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PubMed1994

Abnormal astrocyte differentiation and defective cellular interactions in wobbler mouse spinal cord.

The wobbler mutation is inherited as an autosomal recessive trait and displays a muscular atrophy associated with motoneuron degeneration in early postnatal development. It has been shown that the level of glial fibrillary acidic protein (GFAP) is greatly increased in the spinal cord of wobbler mice. We performed immunocytochemical analyses combined with confocal microscopy to study the developmental distribution of GFAP-positive astrocytes in the spinal cord of wobbler mice during the course of the disease, and in primary cultures of adult wobbler spinal cord astrocytes. Many changes in the number and distribution of astrocytes were observed in the wobbler mice from 1-10 months post-partum. Strongly GFAP-positive astrocytes are present in small number in the anterior horn by 1 month. They increase in number and are observed in the entire spinal cord grey and white matters by 2-10 months. These reactive astrocytes have thick, short, extensively branched processes which contrast with the long, unbranched processes observed in control mice. The wobbler astrocyte processes are oriented perpendicular to the surface of the spinal cord, which contrasts with the normal parallel, concentric orientation. No expansion of astrocyte processes exit from the white matter towards the grey matter. Moreover, the surface of the wobbler spinal cord beneath the meninges displays a dramatic decrease of interdigitating processes, end feet and flattened cell bodies of astrocytes that form a disorganized layer. In vitro, mutant astrocytes have morphological characteristics similar to those in vivo and, in particular, develop short, thick, branched processes. These mutant astrocytes in cultures do not contact one another, whereas normal mature cultures show an increased incidence of cell-cell contacts between long processes. The increase of astrocyte reactivity associated with these modifications in astrocytic process arrangement may reflect an important primary event in the course of the wobbler disease rather than a non-specific response to motoneuronal death.

Animals↗
PubMed1978

[Mathematical model of epidemiology of an anthropozoonose: brucellosis (author's transl)].

We are trying to make a model of the epidemiology of Brucellosis by using a two-type branching process. In this text, we are studying the probability of extinction. A complete study on the theory of process with a finite number of types was made by SEVAST'YANOV (1951) [6]. The study of the behaviour of such a process is made with the help of the generating function of the number of infected animals by a single animal. To express this generating function, we start with the same hypothesis as BARTOSZYNSKI [2] (1965) when he makes an interhuman epidemic model by a simple branching process of GALTON-WATSON. However these hypothesis will be modified in order to adapt them to the types of Brucellosis contamination.

Animals↗
PubMed1999

Expression of c-ret promotes morphogenesis and cell survival in mIMCD-3 cells.

c-Ret, a protein tyrosine kinase receptor, and its ligand glial-derived neurotropic factor (GDNF) are critical for early regulation of ureteric bud development and nephrogenesis. To address whether c-ret directly initiates epithelial cell morphogenesis, the c-ret receptor was expressed in murine inner medullary collecting duct cells (mIMCD-3, a cell line of ureteric bud origin, which has no detectable endogenous c-ret expression). Stable expression of wild-type c-ret was found to yield a constitutively tyrosine-phosphorylated receptor, with no change after the addition of GDNF. Examination of mRNA from these cells demonstrated the message for endogenous GDNF, suggesting that c-ret was potentially being constitutively activated by an autocrine mechanism. When mIMCD-3 cells stably expressing the phosphorylated c-ret receptor were cultured in a type I collagen matrix, they exhibited little GDNF-independent or -dependent branching process formation at early time points compared with the known morphogen hepatocyte growth factor (HGF) (48 h; control, 0.33 +/- 0.33; GDNF, 1.0 +/- 0.58, P = nonsignificant; and HGF, 6.33 +/- 0.33 processes/20 cell clusters, P < 0.001), whereas extended culture (7 days) under serum-free conditions revealed a marked increase in cell survival and the spontaneous development of rudimentary branching process formation. Extended culture (7 days) of c-ret-expressing clones in type I collagen with the epithelial morphogens HGF and/or epidermal growth factor (EGF) resulted in the development of complex three-dimensional spiny cysts, whereas parental mIMCD-3 cells died under these conditions. We conclude that activated c-ret appears to mediate epithelial morphogenesis by prolonging cell survival and, in conjunction with activation of the morphogenic receptors c-met and the EGF receptor, initiates the events required for very early branching morphogenesis.

Animals↗
PubMed1979

Fission models of population variability.

Most models in population genetics are models of allele frequency, making implicit or explicit assumptions of equilibrium or constant population size. In recent papers, we have attempted to develop more appropriate models for the analysis of rare variant data in South American Indian tribes; these are branching process models for the total number of replicates of a variant allele. The spatial distribution of a variant may convey information about its history and characteristics, and this paper extends previous models to take this factor into consideration. A model of fission into subdivisions is superimposed on the previous branching process, and variation between subdivisions is considered. The case where fission is nonrandom and the locations of like alleles are initially positively associated, as would happen were a tribal cluster or village to split on familial lines, is also analyzed. The statistics developed are applied to Yanomama Indian data on rare genetic variants. Due to insufficient time depth, no definitive new inferences can be drawn, but the analysis shows that this model provides results consistent with previous conclusions, and demonstrates the general type of question that may be answered by the approach taken here. In particular, striking confirmation of a higher-than-average growth rate, and hence smaller-than-previously-estimated age, is obtained for the Yan2 serum albumen variant.

Biological Evolution↗
PubMed1995

Immunophenotypic evidence for distinct populations of microglia in the rat hypothalamo-neurohypophysial system.

The morphology, distribution and immunophenotype of microglia throughout the adult rat hypothalamo-neurohypophysial system was examined. Four macrophage-associated antibodies (OX-42, F4/80, ED1 and ED2) were used; the expression of major histocompatibility complex antigens was investigated by use of antibodies against OX-6, OX-17 (MHC class II) and OX-18 (MHC class I). Three distinct types of microglia were identified. The first was located in the magnocellular nuclei; these 'radially branched' ('ramified') microglia had round cell bodies and long branched processes, and were strongly immunoreactive only for OX-42. The second was located outside the blood-brain barrier in the median eminence, pituitary stalk and neurohypophysis often close to blood vessels; these 'compact' microglia had irregular cell bodies and shorter processes, and were strongly labelled by OX-42 and F4/80, weakly labelled by OX-18, and generally unlabelled by ED1, ED2, OX-6 and OX-17. The third type was found in small numbers throughout the system at the surface of the nervous tissue or around blood vessels; these 'perivascular' microglia were elongated cells with no branching processes, and were strongly labelled by ED1, ED2, OX-18, OX-6, OX-17 and F4/80 antibodies but showed variable OX-42 immunoreactivity. Cells in a perivascular location were heterogeneous with respect to their immunophenotype. The presence in the normal adult rat hypothalamo-neurohypophysial system of MHC class-II molecules (OX-6 and OX-17) on a sub-set of perivascular microglia suggests that these cells are capable of presenting antigen to T lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1999

Transduction of cultured oligodendrocytes from normal and twitcher mice by a retroviral vector containing human galactocerebrosidase (GALC) cDNA.

Krabbe disease or globoid cell leukodystropy is a lysosomal disorder caused by a deficiency of galactocerebrosidase (GALC) activity. This results in defects in myelin that lead to severe symptoms and early death in most human patients and animals with this disease. With the cloning of the GALC gene and the availability of the mouse model, called twitcher, it was important to evaluate the effects of providing GALC via a retroviral vector to oligodendrocytes in culture. After differentiation, the untransduced cells from normal mice extended highly branched processes while those from the twitcher mice did not. Oligodendrocytes in culture can be readily transduced to produce much higher than normal levels of GALC activity. Transduced normal and twitcher cells formed clusters when plated at high density. Transduction of twitcher oligodendrocytes plated at lower density, followed by differentiation, resulted in some cells having a completely normal appearance with highly branched processes. Other cells showed retraction and fragmentation. Perhaps over expression of GALC activity may be detrimental to oligodendrocytes. These studies demonstrate that the phenotype of twitcher oligodendrocytes can be corrected by providing GALC via gene transfer, and this could lead the way to future studies to treat this disease.

Animals↗
PubMed1994

Statistical analysis of aberrant crypt assays for colon cancer promotion studies.

The size of aberrant crypt foci in rodent colons is a short-term assay for the promotion of colon carcinogenesis. An approximate model for the number of aberrant crypts in a focus is derived by assuming a branching process with a gamma lifetime. Maximum likelihood estimation can be implemented using symbolic manipulation packages. Although in most cases the model provides a reasonable fit for the distribution of the number of aberrant crypts per focus, there is some evidence of lack of fit. A more robust analysis can be based on summarizing the data for each animal by their mean number of aberrant crypts per focus. The branching process with gamma lifetime is used to obtain expressions for the functional relation between the variance of the number of aberrant crypts per focus and their mean. It is shown that in most cases this relation is quadratic. The quadratic relation also holds for other lifetime distributions. The parameters of the quadratic relation can be estimated by pseudolikelihood. An experiment for assessing the promotional effect of the compound HMF is used for illustration.

Analysis of Variance↗
PubMed1995

Axon-glial relationships in the anterior medullary velum of the adult rat.

The anterior medullary velum is a thin sheet of CNS tissue which roofs the rostral part of the IVth ventricle and contains fascicles of myelinated fibres which, in part, arise from the nucleus of the IVth cranial nerve. This study used histochemical, immunohistochemical, and intracellular dye-injection techniques to describe cellular interrelationships in the velum in whole-mounts and in sections. Rip antibody-stained whole mounts provided a unique description of both oligodendrocyte units (defined as an oligodendrocyte and the complement of myelinated internodal segments it forms), and consecutive myelin sheaths along the same axon. A broad range of unit morphologies was categorised into four arbitrary groups, according to classical criteria, which comprised small cells supporting the short, thin myelin sheaths of 15-30 small diameter axons (Type I), through intermediate types (II & III), to the largest cells forming the long, thick myelin sheaths of 1-3 large diameter axons. Rip antibody and ferric ion-ferrocyanide staining, together with intracellular dye injection, revealed oligodendrocyte process branching patterns and their mode of engagement of myelin sheaths, nodes of Ranvier, and the spatial disposition of the outer cytoplasmic rims of myelin sheaths. The latter formed a conspicuous spiral ridge on the exterior surface of myelin sheaths which connected with the paranodal loops at each heminode. Large bundles of axons decussated through the velum, the bulk of which were IVth nerve fibres which constituted the IVth nerve rootlet. The PNS/CNS transitional zone of the IVth nerve was located 0.25-0.50 mm along the root, where astrocytic end-feet defined an abrupt margin, convex towards the periphery, where the heminodes of central and peripheral myelin were apposed, and where the basal lamina tubes of the Schwann cell units were discontinued. The basal processes of ependymal cells lining the ventricular wall of the velum, passed between axon bundles before abutting on the basal lamina of the pia. Many of these processes branched and ran along the axonal bundles. A monolayer of microglia occupied a subependymal stratum in which the non-overlapping dendritic territories of each cell formed a regular mosaic throughout the velum without any obvious interaction with either axons or other glial cells. Astrocytes were also uniformly distributed; their fine processes made up a dense lattice amongst axons, often running parallel and within the fibre bundles; stouter ones had terminal end-feet which undercoated the basal lamina of both the glia limitans externa and the blood vessels in the velum.

Animals↗
PubMed1998

A reconsideration of Galton's problem (using a two-sex population).

The main purposes of this paper are to promote and expound the bisexual Galton-Watson branching process as a relevant model for the consideration of Francis Galton's problem regarding the extinction of surnames of "men of note." A scheme for adapting the bisexual process to consider Galton's problem is introduced. A necessary and sufficient condition for the certain extinction of a male-induced property in a two-sex species is presented. An approach for calculating the extinction of a male-generated characteristic in the two-sex species is proposed. That approach is then used to find the probability of the extinction of surnames in a bisexual population for Alfred Lotka's data based on an United States Census. Finally, these results are then compared with the classic extinction probabilities (from Lotka) associated with the traditional Galton-Watson branching process using asexual reproduction.

Female↗
PubMed1997

On the genealogy of a rare allele.

The gene genealogy is derived for a rare allele that is descended from a mutant ancestor that arose at a fixed time in the past. Following Thompson (1976, Amer. J. Human Genet. 28, 442-452), the fractional linear branching process is used as a model of the demography of a rare allele. The model does not require the total population size to be constant or the mutant class to be neutral; so long as individuals in the class are selectively equivalent, the class as a whole may have a selective advantage, or disadvantage, relative to other alleles in the population. An exact result is given for the joint probability distribution of the coalescence times among a sample of alleles descended from the mutant. A method is described for rapidly simulating these coalescence times. The relationship between the genealogical structure of a discrete generation branching process and a continuous generation birth-death process is elucidated. The theory may be applied to the problem of estimating the ages of rare nonrecurrent mutations.

Alleles↗
PubMed1986

Immunohistochemical identification of noradrenaline- and adrenaline- synthesizing neurons in the cat ventrolateral medulla.

The distribution and morphology of cell bodies containing the catecholamine biosynthetic enzymes dopamine-beta-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) in the ventrolateral medulla (VLM) of the cat were studied immunohistochemically after intracisternal administration of colchicine. Perikarya immunoreactive to DBH were found throughout the VLM extending from approximately the spinomedullary junction to the level of the superior olivary nucleus. In the caudal VLM DBH neurons were found primarily in the region immediately dorsal to the lateral reticular nucleus (LRN), although a few scattered DBH neurons were also found near the ventral surface of the medulla in and around the parvicellular division of the LRN. These DBH neurons in the caudal VLM were generally fusiform, fusiform-bipolar, or multipolar, with a mean somal area of 422 +/- 32 microns2, and with two to four branching processes. In the rostral VLM DBH neurons formed three distinct groups: one group was found in the nucleus paragigantocellularis lateralis in the region just ventromedial to the retrofacial nucleus (RFN) near the ventrolateral surface of the medulla; the second group was found in the region dorsomedial to the rostral aspects of the nucleus ambiguous and the RFN, and the third group was found in the region along the lateral aspect of the superior olivary nucleus. Perikarya immunoreactive to the adrenaline-synthesizing enzyme PNMT were localized to a more restricted region of the VLM that extended from approximately the rostral aspect of the caudal third of the inferior olivary complex (level of the obex) to the caudal pole of the facial nucleus. These PNMT neurons were fusiform or multipolar with a mean somal area of 273 +/- 21 microns2, and with two to five branching processes. The location, size, shape, and numbers of PNMT-immunoreactive neurons corresponded closely to the rostral groups of DBH neurons, with the exception of the group found along the lateral aspect of the superior olivary nucleus. These data indicate that noradrenaline-synthesizing neurons are primarily found in the caudal VLM and in the region near the superior olivary nucleus, whereas catecholamine neurons in the rostral VLM between these two noradrenergic cell groups synthesize adrenaline. As the VLM has previously been shown to have direct connections with spinal cord, brainstem, and hypothalamic areas implicated in cardiovascular and neuroendocrine regulation, this suggests that DBH- and PNMT-synthesizing neurons are components of neuronal circuits involved in these homeostatic mechanisms.

Adrenergic Fibers↗
PubMed1995

AII amacrine cell population in the rabbit retina: identification by parvalbumin immunoreactivity.

Parvalbumin (PV) is a calcium-binding protein localized to selected neurons in the nervous system, including the retina. This investigation evaluated the distribution of PV immunoreactivity in the rabbit retina using immunohistochemistry with a monoclonal antibody directed to carp PV. In the inner nuclear layer (INL), PV immunoreactivity was present in horizontal and amacrine cells. In the ganglion cell layer, PV immunostaining was confined to somata that are likely to be both displaced amacrine cells and ganglion cells. PV-immunoreactive (IR) amacrine cells were positioned in the proximal INL adjacent to the inner plexiform layer (IPL). These cells usually gave rise to a single primary process, which arborized into two distinct bands in the IPL. In sublamina a, the processes were thin and had large, irregular endings. In sublamina b, multiple processes branched from the primary process and were characterized by varicosities and spines. PV-IR amacrine cell bodies measured from 8 to 10 microns in diameter. Their density was highest in the visual streak and lowest in the periphery of the superior retina. The average number of PV-IR amacrine cells was 464,045 cells per retina (N = 3), and the average regularity index of the PV-IR cell mosaic was 3.23. PV-IR amacrine cells were further characterized by double-label immunofluorescence experiments using antibodies to PV and tyrosine hydroxylase (TH). Varicose TH-IR processes were in close apposition to many PV-IR amacrine cells and often formed "ring structures" around them. Together, these morphological, quantitative, and histochemical observations indicate that PV immunoreactivity in the INL is localized predominantly to AII amacrine cells, and therefore it is a valuable marker for the identification of this cell type.

Animals↗
PubMed1997

The establishment of peripheral sensory arbors in the leech: in vivo time-lapse studies reveal a highly dynamic process.

Pressure-sensitive (P) neurons located in the leech CNS form elaborate terminal arbors in the body wall of the animal during mid-embryogenesis. In the experiments discussed here, arbor development in the target region was studied in intact, unanesthetized leech embryos using time-lapse video microscopy of individual, fluorescently stained P neurons. Analysis of time-lapse recordings made over a period of several days revealed that arbor formation is a very dynamic process. At any particular time, most high-order terminal branches were either extending or retracting, in approximately equal numbers and at very similar rates. Many branches underwent several rounds of extension and retraction every hour. Net arbor growth occurred at a much lower rate than the extension and retraction rates of individual branches. Process retraction sometimes resulted in an apparent change in the topological order of processes. Significantly, the initiation of new branches was restricted to a few locations along the parent process, which were termed "hot spots." Moreover, the capacity to generate high-order branches correlated with parent process stability. The target region of the growing P cell arbor in the body wall was subsequently examined using confocal microscopy in fixed preparations. The arbor expanded between the longitudinal and circular muscle layers, a region occupied by small unidentified cells. Simultaneous imaging of the dye-labeled terminal arbor and the surrounding tissue at two different wavelengths suggested that the high-order processes were navigating around these cells, which sometimes forced the growing processes to assume a bent form. These observations suggest that the formation of the P cell arbor can be best described as a "dynamically unstable" process that is constrained by interactions with its environment.

Animals↗
PubMed1987

Light and electron microscopic localization of a cell surface antigen (NG2) in the rat cerebellum: association with smooth protoplasmic astrocytes.

Immunofluorescence and immunoperoxidase techniques were used to localize a cell surface chondroitin-sulfate proteoglycan antigen, termed NG2, in the developing and adult rat cerebellum. In the adult, both polyclonal and monoclonal anti-NG2 antibodies labeled cells throughout the cerebellar cortex, with the labeled cells being especially prominent in the molecular layer. The labeled cells had small, irregularly shaped cell bodies from which thin highly branched processes radiated in a stellate array. The NG2-labeled cells were not labeled with antibodies against glial fibrillary acidic protein (GFAP), vimentin, or S-100 protein, intracellular markers for astrocytes. However, electron microscopic immunocytochemical analysis of NG2 immunoreactive cells revealed a cell morphology consistent with that of protoplasmic astrocytes. Labeled cell bodies contained a thin rim of organelle-poor cytoplasm surrounding a euchromatic nucleus. Thick processes originating from the cell soma tapered to form thin branches with highly irregular surface contours that extended between adjacent neuronal elements. The labeled processes did not form synapses in the neuropil, and no synaptic profiles onto anti-NG2-labeled cell bodies or processes were observed. Thus, we conclude that the NG2 antigen is a cell surface marker for a class of smooth protoplasmic astrocytes. Immunoreactive cells were seen in the developing cerebellum beginning at embryonic day 16. The number of labeled cells increased during the early stages of cerebellar development, reaching a peak at about postnatal day (PND) 4 or 5 and declining thereafter. In the developing cerebellum, labeled cells lying within the forming molecular layer resembled the cells seen in the adult, whereas cells lying deeper within the folia had an immature appearance with fewer processes and less branching. This apparent gradient of morphological maturation suggests that an interaction with parallel fibers in the developing molecular layer may play a role in the terminal cytodifferentiation of the NG2-labeled smooth protoplasmic astrocytes.

Animals↗
PubMed2003

Epidemiological modeling in a branching population. Particular case of a general SIS model with two age classes.

This paper covers the elaboration of a general class of multitype branching processes for modeling in a branching population, the evolution of a disease with horizontal and vertical transmissions. When the size of the population may tend to infinity, normalization must be carried out. As the initial size tends to infinity, the normalized model converges a.s. to a dynamical system the solution of which is the probability law of the state of health for an individual ancestors line. The focal point of this study concerns the transient and asymptotical behaviors of a SIS model with two age classes in a branching population. We will compare the asymptotical probability of extinction on the scale of a finite population and on the scale of an individual in an infinite population: when the rates of transmission are small compared to the rate of renewing the population of susceptibles, the two models lead to a.s. extinction, giving consistent results, which no longer applies to the opposite situation of important transmissions. In that case the size of the population plays a crucial role in the spreading of the disease.

Age Factors↗