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Scrambled or bisected mouse eggs and the basis of patterning in mammals.

Several findings challenge the notion that specification of cell types and embryonic axes in mammals are rooted entirely in the temporal and spatial relations between cleaving blastomeres. They raise the question as to whether, as in most non-mammalian species, these processes depend on information already present in the egg. However, experiments designed to investigate this possibility directly by perturbing the organization of the zygote or, very recently, by deleting one or other of its polar regions [M. Zernicka-Goetz. Fertile offspring derived from mammalian eggs lacking either animal or vegetal poles. Development 1998;125:4803-4808 (Ref. 1)], have been interpreted to mean that such a role for the egg can be discounted. This conclusion seems premature in view of continuing uncertainty regarding the developmental potential of individual blastomeres in mammals.

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Vitamin carrier proteins during embryonic development in birds and mammals.

Egg maturation in oviparous vertebrates involves the hepatic synthesis, secretion, and deposition in the developing oocyte of several maternal proteins with specific nutrient carrier function. Thus, in the chicken, adequate yolk deposition of riboflavin, thiamin, etc. is obligatorily mediated by carrier proteins specific to each vitamin. Like vitellogenin, these are oestrogen-inducible specific gene products. Despite differences in patterns of embryonic development in mammals vis-à-vis oviparous species, immunologically and biochemically similar maternal vitamin carriers participate in the transplacental transport and fetal accumulation of these vitamins during gestation in the rat. The rodent riboflavin and thiamin carrier proteins are also oestrogen-induced maternal proteins of hepatic origin. Their functional importance in fetal development was established by in vivo passive immunoneutralization of the endogenous proteins, which precipitated fetal wastage leading to pregnancy termination, due to curtailment of the vitamin supply to the fetuses. Similarly, active immunization of female rats with the vitamin carrier proteins led to early fetal resorption without interference with maternal health, cyclicity and fecundity. The discovery of similar gestation-specific carrier proteins in higher mammals and humans suggests that carrier-mediated vitamin delivery mechanisms ensuring embryonic growth have been conserved during evolution.

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The pattern of elastin in the aorta and large arteries of mammals.

The arteries of mammals contain large amounts of elastin arranged in concentric lamellae known as medial lamellar units (MLU). In adult mammals of a variety of species the number of lamellar units is roughly proportional to the radius of the artery and the tension/MLU ratio is roughly constant in all species, but greater in the abdominal than in the thoracic aorta. Re-analysis of these data shows that the number of MLU of the abdominal aorta is linearly related to the pulse pressure, while the number of MLU in both the thoracic and abdominal aorta increases exponentially with stroke volume. Preliminary data are presented showing the decrease in number of MLU along the thoracic aorta of both fetal lambs and sheep, and evidence is provided that some of this elastin may be involved in the formation of the small arteries, such as the intercostals, which arise from the aorta. Scanning electron microscopy showed that the elastin on the intimal side of the media was in the form of fenestrated sheets while that on the adventitial side was a fibrous network. The size and density of the fenestrations was greater in fetal lambs and may play a role in allowing growth of the artery.

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Clonogenic analysis reveals reserve stem cells in postnatal mammals: I. Pluripotent mesenchymal stem cells.

Clonal populations of lineage-uncommitted pluripotent mesenchymal stem cells have been identified in prenatal avians and rodents. These cells reside in the connective tissue matrices of many organs and tissues. They demonstrate extended capabilities for self-renewal and the ability to differentiate into multiple separate tissues within the mesodermal germ line. This study was designed to determine whether such cells are present in the connective tissues of postnatal mammals. This report describes a cell clone derived by isolation from postnatal rat connective tissues, cryopreservation, extended propagation, and serial dilution clonogenic analysis. In the undifferentiated state, this clone demonstrates a high nuclear-to-cytoplasmic ratio and extended capacity for self-renewal. Subsequent morphological, histochemical, and immunochemical analysis after the induction of differentiation revealed phenotypic markers characteristic of multiple cell types of mesodermal origin, such as skeletal muscle, smooth muscle, fat cells, cartilage, and bone. These results indicate that this clone consists of pluripotent mesenchymal stem cells. This report demonstrates that clonal populations of reserve stem cells are present in mammals after birth. Potential roles for such cells in the maintenance, repair, and regeneration of mesodermal tissues are discussed.

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Identification of parvocellular vasopressin and neurophysin neurons in the suprachiasmatic nucleus of a variety of mammals including primates.

The presence of parvocellular vasopressin- and neurophysin-containing neurons in the suprachiasmatic nucleus (SCN) was investigated in 13 mammalian species representing six mammalian orders (marsupials, rodents, lagomorphs, artiodactyls, carnivores, and primates), using specific antisera to vasopressin and neurophysin in the unlabelled antibody=enzyme immunoperoxidase method. In all mammals examined, including man, parvocellular vasopressin and neurophysin neurons were found in the SCN. Only a portion of SCN neurons contain vasopressin and neurophysin, the number varying with species. Cell counts comparing the number of immunoreactive to Nissl-stained neurons showed averages of 17% immunopositive neurons in the rat SCN, and 31% in the human SCN. No oxytocin-containing SCN neurons were observed. These findings suggest that parvocellular vasopressin and neurophysin neurons are widely represented in mammals.

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Craniofacial development in marsupial mammals: developmental origins of evolutionary change.

Biologists have long studied the evolutionary consequences of the differences in reproductive and life history strategies of marsupial and eutherian mammals. Over the past few decades, the impact of these strategies on the development of the marsupial embryo and neonate has received attention. In this review, the differences in development in the craniofacial region in marsupial and eutherian mammals will be discussed. The review will highlight differences at the organogenic and cellular levels, and discuss hypotheses for shifts in the expression of important regulatory genes. The major difference in the organogenic period is a whole-scale shift in the relative timing of central nervous system structures, in particular those of the forebrain, which are delayed in marsupials, relative to the structures of the oral-facial apparatus. Correlated with the delay in development of nervous system structures, the ossification of the bones of the neurocranium are delayed, while those of the face are accelerated. This study will also review work showing that the neural crest, which provides much of the cellular material to the facial skeleton and may also carry important patterning information, is notably accelerated in its development in marsupials. Potential consequences of these observations for hypotheses on constraint, evolutionary integration, and the existence of developmental modules is discussed. Finally, the implications of these results for hypotheses on the genetic modulation of craniofacial patterning are presented.

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Glia:neuron index: review and hypothesis to account for different values in various mammals.

The present paper proposes a hypothesis to account for different values of the glia:neuron index in comparable central nervous system tissues of various mammals. This hypothesis assumes that K+ ions released by active neurons are a mitogenic signal for glial cells. The thicker the tissue (for example, the brain wall), the more difficult is efficient K+ clearance, and more perinatal glial cell proliferation should occur. Thus, this hypothesis accounts for higher glia:neuron indices in mammals with thicker brain walls.

Aging↗

Glial cells in the nerve fiber layer of the main olfactory bulb of embryonic and adult mammals.

This article provides a detailed description of the glial cell types in the nerve fiber layer of the main olfactory bulb during embryonic development, in adult mammals, and at the nerve entry zone of the first cranial nerve. In adult mammals, the glial cell types of the olfactory nerve fiber layer include intrafascicular ensheathing cells, which have the exclusive role of ensheathing the olfactory axons in both the PNS and CNS, and interfascicular astrocytes, which occupy the spaces between adjacent olfactory fascicles. The ensheathing cells are particularly interesting because they possess a mixture of Schwann cell and astrocytic phenotypic features, are more likely to be of placodal than of CNS origin, and have the exclusive role of forming the glia limitans at the PNS-CNS transitional zone. It is proposed that one important function of ensheathing cells is to modulate the growth of olfactory axons within the CNS; this modulation is probably mediated by selective cell adhesion molecules, extracellular matrix molecules, and chemotropic agents.

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Distribution of keratin and associated proteins in the epidermis of monotreme, marsupial, and placental mammals.

The expression of acidic and basic keratins, and of some keratinization marker proteins such as filaggrin, loricrin, involucrin, and trichohyalin, is known for the epidermis of only a few eutherian species. Using light and high-resolution immunocytochemistry, the presence of these proteins has been studied in two monotreme and five marsupial species and compared to that in eutherians. In both monotreme and marsupial epidermis lamellar bodies occur in the upper spinosus and granular layers. Development of the granular layer varies between species and regionally within species. There is great interspecific variation in the size (0.1-3.0 microm) of keratohyalin granules (KHGs) associated with production of orthokeratotic corneous tissues. Those skin regions lacking hairs (platypus web), or showing reduced pelage density (wombat) have, respectively, minute or indiscernible KHGs, associated with patchy, or total, parakeratosis. Ultrastructural analysis shows that monotreme and marsupial KHGs comprise irregular coarse filaments of 25-40 nm that contact keratin filaments. Except for parakeratotic tissues of platypus web, distribution of acidic and basic proteins in monotreme and marsupial epidermis as revealed by anti-keratin antibodies AE1, AE2, and AE3 resembles that of eutherian epidermis. Antibodies against human or rat filaggrins have little or no cross-reactivity with epidermal proteins of other mammals: only sparse areas of wombat and rabbit epidermis show a weak immunofluorescence in transitional cells and in the deepest corneous tissues. Of the available, eutherian-derived antibodies, that against involucrin shows no cross-reactivity with any monotreme and marsupial epidermal tissues and that against trichohyalin cross-reacts only with cells in the inner root sheath and medulla of hairs. These results suggest that if involucrin and trichohyalin are present throughout noneutherian epidermis, they may have species-specific molecular structures. By contrast, eutherian-derived anti-loricrin antibodies show a weak to intense cross-reactivity to KHGs and corneous tissues of both orthokeratotic and parakeratotic epidermis in monotremes and marsupials. High-resolution immunogold analysis of loricrin distribution in immature keratinocytes of platypus parakeratotic web epidermis identifies labeled areas of round or irregular, electron-pale granules within the denser keratohyalin component and keratin network. In the deepest mature tissues, loricrin-like labeling is diffuse throughout the cytoplasm, so that cells lack the preferential distribution of loricrin along the corneous envelope that characterizes mature eutherian keratinocytes. Thus, the irregular distribution of loricrin in platypus parakeratotic tissues more resembles that which has been described for reptilian and avian keratinocytes. These observations on the noneutherian epidermis show that a stratum granulosum is present to different degrees, even discontinuous within one tissue, so that parakeratotic and orthokeratotic areas may alternate: this might imply that parakeratotic monotreme epidermis reflects the primitive pattern of amniote alpha-keratogenesis. Absent from anamniote epidermis and all sauropsid beta-keratogenic tissues, the ubiquitous presence of a loricrin-like protein as a major component of other amniote corneous tissues suggests that this is a primitive feature of amniote alpha-keratogenesis. The apparent lack of specific regionalization of loricin near the plasma membranes of monotreme keratinocytes could be an artifactual result of the immunofluorescence technique employed, or there may be masking of the antigenicity of loricrin-like proteins once they are incorporated into the corneous envelope. Nevertheless, the mechanism of redistribution of such proteins during maturation of monotreme keratinocytes is different from, perhaps more primitive, or less specialized, than that in the epidermis of eutherian mammals.

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Assessment of mercury concentrations in small mammals collected near Las Vegas, Nevada, USA.

Mercury concentrations in liver and hair tissue were determined for five species of small mammals captured near Las Vegas, Nevada, USA. These data were then used to evaluate the suitability of using hair as a noninvasive technique for determining body burdens of mercury. A total of 104 small mammals were captured and analyzed; four main species were examined and included: Dipodomys merriami, Chaetodipus penicillatus, Peromyscus eremicus, and Neotoma lepida. Mean mercury concentrations were highest in N. lepida, followed by D. merriami, C. penicillatus, and P. eremicus respectively. Positive associations were found between hair and liver taken from D. merriami (r = 0.647) and C. penicillatus (r = 0.533) indicating that hair may be a suitable indicator of body burdens in these two species.

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Blood lead concentrations in marine mammals validate estimates of 10(2)- to 10(3)-fold increase in human blood lead concentrations.

Measurements of ultra-low ambient blood lead (PbB) concentrations (mean +/- SD = 0.13 +/- 0.06 micrograms/dL) in Northern elephant seal (Mirounga angustirostris) validate previous estimates of ultra-low PbB levels in preindustrial humans. These estimates had been unsubstantiated, since PbB levels in this range had never been measured in any organisms prior to this study. Similarities in PbB levels among these contemporary and preindustrial mammals are consistent with similarities in their measured and estimated lead exposures, respectively. The marginally higher PbB levels and rates of lead exposure in contemporary marine mammals are, also, consistent with lead isotopic composition analyses that indicate their PbB levels have been elevated from exposure to industrial lead. Consequently, these analyses substantiate concerns that current baseline PbB levels in humans, which are estimated to be two to three orders of magnitude above natural levels, may still constitute public health risks.

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Sequences homologous to the human X- and Y-borne zinc finger protein genes (ZFX/Y) are autosomal in monotreme mammals.

The human zinc finger protein genes (ZFX/Y) were identified as a result of a systematic search for the testis-determining factor gene on the human Y chromosome. Although they play no direct role in sex determination, they are of particular interest because they are highly conserved among mammals, birds, and amphibians and because, in eutherian mammals at least, they have active alleles on both the X and the Y chromosomes outside the pseudoautosomal region. We used in situ hybridization to localize the homologues of the zinc finger protein gene to chromosome 1 of the Australian echidna and to an equivalent position on chromosomes 1 and 2 of the platypus. The localization to platypus chromosome 1 was confirmed by Southern analysis of a Chinese hamster x platypus cell hybrid retaining most of platypus chromosome 1. This localization is consistent with the cytological homology of chromosome 1 between the two species. The zinc finger protein gene homologues were localized to regions of platypus chromosomes 1 and 2 that included a number of other genes situated near ZFX on the short arm of the human X chromosome. These results support the hypothesis that many of the genes located on the short arm of the human X were originally autosomal and have been translocated to the X chromosome since the eutherian-metatherian divergence.

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RNA polymerase I transcription termination: similar mechanisms are employed by yeast and mammals.

Termination of RNA polymerase I (Pol I) transcription requires the interaction of a specific DNA binding factor with terminator elements downstream of the pre-rRNA coding region. Both the terminator elements and the respective termination factors are distinct in yeast and mammals, and differences in the mechanism of transcription termination have been postulated. We have compared in vitro transcription termination of yeast and mouse Pol I using both the murine factor TTF-I, and the yeast homolog Reb1p. We show that, similar to TTF-I, Reb1p was sufficient for pausing of Pol I from either species, but was unable to cause release of the nascent transcripts from the paused ternary complex. The deficiency of Reb1p to mediate transcript release from Pol I of either species was complemented by the recently characterized murine release factor. Thus, both yeast and mouse Pol I termination requires a trans-acting factor that, in conjunction with the T-rich flanking sequence, releases the transcripts and Pol I from the template. The observation that the murine factor causes dissociation of ternary transcription complexes arrested by Reb1p suggests that the mechanism of Pol I termination is highly conserved from yeast to mammals.

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Cutaneous heat flux models do not reliably predict metabolic rates of marine mammals.

Heat flux models have been used to predict metabolic rates of marine mammals, generally by estimating conductive heat transfer through their blubber layer. Recently, Kvadsheim et al. (1997) found that such models tend to overestimate metabolic rates, and that such errors probably result from the asymmetrical distribution of blubber. This problem may be avoided if reliable estimates of heat flux through the skin of the animals are obtained by using models that combine calculations of conductive heat flux through the skin and fur, and convective heat flux from the surface of the animal to the environment. We evaluated this approach based on simultaneous measurements of metabolic rates and of input parameters necessary for heat flux calculations, as obtained from four harp seals (Phoca groenlandica) resting in cold water. Heat flux estimates were made using two free convection models (double-flat-plate and cylindrical geometry) and one forced convection model (single-flat-plate geometry). We found that heat flux estimates generally underestimated metabolic rates, on average by 26-58%, and that small variations in input parameters caused large variations in these estimates. We conclude that cutaneous heat flux models are too inaccurate and sensitive to small errors in input parameters to provide reliable estimates of metabolic rates of marine mammals.

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Can diving-induced tissue nitrogen supersaturation increase the chance of acoustically driven bubble growth in marine mammals?

The potential for acoustically mediated causes of stranding in cetaceans (whales and dolphins) is of increasing concern given recent stranding events associated with anthropogenic acoustic activity. We examine a potentially debilitating non-auditory mechanism called rectified diffusion. Rectified diffusion causes gas bubble growth, which in an insonified animal may produce emboli, tissue separation and high, localized pressure in nervous tissue. Using the results of a dolphin dive study and a model of rectified diffusion for low-frequency exposure, we demonstrate that the diving behavior of cetaceans prior to an intense acoustic exposure may increase the chance of rectified diffusion. Specifically, deep diving and slow ascent/descent speed contributes to increased gas-tissue saturation, a condition that amplifies the likelihood of rectified diffusion. The depth of lung collapse limits nitrogen uptake per dive and the surface interval duration influences the amount of nitrogen washout from tissues between dives. Model results suggest that low-frequency rectified diffusion models need to be advanced, that the diving behavior of marine mammals of concern needs to be investigated to identify at-risk animals, and that more intensive studies of gas dynamics within diving marine mammals should be undertaken.

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Molecular biology of pheromone perception in mammals.

In mammals, olfactory sensory perception is mediated by two anatomically and functionally distinct sensory organs: the main olfactory epithelium (MOE) and the vomeronasal organ (VNO). Pheromones activate the VNO and elicit a characteristic array of innate reproductive and social behaviors, along with dramatic neuroendocrine responses. Recent approaches have provided new insights into the molecular biology of sensory transduction in the vomeronasal organ. Differential screening of cDNA libraries constructed from single sensory neurons from the rat VNO has led to the isolation of a family of genes which are likely to encode mammalian pheromone receptors. The isolation of these receptors from the vomeronasal organ might permit the analysis of the molecular events which translate the bindings of pheromones into innate stereotypic behaviors and help to elucidate the logic of pheromone perception in mammals.

Journal Article↗

Centrosome inheritance after fertilization and nuclear transfer in mammals.

Centrosomes, the main microrubule organizing centers in a cell, are nonmembrane-bound semi-conservative organelles consisting of numerous centrosome proteins that typically surround a pair of perpendicularly oriented cylindrical centrioles. Centrosome matrix is therefore oftentimes referred to as pericentriolar material (PCM). Through their microtubule organizing functions centrosomes are also crucial for transport and distribution of cell organelles such as mitochondria and macromolecular complexes. Centrosomes undergo cell cycle-specific reorganizations and dynamics. Many of the centrosome-associated proteins are transient and cell cycle-specific while others, such as y-tubulin, are permanently associated with centrosome structure. During gametogenesis, the spermatozoon retains its proximal centriole while losing most of the PCM, whereas the oocyte degenerates centrioles while retaining centrosomal proteins. In most mammals including humans, the spermatozoon contributes the proximal centriole during fertilization. Biparental centrosome contributions to the zygote are typical for most species with some exceptions such as the mouse in which centrosomes are maternally inherited and centrioles are assembled de novo during the blastocyst stage. After nuclear transfer in reconstructed embryos, the donor cell centrosome complex is responsible for carrying out functions that are typically fulfilled by the sperm centrosome complex during normal fertilization, including spindle organization, cell cycle progression and development. In rodents, donor cell centrioles are degraded after nuclear transfer, and centrosomal proteins from both donor cell and recipient oocytes contribute to mitotic spindle assembly. However, questions remain about the faithful reprogramming of centrosomes in cloned mammals and its consequences for embryo development. The molecular dynamics of donor cell centrosomes in nuclear transfer eggs need further analysis. The fate and functions of centrosome components in nuclear transfer embryos are being investigated by using molecular imaging of centrosome proteins labeled with specific markers including, but not limited to, green fluorescent protein (GFP).

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Bioavailability of metals and arsenic to small mammals at a mining waste-contaminated wetland.

In support of a baseline ecological risk assessment evaluating the impacts of mining wastes at the Milltown Reservoir Sediments Superfund site in Montana, a food chain transfer analysis was performed for resident small mammals. Deer mice (Peromyscus maniculatus) and meadow voles (Microtus pennsylvanicus) were trapped from a 200 A portion of a mixed upland and palustrine wetland, and concentrations of As, Cd, Cu, Pb, and Zn in carcass, liver, kidney, and testes were quantified. Concurrent to small mammal trapping, samples of grasses, forbs, and soils were collected and analyzed for metal and As residues. Using a linear multimedia food-chain model, assuming a forage base of wetland vegetation from the site, ingestion of local surface water, and incidental ingestion of soils with vegetation, body burdens of the metals and As in the herbivores were estimated. As a means of estimating potential element bioavailability at the site, the modeled body burdens were compared with measured element concentrations in herbivore tissues. The analysis indicates that the bioavailabilities of As, Cd, Cu, Pb, and Zn were equal to or less than 0.2% for internal organs and 0.1% for carcasses, on a microgram/g tissue wet weight basis. Available site data on soil pH, cation exchange capacity, and extractable elements fraction supported the limited release of soil elements. These results suggest that the bioavailable fraction of mining waste metals in riparian wetland soils may be quite small, and, for the Milltown Reservoir site, lower than originally anticipated.

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