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PubMed2001

Persistence patterns of scrapie in a sheep flock.

The epidemiology and transmission dynamics of sheep scrapie is as yet poorly understood. Here we present a theoretical analysis of the transmission dynamics within a sheep flock, concentrating on how persistence properties depend on transmission scenario and flock size. Patterns of disease persistence and extinction are studied analytically using branching-process approximations and numerically using stochastic model simulations. For a given basic reproduction number, disease extinction is most likely when late-stage infected animals are responsible for most of the transmission. This effect can be understood in terms of aggregation in the distribution of the number of secondary infections arising from a single primary infection. The presence of an environmental reservoir reduces the probability of extinction.

Animals↗
PubMed1990

A burst of differentiation in the outer posterior retina of the eleven-week human fetus: an ultrastructural study.

Many studies on human retinal development have cited the third gestational month as a period when the posterior retina undergoes rapid differentiation and maturation, including a lining up of cone precursors. Ultrastructural data on the posterior retina during the third month are very limited, and totally lacking for the cone monolayer. We have examined two human fetal retinas between ten and 11 gestational weeks. Before the appearance of the cone monolayer, the outer neural retina consists of a homogeneous population of undifferentiated neuroblasts. Mitotic figures are still evident, even posteriorally. There is no outer plexiform layer (OPL). The interface of neural retina to retinal pigment epithelium (RPE) is largely featureless. By 11 weeks, the posterior retina has a thin OPL that separates the many rows of cells in the developing inner nuclear layer from the single tier of macular cone precursors. The RPE monolayer consists of cuboidal cells whose apical surface elaborates ridges of cytoplasm and branched processes that project into the subretinal space. The large, cuboidal cones are linked to each other and Müller cells at the outer limiting membrane. They show definitive signs of the structural polarity typical of vertebrate photoreceptors. Their apical cytoplasm contains many organelles common to the inner segment, while the basal cytoplasm has synaptic ribbons and vesicles, and receives invaginating contacts from processes in the OPL neuropil arising from differentiating second-order neurons. Lateral cone surfaces are mutually underlain by large subsurface cisterns.

Cell Differentiation↗
PubMed1992

Immunocytochemical localization of taurine and glial fibrillary acidic protein in human optic nerve.

Taurine immunoreactivity (IR) in 1-microns sections of glutaraldehyde-fixed human optic nerve was observed using light microscopy and an antibody raised in rabbit to taurine conjugated to bovine serum albumin. Throughout the nerve, taurine-IR was prominent in glial cells, in their perinuclear regions, and in their numerous branching processes, some of which extended to the pial septa. The peripheral glial mantle (glia limitans) was densely stained, whereas axons and the pial septa showed relatively little or no taurine-IR. Immunoreactivity for glial fibrillary acidic protein (GFAP), and astrocyte-specific marker, was evaluated on adjacent sections. The pattern of GFAP-IR was highly similar to that for taurine, suggesting that a subset of taurine-immunoreactive glial cells are optic nerve astrocytes. To our knowledge, this is the first localization of taurine and GFAP in human optic nerve.

Adult↗
PubMed2000

Characterization of human RNA splice signals by iterative functional selection of splice sites.

An iterative in vitro splicing strategy was employed to select for optimal 3' splicing signals from a pool of pre-mRNAs containing randomized regions. Selection of functional branchpoint sequences in HeLa cell nuclear extract yielded a sequence motif that evolved from UAA after one round of splicing toward a UACUAAC consensus after seven rounds. A significant part of the selected sequences contained a conserved AAUAAAG motif that proved to be functional both as a polyadenylation signal and a branch site in a competitive manner. Characterization of the branchpoint in these clones to either the upstream or downstream adenosines of the AAUAAAG sequence revealed that the branching process proceeded efficiently but quite promiscuously. Surprisingly, the conserved guanosine, adjacent to the common AAUAAA polyadenylation motif, was found to be required only for polyadenylation. In an independent experiment, sequences surrounding an optimal branchpoint sequence were selected from two randomized 20-nt regions. The clones selected after six rounds of splicing revealed an extended polypyrimidine tract with a high frequency of UCCU motifs and a highly conserved YAG sequence in the extreme 3' end of the randomized insert. Mutating the 3' terminal guanosine of the intron strongly affects complex A formation, implying that the invariant AG is recognized early in spliceosome assembly.

Base Sequence↗
PubMed2002

Light-induced charge redistribution in the retinal chromophore is required for initiating the bacteriorhodopsin photocycle.

Bacteriorhodopsin's photocycle is initiated by the retinal chromophore light absorption. It has usually been assumed that light primarily isomerizes a retinal double bond which in turn induces protein conformational alterations and biological activity. We have studied several artificial pigments derived from retinal analogues tailored to substantially reduce the light-induced chromophore polarization. The lack of chromophore polarization was reflected in an undetectable second harmonic generation (SHG) signal. It was revealed that these artificial pigments did not exhibit any detectable light-induced photocycle nor light acceleration of the hydroxylamine-bleaching reaction. We suggest that light-induced retinal polarization triggers protein polarization which controls the course of the isomerization reaction by determining the relative efficiency of forward versus back-branching processes.

Bacteriorhodopsins↗
PubMed1998

Effects of cyclic AMP on expression of myelin genes in the N20.1 oligodendroglial cell line.

The N20.1 immortalized cell line has several characteristics of differentiating oligodendrocytes (OLs), including expression of the glycolipids galactocerebroside (GalC) and sulfatide, and the myelin proteins CNPase and myelin basic protein (MBP) (1,2). Addition of 1-100 microM forskolin to elevate cyclic AMP (cAMP) levels changed cell morphology from irregular and flattened to a more rounded birefringent cell with multiple branched processes. GalC and sulfatide were detected immunocytochemically after permeabilization in the untreated cells and levels appeared to increase slightly following exposure to forskolin. Further analysis showed that most of the glycolipid was internal, with virtually no detectable levels on the cell surface in untreated cells and a very slight change following treatment with forskolin. Synthesis of the two lipids as measured by [H3]galactose incorporation doubled within 24 hours of treatment with forskolin. Levels of message for UDP-galactose: ceramide galactosyl transferase (CGT), a key enzyme in the synthesis of GalC and sulfatide, were compared with those of MBP and proteolipid protein (PLP), before and after elevation of cAMP. No changes were observed in levels of mRNA for CGT and PLP after 24 hours, with a possible increase by 48 hours. In contrast, levels of MBP message dropped precipitously by 24 hours; this was accompanied by an increase in levels of message for suppressed cAMP-inducible POU (SCIP). Thus CGT transcription is regulated independently of MBP and SCIP in N20.1 cells. Analysis of MBP levels by immunocytochemistry and Western blot showed little or no change in protein levels at 24 and 48 hours, in contrast to the sharp decrease in message levels by 24 hours, indicating a relatively long half life for MBP in this cell line. Thus, the N20.1 cells are an informative model for examining regulation of expression of myelinotypic proteins and GalC, as well as the transport of this lipid to the plasma membrane.

Animals↗
PubMedDate not supplied

Synantocytes: new functions for novel NG2 expressing glia.

In the adult CNS, antibodies to the NG2 chondroitin sulphate proteoglycan (CSPG) label a large population of glia that have the antigenic phenotype of oligodendrocyte progenitor cells (OPC). However, NG2 expressing glia have the morphological phenotype of astrocytes, not OPC. We propose adult NG2 expressing glia are a distinct mature glial type, which we have called syantocytes or synantoglia after the Greek 'to contact', because they specifically contact neurons and axons at synapses and nodes of Ranvier, respectively. Synantocytes are highly complex cells that elaborate multiple branching processes and are an equally significant population in both white and grey matter. We provide evidence that phenotypically distinct synantocytes develop postnatally and that neither postnatal nor adult synantocytes depend on axons for their survival, indicating they respond with markedly different behaviours to the environmental cues and axonal signals that control the differentiation of OPC into oligodendrocytes. The primary response of synantocytes to changes in the CNS environment is a rapid and localised reactive gliosis. Reactive synantocytes interact intimately with astrocytes and macrophages at lesion sites, consistent with them playing a key role in the orchestration of scar formation that protects the underlying neural tissue. It is our hypothesis that synantocytes are specialised to monitor and respond to changes in the integrity of the CNS, by way of their cellular contacts, repertoire of plasmalemmal receptors and the NG2 molecule itself. To paraphrase Del Rio Hortega, we propose that synantocytes are the fifth element in the CNS, in addition to neurons, astrocytes, oligodendrocytes and microglia.

Animals↗
PubMed1988

Selective responses of visual cortical cells do not depend on shunting inhibition.

Theoretical analyses of the electrical behaviour of the highly branched processes of nerve cells has focused attention on the possibility that single cells perform complex logical operations rather than simply summing their synaptic inputs. In particular, it has been suggested that the orientation and direction selectivity of cells in the visual cortex results from the action of a nonlinear 'shunting' inhibition that emulates an AND-NOT logical operation. The characteristic biophysical feature of this proposed inhibitory mechanism is that it evokes a large and relatively sustained increase in the conductance of the neuronal membrane while leaving the membrane potential unaffected. This shunting mechanism contrasts with linear 'summative' inhibition in which conductance changes are less prominent, and inhibition is achieved by hyperpolarization of the membrane potential. In a direct experimental test of the hypothesis that the selectivity of visual cortical neurons depends on shunting inhibition we found no evidence for the large conductance changes predicted by the theory.

Action Potentials↗
PubMed1986

Mitochondrial DNA variability in natural populations of Hawaiian Drosophila. I. Methods and levels of variability in D. silvestris and D. heteroneura populations.

We describe techniques by which mitochondrial DNA (mtDNA) restriction site information can be obtained for up to 16 different restriction endonucleases on individual Hawaiian Drosophila, in particular D. silvestris and D. heteroneura. We have constructed mtDNA restriction site maps for a total of forty-eight wild caught genomes of both species, from seven major collecting sites on the island of Hawaii. Levels of variability are, in general, high in D. silvestris (p of Ewens et al., 1981 = 0.0486) and lower in D. heteroneura (p = 0.0327). Measures of population subdivision using Nei's Gst indicate that about 50-60 per cent of the observed variability is due to interdemic subdivision. In accordance, populations within a species show a much lower level of variability, however some populations harbour individuals that are very divergent from the rest of their conspecifics at the same locality. We review two possible mechanisms that could explain the presence of these divergent individuals, hybridisation and the effects of stochastic branching processes.

Animals↗
PubMed1995

The GATA-3 gene is expressed during human kidney embryogenesis.

GATA-3 is a transcription factor involved in the differentiation of T lymphocytes and additionally expressed in several chicken and mouse embryonic tissues. Using in situ hybridization, we found that the human GATA-3 gene is selectively expressed in the developing kidney. GATA-3 mRNA is first detected in the Wolffian duct from the time of its emergence in the embryonic intermediate mesoderm and further expressed in the collecting ducts of the mesonephros until its involution. In the metanephros, GATA-3 is expressed in the ureteric bud where it is constitutively transcribed, throughout development, along the branching process that gives rise to the whole collecting system of the definitive kidney. Besides the Wolffian duct and derivatives, we also report the expression of GATA-3 gene in the glomerular mesangium and adjacent endocapillary cells, in both meso- and metanephros. This early and specific expression of the GATA-3 gene suggests a role for this transcription factor in the differentiation of the human kidney.

DNA-Binding Proteins↗
PubMed2002

Morphological characteristics of C1 and C2 adrenergic neurone groups in marmoset monkey brainstem by using antibody against phenylethanolamine-N-methyltransferase.

This work describes a mapping study of phenylethanolamine-N-methyltransferase (PNMT) immunoreactive neurones and fibres in the medulla oblongata of the marmoset monkey, Callithrix jacchus. Two groups of PNMT-immunoreactive neurones were found in the marmoset monkey medulla oblongata: a ventrolateral (C1 group) and a dorsomedial PNMT-immunoreactive cells group (C2 group). The PNMT-immunoreactive cells in the ventrolateral group C1 were found to be located around the lateral reticular nucleus. The PNMT-immunoreactive somata within the ventrolateral medulla are round to oval, and mostly multipolar with branched processes. In the dorsomedial group C2, PNMT-immunoreactive cell bodies appeared near the obex. The majority of the dorsomedial PNMT-immunoreactive neurones were observed in the nucleus tractus solitarius; although some were present in the dorsal motor nucleus of the vagus. The PNMT-immunoreactive somata in the dorsomedial medulla were small and round or ovoid. These results provide information upon the adrenergic system in the medulla oblongata of a species that presents a useful model of a small primate brain, the marmoset monkey.

Adrenergic Fibers↗
PubMed2000

Glutamate receptor subunit delta2 is highly expressed in a novel population of glial-like cells in rat pineal glands in culture.

The mammalian pineal gland uses L-glutamate as an intercellular chemical transmitter to regulate negatively melatonin synthesis. To receive glutamate signals, pinealocytes express at least three kinds of glutamate receptors: metabotropic receptor types 3 and 5 and an ionotropic receptor, GluR1. In this study, we examined whether or not the fourth class of ionotropic receptor, delta, which is known for its nondefinitive molecular function and its unique expression pattern in brain, is expressed in pineal gland. RT-PCR analyses with specific probes indicated the expression of mRNA of delta2 but not that of delta1 in pineal gland and cultured pineal cells. Western blotting analysis with polyclonal antibodies specific to the carboxyl-terminal region of the delta2 receptor recognized a single 110-kDa polypeptide of cerebellar membranes and specifically immunostained Purkinje cells. The delta2 antibodies recognized a 110-kDa polypeptide of pineal membranes and specifically immunostained huge glial-like cells with the occasional presence of several long, branching processes in a pineal cell culture. delta2 is not uniformly distributed throughout the cells and is relatively abundant at the periphery of the cell bodies and long processes, where the terminals of synaptophysin-positive processes of pinealocytes, a site for glutamate secretion, are frequently present. The delta2-positive cells constitute a very minor population among total pineal cells (approximately 0.03%). Double immunolabeling with delta2 antibodies and antibodies against marker proteins for pineal interstitial cells clearly distinguishes delta2-positive pineal cells and other known interstitial cells, including glial fibrillary acidic protein- or vimentin-positive glial-like cells. These results indicated that the delta2 glutamate receptor is expressed in a novel subpopulation of pineal glial-like cells in culture and suggest the presence of a glutamate-mediated intercellular signal transduction mechanism between pinealocytes and delta2-expressing cells. The pineal cells may provide a good experimental system for studies on the function of glutamate receptor delta2.

Amino Acid Sequence↗
PubMed1989

Reversible inhibition of oligodendrocyte progenitor differentiation by a monoclonal antibody against surface galactolipids.

We have hypothesized that oligodendrocyte (OL) surface glycolipids, specifically galactocerebroside and sulfatide, play a role in the regulation of OL development by acting as sensors/transmitters of environment information. In support of this hypothesis we report here a reversible inhibition of OL progenitor cell differentiation by a monoclonal antibody [Ranscht mAb (R-mAb); Ranscht, B., Clapshaw, P. A. & Seifert, W. (1982) Proc. Natl. Acad. Sci. USA 79, 2709-2713] that reacts with these glycolipids. When isolated OL progenitors or mixed primary cultures are grown in the presence of the antibody, myelinogenic development is blocked in a dose-dependent manner at concentrations as low as 2 micrograms of IgG per ml. The inhibited cells express the OL progenitor markers O4 and vimentin but are negative for galactosylcerebroside, sulfatide, 2',3'-cyclic nucleotide 3'-phosphohydrolase, myelin basic protein, and myelin basic protein RNA expression. In contrast, the levels of total cellular protein and the expression of astrocytic glial fibrillary acidic protein in mixed cultures are not affected. Antibody-blocked cells have a distinctive morphology in which long, sparsely branched processes emanate from round cell bodies. Upon removing the perturbing antibody, the cells rapidly resume differentiation. Reverted mixed primary cultures, in which OL progenitors of several sequential developmental stages are present at the time of plating, differentiate more rapidly than control cultures, suggesting that the antibody-induced block results in a synchronization of developmental progression along the OL lineage by accumulating cells at the inhibition point. However, the normal temporal sequence of marker expression is maintained. Control studies with several other antibodies recognizing OL cell surface antigens, including HNK-1, neural cellular adhesion molecule (N-CAM), 1A9, anticholesterol, and O1, did not inhibit development. Since the inhibition occurs in highly enriched populations of OL progenitors, the inhibition does not involve cell-cell interactions between OLs and other cell types but concerns interactions of OLs with themselves, soluble factors, or OL extracellular matrix molecules and adhesion factors that provide essential environmental signals required for normal myelinogenic development.

Animals↗
PubMed1996

Population dynamics of flaviviruses revealed by molecular phylogenies.

The phylogeny of 123 complete envelope gene sequences was reconstructed in order to understand the evolution of tick- and mosquito-borne flaviviruses. An analysis of phylogenetic tree structure reveals a continual and asymmetric branching process in the tick-borne flaviviruses, compared with an explosive radiation in the last 200 years in viruses transmitted by mosquitoes. The distinction between these two viral groups probably reflects differences in modes of dispersal, propagation, and changes in the size of host populations. The most serious implication of this work is that growing human populations are being exposed to an expanding range of increasingly diverse viral strains.

Animals↗
PubMed1998

Social transmission of reproductive behavior increases frequency of inherited disorders in a young-expanding population.

The observation of high frequencies of certain inherited disorders in the population of Saguenay-Lac Saint Jean can be explained in terms of the variance and the correlation of effective family size (EFS) from one generation to the next. We have shown this effect by using the branching process approach with real demographic data. When variance of EFS is included in the model, despite its profound effect on mutant allele frequency, any mutant introduced in the population never reaches the known carrier frequencies (between 0.035 and 0.05). It is only when the EFS correlation between generations is introduced into the model that we can explain the rise of the mutant alleles. This correlation is described by a c parameter that reflects the dependency of children's EFS on their parents' EFS. The c parameter can be considered to reflect social transmission of demographic behavior. We show that such social transmission dramatically reduces the effective population size. This could explain particular distributions in allele frequencies and unusually high frequency of certain inherited disorders in some human populations.

Female↗
PubMed1999

Declining survival probability threatens the North Atlantic right whale.

The North Atlantic northern right whale (Eubalaena glacialis) is considered the most endangered large whale species. Its population has recovered only slowly since the cessation of commercial whaling and numbers about 300 individuals. We applied mark-recapture statistics to a catalog of photographically identified individuals to obtain the first statistically rigorous estimates of survival probability for this population. Crude survival decreased from about 0.99 per year in 1980 to about 0.94 in 1994. We combined this survival trend with a reported decrease in reproductive rate into a branching process model to compute population growth rate and extinction probability. Population growth rate declined from about 1. 053 in 1980 to about 0.976 in 1994. Under current conditions the population is doomed to extinction; an upper bound on the expected time to extinction is 191 years. The most effective way to improve the prospects of the population is to reduce mortality. The right whale is at risk from entanglement in fishing gear and from collisions with ships. Reducing this human-caused mortality is essential to the viability of this population.

Animals↗
PubMed1995

Regulation of Wnt5a mRNA expression in human mammary epithelial cells by cell shape, confluence, and hepatocyte growth factor.

The Wnts are a family of genes with a role in cell fate and morphological development in numerous embryonic and adult tissues. In mouse mammary tissue a subset of the Wnts have a function in the normal development of the gland, and aberrant expression of Wnts normally silent in this tissue causes mammary carcinomas. We have previously shown that Wnt5a expression is elevated in the epithelial component of proliferative lesions of human breast and have therefore examined the regulation of Wnt5a mRNA expression in the human mammary epithelial cell line HB2, which has a luminal phenotype and thus represents the most commonly transformed cell type in human breast cancer. Wnt5a was up-regulated 30-fold at confluence. This up-regulation was induced specifically by confluence and not by the growth arrest that accompanied it. In addition, Wnt5a was down-regulated 3-fold by changes in cell shape associated with the transition from growth on a two-dimensional surface (flat cell morphology) to growth in three-dimensional gels (spherical cell morphology). Cytoskeletal disruption with non-toxic doses of colchicine also induced a spherical morphology and brought about a dose-dependent down-regulation of Wnt5a. Wnt5a was also down-regulated 10-fold during the hepatocyte growth factor-induced branching of HB2 cell aggregates in collagen gels. The down-regulation of Wnt5a preceded the branching process. A similar result was obtained with primary human breast epithelial populations and the breast cancer cell line MDA468. We conclude that regulation of Wnt5a expression is a down-stream effect of signaling by hepatocyte growth factor. These results are consistent with a role for Wnt5a in mammary epithelial cell motility and are in accord with Xwnt5a's function in embryonal cell migration. If Wnt5a's function in human mammary epithelial cells is similar to that of Xwnt5a, its up-regulation at confluence may be a mechanism for inhibition of cell migration beyond confluence.

Breast↗
PubMed2005

Extinction probabilities and times to extinction for populations of tsetse flies Glossina spp. (Diptera: Glossinidae) subjected to various control measures.

A stochastic branching process was used to derive equations for the mean and variance of the probability of, and time to, extinction in tsetse populations. If the remnant population is a single inseminated female, the extinction probability increases linearly with adult mortality and is always certain if this mortality >3.5% per day even for zero pupal mortality. If the latter mortality is 4% per day, certain extinction is only avoided if adult mortality <1.5% per day. For remnant female populations >1, the extinction probability increases in a non-linear manner with adult mortality. Extinction is still certain for adult mortality >3.5% per day but, when the remnant population is >16, extinction is highly unlikely for adult mortality <2.5% per day if all females are inseminated. Extinction probability increases with increasing probability of sterile mating in much the same way as it does with increasing adult mortality. Extinction is assured if the probability of insemination can be reduced to 0.1. The required reduction decreases with increasing adult mortality. For adult mortality = 6-8% per day, the time to extinction increases only by one generation per order of magnitude increase in the starting population. Time to extinction is less sensitive to changes in the pupal than in the adult mortality. Reductions in the probability of insemination only become important when adult mortality is small; if the adult mortality is 8% per day, reducing the insemination probability from 1 to 0.1 only reduces the expected time to extinction by two generations. Conversely, increases in adult mortality produce important reductions in the required time even when the probability of insemination is 0.1. The practical, economic implication for the sterile insect technique is that the low-tech methods used to suppress tsetse populations should not be halted when the release of sterile males is initiated. The sterile insect technique should only be contemplated when it has been demonstrated that the low-tech methods have failed to effect eradication. The theory is shown to be in good accord with the observed results of tsetse control campaigns involving the use of odour-baited targets in Zimbabwe and the sterile insect technique on Unguja Island, Zanzibar.

Animals↗