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PubMed1997

The binding potential of commercial antibody conjugates with sera of various small terrestrial mammals.

Infectious diseases of wild animals are of increasing importance, both from an economic viewpoint and because several of these diseases are pathogenic to man. However, serosurveys to determine the circulation of infectious organisms in wildlife are complicated by the fact that antibodies to species-specific immunoglobulins are not available for use in serological assays such as enzyme-linked immunosorbent assays (ELISAs) or immunofluorescence assays. To determine the binding potential of four commercially available antibody conjugates with the sera of wild animals, sera from 27 species of small terrestrial mammals were allowed to react with alkaline phosphatase-labelled protein A, anti-rabbit IgG, anti-mouse IgG and anti-human IgG by by the use of an ELISA. It was found that sera from some species of the order Lagomorpha bound optimally to anti-rabbit IgG, while anti-mouse IgG could be used for most species of Rodentia. For all Carnivora, Insectivora, Macroscelidea, Hyracoidea and other Rodentia, staphylococcal protein A demonstrated optimal binding. None of the sera that was tested bound to anti-human IgG. These results demonstrate that commercial conjugates can be used in serological assays in which wild animal sera are used, and should be useful for future serosurveys to determine the circulation of infectious agents in small terrestrial mammals.

Animals↗
PubMed2005

Gas chromatographic quantification of free D-amino acids in higher vertebrates.

D-amino acids were determined in brain, body fluids (urine, blood coagulate, serum, plasma) and faeces of animals belonging to nine out of 11 taxonomic orders of vertebrates (Artiodactyla, Aves, Carnivora, Lagomorpha, Marsupalia, Osteichthyes, Primates, Rodentia, Tubilidentata). Free amino acids were isolated by means of cation exchangers and converted into volatile N(O)-perfluoroacylamino acid propyl esters. Derivatives of amino acids were separated into D- and L-enantiomers using Chirasil-L-Val capillary columns and detected by selected ion monitoring mass spectrometry. Quantification of amino acids was achieved by comparison of analytes with amino acid standards using L-norleucine as internal standard. Large relative amounts of D-serine were determined in brains of mammals but not of birds. In body fluids the D-enantiomers of most proteinogenic L-amino acids were detected, largest absolute and relative amounts were found in urine. Therein quantities of D-Ala and D-Ser exceeded 50% relative to the L-enantiomers in many instances. Feeding animals with diet fortified with DL-Met resulted in excretion of almost racemic Met in urine. D-Amino acids were also abundant in faeces of rodents. The data confirm that d-amino acids are common in body fluids and certain tissues of vertebrates.

Amino Acids↗
PubMed1990

Descending pathways to the spinal cord, III: Sites of origin of the corticospinal tract.

The somata of corticospinal neurons were labeled with horseradish peroxidase that had been applied to a hemisection of the spinal cord at the C1-C2 junction in 22 species of mammals. After tetramethylbenzidine processing, with and without counterstaining with cresyl violet or neutral red, the labeled cells in systematic sets of sections throughout the cerebral cortex were plotted and counted. Several morphological features of the corticospinal cells were examined including their cell type, number, density, concentration, laminar distribution, and their distribution across the cortical surface. The results show that the labeled corticospinal neurons were invariably layer V pyramidal cells. However, in many mammals they were found to be stacked one above the other within layer V, sometimes many neurons deep. Despite the concentration of corticospinal neurons within layer V, many unlabeled neurons were also present within the layer throughout the extent of the labeled region. The results also indicate that at least two spatially distinct regions of neocortex originate corticospinal fibers in each of the animals in the sample. In addition to these two regions, a third segregated region is present in the cortex of primates and an apparently different third region is present in the cortex of Glires (Rodentia and Lagomorpha). The third region of corticospinal cortex in primates is located on the lateral surface of the cortex in prosimians and New World monkeys and is buried in the caudal bank of the inferior arcuate sulcus in Old World monkeys. The results also show a predominantly contralateral corticospinal tract in all but 4 of the 22 mammals in the sample. Although these 4 mammals are each members of the order Insectivora, a less modified member of the same order possessed the predominantly contralateral projection of most mammals, hence denying the notion that a predominantly ipsilateral tract is a characteristic of Insectivora.

Animals↗
PubMed1989

The IgA heavy-chain gene family in rabbit: cloning and sequence analysis of 13 C alpha genes.

Southern analysis has previously shown that the rabbit genome contains multiple genes coding for the constant regions of IgA heavy chains. In the present study, clones containing these C alpha genes have been isolated from cosmid and phage libraries. Restriction mapping and Southern analysis of the clones identified 13 non-allelic C alpha genes; 11 of the genes were clustered in individual or overlapping clones. The clustered genes are separated by 8-18 kb, and in total, the C alpha genes span a minimum of 160 kb of DNA. Southern analysis has shown that all genes within a cluster have the same transcriptional orientation, and that switch sequences are present 5' of at least 12 of the 13 genes. The nucleotide sequence of each C alpha gene was determined, and it appears that all genes are functional; thus, rabbit may have as many as 13 IgA isotypes. Comparisons of the protein sequences encoded by the 13 C alpha genes showed that the CH2 and CH3 domains of the alpha-chains are highly conserved, whereas the CH1 and hinge regions are highly diverse. Southern analysis of genomic DNA samples from other species within the order Lagomorpha showed that all samples had multiple C alpha hybridizing fragments. Thus, it is likely that all lagomorphs have multiple IgA isotypes and hence complex secretory immune systems.

Amino Acid Sequence↗
PubMed1977

Immunofluorescent localization of acrosin in mammalian spermatozoa (1).

Acrosin was detected by immunofluorescence in the spermatozoan acrosomes of artiodactyla (bull, ram and boar), perissodactyla (horse), carnivora (dog and cat), lagomorpha (rabbit) and primates (human) using anti-bovine acrosin immunoglobulins. The results indicate that the acrosin molecules of several mammalian species possess antigenic similarities.

Acrosin↗
PubMed1998

The complete mitochondrial DNA sequence of the rabbit, Oryctolagus cuniculus.

The nucleotide sequence of the complete mitochondrial DNA (mtDNA) molecule of the rabbit (Oryctolagus cuniculus, order Lagomorpha) was determined. The length of the molecule is 17,245 nt, but the length is not absolute due to the presence of different numbers of repeated motifs in the control region. The organization and gene contents of the mtDNA of the rabbit conform to those of other eutherian species. The putative secondary structures of the tRNAs of the rabbit have been described. These structures as well as the structure of the L-strand origin of replication comply with those characteristic for eutherians in general. The compositional differences between the two mtDNA strands have also been detailed.

Animals↗
PubMedDate not supplied

Biochronological implications of the Arvicolidae (Rodentia, Mammalia) from the Lower Pleistocene hominid-bearing level of Trinchera Dolina 6 (TD6, Atapuerca, Spain).

Level TD6 of the Trinchera Dolina Section in the railway cutting of the Sierre de Atapuerca (Trinchera del Ferrocarril) has yielded a rich small mammal assemblage (26 species) in association with fossil human remains of Homo antecessor. The arvicolids of TD6 are identified as: Mimomys savini, Microtus seseae, Stenocranius gregaloides, Terricola arvalidens, Iberomys huescarensis, Allophaiomys chalinei, and Pliomys episcopalis. The rodent association also includes large rodents (i.e., Castor fiber, Marmota sp., and Hystrix refossa) and the small Allocricetus sp., Eliomys helleri, Micromys minutus, and Apodemus aff. flavicollis. The small vertebrate remains also include Insectivora (Beremendia fissidens, Sorex sp, Neomys sp., Crocidura sp., Galemys sp., Talpa sp., Erinaceus sp.), Chiroptera (Miniopterus schreibersii, Myotis sp., Rhinolophus sp.), and Lagomorpha (Oryctolagus sp., Lepus sp.), as well as lizards, birds and amphibians. The H. antecessor remains are derived from a 15 cm thick layer at the top of TD6 (TD6-T36-43), where A. chalinei, H. refossa and Marmota sp. do not occur. The paleomagnetic Matuyama/Brunhes boundary is found in the overlying level TD7 of the Gran Dolina Section. On the basis of the arvicolids, TD6 can be referred to the Biharian biochron. The Matuyama/Brunhes boundary is fixed in the late Biharian (Microtus-Mimomys rodent Superzone). The species M. savini (without M. pusillus), as well as the evolutionary stage of Microtus s.l., are characteristic of the Late Biharian. The evolutionary level of the species M. savini, T. arvalidens, S. gregaloides indicates that TD6 is older than West Runton (type Cromerian). In the Trinchera Dolina Section we are able to calibrate, for the first time, the evolutionary level of important biochronological markers with magnetostratigraphy. We propose that a radiation of Microtus s.l., along with the first appearance of primitive S. gregaloides, T. arvalidens and Iberomys, took place just before the Matuyama/Brunhes boundary. These species can be considered as characteristic elements of early Pleistocene faunas.

Animals↗
PubMed1996

A congruence study of molecular and morphological data for eutherian mammals.

The four orders of eutherian mammals which are traditionally placed in the superorder Archonta [Chiroptera (microbats and megabats), Dermoptera (flying lemurs), Primates (primates), and Scandentia (tree shrews)] are among the best-studied taxa of their infraclass from both the molecular and morphological perspectives. Nevertheless, the ordinal relationships of archontans remain unresolved. While morphological studies favor their monophyly, molecular investigations do not. To evaluate these opposing conclusions, parsimony analyses were conducted with three separate sets of DNA sequences from both the nuclear and mitochondrial genomes and one file of morphological data for archontans and other eutherian mammals. Statistical tests of character support and ordinal branching pattern differences documented that the three sets of DNA sequences and their results were homogeneous and congruent, thereby allowing for the combination of these data into one large matrix for further phylogenetic analysis. In contrast, these same tests revealed that the combined sequence and morphological files and their topologies were in strong conflict. Archontan monophyly was supported by the morphological evidence, but this arrangement was strongly rejected by the combined DNA sequences that favored instead a grouping of Dermoptera, Primates, and Scandentia with Lagomorpha (rabbits) and Rodentia (rodents). Resolution of these significant differences will require further evaluations about the homologies and evolutionary properties of the molecular and morphological characters and about the appropriateness of the chosen phylogenetic methods, as well as the incorporation of new comparative data from both sources.

Animals↗
PubMed1998

Higher taxonomic relationships among extant mammals based on morphology, with selected comparisons of results from molecular data.

Until a few decades ago, phylogenetic relationships among placental orders were ambiguous and usually depicted to radiate as an unresolved "bush." Resolution of this bush by various workers has been progressing slowly, but with promising results corroborated by nondental, dental, and molecular characters. In this study we continue to seek resolution. A total of 258 nondental and 2 dental characters was analyzed by PAUP and MacClade on 39 vertebrate taxa (3 reptiles, 1 nonmammalian therapsid, and 35 mammals; 20 of the mammals are extant and 15 are extinct) to study higher taxonomic relationships with emphasis on Placentalia (Eutheria). About two-thirds of the characters are osteological, the rest concern soft tissues, including myological but excluding molecular characters (most are our data, the rest are from the literature). Cladistic analysis included all 39 taxa (fossil taxa help to evaluate polarities of characters) and all characters were given equal weight. Extant Mammalia are divided into Prototheria and Theria, the latter into Marsupialia and Placentalia. Placentalia comprises Xenarthra and Epitheria. Within Epitheria, Lipotyphla and Preptotheria (emended) are sister-taxa. Preptotherian taxa group into: ungulate-related taxa and various nonungulates. The former include Carnivora, Pholidota, Tubulidentata, Artiodactyla, Cetacea, Perissodactyla, Hyracoidea, Proboscidea, and Sirenia. A possible association to embrace Lagomorpha, Rodentia, Macroscelidea, Scandentia, Primates, Chiroptera, and Dermoptera is suggested. Significant differences between our findings and those of recent investigators include the dissociation of Pholidota from Xenarthra and the plesiomorphous position of Lipotyphla within Epitheria. Congruence between morphological and molecular results is closer than previously reported.

Animals↗
PubMed2001

Molecular evidence of an African Phiomorpha-South American Caviomorpha clade and support for Hystricognathi based on the complete mitochondrial genome of the cane rat (Thryonomys swinderianus).

The complete mitochondrial genome of an African cane rat, Thryonomys swinderianus (Rodentia, Hystricognathi), was included in a phylogenetic analysis along with 4 rodents, 14 additional eutherians, and 3 noneutherian outgroups. Monophyly of the suborder Hystricognathi, represented by the cane rat and the South American guinea pig, Cavia porcellus, was strongly supported by maximum-parsimony, neighbor-joining, and maximum-likelihood methods. The molecular-based estimate of the divergence time of Old and New World Hystricognathi (approximately 85 million years before present, MYBP) is consistent with an hypothesis of vicariance divergence due to the rifting of the African and South American continents 86-100 MYBP. Monophyly of Rodentia or the superordinal clade Glires (Rodentia and Lagomorpha) were not supported.

Africa↗
PubMed2001

Phylogenetic position of Eulipotyphla inferred from the cDNA sequences of pepsinogens A and C.

Although to date the phylogenetic position of the provisional order Eulipotyphla has been assessed by various molecular markers, it has not been conclusively clarified due to low statistical supporting values and inconsistent results. To clarify the phylogenetic position of Eulipotyphla, we cloned cDNAs for pepsinogens A and C from five mammalian species belonging to four different orders and determined their nucleotide sequences. Molecular phylogenetic analysis based on the 1st and 2nd codon positions of the protein-coding region of cDNA sequences strongly supported the close relationship between Eulipotyphla and Chiroptera. Carnivora was found to be a sister group to these two orders. The monophyly of the order Rodentia and that of the cohort Glires (Rodentia and Lagomorpha) was also shown by the present phylogenetic trees of pepsinogens.

Amino Acid Sequence↗
PubMed1983

The fibre-type composition of the first branchial arch muscles in Carnivora and Primates.

A combination of standard histochemical techniques and immunohistochemical staining using myosin type-specific antisera was used to determine the fibre-type composition of the muscles of first branchial arch origin (that is, masseter, temporalis, pterygoideus medialis and lateralis, tensor veli palatini, tensor tympani, anterior digastricus and mylohyoideus) in a wide range of the Carnivora and the Primates. The rare IIM fibre type was found in the first branchial arch muscles of most of the species examined, but never in the limb muscles used as controls for this study. The jaw-closer muscles (masseter, temporalis and pterygoideus medialis) were found to contain IIM fibres in all the Carnivora except the lesser panda and in all the Primates except man. When present, the IIM fibres were usually the predominant fibre type, and the only other fibre types present were types I, II or IIC. The presence of IIM fibres in the jaw-closer muscles of most of the Carnivora and the Primates seems to be associated with an aggressive bite which is required for predation by the former and defence by the latter. In both groups of species there was the member which does not have an aggressive bite, the lesser panda and man, respectively, and these (like all other orders of mammals such as Lagomorpha, Rodentia, etc.) were found to have no IIM fibres in the jaw-closer muscles. The two muscles of the first branchial arch group which are derived from the ventral constrictor muscles of the (phylogenetically) original mandibular arch never contained IIM fibres, and were composed of type I and II fibres similar to those found in the control muscles of the limb. Tensor veli palatini and tensor tympani showed species-dependent variations in fibre-type composition and did not always reflect the composition of the jaw-closer muscles. Thus their common origin with the jaw-closers cannot be responsible for the occurrence of IIM fibres in tensor veli palatini and tensor tympani in some species. Furthermore, in tensor tympani but not in tensor veli palatini, the presence of IIM fibres was always accompanied by immunohistochemically slow-tonic fibres. Finally, the regard to the association of oxidative activity with the fibre type as defined by the myofibrillar ATPase method and by the isoform of myosin present, we suggest that in the first branchial arch muscles this is probably not directly comparable to the situation in the typical limb muscle.

Animals↗
PubMed1991

The branching order of mammals: phylogenetic trees inferred from nuclear and mitochondrial molecular data.

In order to clarify some controversial phylogenies such as those regarding the triplet of human, rodent, and cow and the evolutionary position of Lagomorpha with respect to other mammals, we have analyzed both nuclear and mitochondrial genes using the stationary Markov model developed in our laboratory. We found that the two sets of genes give different results. In particular the mitochondrial tree showed rabbit linked first to rodents and the rabbit-rodents branch linked to artiodactyls with human as the outgroup. The most favorite nuclear tree showed human linked first to artiodactyls and the human-artiodactyls branch linked to rabbit with rodents as the outgroup. The obvious questions, (1) which tree is the correct one, or (2) both trees can be incorrect, and (3) how can we explain such an evolutionary pattern, are discussed on the basis of our limited knowledge of factors that influence the clocklike behavior of biological macromolecules.

Animals↗
PubMed1999

Evolution of Mhc-DRB introns: implications for the origin of primates.

Introns are generally believed to evolve too rapidly and too erratically to be of much use in phylogenetic reconstructions. Few phylogenetically informative intron sequences are available, however, to ascertain the validity of this supposition. In the present study the supposition was tested on the example of the mammalian class II major histocompatibility complex (Mhc) genes of the DRB family. Since the Mhc genes evolve under balancing selection and are believed to recombine or rearrange frequently, the evolution of their introns could be expected to be particularly rapid and subject to scrambling. Sequences of intron 4 and 5 DRB genes were obtained from polymerase chain reaction-amplified fragments of genomic DNA from representatives of six eutherian orders-Primates, Scandentia, Chiroptera, Dermoptera, Lagomorpha, and Insectivora. Although short stretches of the introns have indeed proved to be unalignable, the bulk of the intron sequences from all six orders, spanning >85 million years (my) of evolution, could be aligned and used in a study of the tempo and mode of intron evolution. The analysis has revealed the Mhc introns to evolve at a rate similar to that of other genes and of synonymous sites of non-Mhc genes. No evidence of homogenization or large-scale scrambling of the intron sequences could be found. The Mhc introns apparently evolve largely by point mutations and insertions/deletions. The phylogenetic signals contained in the intron sequences could be used to identify Scandentia as the sister group of Primates, to support the existence of the Archonta superorder, and to confirm the monophyly of the Chiroptera.

Animals↗
PubMed2002

MHC class I genes of the tree shrew Tupaia belangeri.

Two MHC class I cDNA sequences from the tree shrew (Tupaia belangeri), Tube-W01 and Tube-W02, have been isolated which are probably derived from classical class I genes. Expression of the tupaia class I genes was investigated in several organs, in particular the brain, in which slightly different amounts of class I transcripts are detectable in different areas. Gene tree analysis performed with Tube-W01 and Tube-W02, and including class I sequences derived from other orders, indicated that the tupaia sequences cluster differently from Primates, Carnivora, Artiodactyla, Perissodactyla, and Rodentia, but might be related to Lagomorpha class I genes.

Amino Acid Sequence↗
PubMed2005

Evolutionary implications of multiple SINE insertions in an intronic region from diverse mammals.

An analysis of the nuclear beta-fibrinogen intron 7 locus from 30 taxa representing 12 placental orders of mammals reveals the enriched occurrences of short interspersed element (SINE) insertion events. Mammalian-wide interspersed repeats (MIRs) are present at orthologous sites of all examined species except those in the order Rodentia. The higher substitution rate in mouse and a rare MIR deletion from rat account for the absence of MIR in the rodents. A minimum of five lineage-specific SINE sequences are also found to have independently inserted into this intron in Carnivora, Artiodactyla and Lagomorpha. In the case of Carnivora, the unique amplification pattern of order-specific CAN SINE provides important evidence for the "pan-carnivore" hypothesis of this repeat element and reveals that the CAN SINE family may still be active today. Particularly interesting is the finding that all identified lineage-specific SINE elements show a strong tendency to insert within or in very close proximity to the preexisting MIRs for their efficient integrations, suggesting that the MIR element is a hot spot for successive insertions of other SINEs. The unexpected MIR excision as a result of a random deletion in the rat intron locus and the non-random site targeting detected by this study indicate that SINEs actually have a greater insertional flexibility and regional specificity than had previously been recognized. Implications for SINE sequence evolution upon and following integration, as well as the fascinating interactions between retroposons and the host genomes are discussed.

Animals↗
PubMed1999

Defining the ancestral karyotype of all primates by multidirectional chromosome painting between tree shrews, lemurs and humans.

We used multidirectional chromosome painting with probes derived by bivariate fluorescence-activated flow sorting of chromosomes from human, black lemur (Eulemur macaco macaco) and tree shrew (Tupaia belangeri, order Scandentia) to better define the karyological relationship of tree shrews and primates. An assumed close relationship between tree shrews and primates also assists in the reconstruction of the ancestral primate karyotype taking the tree shrew as an "outgroup" species. The results indicate that T. belangeri has a highly derived karyotype. Tandem fusions or fissions of chromosomal segments seem to be the predominant mechanism in the evolution of this tree shrew karyotype. The 22 human autosomal painting probes delineated 40 different segments, which is in the range found in most mammals analyzed by chromosome painting up to now. There were no reciprocal translocations that would distinguish the karyotype of the tree shrew from an assumed primitive primate karyotype. This karyotype would have included the chromosomal forms 1a, 1b, 2a, 2b, 3/21, 4-11, 12a/22a, 12b/22b, 13, 14/15, 16a, 16b, 17, 18, 19a, 19b, 20 and X and Y and had a diploid chromosome number of 2n=50. Of these forms, chromosomes 1a, 1b, 4, 8, 12a/22a, and 12b/22b may be common derived characters that would link the tree shrew with primates. To define the exact phylogenetic relationships of the tree shrews and the genomic rearrangements that gave rise to the primates and eventually to humans further chromosome painting in Rodentia, Lagomorpha, Dermoptera and Chiroptera is needed, but many of the landmarks of genomic evolution are now known.

Animals↗