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Spatial segregation of odorant receptor expression in the mammalian olfactory epithelium.

The signal elicited by the interaction of odorous ligands with receptors on olfactory sensory neurons must be decoded by the brain to determine which of the numerous receptors have been activated. We have examined the patterns of odorant receptor expression in the rat olfactory epithelium to determine whether the mammalian olfactory system employs spatial segregation of sensory input to encode the identity of an odorant stimulus. In situ hybridization experiments with probes for 11 different odorant receptors demonstrate that sensory neurons expressing distinct receptors are topologically segregated into a small number of broad, yet circumscribed, zones within the olfactory epithelium. Within a given zone, however, olfactory neurons expressing a specific receptor appear to be randomly distributed, rather than spatially localized. The complex mammalian olfactory system may therefore compartmentalize the epithelium into anatomically and functionally discrete units, such that each zone expresses only a subset of the entire receptor repertoire.

Animals↗

Segregational instability of pUB110-derived recombinant plasmids in Bacillus subtilis.

To study plasmid instability in Bacillus subtilis the pUB110-derived hybrid plasmid pLB2 (3.6 kb) and the bifunctional replicon pLB5 (5.9 kb), able to replicate in B. subtilis and Escherichia coli, were constructed. In both vectors homologous B. subtilis, or heterologous E. coli DNA fragments of various lengths were inserted. Irrespective of the source of the cloned DNA, the segregational stability of the recombinant plasmids in B. subtilis was severely affected by the DNA inserts. In contrast, no instability was observed in E. coli. In B. subtilis a steep inverse relationship existed between the size of the inserts and the level of stability. Increased size of the pLB plasmids resulted in strongly reduced copy numbers. This seems to be the primary cause of the size-dependent segregational instability.

Bacillus subtilis↗

Kinetic and segregational analysis of mitochondrial DNA recombination in yeast.

A pair of yeast strains of opposite mating type was constructed to contain polymorphisms at three loci on the mitochondrial genome--the 21 S rRNA gene, var1, and cob--such that parental and recombinant forms of these genes could be easily detected by Southern blot analysis. These polymorphisms were used to measure in a single cross gene conversions at the 21 S rRNA and var1 loci and a reciprocal recombination at cob. For all three loci, recombination initiates at about the same time, 4 to 6 h after mixing cells, and increases with similar kinetics over a 24-h period. The segregation of parental and recombinant forms of these genes was then followed by pedigree analysis. The results, which show a high variance in the distribution of parental and recombinant forms of all three genes in cells derived from both the first bud and the mother zygote, are consistent with the segregation of a small number of mitochondrial DNA molecules from the zygote to diploid buds. Based on these results and previous experiments of this type, a limited "zone of mixing" of parental mitochondrial DNA molecules probably exists in the zygote. The extent of sampling from this zone, together with the intrinsic properties of the recombination events themselves, is likely to determine the observed pattern of recombination of mitochondrial DNA sequences at the population level.

Alleles↗

Plasmid pTB913 derivatives are segregationally stable in Bacillus subtilis at elevated temperatures.

We studied the effects of temperature on the segregational stability of derivatives of the rolling-circle-type plasmid pTB913 in Bacillus subtilis. This 4.5-kb plasmid is a deletion derivative of pTB19, which was originally isolated from a thermophilic Bacillus. pTB913 derivatives carrying large inserts or lacking the minus origin for complementary strand synthesis were segregationally unstable at 37 degrees C. In contrast, at 47 degrees C all pTB913 derivatives tested were stably maintained in B. subtilis. The increased stability at 47 degrees C was attributed, at least partly, to increased copy numbers at this temperature. Although considerable amounts of single-stranded and high-molecular-weight plasmid DNA were formed at 47 degrees C, these products did not reduce plasmid stability at this temperature. The increased stability and increased copy number of pTB913 at elevated temperatures extend the use of this plasmid as a cloning vector in B. subtilis and other bacilli.

Bacillus subtilis↗

Lack of effect of acute acetaldehyde treatment on X chromosome segregation in Drosophila melanogaster females.

The effect of acute acetaldehyde treatments on X chromosome segregation was tested in germinal cells of Drosophila melanogaster females. The experiments were carried out using a test system where the nondisjunctional females (XXY) and only 1/4 of the expected regular progeny are viable. 24 h old virgin females were exposed for 60 min to 3, 4 and 5% acetaldehyde solutions by means of soaked tissue paper placed at the bottom of regular culture vials. After mating the females were brooded daily. Two additional experiments were performed with 0-2 h old and 4-5 day old virgin females using a 4% acetaldehyde solution. The results obtained show that acetaldehyde did not affect X chromosomal segregation in oocytes. This lack of effect could result from the highly efficient ADH-ALDH dependent detoxifying mechanism operating in Drosophila melanogaster.

Acetaldehyde↗

Exogenous monoamines affect the segregation of retinogeniculate fibers in developing rats.

The development of retinogeniculate projections was examined in rats which had norepinephrine levels altered by subcutaneous administration of 6-hydroxydopamine (6-OHDA) or exogenous norepinephrine (NE) during early postnatal life. NE, but not 6-OHDA, treatment resulted in an abnormal segregation of crossed and uncrossed axons at postnatal day 10, such that projections from the two eyes occupied extensively overlapping territory. This effect is at least partially reversible since in animals examined 30 days after cessation of NE treatment the retinogeniculate projections ultimately became segregated.

Animals↗

Cheaters sometimes prosper: distortion of mendelian segregation by meiotic drive.

Two of Mendel's three laws were quickly discarded as information on the organization and transmission of genes accumulated at the beginning of this century, but his law of segregation has shown remarkable staying power. In fact, within most of population genetic theory for sexual diploids is buried the tacit assumption that heterozygous alleles are represented in gametes in a 1:1 ratio. Nevertheless, there is a small, but important, group of genetic systems that subvert the law of segregation, and show 'meiotic drive'.

Animals↗

HLA-linked B cell alloantigens of a new segregant series: population and family studies of the SB antigens.

In order to define the new human histocompatibility antigens, we have generalized primed lymphocytes using responder and stimulator cells matched for all recognized HLA-linked histocompatibility antigens (A,B,C,D,DR,MB). Many such primed lymphocytes give highly discriminatory proliferative responses specific for antigens which differ between HLA-A,B,C,D,DR, and MB matched restimulating cells. Five distinct antigens have been defined which appear to be part of a single segregant series (designated "SB"). Studies in a DR/GLO recombinant family indicate that the antigens are coded by an HLA-linked gene telomeric to GLO. Family studies of 57 HLA haplotypes provide an estimate of genotype frequency which is 12% or less for four of the SB alleles but approximately 50% for the most common (SB4, which may be a "public: determinant); approximately 25% of haplotypes are blank. Population studies of one of the SB antigens (SB1) suggest that it is in linkage disequilibrium with the SB antigens are part of a highly polymorphic new segregant series of B cell alloantigens encoded by a gene that maps between HLA-B and GLO.

B-Lymphocytes↗

Studies on transformation markers and tumorigenicity in segregant clones from a human hybrid line.

Thirty segregant clones were back-selected in 8AG or 5BUdR media from a non-tumorigenic human intraspecific hybrid line (HeLa TK- X fibroblasts HPRT-) displaying a high plasminogen activator (PA) level, a disorganized fibronectin (FN) matrix and anchorage-independence. These clones exhibited a widely modulated expression of the above markers concomitantly with different degrees of chromosome loss. Out of six representative segregant clones tested in nude mice, two were found to re-express tumorigenicity. No significant correlation was observed between PA or FN levels and anchorage-independence, as well as between these markers and tumorigenicity.

Animals↗

Influence of food intake and sexual segregation on longevity, organ weights and the incidence of non-neoplastic and neoplastic diseases in rats.

In a small-scale experiment the effects of four variables were investigated in male and female Wistar rats fed on a standard laboratory chow and not deliberately exposed to any carcinogen. The variables investigated were (i) diet restriction by limiting access to food to 6.5 hr/day instead of 24 hr/day; (ii) housing males in a single-sex room as distinct from a mixed-sex room; (iii) life-long segregation of males from females, as distinct from access to one virgin female for 5 days during each alternate week; and (iv) uniparity in females versus life-long virginity. The endpoints compared were body-weight gain, longevity, visible and palpable swellings, absolute and relative organ weights, microscopically confirmed malignant neoplasms that contributed to death before the end of the study at 2 yr, incidence of other neoplasms in decedents and rats killed at the end of the study, and the incidence of various non-neoplastic conditions including myocardial inflammation and fibrosis, chronic progressive nephropathy, liver glycogen storage and fatty degeneration, mammary gland hyperplasia and secretory activity, pancreatic polyarteritis, radiculoneuropathy and certain testicular changes. The results indicated major beneficial effects of dietary restriction on most of the endpoints. By comparison the other variables had only marginal effects. Leydig-cell hyperplasia and neoplasia occurred at significantly higher incidences in males housed intermittently with females than in permanently segregated males. No convincing differences were seen between females that littered once and those that remained virgins. The relevance of these findings to the prediction of cancer mortality risk in man and to the design of rodent carcinogenicity studies is discussed.

Animals↗

Segregation of optic fibre projections into eye-specific bands in dually innervated tecta in Xenopus.

The restoration of the retinotectal projections following metamorphic bilateral optic nerve sections was studied in Xenopus 20 weeks after operation, using [3H]proline autoradiography and electrophysiological mapping. Optic fibre projections from each eye to both tecta segregated into eye-specific termination bands with some overlap at the borders. The bands were arranged rostrocaudally in the tecta such that each eye had an alternating representation. Despite the segregation of the optic fibres, each eye had a complete visual (retinal) projection with the normal nasotemporal and dorsoventral polarities of the visual field maintained. The results suggest competition for preferred terminal sites between the two sets of optic fibres in dually innervated tecta.

Animals↗

Segregation of cerebrorubral and cerebellorubral synaptic inputs on rubrospinal neurons of fetal cats as demonstrated by intracellular recording.

Cerebrorubral and cerebellorubral inputs are localized to distal dendrites and somata of red nucleus neurons in adult cats, respectively. To examine if this segregation is established early in development, we performed intracellular recording from rubrospinal neurons of fetal cats aged from embryonic day 58 to 65. Stimulation of the contralateral cerebellar nuclei evoked excitatory postsynaptic potentials (EPSPs). EPSPs were also induced by stimulation of the ipsilateral pericruciate cortext but they were much slower in time course and smaller in amplitude compared to cerebellar ones. We suggest that cerebrorubral and cerebellorubral synapses are segregated on soma-dendritic membrane of rubrospinal neurons early in development.

Animals↗

Topographical organization of the nigrotectal projection in rat: evidence for segregated channels.

Recent evidence suggests that projections from the superior colliculus to the brainstem in rat are organized into a series of anatomically segregated output channels. To understand how collicular function may be modified by the basal ganglia it is important to know whether particular output modules of the superior colliculus can be selectively influenced by input from substantia nigra. The purpose of the present study was, therefore, to examine in more detail topography within the nigrotectal system in the rat. Small injections (10-50 nl) of a 1% solution of wheatgerm agglutinin conjugated with horseradish peroxidase were made at different locations within substantia nigra and surrounding structures. A discontinuous puff-like pattern of anterogradely transported label was found in medial and caudal parts of the ipsilateral intermediate layers of the superior colliculus. In contrast, the rostrolateral enlargement of the intermediate layers contained a greater density of more evenly distributed terminal label. Injection sites associated with this dense pattern of laterally located label were concentrated in lateral pars reticulata, while the puff-like pattern was produced by injections into ventromedial pars reticulata. Retrograde tracing experiments with the fluorescent dyes True Blue and Fast Blue revealed that injections involving the rostrolateral intermediate layers were consistently associated with a restricted column of labelled cells in the dorsolateral part of ipsilateral pars reticulata. Comparable injections into medial and caudal regions of the superior colliculus produced retrograde labelling in ventral and medial parts of the rostral two-thirds of pars reticulata. Both anterograde and retrograde tracing data indicated that contralateral nigrotectal projections arise from cells located in ventral and medial pars reticulata. The present results suggest that the main ipsilateral projection from substantia nigra pars reticulata to the superior colliculus comprises two main components characterized by regionally segregated populations of output cells and spatially separated zones of termination. Of particular interest is the apparent close alignment between terminal zones of the nigrotectal channels and previously defined populations of crossed descending output cells in the superior colliculus. Thus, the rostrolateral intermediate layers contain a concentration of terminals specifically from dorsolateral pars reticulata and output cells which project to the contralateral caudal medulla and spinal cord. Conversely, the medial and caudal intermediate layers receive terminals from ventral and medial pars reticulata and contain cells which project specifically to contralateral regions of the paramedian pontine and medullary reticular formation.(ABSTRACT TRUNCATED AT 400 WORDS)

Amidines↗

Subsynaptic segregation of metabotropic and ionotropic glutamate receptors as revealed by immunogold localization.

Glutamate is a major neurotransmitter in the brain that acts both through fast ionotropic receptors and through slower metabotropic receptors coupled to G proteins. Both receptors are present throughout the somatodendritic domain of neurons as shown by immunohistochemical and patch clamp recording studies. Immunogold labelling revealed a concentration of metabotropic receptors at the edge, but not within the main body of anatomically defined synapses, raising the possibility that ionotropic and metabotropic receptors are segregated. We applied double immunogold labelling to study glutamatergic parallel and climbing fibre synapses in the cerebellar cortex. The ionotropic AMPA type receptors occupy the membrane opposite the release site in the main body of the synaptic junction, whereas the metabotropic receptors are located at the periphery of the same synapses. Furthermore, immunoreactivity for AMPA receptors is at least twice as high in the parallel fibre synapses as in glutamatergic mossy fibre synapses. We suggest that the spatial segregation of ionotropic and metabotropic glutamate receptors permits the differential activation of these receptors according to the amount of glutamate released presynaptically, whereas the different densities of the ionotropic receptor at distinct synapses could allow the same amount of glutamate to evoke fast responses of different magnitude.

Amino Acid Sequence↗

Mercury-induced segregational errors of chromosomes in human lymphocytes and in Indian muntjac cells.

Segregational errors of chromosomes were studied in human lymphocytes and in Indian muntjac fibroblasts exposed to methylmercury chloride (CH3HgCl) or mercury chloride (HgCl2). The cells were exposed to the mercury compounds only during a limited period of the pre-DNA synthetic stage of the cell cycle or from that stage up to mitosis. In the lymphocytes we observed a clear increase of C-mitotic figures for both mercury compounds and for both exposure times. Segregational errors were, however, much more important after the shorter exposure period. Muntjac fibroblasts appear to be more sensitive to the mercury than are the lymphocytes so that their suitability for the study of C-mitosis may be questionable. The muntjac cells may be an important tool for the study of polyploidy induction.

Aneuploidy↗

Lipid segregation and IgE receptor signaling: a decade of progress.

Recent work to characterize the roles of lipid segregation in IgE receptor signaling has revealed a mechanism by which segregation of liquid ordered regions from disordered regions of the plasma membrane results in protection of the Src family kinase Lyn from inactivating dephosphorylation by a transmembrane tyrosine phosphatase. Antigen-mediated crosslinking of IgE receptors drives their association with the liquid ordered regions, commonly called lipid rafts, and this facilitates receptor phosphorylation by active Lyn in the raft environment. Previous work showed that the membrane skeleton coupled to F-actin regulates stimulated receptor phosphorylation and downstream signaling processes, and more recent work implicates cytoskeletal interactions with ordered lipid rafts in this regulation. These and other results provide an emerging view of the complex role of membrane structure in orchestrating signal transduction mediated by immune and other cell surface receptors.

Animals↗

Age-related changes in neural activity associated with concurrent vowel segregation.

Older adults exhibit degraded speech comprehension in complex sound environments, which may be related to overall age-related declines in low-level sound segregation. This hypothesis was tested by measuring event-related potentials (ERPs) while listeners identified two different vowels presented simultaneously. Older adults were less accurate than young adults at identifying the two vowels, although both groups improved similarly with increasing fundamental frequency differences (Deltaf0) between vowels. Reaction time data showed that older adults took more time to process stimuli, especially those with smaller Deltaf0. A negative ERP wave indexing the automatic registration of Deltaf0 (the object-related negativity) was reduced in older adults. In contrast, young and older adults showed a similar pattern of neural activity indexing attentive processing of Deltaf0. The results suggest that aging affects the ability to automatically segregate speech sounds.

Adult↗