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PubMed1993

Culture of rat brain preoptic area neurons: effects of sex steroids.

The aim of the present study was to investigate the direct effects of testosterone (T), estradiol-17 alpha (E2) and 5 alpha-dihydrotestosterone (5 alpha-DHT) on the survival of neurons and the morphology of neuronal processes (the total process length, the number of processes and the number of process branchings) of the cells derived from neonatal rat preoptic area (POA) to analyze the bases of morphological sexual dimorphism of the POA. The POA cells were exposed to sex steroids during 2-14 days of culture in serum-free medium. Neuronal survival was significantly greater in cultures treated with 1.0 nM T than controls. Although T at any concentration tested (0.1, 1.0 or 10.0 nM) failed to affect the mean value of the process length, a significant difference was observed in the frequency distribution of total process length between 1.0 nM T-treated cultures and control cultures. By 1.0 nM T, the number of process branchings was also significantly increased. In contrast, the number of neurons and the total process length of neurons were not affected by 5 alpha-DHT. However, the number of process branchings was significantly increased by 0.1 nM 5 alpha-DHT. E2 had no significant effect on any of these parameters. These results suggest that T directly enhances the survival of neurons and increases the dendritic and/or axonic field of the POA, which serves, at least partly, for the development of morphological sexual dimorphism in the rat.

Androgens↗
PubMed1998

Waiting with and without recombination: the time to production of a double mutant.

R.A. Fisher and H.J. Muller argued in the 1930s that a major evolutionary advantage of recombination is that it allows favorable mutations to be combined within an individual even when they first appear in different individuals. This effect is evaluated in a two-locus, two-allele model by calculating the average waiting time until a new genotypic combination first appears in a haploid population. Three approximations are developed and compared with Monte Carlo simulations of the Wright-Fisher process of random genetic drift in a finite population. First, a crude method, based on the deterministic accumulation of single mutants, produces a waiting time of 1/square root of N mu(2) with no recombination and [formula: see text] with recombination between the two loci, where mu is the mutation rate, N is the haploid population size, and R is the recombination rate. Second, the waiting time is calculated as the expected value of a heterogeneous geometric distribution obtained from a branching process approximation. This gives accurate estimates for small values of N mu large. The estimates for small values of N mu are considerably lower than the simulated values. Finally, diffusion analysis of the Wright-Fisher process provides accurate estimates for N mu small, and the time scales of the diffusion process show a difference between R = 0 and for R >> 0 of the same order of magnitude as seen in the deterministic analysis. In the absence of recombination, accurate approximations to the waiting time are obtained by using the branching process for high N mu and the diffusion approximation for low N mu. For low N mu the waiting time is well approximated by 1/the square root of 8N2 mu(3). With R >> 0, the following dependence on N mu is observed: For N mu > 1 the waiting time is virtually independent of recombination and is well described by the branching process approximation. For N mu approximately equal to 1 the waiting time is well described by a simplified diffusion approximation that assumes symmetry in the frequencies of single mutants. For N mu << 1 the waiting time is well described by the diffusion approximation allowing asymmetry in the frequencies of single mutants. Recombination lowers the waiting time until a new genotypic combination first appears, but the effect is small compared to that of the mutation rate and population size. For large N mu, recombination has a negligible effect, and its effect is strongest for small N mu, in which case the waiting time approaches a fixed fraction of the waiting time for R = 0. Free recombination lowers the waiting time to about 45% of the waiting time for absolute linkage for small N mu. Selection has little effect on the importance of recombination in general.

Biological Evolution↗
PubMed1986

Quantitation and morphological characterization of rapid axon and dendritic growth from single cerebral hemispheric neurons in hydrated collagen lattice culture.

Quantitative and qualitative data are reported for single cerebral hemispheric neurons in a 3-dimensional hydrated collagen lattice (HCL) culture system. Individual neurons not in contact with other cells or cell processes, including synapses, rapidly displayed two morphologically distinct classes of processes that could be traced from origin to termination: long thin processes interpreted as being axons, and shorter tapering and sometimes branched processes interpreted as being dendrites. Axons and dendrites of single neurons that had at least one process longer than the cell body were measured on each of 3 days after plating using an image analysis system coupled to a phase-contrast microscope and a microcomputer. Mean lengths of axons and dendrites alone or combined as total new growth per neuron, increased 3- to 5-fold and were as high as 745, 694 and 1226 microns respectively after 3 days in HCL, although some individual axons measured over 1500 microns. Other indices of neuron growth and differentiation increased 1- to 5-fold including the number of primary processes, branch points, segments and growth cones. Phase-contrast microscopy, staining with Nissl and silver, and scanning and transmission electron microscopy demonstrated many single multipolar and other neurons with axons, dendrites and well-differentiated properties. The data show that individual central nervous system neurons have an inherent capacity to quickly express characteristic differentiated features and also to grow rapidly in HCL.

Animals↗
PubMed1980

[Dynamics of systems with induced cell proliferation within the framework of a branching stochastic process model. I. The number of cell generations induced to proliferate].

The probabilistic description of cell generation numbers in the populations induced to proliferate is considered on the basis of the model of branching age-dependent stochastic process. The recurrent formulas for generating functions (and moments) are derived for the following process which are of interest for the analysis of induced proliferation in closed cell populations: the number of cells in n-th generation horn up to the moment of observation, and the number of cell in n-th generation existing at a given moment.

Cell Division↗
PubMed1999

A simple explanation for taxon abundance patterns.

For taxonomic levels higher than species, the abundance distributions of the number of subtaxa per taxon tend to approximate power laws but often show strong deviations from such laws. Previously, these deviations were attributed to finite-time effects in a continuous-time branching process at the generic level. Instead, we describe herein a simple discrete branching process that generates the observed distributions and find that the distribution's deviation from power law form is not caused by disequilibration, but rather that it is time independent and determined by the evolutionary properties of the taxa of interest. Our model predicts-with no free parameters-the rank-frequency distribution of the number of families in fossil marine animal orders obtained from the fossil record. We find that near power law distributions are statistically almost inevitable for taxa higher than species. The branching model also sheds light on species-abundance patterns, as well as on links between evolutionary processes, self-organized criticality, and fractals.

Animals↗
PubMed1987

Limit theorems for the population size of a birth and death process allowing catastrophes.

The linear birth and death process with catastrophes is formulated as a right continuous random walk on the non-negative integers which evolves in continuous time with an instantaneous jump rate proportional to the current value of the process. It is shown that distributions of the population size can be represented in terms of those of a certain Markov branching process. The ergodic theory of Markov branching process transition probabilities is then used to develop a fairly complete understanding of the behaviour of the population size of the birth-death-catastrophe process.

Animals↗
PubMed2006

Intramembrane receptor-receptor interactions: a novel principle in molecular medicine.

In 1980/81 Agnati and Fuxe introduced the concept of intramembrane receptor-receptor interactions and presented the first experimental observations for their existence in crude membrane preparations. The second step was their introduction of the receptor mosaic hypothesis of the engram in 1982. The third step was their proposal that the existence of intramembrane receptor-receptor interactions made possible the integration of synaptic (WT) and extrasynaptic (VT) signals. With the discovery of the intramembrane receptor-receptor interactions with the likely formation of receptor aggregates of multiple receptors, so called receptor mosaics, the entire decoding process becomes a branched process already at the receptor level in the surface membrane. Recent developments indicate the relevance of cooperativity in intramembrane receptor-receptor interactions namely the presence of regulated cooperativity via receptor-receptor interactions in receptor mosaics (RM) built up of the same type of receptor (homo-oligomers) or of subtypes of the same receptor (RM type1). The receptor-receptor interactions will to a large extent determine the various conformational states of the receptors and their operation will be dependent on the receptor composition (stoichiometry), the spatial organization (topography) and order of receptor activation in the RM. The biochemical and functional integrative implications of the receptor-receptor interactions are outlined and long-lived heteromeric receptor complexes with frozen RM in various nerve cell systems may play an essential role in learning, memory and retrieval processes. Intramembrane receptor-receptor interactions in the brain have given rise to novel strategies for treatment of Parkinson's disease (A2A and mGluR5 receptor antagonists), schizophrenia (A2A and mGluR5 agonists) and depression (galanin receptor antagonists). The A2A/D2, A2A/D3 and A2A/mGluR5 heteromers and heteromeric complexes with their possible participation in different types of RM are described in detail, especially in the cortico-striatal glutamate synapse and its extrasynaptic components, together with a postulated existence of A2A/D4 heteromers. Finally, the impact of intramembrane receptor-receptor interactions in molecular medicine is discussed outside the brain with focus on the endocrine, the cardiovascular and the immune systems.

Animals↗
PubMed2000

Phylogenetic analysis under reticulate evolution.

The usual assumption that species have evolved from a common ancestor by a simple branching process--where each branch is genetically isolated--has been challenged by the observation of frequent hybridization between species in natural populations. In fact, most plant species are thought to have hybrid origins. This reticulate pattern of species evolution has posed problems in the definition of speciation and in phylogenetic reconstruction, especially when molecular data are used. As a result, hybridization has been largely treated as an evolutionary accident or statistical error in phylogenetic analysis. In this paper, I explicitly incorporate hybridization as an evolutionary occurrence and then conduct phylogenetic reconstruction. I first examine the reticulate evolution under a pure drift model, and then extend the theory to fit a mutation model. A least-squares method is developed for reconstructing a reticulate phylogeny using gene frequency data. The efficacy of the method under the pure drift model is verified via Monte Carlo simulations.

Evolution, Molecular↗
PubMed2004

Genetic control of cell intercalation during tracheal morphogenesis in Drosophila.

BACKGROUND: Branching morphogenesis transforms an epithelial sheet into a tubular network with distinct features regarding the length and diameter of individual tubes. Branching is controlled by several signaling pathways, but the molecular consequences of these pathways in the responding cells are poorly understood. RESULTS: We have undertaken a detailed characterization of cell rearrangements during tracheal branching morphogenesis in Drosophila embryos with a GFP fusion protein labeling the adherens junctions (AJs) and high-resolution live imaging. To analyze the branching process at the cellular level, we further developed an imaging approach that allows us to follow single cells during the branching process. We find that controlled cell intercalation, which requires extensive AJ remodeling, is key to the formation of tracheal branches of different cellular complexities. In particular, most branches consist of tubes with individual cells wrapped around the lumen. These branches form through cell intercalation, which requires the transformation of most of the initial intercellular AJs into autocellular AJs. We propose a step-wise model explaining how this AJ remodeling occurs and use this model to better understand defects in various mutants. We find that Dpp and Wnt signaling control cell intercalation by regulating the expression of Spalt, a zinc finger transcription factor; Spalt inhibits intercalation, leading to the formation of large, multicellular tubes. CONCLUSION: Tracheal morphogenesis is regulated by an interplay of different signaling systems that control cell migration and cell intercalation, respectively. Only the combined action of these signaling systems allows efficient branch elongation and the formation of morphologically distinct branches.

Adherens Junctions↗
PubMed1992

Asymptotic rates of growth of the extinction probability of a mutant gene.

We prove that a result of Haldane (1927) that relates the asymptotic behaviour of the extinction probability of a slightly supercritical Poisson branching process to the mean number of offspring is true for a general Bienaymé-Galton-Watson branching process, provided that the second derivatives of the probability-generating functions converge uniformly to a non-zero limit. We show also by examples that such a result is true more widely than our proof suggests and exhibit some counter-examples.

Gene Frequency↗
PubMed1992

The survival probability of a mutant in a multidimensional population.

We prove a general result about the asymptomatic behaviour of the survival probability of a slightly supercritical multitype Bienaymé-Galton-Watson branching process. This is the complete analogue of a result which Ewens (1968) obtained for a Poisson branching process.

Genetics, Population↗
PubMed1996

Processing of branched DNA intermediates by a complex of human FEN-1 and PCNA.

In eukaryotic cells, a 5' flap DNA endonuclease activity and a ds DNA 5'-exonuclease activity exist within a single enzyme called FEN-1 [flap endo-nuclease and 5(five)'-exo-nuclease]. This 42 kDa endo-/exonuclease, FEN-1, is highly homologous to human XP-G, Saccharomyces cerevisiae RAD2 and S.cerevisiae RTH1. These structure-specific nucleases recognize and cleave a branched DNA structure called a DNA flap, and its derivative called a pseudo Y-structure. FEN-1 is essential for lagging strand DNA synthesis in Okazaki fragment joining. FEN-1 also appears to be important in mismatch repair. Here we find that human PCNA, the processivity factor for eukaryotic polymerases, physically associates with human FEN-1 and stimulates its endonucleolytic activity at branched DNA structures and its exonucleolytic activity at nick and gap structures. Structural requirements for FEN-1 and PCNA loading provide an interesting picture of this stimulation. PCNA loads on to substrates at double-stranded DNA ends. In contrast, FEN-1 requires a free single-stranded 5' terminus and appears to load by tracking along the single-stranded DNA branch. These physical constraints define the range of DNA replication, recombination and repair processes in which this family of structure-specific nucleases participate. A model explaining the exonucleolytic activity of FEN-1 in terms of its endonucleolytic activity is proposed based on these observations.

Antibodies, Monoclonal↗
PubMed1999

Vesicular monoamine transporter 1 is responsible for storage of 5-hydroxytryptamine in rat pinealocytes.

Vesicular monoamine transporters (VMATs) are involved in chemical transduction in monoaminergic neurons and various endocrine cells through the storage of monoamines in secretory vesicles. Mammalian pinealocytes contain more 5-hydroxytryptamine (5-HT) than any other cells and are expected to contain VMAT, although no information is available so far. Upon the addition of ATP, radiolabeled 5-HT was taken up by a particulate fraction prepared from cultured rat pinealocytes. The 5-HT uptake was inhibited significantly by bafilomycin A1 (an inhibitor of vacuolar H+-ATPase), 3,5-di-tert-butyl-4-hydroxybenzylidenemalononitrile (a proton conductor), or reserpine (an inhibitor of VMAT). RT-PCR analysis suggested that VMAT type 1 (VMAT1), but not type 2, is expressed. Antibodies against VMAT1 recognized a single polypeptide with an apparent molecular mass of approximately 55 kDa, and specifically immunostained pinealocytes. VMAT1 immunoreactivity was high in the vesicular structures in the varicosities of long branching processes and was associated with 5-HT, but not with synaptophysin, a marker protein for microvesicles. The 5-HT immunoreactivity in the long branching processes disappeared upon incubation with reserpine. These results indicate that 5-HT, at least in part, is stored in vesicles other than microvesicles in pinealocytes through a mechanism similar to that of various secretory vesicles.

Adenosine Triphosphate↗
PubMed2005

Linking population-level models with growing networks: a class of epidemic models.

We introduce a class of growing network models that are directly applicable to epidemiology. We show how to construct a growing network model (individual-level model) that generates the same epidemic-level outcomes as a population-level ordinary differential equation (ODE) model. For concreteness, we analyze the susceptible-infected (SI) ODE model of disease invasion. First, we give an illustrative example of a growing network whose population-level variables are compatible with those of this ODE model. Second, we demonstrate that a growing network model can be found that is equivalent to the Crump-Mode-Jagers (CMJ) continuous-time branching process of the SI ODE model of disease invasion. We discuss the computational advantages that our growing network model has over the CMJ branching process.

Animals↗
PubMed2005

Process outgrowth in oligodendrocytes is mediated by CNP, a novel microtubule assembly myelin protein.

Oligodendrocytes (OLs) extend arborized processes that are supported by microtubules (MTs) and microfilaments. Little is known about proteins that modulate and interact with the cytoskeleton during myelination. Several lines of evidence suggest a role for 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in mediating process formation in OLs. In this study, we report that tubulin is a major CNP-interacting protein. In vitro, CNP binds preferentially to tubulin heterodimers compared with MTs and induces MT assembly by copolymerizing with tubulin. CNP overexpression induces dramatic morphology changes in both glial and nonglial cells, resulting in MT and F-actin reorganization and formation of branched processes. These morphological effects are attributed to CNP MT assembly activity; branched process formation is either substantially reduced or abolished with the expression of loss-of-function mutants. Accordingly, cultured OLs from CNP-deficient mice extend smaller outgrowths with less arborized processes. We propose that CNP is an important component of the cytoskeletal machinery that directs process outgrowth in OLs.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1986

Cell population dynamics during the differentiative phase of tissue development.

The dynamics of a cell population whose numbers are growing exponentially have been described well by a mathematical model based on the theory of age-dependent branching processes. Such a model, however, does not cover the period following exponential growth when cell differentiation curtails population size. This paper offers an extension to the branching process model to remedy this deficiency. The extended model is ideal for describing embryonic growth; its use is illustrated with data from embryonic retina. The model offers a better computational framework for the interpretation of a variety of data (growth curves of cell numbers, DNA histograms, thymidine labelling indices, FLM curves, BUdR-labelled mitoses curves) because age-distributions can be calculated at any stage of development, not just during exponential growth. Proportions of cells in the various phases of the cell cycle can be computed as growth slows. Such calculations show the gradual transition from a population dominated by cells which are young with respect to cell cycle age to one dominated by those which are old, and the effects such biases have on the proportions of cells in each phase.

Age Factors↗
PubMed2005

Distribution of microglial cells in the cerebral hemispheres of embryonic and neonatal chicks.

The distribution, morphology and morphometry of microglial cells in the chick cerebral hemispheres from embryonic day 4 (E4) to the first neonatal day (P1) were studied by histochemical labeling with a tomato (Lycopersicon esculentum) lectin. The histochemical analysis revealed lectin-reactive cells in the nervous parenchyma on day E4. Between E4 (5.7 +/- 1.35 mm length) and E17 (8.25 +/- 1.2 mm length), the lectin-reactive cells were identified as ameboid microglia and observed starting from the subventricular layer, distributed throughout the mantle layer and in the proximity of the blood vessels. After day E13, the lectin-reactive cells exhibited elongated forms with small branched processes, and were considered primitive ramified microglia. Later, between E18 (5.85 +/- 1.5 mm cell body length) and P1 (3.25 +/- 0.6 mm cell body length), cells with more elongated branched processes were observed, constituting the ramified microglia. Our findings provide additional information on the migration and differentiation of microglial cells, whose ramified form is observed at the end of embryonic development. The present paper focused on the arrangement of microglial cells in developing cerebral hemispheres of embryonic and neonatal chicks, which are little studied in the literature. Details of morphology, morphometry and spatial distribution of microglial cells contributed to the understanding of bird and mammal central nervous system ontogeny. Furthermore, the identification and localization of microglial cells during the normal development could be used as a morphological guide for embryonic brain injury researches.

Animals↗