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Description of a cranial endocast from a fossil platypus, Obdurodon dicksoni (Monotremata, Ornithorhynchidae), and the relevance of endocranial characters to monotreme monophyly.

A digital cranial endocast of the Miocene platypus Obdurodon dicksoni was extracted from high-resolution X-ray computed tomography scans. This endocast represents the oldest from an unequivocal member of either extant monotreme lineage and is therefore important for inferring character support for Monotremata, a clade that is not well diagnosed. We describe the Obdurodon endocast with reference to endocasts extracted from skulls of the three species of extant monotremes, particularly Ornithorhynchus anatinus, the duckbill platypus. We consulted published descriptions and illustrations of whole and sectioned brains of monotremes to determine which external features of the nervous system are represented on the endocasts. Similar to Ornithorhynchus, well-developed parafloccular casts and reduced olfactory bulb casts are present in the Obdurodon endocast. Reduction of the olfactory bulbs in comparison with tachyglossids and therian mammals is a potential apomorphy for Ornithorhynchidae. The trigeminal nuclei, ganglia, and nerves (i.e., trigeminal complex) are enlarged in Obdurodon, as evidenced by their casts on the endocast, as is the case in the extant platypus. The visibility of enlarged trigeminal nucleus casts on the endocasts of Obdurodon and Ornithorhynchus is a possible synapomorphy of Ornithorhynchidae. Electroreception and enlargement of the trigeminal complex are possible synapomorphies for Monotremata.

Animals↗

The complete mitochondrial genome of the wallaroo (Macropus robustus) and the phylogenetic relationship among Monotremata, Marsupialia, and Eutheria.

The complete mitochondrial DNA (mtDNA) (16,896 nt) of the wallaroo (Macropus robustus) was sequenced. The concatenated amino acid sequences of 12 mitochondrial protein-coding genes of the wallaroo plus those of a number of other mammals were included in a phylogenetic study of early mammalian divergences. The analysis joined monotremes and marsupials (the Marsupionta hypothesis) to the exclusion of eutherians. The analysis rejected significantly the commonly acknowledged Theria hypothesis, according to which Marsupialia and Eutheria are grouped together to the exclusion of Monotremata. The region harboring the gene for lysine tRNA (tRNA-Lys) in the mtDNA of other vertebrates is in the wallaroo occupied by a sequence (tRNA-Lys) that lacks both an anticodon loop as well as the anticodon for the amino acid lysine. An alternative tRNA-Lys gene was not identified in any other region of the mtDNA of the wallaroo, suggesting that a tRNA-Lys of nuclear origin is imported into marsupial mitochondria. Previously described RNA editing of tRNA-Asp and rearrangement of some tRNA genes were reconfirmed in the mtDNA of the wallaroo. The divergence between Monotremata/Marsupialia and Eutheria was timed to approximately 130 million years before present (MYBP). The same calculations suggested that Monotremata and Marsupialia diverged approximately 115 MYBP and that Australian and American marsupials separated approximately 75 MYBP. The findings also show that many, probably most, extant eutherian orders had their origin in middle to late Cretaceous times, 115-65 MYBP.

Animals↗

Karyotypic conservation in the mammalian order monotremata (subclass Prototheria).

The order Monotremata, comprising the platypus and two species of echidna (Australian and Nuigini) is the only extant representative of the mammalian subclass Prototheria, which diverged from subclass Theria (marsupials and placental mammals) 150-200 million years ago. The 2n = 63 male, 64 female karyotype (newly described here) of the Nuigini echidna is almost identical in morphology and G-band pattern to that of the Australian echidna, from which it diverged about a million years ago. The karyotype of the platypus (2n = 52) has several features in common with those of the echidna species; six pairs of large autosomes, many pairs of small (but not micro-) chromosomes, and a series of small unpaired chromosomes which form a multivalent at meiosis. Comparison of the G-band patterns of platypus and echidna autosomes reveals considerable homology. Chromomycin banding demonstrates GC-rich heterochromatin at the centromeres of many platypus and echidna chromosomes, and at the nucleolar organizing regions; some of this heterochromatin C-bands weakly in platypus (but not echidna) spreads. Late replication banding patterns resemble G-banding patterns and confirm the homologies between the species. Striking heteromorphism between chromosomes of some of the large autosomal pairs can be accounted for in the echidna by differences in amount of chromomycin-bright, late replicating heterochromatin. The sex chromosomes in all three species also bear striking homology, despite the difference in sex determination mechanism between platypus (XX/XY) and the echidna species (X1X1X2X2/X1X2Y). The platypus X and echidna X1 each represent about 5.8% of haploid chromosome length, and are G-band identical. Y chromosomes are similar between species, and are largely homologous to the X (or X1).

Animals↗

The lamina cribrosa of Ornithorhynchus (Monotremata, Mammalia).

A vestigial and transitory lamina cribrosa was found in nestling platypus (Ornithorhynchus anatinus). The heads of two nest-young (180 and 333 mm length), one subadult and one adult Ornithorhynchus were serially sectioned and studied with special reference to the development of the nasal region. In nest-young Ornithorhynchus an irregularly shaped bar of cartilage develops at the foramen olfactorium advehens. In the subadult it is largely resorbed, and in the osseous skull of the adult it is completely lacking. Ontogeny and topographical relationships of this bar of cartilage indicate that it is part of a lamina cribrosa. It embraces the ramus medialis of the nervus ethmoidalis and the arteria ethmoidalis, as do the corresponding parts of the lamina cribrosa of Tachyglossus. Compared to other parts of the chondrocranium this bar develops late in ontogeny, as does the lamina cribrosa of other mammals. Therefore, it can be concluded that part of the lamina cribrosa is present for a short period during the ontogeny of Ornithorhynchus, contrary to earlier reports. As in many other water-adapted mammals, the olfactory system of Ornithorhynchus is reduced. This suggests that the rest of the lamina cribrosa of Ornithorhynchus is secondarily reduced. The common ancestor of Ornithorhynchus and Tachyglossidae most probably possessed a lamina cribrosa which can be traced back to the common mammalian stock. The lamina cribrosa developed only once in the phylogeny of mammals. Its lack in the adult Ornithorhynchus is not a "reptilian" character.

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Two monotreme cell lines, derived from female platypuses (Ornithorhynchus anatinus; Monotremata, Mammalia).

Two diploid platypus cell lines, designated Oa-1F and Oa-2F, have been derived from the toe webs of two females. The development and growth characteristics of the lines are described and G-banded karyotypes presented (the first reported for the platypus). The availability of these lines will greatly facilitate chromosome and gene mapping studies of the platypus and permit the extension of comparative studies of mammalian chromosome and genome evolution to the monotremes.

Animals↗

Fine structure of the secretion granules in the mandibular gland of the echidna, Tachyglossus aculeatus (Monotremata).

The cells of the secretory tubules in the mandibular gland of the echidna are packed with fairly large birefringent granules, which show a lamellated structure consisting of alternating thin and thick layers or shells of protein. This presumably rigid substructure collapses during exocytosis and the shells unravel as sheets that form a tangled mass in the lumen of the secretory tubule. Relatively pure fractions were obtained of the relevant granules and protein sheets, which should allow a further study to be made on the secretory proteins in this gland.

Animals↗

Descriptive study of the diaphragm and lungs in the short-nosed echidna, Tachyglossus aculeatus (Mammalia: monotremata).

In this descriptive study, we characterize the diaphragm and lungs of the short-nosed echidna, Tachyglossus aculeatus, using a combination of gross anatomical, light-microscopic, electron microscopic, and morphometric techniques, including airway casting. The diaphragm is inclined from ventro-cranial to dorso-caudal and possesses a large central tendon (centrum tendineum). The crural and costal muscle groups and the associated trigoni are located in the same positions as in other mammals. The bronchial branching pattern reveals cranially broad, tapering stem bronchi and an unusually small number of first order bronchi. The asymmetrical primary branching pattern and possibly also the asymmetry of right and left lungs are plesiomorphic within the Mammalia. The histology and ultrastructure of the airways and lung parenchyma reveal no unusual features: alveolar type 1 and type 2 cells in the parenchyma; type 2 cells, exocrine bronchiolar cells (Clara cells), ciliated cells, and goblet cells in the terminal airways and the latter two cell types in the bronchi. Both a double and a single capillary net are found on the interalveolar septa. The high capillary loading of the double net may be of selective advantage because of long apneas and low metabolic rate in the echidna.

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Morphology of the kidney of the platypus (Ornithorhynchus anatinus: Monotremata).

The platypus kidney shows morphological similarities to those of other mammals. Macroscopically, the cortex is easily distinguishable from the fairly wide medulla. Within the medulla, no clear border is observed between the inner and outer zones. Light and transmission electron microscopically, the glomeruli show quite similar architecture to those of other mammals; however, the glomerular lobulation is very clear. The glomerular tufts are rather simple, but capillary lumen varies widely in size, which is one of the unique features of the platypus kidney. The urinary tubule is generally similar to that of human and other mammals in shape and segmentation; however, the staining specificities of histochemical reactions and the shape of epithelial cells of the Henle's loop differ from those of other mammals. The most conspicuous features are: 1) although no protein casts are found in the tubular lumina, epithelial cells of the proximal convoluted tubule (PCT) have numerous electron-dense vesicles as in human nephrotic kidneys; and 2) the platypus Henle's loop consists of the thick epithelial cells similar to the mammalian type nephron of birds. As compared to those of other mammals such as humans and rats, our observations suggest that the platypus kidney is less developed, in terms of evolution.

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Evolution and molecular characterization of a beta-globin gene from the Australian Echidna Tachyglossus aculeatus (Monotremata).

Coinciding with a period in evolution when monotremes, marsupials, and eutherians diverged from a common ancestor, a proto-beta-globin gene duplicated, producing the progenitors of mammalian embryonic and adult beta-like globin genes. To determine whether monotremes contain orthologues of these genes and to further investigate the evolutionary relationships of monotremes, marsupials, and eutherians, we have determined the complete DNA sequence of an echidna (Tachyglossus aculeatus) beta-like globin gene. Conceptual translation of the gene and sequence comparisons with eutherian and marsupial beta-like globin genes and echidna adult beta-globin indicate that the gene is adult expressed. Phylogenetic analyses do not clearly resolve the branching pattern of mammalian beta-like globin gene lineages and it is therefore uncertain whether monotremes have orthologues of the embryonic beta-like globin genes of marsupials and eutherians. Four models are proposed that provide a framework for interpreting further studies on the evolution of beta-like globin genes in the context of the evolution of monotremes, marsupials, and eutherians.

Amino Acid Sequence↗

Cone photoreceptors lacking oil droplets in the retina of the echidna, Tachyglossus aculeatus (Monotremata).

The echidna, Tachyglossus aculeatus, a monotreme mammal, is thought to possess an all-rod retina (O'Day, 1952). This study provides anatomical evidence for the presence of cone-like photoreceptors in the retina of the echidna. The cones, which constitute 10-15% of the photoreceptors, have all of the ultrastructural characteristics previously shown in the cones of placental mammals, and, like cones of other animals (Blanks & Johnson, 1984), they bind peanut agglutinin. Unlike the cones of another monotreme, the platypus, the cones of the echidna retina do not possess oil droplets. Twin cones, pairs of cones in which there is no obvious difference in the size, shape, or ultrastructural features of the members of a pair, are common. The density of cones varies from 9000 cells/mm2 in the superior periphery to 22,000 cells/mm2 in the central retina. Nearest-neighbor analysis suggests that the cone mosaic in the echidna retina results from the presence of single and twin cones in a relatively regular array.

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Foetal genital development in Hyrax capensis, a species with primary testicondia: proposal for the evolution of Hunter's gubernaculum.

BACKGROUND: Control of testicular descent is poorly understood. There are a number of mammalian species in which testis descent does not occur, and the phenomenon is called testicondia. Analysis of foetal development of such species could contribute to a better understanding of the key events in anatomical development underlying testis descent. Specific attention is to be given to the development of the so-called gubernaculum of Hunter: a structure of complex architecture and composition, which extends from the caudal pole of the mesonephric remnants into the inguinal abdominal wall and which is present in most mammals but not in submammalian vertebrates. METHODS: Serially sectioned male and female foetuses of Hyrax capensis, a species in which testes remain close to the caudal pole of the kidneys throughout life, were analysed for architecture of the developing genital apparatus and the immediately surrounding structures. The results of this analysis were compared with those from a similar analysis of reptiles, of monotremata without testis descent, and of mammals with testis descent. RESULTS: Reptiles and Monotremata showed no gubernacular structures. Mammals with testis descent showed gubernacular growth and differentiation which varied between the sexes and with the stage of foetal development. Hyrax capensis foetuses showed the development of only one component of the gubernaculum: the gubernacular cord as a part of the mesonephric mesentery and extending between the caudal pole of the mesonephros (or mesonephric remnants in older foetuses) and the lateral bladder ligament. No part developed in the inguinal abdominal wall components, which are the primordia of the cremaster sacs in species with testis descent. CONCLUSION: Hyrax capensis shows only partial development of the gubernacular structures, and, specifically, the primordia of the cremaster sacs remain absent. Thus, this species lacks a key anatomical condition for testis descent. Gubernacular architecture in Hyrax capensis seems of a degree intermediate between egg laying monotremata mammals and mammals with testis descent. A model is proposed within which to understand the development of the gubernacular components within the mammalian class.

Animals↗

"Not proper mammals": immunity in monotremes and marsupials.

Immune systems and responses in Monotremata and Marsupialia are reviewed. The Monotremata (Prototheria) are egg-laying mammals. Few studies have been carried out on monotremes. The structure of the lymph nodules of Tachyglossus aculeatus is unusual, and the occurrence of IgG in this species is noteworthy: IgG has not yet been found in any non-mammal. A number of Marsupialia (Metatheria) species have been used as immunological models. Generally immune responses are somewhat slower and less accentuated than in placental (eutherian) mammals. Of interest is the presence of cervical and thoracic thymuses in several marsupials. Marsupials are born very immature and possess rather rudimentary immune responses at birth: the neonate may provide a helpful model for immune ontogenesis. Marsupials have a full repertoire of immunoglobulin classes. MHC Class II (but not Class I) gene polymorphism may be limited. Studies using molecular biology techniques are awaited to elucidate the structural organization of the immune components and to determine similarities and differences between marsupials' and other animals' immune systems.

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The platypus is in its place: nuclear genes and indels confirm the sister group relation of monotremes and Therians.

Morphological data supports monotremes as the sister group of Theria (extant marsupials + eutherians), but phylogenetic analyses of 12 mitochondrial protein-coding genes have strongly supported the grouping of monotremes with marsupials: the Marsupionta hypothesis. Various nuclear genes tend to support Theria, but a comprehensive study of long concatenated sequences and broad taxon sampling is lacking. We therefore determined sequences from six nuclear genes and obtained additional sequences from the databases to create two large and independent nuclear data sets. One (data set I) emphasized taxon sampling and comprised five genes, with a concatenated length of 2,793 bp, from 21 species (two monotremes, six marsupials, nine placentals, and four outgroups). The other (data set II) emphasized gene sampling and comprised eight genes and three proteins, with a concatenated length of 10,773 bp or 3,669 amino acids, from five taxa (a monotreme, a marsupial, a rodent, human, and chicken). Both data sets were analyzed by parsimony, minimum evolution, maximum likelihood, and Bayesian methods using various models and data partitions. Data set I gave bootstrap support values for Theria between 55% and 100%, while support for Marsupionta was at most 12.3%. Taking base compositional bias into account generally increased the support for Theria. Data set II exclusively supported Theria, with the highest possible values and significantly rejected Marsupionta. Independent phylogenetic evidence in support of Theria was obtained from two single amino acid deletions and one insertion, while no supporting insertions and deletions were found for Marsupionta. On the basis of our data sets, the time of divergence between Monotremata and Theria was estimated at 231-217 MYA and between Marsupialia and Eutheria at 193-186 MYA. The morphological evidence for a basal position of Monotremata, well separated from Theria, is thus fully supported by the available molecular data from nuclear genes.

Amino Acid Sequence↗