Genetic interactions among cytoplasmic dynein, dynactin, and nuclear distribution mutants of Neurospora crassa.

Cytoplasmic dynein is a multisubunit, microtubule-associated, mechanochemical enzyme that has been identified as a retrograde transporter of various membranous organelles. Dynactin, an additional multisubunit complex, is required for efficient dynein-mediated transport of vesicles in vitro. Recently, we showed that three genes defined by a group of phenotypically identical mutants of the filamentous fungus Neurospora crassa encode proteins that are apparent subunits of either cytoplasmic dynein or dynactin. These mutants, designated ropy (ro), display abnormal hyphal growth and are defective in nuclear distribution. We propose that mutations in other genes encoding dynein/dynactin subunits are likely to result in a ropy phenotype and have devised a genetic screen for the isolation of additional ro mutants. Cytoplasmic dynein/dynactin is the largest and most complex of the cytoplasmic motor proteins, and the genetic system described here is unique in its potentiality for identifying mutations in undefined genes encoding dynein/dynactin subunits or regulators. We used this screen to isolate > 1000 ro mutants, which were found to define 23 complementation groups. Unexpectedly, interallelic complementation was observed with some allele pairs of ro-1 and ro-3, which are predicted to encode the largest subunits of cytoplasmic dynein and dynactin, respectively. The results suggest that the Ro1 and Ro3 polypeptides may consist of multiple, functionally independent domains. In addition, approximately 10% of all newly isolated ro mutantsdisplay unlinked noncomplementation with two or more of the mutants that define the 23 complementation groups. The frequent appearance of ro mutants showing noncomplementation with multiple ro mutants having unlinked mutations suggests that nuclear distribution in filamentous fungi is a process that is easily disrupted by affecting either dosage or activity of cytoplasmic dynein, dynactin, and perhaps other cytoskeletal proteins or regulators.

Distinct pharmacological properties and distribution in neurons and endocrine cells of two isoforms of the human vesicular monoamine transporter.

A second isoform of the human vesicular monoamine transporter (hVMAT) has been cloned from a pheochromocytoma cDNA library. The contribution of the two transporter isoforms to monoamine storage in human neuroendocrine tissues was examined with isoform-specific polyclonal antibodies against hVMAT1 and hVMAT2. Central, peripheral, and enteric neurons express only VMAT2. VMAT1 is expressed exclusively in neuroendocrine, including chromaffin and enterochromaffin, cells. VMAT1 and VMAT2 are coexpressed in all chromaffin cells of the adrenal medulla. VMAT2 alone is expressed in histamine-storing enterochromaffin-like cells of the oxyntic mucosa of the stomach. The transport characteristics and pharmacology of each VMAT isoform have been directly compared after expression in digitonin-permeabilized fibroblastic (CV-1) cells, providing information about substrate feature recognition by each transporter and the role of vesicular monoamine storage in the mechanism of action of psychopharmacologic and neurotoxic agents in human. Serotonin has a similar affinity for both transporters. Catecholamines exhibit a 3-fold higher affinity, and histamine exhibits a 30-fold higher affinity, for VMAT2. Reserpine and ketanserin are slightly more potent inhibitors of VMAT2-mediated transport than of VMAT1-mediated transport, whereas tetrabenazine binds to and inhibits only VMAT2. N-methyl-4-phenylpyridinium, phenylethylamine, amphetamine, and methylenedioxymethamphetamine are all more potent inhibitors of VMAT2 than of VMAT1, whereas fenfluramine is a more potent inhibitor of VMAT1-mediated monamine transport than of VMAT2-mediated monoamine transport. The unique distributions of hVMAT1 and hVMAT2 provide new markers for multiple neuroendocrine lineages, and examination of their transport properties provides mechanistic insights into the pharmacology and physiology of amine storage in cardiovascular, endocrine, and central nervous system function.

Distribution of endogenous type B and type D sheep retrovirus sequences in ungulates and other mammals.

The jaagsiekte sheep retrovirus (JSRV), which appears to be a type B/D retrovirus chimera, has been incriminated as the cause of ovine pulmonary carcinoma. Recent studies suggest that the sequences related to this virus are found in the genomes of normal sheep and goats. To learn whether there are breeds of sheep that lack the endogenous viral sequences and to study their distribution among other groups of mammals, we surveyed several domestic sheep and goat breeds, other ungulates, and various mammal groups for sequences related to JSRV. Probes prepared from the envelope (SU) region of JSRV and the capsid (CA) region of a Peruvian type D virus related to JSRV were used in Southern blot hybridization with genomic DNA followed by low- and high-stringency washes. Fifteen to 20 CA and SU bands were found in all members of the 13 breeds of domestic sheep and 6 breeds of goats tested. There were similar findings in 6 wild Ovis and Capra genera. Within 22 other genera of Bovidae including domestic cattle, and 7 other families of Artiodactyla including Cervidae, there were usually a few CA or SU bands at low stringency and rare bands at high stringency. Among 16 phylogenetically distant genera, there were generally fewer bands hybridizing with either probe. These results reveal wide-spread phylogenetic distribution of endogenous type B and type D retroviral sequences related to JSRV among mammals and argue for further investigation of their potential role in disease.

The chromosomal distribution of mouse odorant receptor genes.

Odorant receptors (ORs) on nasal olfactory sensory neurons are encoded by a large multigene family. Each member of the family is expressed in a small percentage of neurons that are confined to one of several spatial zones in the nose but are randomly distributed throughout that zone. This pattern of expression suggests that when the sensory neuron selects which OR gene to express it may be confined to a particular zonal gene set of several hundred OR genes but select from among the members of that set via a stochastic mechanism. Both locus-dependent and locus-independent models of OR gene choice have been proposed. To investigate the feasibility of these models, we determined the chromosomal locations of 21 OR genes expressed in four different spatial zones. We found that OR genes are clustered within multiple loci that are broadly distributed in the genome. These loci lie within paralogous chromosomal regions that appear to have arisen by duplications of large chromosomal domains followed by extensive gene duplication and divergence. Our studies show that OR genes expressed in the same zone map to numerous loci; moreover, a single locus can contain genes expressed in different zones. These findings raise the possibility that OR gene choice may be locus-independent or involve consecutive stochastic choices.

Distribution of parthenogenetic cells in the mouse brain and their influence on brain development and behavior.

A systematic analysis of parthenogenetic (PG) cell fate within the central nervous system (CNS) was made throughout fetal development and neonatal and adult life. Chimeras were made between PG embryos carrying a ubiquitously expressed lacZ transgene and normal fertilized embryos. After detailed histological analysis, we find that the developmental potential of PG cells is spatially restricted to certain parts of the brain. PG cells are prevalent in telencephalic structures and are largely excluded from diencephalic structures, especially the hypothalamus. These spatial restrictions are established early in development. Behavioral studies with chimeras identified an increase in male aggression when the proportion of PG cells in the brain was high. These studies demonstrate that imprinted genes play key roles in development of the CNS and may be involved in behavior.

min K channels form by assembly of at least 14 subunits.

Injection of min K mRNA into Xenopus oocytes results in expression of slowly activating voltage-dependent potassium channels, distinct from those induced by expression of other cloned potassium channels. The min K protein also differs in structure, containing only a single predicted transmembrane domain. While it has been demonstrated that all other cloned potassium channels form by association of four independent subunits, the number of min K monomers which constitute a functional channel is unknown. In rat min K, replacement of Ser-69 by Ala (S69A) causes a shift in the current-voltage (I-V) relationship to more depolarized potentials; currents are not observed at potentials negative to 0 mV. To determine the subunit stoichiometry of min K channels, wild-type and S69A subunits were coexpressed. Injections of a constant amount of wild-type mRNA with increasing amounts of S69A mRNA led to potassium currents of decreasing amplitude upon voltage commands to -20 mV. Applying a binomial distribution to the reduction of current amplitudes as a function of the different coinjection mixtures yielded a subunit stoichiometry of at least 14 monomers for each functional min K channel. A model is presented for how min K subunits may form a channel.

Paleomagnetic and stable isotope measurements and distribution of benthic foraminifera in sediment core PS1388-3 from the Antarctic continental margin in the eastern Weddell Sea

A stable isotope record from the eastern Weddell Sea from 69°S is presented. For the first time, a 250,000-yr record from the Southern Ocean can be correlated in detail to the global isotope stratigraphy. Together with magnetostratigraphic, sedimentological and micropalaeontological data, the stratigraphic control of this record can be extended back to 910,000 yrs B.P. A time scale is constructed by linear interpolation between confirmed stratigraphic data points. The benthic d18O record (Epistominella exigua) reflects global continental ice volume changes during the Brunhes and late Matuyama chrons, whereas the planktonic isotopic record (Neogloboquadrina pachyderma) may be influenced by a meltwater lid caused by the nearby Antarctic ice shelf and icebergs. The worldwide climatic improvement during deglaciations is documented in the eastern Weddell Sea by an increase in production of siliceous plankton followed, with a time lag of approximately 10,000 yrs, by planktonic foraminifera production. Peak values in the difference between planktonic and benthic d13C records, which are 0.5 per mil higher during warm climatic periods than during times with expanded continental ice sheets, also suggest increased surface productivity during interglacials in the Southern Ocean.

Distribution of grain size and clay minerals in surface sediments of the Kara Sea (Fig 4, 5)

In this paper, we summarize data on terrigenous sediment supply in the Kara Sea and its accumulation and spatial and temporal variability during Holocene times. Sedimentological, organic-geochemical, and micropaleontological proxies determined in surface sediments allow to characterize the modern (riverine) terrigenous sediment input. AMS-14C dated sediment cores from the Ob and Yenisei estuaries and the adjacent inner Kara Sea were investigated to determine the terrigenous sediment fluxes and their relationship to paleoenvironmental changes. The variability of sediment fluxes during Holocene times is related to the post-glacial sea-level rise and changes in river discharge and coastal erosion input. Whereas during the late/middle Holocene most of the terrigenous sediments were deposited in the estuaries and the areas directly off the estuaries, huge amounts of sediments accumulated on the Kara Sea shelf farther north during the early Holocene before about 9 cal kyr BP. The maximum accumulation at that time is related to the lowered sea level, increased coastal erosion, and increased river discharge. Based on sediment thickness charts, echograph profiles and sediment core data, we estimate an average Holocene (0-11 cal kyr BP) annual accumulation of 194,106 t/yr of total sediment for the whole Kara Sea. Based on late Holocene (modern) sediment accumulation in the estuaries, probably 12,106 t/yr of riverine suspended matter (i.e. about 30% of the input) may escape the marginal filter on a geological time scale and is transported onto the open Kara Sea shelf. The high-resolution magnetic susceptibility record of a Yenisei core suggests a short-term variability in Siberian climate and river discharge on a frequency of 300-700 yr. This variability may reflect natural cyclic climate variations to be seen in context with the interannual and interdecadal environmental changes recorded in the High Northern Latitudes over the last decades, such as the NAO/AO pattern. A major decrease in MS values starting near 2.5 cal kyr BP, being more pronounced during the last about 2 cal kyr BP, correlates with a cooling trend over Greenland as indicated in the GISP-2 Ice Core, extended sea-ice cover in the North Atlantic, and advances of glaciers in western Norway. Our still preliminary interpretation of the MS variability has to be proven by further MS records from additional cores as well as other high-resolution multi-proxy Arctic climate records.

Quantitative composition and distribution of the silt fraction in surface sediments of the North-West African continental margin

Surface sediments from the continental slope and rise of North-West Africa between the Canary lslands and the Cape Verde Islands are mainly composed of silt-sized material (2-63 µm). A number of sampling profiles were run normal to the coast and the composition of the silt fraction was determined quantitatively by scanning electron microscope analysis. The carbonate portion of the sediment was found to be nearly exclusively of biogenic origin. The most important contributors are planktonic foraminifers and coccoliths with minor contributions derived from pteropods. Plankton-produced biogenic opal such as diatoms and radiolarians play a very minor role. The high production rates of opal-silica plankton which exists in the surface waters of the NW-African upwelling system does not give rise to corresponding increases of opal accumulation in the bottom sediment. Benthic producers consist mainly of foraminifers and molluscs but the entire input from benthic producers is extremely small. An exception to this occurs in the prodelta sediments of the Senegal river. Downslope particle transport is indicated by the occurrence of shallow-water coralline algae, ascidian sclerites and cliona boring chips and can be traced as far down as the continental rise. The non-carbonate silt fraction mostly consists of quartz which is derived as eolian dust from the Sahara desert by the Harmattan and the NE-Trade-wind system. The percentage of carbonate in the surface sediments directly indicates the relative proportions of autochthonous biogenic components and terrigenous allochthonous quartz particles.

Distribution of benthic foraminifera in surface sediments of the Portuguese continental margin

Living and dead benthic Foraminifera of 26 sediment surface samples from the East Atlantic continental margin (off Portugal) are studied. The stations are located on two profiles off Cape Mondego and off Cape Sines, ranging in water depth from 45 to 3905 meters. The highest values of standing crop are on the shelf (200 m) (up to 420 specimens/10 cm**3). Below 1000 m water depth standing crop is low (5 -24 specimens/10 cm**3). 151species and species groups are distinguished. Most of the living species do occur in a wide depth range. Faunal depth boundaries are at 50/100m, at 600/800 m, and at 1000 m. Results published from the North Atlantic and the East Mediterranean do not differ from those obtained in samples off Portugal. Depth of water (e.g. hydrostatic pressure) or another factor being controlled by depth (e.g. limitation of food supply) seems to be the most important factor of the benthic foraminiferal distribution.

(Table 1) Vertical distribution of relative contents of the main planktonic groups in the water column of the Kuril-Kamchatka region at Station VITYZ5635

Determinations were made of contents of carbon, lipids, nitrogen and, in some material, protein, carbohydrates, elementary composition of lipids and their spectral composition in total plankton samples from different depths (from the surface to 3000 m) and in several species of macroplanktonic deep-water crustaceans (decapods and mysids) living at different depths. Content of organic carbon and lipids in total plankton is high (40 to 60 and 35 to 70% of dry weight, respectively) and it does not change significantly with increasing depth. Deep-water macroplanktonic crustaceans have extremely high content of organic carbon and lipids, but there are no significant differences in this respect between species that live in different layers of the deep-water zone. Elementary composition of lipids indicates that they are highly saturated, with a marked predominance of unsaponifiable fraction, about 20% of which consists of methane hydrocarbons.

Distribution of Uvigerina proboscidea in Late Neogene sediments of DSDP Hole 22-214 in the northern Indian Ocean (Table 1)

This study attempts to understand the significance of Uvigerina proboscidea in paleoceanographic reconstructions at the northern (tropical) Indian Ocean DSDP Site 214 from the Late Miocene through the Pleistocene. In this interval at this site, U. proboscidea is the most abundant species of the benthic assemblage and shows abrupt frequency changes (about 1-74%). Based on relative percentages of U. proboscidea calibrated with oxygen and carbon isotope record and the sediment accumulation rates, the modern distribution of the species in the Indian Ocean, and other evidence, the peaks of abundance of U. proboscidea are inferred to represent times of high-surface productivity, This productivity is related to intensified trade winds during strong southwest (SW) Indian monsoons, causing widespread upwelling along equatorial divergemce in the Indian Ocean. The sudden increase of U. proboscidea abundance at approximately 8.5-7.5 Ma reflects significant upwelling at the equatorial divergence. This event corresponds to the permanent build-up of West Antarctic ice sheets, and a major increase in SW Indian monsoons related upwelling in the northwestern Indian Ocean. The Chron-6 carbon shift at approximately 6.2 Ma is marked by another peak of abundance, reflecting widespread ocean fertility. The highest abundances of U. proboscidea and highest sediment accumulation rates occur between 5.8 and 5.1 Ma, which coincidies with the greatest development of Antarctic ice sheets and strong southwest monsoons. The higher percentages at 3.2-3.1 Ma, approximately 2.4 Ma, and 1.6 Ma all represent phases of high productivity at the equatorial divergence.

Distribution of molluscs on the Atlantic continental shelf off Southern Spanish Sahara, West Africa

Two hundred and seventy five mollusc species from the continental shelf off Southern Spanish Sahara (depth: 32-60 m) were identified. Their distribution pattern is strongly influenced by the nature of the bottom (firm substrate, shelter, stability of sediment) rather than other factors at that depth interval. This faunal assemblage shows great affinity to the Mediterranean and Lusitanian faunas, and comprises only few (22 %) exclusively Senegalese and species living south of Senegal.