from publications of other authors, were compared by a cladistic method. Fish, toad and bird protamines were used for the pertinent "outgroup comparisons", i.e. they provided relevant data for the comparative alignment of the sequences and for the recognition of evolutionary trends. In the sequence positions compared, each amino acid was individually assigned as a plesiomorphic or apomorphic character state (qualitative treatment). The resulting phylogenetic tree (Fig. 2) is only partially in accordance with common ideas on eutherian phylogeny. Disagreements refer to the branching points of Perissodactyla, Lagomorpha and Rodentia.

Amino Acid Sequence↗

Tension fibroblasts and the connective tissue matrix of the spiral ligament.

Fibroblasts with stress fibers (tension fibroblasts) have previously been described in the marginal region of the spiral ligament of bats and mice (Henson et al., 1984, 1985). The location, structure and attachments of these cells and the fact that they contain contraction associated proteins, have suggested a role in the generation of tension within the basilar membrane-spiral ligament complex. In this study the structure of these fibroblasts and their relationships to different types of connective tissue matrices were examined in representative Marsupalia, Insectivora, Chiroptera, Rodentia, Lagomorpha, Carnivora and Primates. Tension fibroblasts occur in all species but they are remarkably different in their actin filament content, their structure and distribution, and in their association with the extracellular matrix and otic capsule. Six types of matrices are described (dense filamentous, bundled, laminated, honeycombed, trabeculated and loose filamentous). The configurations of the cells and fibers indicate that tension on the basilar membrane may be accomplished in different ways and to different degrees in different mammals. The arrangement of the matrix and cells in some animals is markedly different in the parts of the cochlea that respond to high, middle and low frequencies.

Actins↗

The linear allometric relationship between total metabolic energy per life span and body mass of mammals.

The aim of this study is to establish and calculate the exact allometric relationship between the total metabolic energy per life span and the body mass in a wide range of mammals with about six orders of magnitude variation of the body mass of animals. The study shows that it exists a linear relationship between the total metabolic energy per life span PT(ls) (kJ) and the body mass M (kg) of 95 mammals (3 monotremes, Subclass Prototheria, 16 marsupialis (Subclass Theria, Infraclass Metatheria) and 76 placentals (Subclass Theria, Infraclass Eutheria)) from type: PT(ls)=A(ls)(+)M(1.0511), where P (kJ/day) is the basal rate of metabolism and T(ls) (days) is the mean life span of animals. The linear coefficient A(ls)(+)=7.158x10(5) kJ/kg is the total metabolic energy, exhausted during the life span per 1 kg body mass of the animals. The mean values of the total metabolic energy per life span, per unit body mass (A(ls)) for orders from Subclass Prototheria and Theria (Infraclass Metatheria) and orders Xenarthra, Pholidota, Soricomorpha, Rodentia (Infraclass Eutheria) varied negligible in interval (4.656-5.80)x10(5) kJ/kg. The coefficient A(ls) grows from (7.68-8.36)x10(5) kJ/kg in Lagomorpha and Artiodactyla (Eutheria) to (10.58-12.64)x10(5) kJ/kg in orders Carnivora, Pinnipeda and Chiroptera (Eutheria). A(ls) grows maximum to 18.5x10(5) kJ/kg in Primates. Thus, the values of coefficient A(ls) differ maximum four-fold in all orders. Across the all species the values of A(ls) are changes about one order of magnitude. Consequently, our survey shows that the changes of the body mass, basal metabolic rate and the life span of animals are three mutually related parameters, so that the product A(ls)=(PT(ls))/M remains relatively constant in comparison to 1 million fold difference in body mass and total metabolic energy per life span between mammals.

Animals↗

Lagomorphs (rabbits, pikas and hares) do not use telomere-directed replicative aging in vitro.

Telomere shortening is used for replicative aging in primates and ungulates but not rodents. We examined telomere biology in rabbits to expand the comparative biology of telomere-directed replicative senescence within mammals. The order Lagomorpha consists of two families; Leporidae and Ochotonidae. We examined telomere biology in species representing three leporid genera (European White Rabbit, Black-tailed Jack Rabbit, and Swamp Rabbit) and the monotypic ochotonid genus (North American Pika). Of the leporids one species was a laboratory strain and the others were wild caught. The leporids neither exhibited cellular senescence after sustained periods in culture nor displayed detectable telomerase activity. Continued culture was possible because of their extremely long telomeric arrays. Immunofluorescence showed robust telomere signals at chromosome ends and significant internal chromosomal staining in some instances. Pika was unique in displaying endogenous telomerase activity throughout time in culture. These results show that it is unlikely that lagomorphs use telomere shortening and replicative senescence as a tumor protective mechanism.

Animals↗

The broad spectrum of Trichinella hosts: from cold- to warm-blooded animals.

In recent years, studies on Trichinella have shown that the host range is wider than previously believed and new Trichinella species and genotypes have been described. Three classes of vertebrates are known to act as hosts, mammals, birds and reptiles, and infected vertebrates have been detected on all continents but Antarctica. Mammals represent the most important hosts and all Trichinella species are able to develop in this vertebrate class. Natural infections with Trichinella have been described in more than 150 mammalian species belonging to 12 orders (i.e., Marsupialia, Insectivora, Edentata, Chiroptera, Lagomorpha, Rodentia, Cetacea, Carnivora, Perissodactyla, Artiodactyla, Tylopoda and Primates). The epidemiology of the infection greatly varies by species relative to characteristics, such as diet, life span, distribution, behaviour, and relationships with humans. The non-encapsulated species Trichinella pseudospiralis, detected in both mammals (14 species) and birds (13 species), shows a cosmopolitan distribution with three distinguishable populations in the Palearctic, Nearctic and Australian regions. Two additional non-encapsulated species, Trichinella papuae, detected in wild pigs and saltwater crocodiles of Papua New Guinea, and Trichinella zimbabwensis, detected in farmed Nile crocodiles of Zimbabwe, can complete their life cycle in both mammals and reptiles. To the best of our knowledge, T. papuae and T. zimbabwensis are the only two parasites known to complete their entire life cycle independently of whether the host is warm-blooded or cold-blooded. This suggests that these two Trichinella species are capable of activating different physiological mechanisms, according to the specific vertebrate class hosting them.

Animals↗

Amblyomma tigrinum (Acari: Ixodidae): new data on hosts and biology of immature stages and on DNA composition.

Biological data of immature stages of Amblyomma tigrinum were obtained from larvae and nymphs both fed on rats and rabbits. Data from nymphs recovered from a wild rodent (Galea musteloides) are also reported. Additional results in DNA composition of males moulted from nymphs fed on laboratory and wild hosts are presented. The ticks were maintained in darkness at 27 +/- 1 degrees C and 83-86% RH. The mean recovery rates were 49.1% and 43.6% with a moulting success of 96.2% and 90.8% in larvae fed on rats and rabbits, respectively. The engorgement weights were almost identical in larvae recovered from both hosts. The mean recovery rates of nymphs were 37.3% and 69.9% in specimens fed on rats and rabbits, respectively. The moulting success was 94.5%, 100% and 98.1% in nymphs fed on rats, rabbits and G. musteloides, respectively. Nymphs from all hosts moulting to females were significantly heavier (P < 0.01) than those moulting to males despite their range of engorgement weight showed overlap. A higher proportion (>or=61.5%) of nymphs from all hosts moulted to females. Present results suggest that members of Rodentia and Lagomorpha are suitable hosts for the immature stages of A. tigrinum, contrasting with previous results from Brazilian colonies of this tick. The DNA sequence from ticks fed on G. musteloides showed 99.7% identity with that from ticks fed on rabbits and also with the DNA sequence already available (GenBank AY498562 ) for A. tigrinum.

Animals↗

Rabbits, if anything, are likely Glires.

Rodentia (e.g., mice, rats, dormice, squirrels, and guinea pigs) and Lagomorpha (e.g., rabbits, hares, and pikas) are usually grouped into the Glires. Status of this controversial superorder has been evaluated using morphology, paleontology, and mitochondrial plus nuclear DNA sequences. This growing corpus of data has been favoring the monophyly of Glires. Recently, Misawa and Janke [Mol. Phylogenet. Evol. 28 (2003) 320] analyzed the 6441 amino acids of 20 nuclear proteins for six placental mammals (rat, mouse, rabbit, human, cattle, and dog) and two outgroups (chicken and xenopus), and observed a basal position of the two murine rodents among the former. They concluded that "the Glires hypothesis was rejected." We here reanalyzed [loc. cit.] data set under maximum likelihood and Bayesian tree-building approaches, using phylogenetic models that take into account among-site variation in evolutionary rates and branch-length variation among proteins. Our observations support both the association of rodents and lagomorphs and the monophyly of Euarchontoglires (=Supraprimates) as the most likely explanation of the protein alignments. We conducted simulation studies to evaluate the appropriateness of lissamphibian and avian outgroups to root the placental tree. When the outgroup-to-ingroup evolutionary distance increases, maximum parsimony roots the topology along the long Mus-Rattus branch. Maximum likelihood, in contrast, roots the topology along different branches as a function of their length. Maximum likelihood appears less sensitive to the "long-branch attraction artifact" than is parsimony. Our phylogenetic conclusions were confirmed by the analysis of a different protein data set using a similar sample of species but different outgroups. We also tested the effect of the addition of afrotherian and xenarthran taxa. Using the linearized tree method, [loc. cit.] estimated that mice and rats diverged about 35 million years ago. Molecular dating based on the Bayesian relaxed molecular clock method suggests that the 95% credibility interval for the split between mice and rats is 7-17 Mya. We here emphasize the need for appropriate models of sequence evolution (matrices of amino acid replacement, taking into account among-site rate variation, and independent parameters across independent protein partitions) and for a taxonomically broad sample, and conclude on the likelihood that rodents and lagomorphs together constitute a monophyletic group (Glires).

Animals↗

Cross reactivities among some mammalian haptoglobins studied by a monoclonal antibody.

Mouse monoclonal antibody antihuman-Hp, product of clone 2.36.71.41 was found to recognize, however, with different affinities, the immunological determinant on haptoglobin of some mammals (goat, sheep, cow, horse, rabbit). The following differences in immunological cross reactivities were noticed: (i) haptoglobins present in sera of goat, sheep and cow (Artiodactyla, Bovidae) react in enzyme-linked immunosorbent assay (ELISA) and form precipitates in agarose gel; (ii) horse (Perissodactyla) and rabbit (Lagomorpha) haptoglobins react in ELISA, but they do not form precipitates; and (iii) haptoglobins of dog, fox, cat (Carnivora) and pig (Artiodactyla, Suidae) are not recognized by the tested monoclonal antibody either in ELISA or in agarose gel. This study suggests that monoclonal antibody, clone 2.36.71.41, recognizes the structure on haptoglobin around the disulphide bond linking two alpha chains. Antigenic structure of Bovidae haptoglobins is rather similar to human haptoglobin 2-2 (circular polymers) than to 2-1 type (linear polymers). Monoclonal antibody 2.36.71.41 could be used for classification of mammalian haptoglobins by epitope structure. It can distinguish polymeric haptoglobins similar to human type 2-2 from other mammalian haptoglobins.

Animals↗

Maternal separation and social isolation modulate the postnatal development of synaptic composition in the infralimbic cortex of Octodon degus.

We analysed the influence of preweaning periodic maternal separation followed by postweaning chronic social isolation on the development of synaptic composition in the infralimbic cortex of Octodon degus, a South American species formerly classified as a caviomorph rodent but now considered to belong to Lagomorpha (rabbits). Three groups of animals were analysed: (1) control pups which remained undisturbed with their families; (2) pups which were exposed to individual periodic maternal deprivation [postnatal day 1 (P1) until P21], followed by social isolation (P22 until P45); and (3) pups which were handled daily without being removed from the families (P1 until P21) and thereafter remained undisturbed with the family (P22 until P45). The mean synaptic density and mean projected height of synapses were quantified using the "dissector" method. In the deprived group, significantly higher (up to 137.8%) mean synaptic densities were found in layer II of the infralimbic cortex compared to normal control animals. In handled pups, asymmetric shaft synapses were significantly decreased (down to 54%) compared to the control group.These results indicate that early postnatal changes in the socio-emotional environment change the number of synaptic connections in the infralimbic cortex. Since this subregion of the medial prefrontal cortex is involved in a variety of emotional behaviors and plays a role in associative learning tasks, these environmentally induced synaptic changes may be indicative, and perhaps the cause, of alterations of behavioral and cognitive capacities.

Animals↗

The "human"-like morphology of the swine papilla ilealis.

The morphology of the ileocecocolic transition, between the small and large intestine was studied in 33 male, adult pigs of unknown breed. The results were analyzed by the Wilcoxon Test (alpha < or = 0.05). The ileum opened into the large intestine, in all of the cases, at the level of the cecocolic junction in the form of a conical papilla (ileal papilla) turned towards the large intestine, with an oblique caudal and lateral direction, pointing to the cecal lumen. The cranial contour is higher than the caudal one, with average values of 3.0 cm, and 1.5 cm in the fresh specimens and 2.8 cm and 1.4 cm after fixation of the specimens. The average diameter of the papilla apex was 1.5 cm, both in the fresh specimens and in the fixed ones. In 100% of the cases the ileal papilla appeared with a star-shaped orifice in its apex in both the fresh and in the fixed specimens. Our studies showed that the transition between the small and large intestine of the swine is ileocecocolic, thus similar to humans and different from the ileocecal (Perissodactyla, Rodentia, Lagomorpha) and, according to most descriptions, ileocolic (dog) transitions found in other animals.

Animals↗

Evolution of genome organizations of squirrels (Sciuridae) revealed by cross-species chromosome painting.

With complete sets of chromosome-specific painting probes derived from flow-sorted chromosomes of human and grey squirrel (Sciurus carolinensis), the whole genome homologies between human and representatives of tree squirrels (Sciurus carolinensis, Callosciurus erythraeus), flying squirrels (Petaurista albiventer) and chipmunks (Tamias sibiricus) have been defined by cross-species chromosome painting. The results show that, unlike the highly rearranged karyotypes of mouse and rat, the karyotypes of squirrels are highly conserved. Two methods have been used to reconstruct the genome phylogeny of squirrels with the laboratory rabbit (Oryctolagus cuniculus) as the out-group: (1) phylogenetic analysis by parsimony using chromosomal characters identified by comparative cytogenetic approaches; (2) mapping the genome rearrangements onto recently published sequence-based molecular trees. Our chromosome painting results, in combination with molecular data, show that flying squirrels are phylogenetically close to New World tree squirrels. Chromosome painting and G-banding comparisons place chipmunks (Tamias sibiricus ), with a derived karyotype, outside the clade comprising tree and flying squirrels. The superorder Glires (orde Rodentia + order Lagomorpha) is firmly supported by two conserved syntenic associations between human chromosomes 1 and 10p homologues, and between 9 and 11 homologues.

Animals↗

Cross-species characterization of the promoter region of the cystic fibrosis transmembrane conductance regulator gene reveals multiple levels of regulation.

The cystic fibrosis transmembrane conductance regulator (CFTR) gene is highly conserved within vertebrate species. Its pattern of expression in vivo seems to be tightly regulated both developmentally and in a tissue-specific manner, but shows differences with species. To identify transcriptional regulatory elements in the CFTR promoter region, we have used a combined approach based both on the analysis of the chromatin structure in vivo in rat tissues and on evolutionary clues (i.e. phylogenetic footprinting). In CFTR-expressing tissues, 15 DNase I-hypersensitive sites were identified within a 36 kb region encompassing exon 1. Eleven of them are clustered in a 3.5 kb region that exhibits eleven phylogenetic footprints observed when comparing sequences from eight mammalian species representing four orders (Primates, Artiodactylia, Lagomorpha and Rodentia). Comparison of the two sets of data allows the identification of two types of regulatory elements. Some are conserved between species, such as a non-consensus cAMP response element (CRE) and a PMA-responsive element (TRE) located respectively at positions -0.1 and -1.3 kb relative to ATG. Some are species-specific elements such as a 300 bp purine.pyrimidine (Pu.Py) stretch that is present only in rodents. Analysis of protein/DNA interactions in vitro with rat tissue protein extracts on the conserved elements revealed that the TRE site binds a specific heterodimeric complex composed of Fra-2, Jun D and a protein immunologically related to Jun/CRE-binding protein in the duodenum, whereas the CRE-like site binds ATF-1 ubiquitously. Functional analysis in Caco-2 cells showed that the CRE-like site supports a high basal transcriptional activity but is not able by itself to induce a response to cAMP, whereas the TRE site acts as a weak transactivator stimulated by PMA. Lastly, we found that the rodent-specific Pu.Py stretch confers nuclease S1 hypersensitivity under conditions of acidic pH and supercoiling. This indicates a non-B DNA conformation and thus reinforces the biological significance of non-random Pu.Py strand asymmetry in the regulation of transcription. Thus the tight transcriptional regulation of CFTR expression involves the combination of multiple regulatory elements that act in the chromatin environment in vivo. Some of them are conserved throughout evolution, such as the CRE-like element, which is clearly involved in the basal level of transcription; others are species-specific.

Activating Transcription Factor 1↗

Hormonal and immunological aspects of the phylogeny of sex steroid binding plasma protein.

Sex steoid binding plasma protein (Sbp) in man and in monkeys binds the androgens dihydrotestosterone and testosterone and the estrogen estradiol with high affinity (Kd approximately 0.5, 1, and 2 nM, respectively). Detailed studies of steroid binding specificity give the same results in all primates, except that in humans and chimpanzees estrone does not compete for dihydrotestosterone binding. In other mammals, Sbps of Artiodactyla and Lagomorpha have the same range of affinities for androgens but they do not bind estradiol to any significant extent (Kd > 280 nM). The dog has an unusual Sbp (Kd for dihydrotestosterone, 7.1 nM; for estradiol, 125 nM), and rodents do not have a specific dihydrotestosterone-binding plasma protein. Gel filtration and immunoelectrophoretic experiments have been performed with a monospecific antiserum against human Sbp. The results indicate variable crossreactivities with Sbps of primates (from complete in chimpanzee and gorilla to weak in Prosimii). No crossreaction was observed with specific androgen-binding plasma proteins of other species. These results suggest the evolutionary emergence of bifunctional Sbp.

Animals↗

Evolutionary and developmental aspects of two hemoglobin beta-chain genes (epsilon M and beta M) of opossum.

A series of gene duplications that began in a stem species of Mammalia and led to five developmentally regulated hemoglobin beta-chain loci (epsilon, gamma, eta, delta, and beta) in a common ancestor of eutherian orders Artiodactyla, Rodentia, Lagomorpha, and Primates had important consequences in mammalian evolution. Findings reported here indicate that two progenitors of the five linked genes existed by the time of the eutherian (placental mammal)-metatherian (marsupial mammal) split and that these two genes were already differentiated with respect to their promoter regions and developmental expression. Southern blot and sequence analyses of the hemoglobin beta-chain genes of the opossum (Didelphis virginiana) revealed only two genes, one with coding and promoter sequences similar to eutherian prenatally expressed epsilon, gamma, and eta genes and the other coding for adult opossum hemoglobin beta-chains and having eutherian adult beta-type promoters. The most parsimonious arrangement of greater than 80 beta-globin exon sequences depicts the opossum embryonic-type gene as orthologously related to eutherian epsilon, gamma, and eta genes and the opossum adult-type gene as orthologously related to delta and beta genes. These data further indicate that after the initial beta duplication in the stem of Mammalia, the locus that became developmentally delayed in its expression evolved at a faster rate than the locus that became embryonically expressed.

Amino Acid Sequence↗

Neural BC1 RNA as an evolutionary marker: guinea pig remains a rodent.

The traditional morphologically grounded placement of South American guinea pig-like rodents (Caviomorpha) within one of the two rodent suborders, Hystricognathi, has been disputed by recent analysis of protein and nucleic acid sequence data. The Caviomorpha and possibly all Hystricognathi would be considered a separate order, distinct from the other rodent suborder, Sciurognathi, and thus of the order Rodentia, and would be placed closer phylogenetically to other mammals [Graur, D., Hide, W. A. & Li, W.-H. (1991) Nature (London) 351, 649-652]. To address the discrepancy between morphological comparisons and sequence analyses, we have applied an alternative form of molecular analysis. We demonstrate that BC1 RNA, a neural-specific small cytoplasmic RNA that is the product of a retropositionally generated gene (a gene derived by reverse transcription of RNA followed by insertion of the DNA copy into the genome), is present in Sciurognathi and guinea pig but not in other mammalian orders including Lagomorpha, Artiodactyla, and Primates. The species-confined, tissue-specific expression of a retroposed sequence therefore supports the morphological evidence for monophyly of Rodentia inclusive of guinea pig and demonstrates the usefulness of such molecular genetic markers. Furthermore, the conservation and tissue-specific expression of the BC1 RNA gene in the two divergent rodent suborders suggests that this macromolecule has been exapted into a functional role (i.e., coopted into a variant or novel function) in the rodent nervous system.

Animals↗

Phylogenetic assessment of molecular and morphological data for eutherian mammals.

The interordinal relationships of eutherian (placental) mammals were evaluated by a phylogenetic analysis of four published data sets (three sequences and one morphological). The nature and degree of support and conflict for particular groups were assessed by separate bootstrap and homogeneity tests, which were followed by combined analyses of the sequence and morphological data. Between orders, strong support (i.e., > or = 95% bootstrap scores) was found for a paraphyletic Artiodactyla (relative to Cetacea) and a monophyletic Cetartiodactyla (Artiodactyla and Cetacea) and Paenungulata (Hyracoidea, Proboscidea, and Sirenia). In turn, some reasonable to strong evidence (> or = 85%) was obtained for Hyracoidea with Sirenia, Dermoptera with Scandentia, Glires (Lagomorpha with Rodentia), and Afrotheria (Amblysomus, Macroscelidea, Paenungulata, and Tubulidentata). Otherwise, no other interordinal clades were supported at these reasonable to strong levels. This overall lack of resolution for eutherian interordinal clusters agrees with other studies that suggest further progress will continue to be slow and difficult. Further resolution will require the integration of more recently published data, the continued sampling of taxa and characters, and the use of more powerful methods of data analysis.

Animals↗