Ephrin-dependent growth and pruning of hippocampal axons

Neuronal connections are arranged topographically such that the spatial organization of neurons is preserved by their termini in the targets. During the development of topographic projections, axons initially explore areas much wider than the final targets, and mistargeted axons are pruned later. The molecules regulating these processes are not known. We report here that the ligands of the Eph family tyrosine kinase receptors may regulate both the initial outgrowth and the subsequent pruning of axons. In the presence of ephrins, the outgrowth and branching of the receptor-positive hippocampal axons are enhanced. However, these axons are induced later to degenerate. These observations suggest that the ephrins and their receptors may regulate topographic map formation by stimulating axonal arborization and by pruning mistargeted axons.

EphA4 (Sek1) receptor tyrosine kinase is required for the development of the corticospinal tract

Members of the Eph family of tyrosine kinase receptors have been implicated in the regulation of developmental processes and, in particular, axon guidance in the developing nervous system. The function of the EphA4 (Sek1) receptor was explored through creation of a null mutant mouse. Mice with a null mutation in the EphA4 gene are viable and fertile but have a gross motor dysfunction, which is evidenced by a loss of coordination of limb movement and a resultant hopping, kangaroo-like gait. Consistent with the observed phenotype, anatomical studies and anterograde tracing experiments reveal major disruptions of the corticospinal tract within the medulla and spinal cord in the null mutant animals. These results demonstrate a critical role for EphA4 in establishing the corticospinal projection.

Implications of EPHB6, EFNB2, and EFNB3 expressions in human neuroblastoma

Neuroblastoma (NB) is a common pediatric tumor that exhibits a wide range of biological and clinical heterogeneity. EPH (erythropoietin-producing hepatoma amplified sequence) family receptor tyrosine kinases and ligand ephrins play pivotal roles in neural and cardiovascular development. High-level expression of transcripts encoding EPHB6 receptors (EPHB6) and its ligands ephrin-B2 and ephrin-B3 (EFNB2, EFNB3) is associated with low-stage NB (stages 1, 2, and 4S) and high TrkA expression. In this study, we showed that EFNB2 and TrkA expressions were associated with both tumor stage and age, whereas EPHB6 and EFNB3 expressions were solely associated with tumor stage, suggesting that these genes were expressed in distinct subsets of NB. Kaplan-Meier and Cox regression analyses revealed that high-level expression of EPHB6, EFNB2, and EFNB3 predicted favorable NB outcome (P < 0.005), and their expression combined with TrkA expression predicted the disease outcome more accurately than each variable alone (P < 0.00005). Interestingly, if any one of the four genes (EPHB6, EFNB2, EFNB3, or TrkA) was expressed at high levels in NB, the patient survival was excellent (>90%). To address whether a good disease outcome of NB was a consequence of high-level expression of a “favorable NB gene,” we examined the effect of EPHB6 on NB cell lines. Transfection of EPHB6 cDNA into IMR5 and SY5Y expressing little endogenous EPHB6 resulted in inhibition of their clonogenicity in culture. Furthermore, transfection of EPHB6 suppressed the tumorigenicity of SY5Y in a mouse xenograft model, demonstrating that high-level expressions of favorable NB genes, such as EPHB6, can in fact suppress malignant phenotype of unfavorable NB.

Regulation of topographic projection in the brain: Elf-1 in the hippocamposeptal system.

The hippocampus and septum play central roles in one of the most important spheres of brain function: learning and memory. Although their topographic connections have been known for two decades and topography may be critical for cognitive functions, the basis for hippocamposeptal topographic projection is unknown. We now report for the first time that Elf-1, a membrane-bound eph family ligand, is a candidate molecular tag for the genesis of the hippocamposeptal topographic projection. Elf-1 is expressed in an increasing gradient from dorsal to ventral septum. Furthermore, Elf-1 selectively allows growth of neurites from topographically appropriate lateral hippocampal neurons, while inhibiting neurite outgrowth by medial hippocampal neurons. Complementary to the expression of Elf-1, an eph family receptor, Bsk, is expressed in the hippocampus in a lateral to medial gradient, consistent with a function as a receptor for Elf-1. Further, Elf-1 specifically bound Bsk, eliciting tyrosine kinase activity. We conclude that the Elf-1/Bsk ligand-receptor pair exhibits traits of a chemoaffinity system for the organization of hippocamposeptal topographic projections.

A tyrosine kinase profile of prostate carcinoma.

Tyrosine kinases play central roles in the growth and differentiation of normal and tumor cells. In this study, we have analyzed the general tyrosine kinase expression profile of a prostate carcinoma (PCA) xenograft, CWR22. We describe here an improved reverse transcriptase-PCR approach that permits identification of nearly 40 different kinases in a single screening; several of these kinases are newly cloned kinases and some are novel. According to this, there are 11 receptor kinases, 9 nonreceptor kinases, and at least 7 dual kinases expressed in the xenograft tissue. The receptor kinases include erbB2, erbB3, Ret, platelet-derived growth factor receptor, sky, nyk, eph, htk, sek (eph), ddr, and tkt. The nonreceptor kinases are lck, yes, abl, arg, JakI, tyk2, and etk/bmx. Most of the dual kinases are in the mitogen-activating protein (MAP) kinase-kinase (MKK) family, which includes MKK3, MKK4, MEK5, and a novel one. As a complementary approach, we also analyzed by specific reverse transcriptase-PCR primers the expression profile of erbB/epidermal growth factor receptor family receptors in a variety of PCA specimens, cell lines, and benign prostatic hyperplasia. We found that erbB1, -2, and -3 are often coexpressed in prostate tissues, but not in erbB4. The information established here should provide a base line to study the possible growth and oncogenic signals of PCA.

Contribution différentielle de Neuroligine‐1 et d’EphA4 à la régulation du sommeil

Le sommeil est un besoin vital et le bon fonctionnement de l’organisme dépend de la quantité et de la qualité du sommeil. Le sommeil est régulé par deux processus : un processus circadien qui dépend de l’activité des noyaux suprachiasmatiques de l’hypothalamus et qui régule le moment durant lequel nous allons dormir, et un processus homéostatique qui dépend de l’activité neuronale et se reflète dans l’intensité du sommeil. En effet, le sommeil dépend de l’éveil qui le précède et plus l’éveil dure longtemps, plus le sommeil est profond tel que mesuré par des marqueurs électroencéphalographiques (EEG). Des études ont montré que le bon fonctionnement de ces deux processus régulateurs du sommeil dépend de la plasticité synaptique. Ainsi, les éléments synaptiques régulant la communication et la force synaptique sont d’importants candidats pour agir sur la physiologie de la régulation du sommeil. Les molécules d’adhésion cellulaire sont des acteurs clés dans les mécanismes de plasticité synaptique. Elles régulent l’activité et la maturation des synapses. Des études ont montré que leur absence engendre des conséquences similaires au manque de sommeil. Le but de ce projet de thèse est d’explorer l’effet de l’absence de deux familles de molécule d’adhésion cellulaire, les neuroligines et la famille des récepteur Eph et leur ligand les éphrines dans les processus régulateurs du sommeil. Notre hypothèse est que l’absence d’un des membres de ces deux familles de molécule affecte les mécanismes impliqués dans le processus homéostatique de régulation du sommeil. Afin de répondre à notre hypothèse, nous avons étudié d’une part l’activité EEG chez des souris mutantes n’exprimant pas Neuroligine‐1 (Nlgn1) ou le récepteur EphA4 en condition normale et après une privation de sommeil. D’autre part, nous avons mesuré les changements moléculaires ayant lieu dans ces deux modèles après privation de sommeil. Au niveau de l’activité EEG, nos résultats montrent que l’absence de Nlgn1 augmente la densité des ondes lentes en condition normale et augment l’amplitude et la pente des ondes lentes après privation de sommeil. Nlgn1 est nécessaire au fonctionnement normal de la synchronie corticale, notamment après une privation de sommeil, lui attribuant ainsi un rôle clé dans l’homéostasie du sommeil. Concernant le récepteur EphA4, son absence affecte la durée du sommeil paradoxal ainsi que l’activité sigma qui dépendent du processus circadien. Nos résultats suggèrent donc que ce récepteur est un élément important dans la régulation circadienne du sommeil. Les changements transcriptionnels en réponse à la privation de sommeil des souris n’exprimant pas Nlgn1 et EphA4 ne sont pas différents des souris sauvages. Toutefois, nous avons montré que la privation de sommeil affectait la distribution des marques épigénétiques sur le génome, tels que la méthylation et l’hydroxyméthylation, et que l’expression des molécules régulant ces changements est modifiée chez les souris mutantes pour le récepteur EphA4. Nos observations mettent en évidence que les molécules d’adhésion cellulaire, Nlgn1 et le récepteur EphA4, possèdent un rôle important dans les processus homéostatique et circadien du sommeil et contribuent de manière différente à la régulation du sommeil.

Pollen abundance in Late Quaternary sediment cores off West Africa

The pollen record of three marine late Quaternary cores off Senegal shows a juxtaposition of Mediterranean, Northern Saharan, Central Saharan elements, which are considered transported by the trade winds from a winter-rainfall area, and Sahelian, Soudanese, Soudano-Guinean elements, considered transported both by winds and mostly by the Senegal River, and coming from the monsoonal, summer tropical rainfall area of southern West Africa. Littoral vegetation is either the edaphically dry and saline Chenopodiaceae from sebkhas at the time of the main regression, or the warm tropical humid mangrove with Rhizophora during the humid optimum period. Four stratigraphic zones reflect, from basis to top: Zone 4. A semi-arid period with a balanced pollen input. Zone 3. A very arid period with the disappearance of monsoonal pollen, probably from the disappearance of the Senegal River, a very saline littoral plain with Chenopodiaceae, a larger input of northern Saharan pollen from intensified trade winds. Zone 2. A quite humid period, much more so than today, very suddenly established, with a northward extension of the monsoonal areas, a rich littoral mangrove, and weakening of the trade winds. Zone l. A slow and steady evolution toward the present semi-humid conditions with regression of the mangrove, and of the monsoonal areas toward the south. Tentative datations and correlations with the Tchad area suggested: zone 4: 22,500 to 19,000 years BP; zone 3: 19,000 to 12,500 years BP; zone 2: 12,500 to 5,500 years BP; zone 1: 5,500 years BP to top of core. Dinoflagellate cysts display a tropical assemblage with mostly estuarine neritic elements and also a weak oceanic component, mostly in the lower slope core 47. Cosmopolitan taxa dominate the assemblage and only a few species point to more specialized environments. Quantitative variations of the assemblage are the basis of stratigraphy which is not similar to the pollen stratigraphy, and an inshore-outshore gradient has to be taken into account to correlate the three cores.

Pollen record and age determiation of sediments from the Labaz Lake area

Pollen records from perennially frozen sequences provide vegetation and climate reconstruction for the last 48,000 14C years in the central part of Taymyr Peninsula. Open larch forest with Alnus fruticosa and Betula nana grew during the Kargin (Middle Weichselian) Interstade, ca. 48,000-25,000 14C yr B.P. The climate was generally warmer and wetter than today. Open steppe-like communities with Artemisia, Poaceae, Asteraceae, and herb tundralike communities with dwarf Betula and Salix dominated during the Sartan (Late Weichselian) Stade, ca. 24,000-10,300 14C yr B.P. The statistical information method used for climate reconstruction shows that the coldest climate was ca. 20,000-17,000 14C yr B.P. A warming (Allerød Interstade?) with mean July temperature ca. 1.5°C warmer than today occurred ca. 12,000 14C yr B.P. The following cooling with temperatures about 3°-4°C cooler than present and precipitation about 100 mm lower corresponds well with the Younger Dryas Stade. Tundra-steppe vegetation changed to Betula nana-Alnus fruticosa shrub tundra ca. 10,000 14C yr B.P. Larch appeared in the area ca. 9400 14C yr B.P. and disappeared after 2900 14C yr B.P. Cooling events ca. 10,500, 9600, and 8200 14C yr B.P. characterized the first half of the Holocene. A significant warming occurred ca. 8500 14C yr B.P., but the Holocene temperature maximum was at about 6000-4500 14C yr B.P. The vegetation cover approximated modern conditions ca. 2800 14C yr B.P. Late Holocene warming events occurred at ca. 3500, 2000, and 1000 14C yr B.P. A cooling (Little Ice Age?) took place between 500 and 200 14C yr ago.

The late Pliocene pollen record of ODP Site 108-658 off Cape Blanc, Atlantic Ocean

A 200 m long marine pollen record from ODP Site 658 (21°N, 19°W) reveals cyclic fluctuations in vegetation and continental climate in northwestern Africa from 3.7 to 1.7 Ma. These cycles parallel oxygen isotope stages. Prior to 3.5 Ma, the distribution of tropical forests and mangrove swamps reached Cape Blanc, 5°N of the present distribution. Between 3.5 and 2.6 Ma, forests occurred at this latitude during irregular intervals and nearly disappeared afterwards. Likewise, a Saharan paleoriver flowed continuously until isotope Stage 134 (3.35 Ma). When river discharge ceased, wind transport of pollen grains prevailed over fluvial transport. Pollen indicators of trade winds gradually increased between 3.3 and 2.5 Ma. A strong aridification of the climate of northwestern Africa occurred during isotope Stage 130 (3.26 Ma). Afterwards, humid conditions reestablised followed by another aridification around 2.7 Ma. Repetitive latitudinal shifts of vegetation zones ranging from wooded savanna to desert flora dominated for the first time between between 2.6 and 2.4 Ma as a response to the glacial stages 104, 100 and 98. Although climatic conditions, recorded in the Pliocene, were not as dry as those of the middle and Late Pleistocene, latitudinal vegetation shifts near the end of the Pliocene resembled those of the interglacial-glacial cycles of the Brunhes chron.

Results of a sediment profile and water analyses from Lake Hancza in northeastern Poland

We investigated the sedimentary record of Lake Hancza (northeastern Poland) using a multi-proxy approach, focusing on early to mid-Holocene climatic and environmental changes. AMS 14C dating of terrestrial macrofossils and sedimentation rate estimates from occasional varve thickness measurements were used to establish a chronology. The onset of the Holocene at c. 11600 cal. a BP is marked by the decline of Lateglacial shrub vegetation and a shift from clastic-detrital deposition to an autochthonous sedimentation dominated by biochemical calcite precipitation. Between 10000 and 9000 cal. a BP, a further environmental and climatic improvement is indicated by the spread of deciduous forests, an increase in lake organic matter and a 1.7% rise in the oxygen isotope ratios of both endogenic calcite and ostracod valves. Rising d18O values were probably caused by a combination of hydrological and climatic factors. The persistence of relatively cold and dry climate conditions in northeastern Poland during the first one and a half millennia of the Holocene could be related to a regional eastern European atmospheric circulation pattern. Prevailing anticyclonic circulation linked to a high-pressure cell above the retreating Scandinavian Ice Sheet might have blocked the influence of warm and moist Westerlies and attenuated the early Holocene climatic amelioration in the Lake Hancza region until the final decay of the ice sheet.

Changes in the vegetation and trade winds in equatorial Northwest Africa 140,000-70,000 yr B.P., as deduced from two marine pollen records

The deep-sea cores M 16415-2 and M 16416-2 at about 9°N off Sierra Leone were analysed palynologically for the time interval 140,000-70,000 yr B.P. Results were presented in absolute (pollen concentration and pollen influx) and relative diagrams (pollen percentage). In a previous study it was evidenced that in northwest Africa pollen is mainly transported to the Atlantic by wind, so that the efficiency of aeolian pollen transport (pollen flux) could be used to evaluate changes in the intensity of the northeast trade winds. The glacial episodes (represented by the oxygen isotope stages 6 and 4) are characterized by strong northeast trade winds, whereas the last interglacial (stage 5) is characterized by weak trade winds. The pollen influx diagram shows that the intensity of the trade winds increased slightly during the relatively cool intervals of stage 5 (viz. 5.4 and 5.2). Tropical forest had maximally expanded around 124,000 yr B.P. (stage 5.5), around 98,000 yr B.P. (transition of stage 5.3 to 5.2), and around 70,000 yr B.P. (first part of stage 4): an increasing delay of the response of tropical forest to global intervals with maximum temperature is apparent during the last interglacial. As tropical forests need continuous humidity, the record of tropical forest monitors changes in climatic humidity south of the Sahara. During the last interglacial, the southern boundary of the Sahara shifted only little: expansions and contractions of the tropical forest area are correlated with contra-oscillations of the grass-dominated savanna zone. Great latitudinal shifts of the desert savanna boundary, on the contrary, occurred during the penultimate glacial interglacial transition (around 128,000 yr B.P.) to the north, and during the last interglacial-glacial transition (around 65,000 yr B.P.) to the south.

Palynology on lake sediments from the Pyrenees

A multi-proxy palaeoecological investigation including pollen, plant macrofossil, radiocarbon and sedimentological analyses, was performed on a small mountain lake in the Eastern Pyrenees. This has allowed the reconstruction of: (1) the vegetation history of the area based on five pollen diagrams and eight AMS14C dates and (2) the past lake-level changes, based on plant macrofossil, lithological and pollen analysis of two stratigraphical transects correlated by pollen analysis. The palaeolake may have appeared before the Younger Dryas; the lake-level was low and the vegetation dominated by cold steppic grasslands. The lake-level rose to its highest level during the Holocene in the Middle Atlantic (at ca. 5060±45 b.p.). Postglacial forests (Quercetum mixtum and Abieto-Fagetum) developed progressively in the lower part of the valley, while dense Pinus uncinata forests rapidly invaded the surroundings of the mire and remained the dominant local vegetation until present. The observed lowering of the lake levels during the Late Atlantic and the Subboreal (from 5060 ± B.P. to 3590±40 b.p.) was related to the overgrowth of the mire. The first obvious indications of anthropogenic disturbances of the vegetation are recorded at the Atlantic/Subboreal boundary as a reduction in the forest component, which has accelerated during the last two millennia.

Pollen profiles from a Late Palaeolithic site in the community of Bad Buchau-Kappel, Baden-Wuerttemberg, Germany

Excavations were carried out in a Late Palaeolithic site in the community of Bad Buchau-Kappel between 2003 and 2007. Archaeological investigations covered a total of more than 200 m**2. This site is the product of what likely were multiple occupations that occurred during the Late Glacial on the Federsee shore in this location. The site is situated on a mineral ridge that projected into the former Late Glacial lake Federsee. This beach ridge consists of deposits of fine to coarse gravel and sand and was surrounded by open water, except for a connection to the solid shore on the south. A lagoon lay between the hook-shaped ridge and the shore of the Federsee. This exposed location provided optimal access to the water of the lake. In addition, the small lagoon may have served as a natural harbor for landing boats or canoes. Sedimentological and palynological investigations document the dynamic history of the location between 14,500 and 11,600 years before present (cal BP). Evidence of the deposition of sands, gravels and muds since the Bølling Interstadial is provided by stratigraphic and palynological analyses. The major occupation occurred in the second half of the Younger Dryas period. Most of the finds were located on or in the sediments of the ridge; fewer finds occurred in the surrounding mud, which was also deposited during the Younger Dryas. Direct dates on some bone fragments, however, demonstrate that intermittent sporadic occupations also took place during the two millennia of the Meiendorf, Bølling, and Allerød Interstadials. These bones were reworked during the Younger Dryas and redeposited in the mud. A 14C date from one bone of 11,600 years ago (cal BP) places the Late Palaeolithic occupation of the ridge at the very end of the Younger Dryas, which is in agreement with stratigraphic observations. Stone artifacts, numbering 3,281, comprise the majority of finds from the site. These include typical artifacts of the Late Palaeolithic, such as backed points, short scrapers, and small burins. There are no bipointes or Malaurie-Points, which is in accord with the absolute date of the occupation. A majority of the artifacts are made from a brown chert that is obtainable a few kilometers north of the site in sediments of the Graupensandrinne. Other raw materials include red and green radiolarite that occur in the fluvioglacial gravels of Oberschwaben, as well as quartzite and lydite. The only non-local material present is a few artifacts of tabular chert from the region near Kelheim in Bavaria. A unique find consists of two fragments of a double-barbed harpoon made of red deer antler, which was found in the Younger Dryas mud. It is likely, but not certain, that this find belongs to the same assemblage as the numerous stone artifacts. Although not numerous, animal bones were also found in the excavations. Most of them lay in sediments of the Younger Dryas, but several 14C dates place some of these bones in earlier periods, including the Meiendorf, Bølling, and Allerød Interstadials. These bones were reworked by water and redeposited in mud sediments during the Younger Dryas. As a result, it is difficult to attribute individual bones to particular chronological positions without exact dates. Species that could be identified include wild horse (Equus spec.), moose or elk (Alces alces), red deer (Cervus elaphus), roe deer (Capreolus capreolus), aurochs or bison (Bos spec.), wild boar (Sus scrofa), as well as birds and fish, including pike (Esox Lucius).

Palynological and stratigraphic record from Löddigsee in Mecklenburg-Vorpommern, Germany

A Late Pleistocene and Holocene sediment core from the nowadays terrestrialised portion of the Löddigsee in Southern Mecklenburg, Germany was palynologically investigated. The lake is situated in the rarely investigated Young moraine area at the transition from the Weichselian to the Saalian glaciation. The high-resolution pollen diagram contributes to the establishment of the north-eastern German Late Pleistocene pollen stratigraphy. The vegetation distribution pattern after the end of the Weichselian is in good agreement with other studies from North-eastern Germany, but also has its own characteristics. The Holocene vegetation development reveals features from the north-eastern and north-western German lowlands. A special focus was laid on the environmental history of the two settlements on an island within the lake (Late Neolithic and Younger Slavic period), which were preserved under moist conditions. Both settlements were constructed during a period of low lake level. Although there is evidence of agriculture in the area during the respective periods, the two island settlements seem to have served other purposes.

Palynology of ODP Leg 127 sites

This initial survey of pollen from 192 samples from Hole 794A, supplemented by 189 samples from Hole 795 and 797B, suggests that marine pollen assemblages from the southwestern Sea of Japan provide a consistent Neogene pollen stratigraphy and a solid basis for regional paleoenvironmental reconstructions. Late Miocene vegetation inferred from these pollen data, a mix of conifer and broad-leaf elements with now-extinct Tertiary types well represented, appears similar to Aniai-type floras of Japan. During the late Miocene through early Pliocene, as Tertiary types declined, conifers (including the Sequoia/Cryptomeria group) became more prominent than broad-leaf elements, and herbs played an increasing role in the vegetation. Middle Pliocene pollen assemblages imply significant changes in forest composition. In a 500,000-yr interval centered at ~4 m.y., Tertiary and warm-temperate deciduous types re-expanded and were comparable to or greater than middle-late Miocene levels. Temperate and cold-temperate conifers {Picea, Abies, Tsuga) were minimal. Subsequently, Tertiary and deciduous forest components (including Quercus) decreased, Picea, Tsuga, and Abies were again prominent, and herbs formed an increasingly larger part of the vegetation. Between ~3 m.y. and -2.5 m.y., conifers, except for Cryptomeria types, were prominent, Quercus continued to decline, and other broad-leaf trees were minor. Over the last 2 Ma, the very large and frequent changes in forest composition inferred from pollen in the Sea of Japan correspond to forest dynamics inferred from changes in pollen and floral assemblages throughout Japan. Given present vegetation/climate relationships, broad trends in Neogene climate inferred from these preliminary pollen data include decreasing temperatures, increasing seasonality in temperatures and precipitation, and increasing amplitude and frequency of climatic change. Two significant events, centered at ~9 m.y. and ~4 m.y., punctuate the gradual deterioration of the equable warm, humid subtropical/warm temperate late Miocene and early Pliocene climates. The first indication of cold-temperate conditions comparable to those of Pleistocene glacial intervals occurs ~3 m.y. Subsequently, regional climates oscillated rapidly between temperate and cold-temperate regimes that supported conifer and mixed broad-leaf forests; however, climatic extremes were apparently never great enough to displace warm-temperate and temperate forests from Honshu nor to produce arctic climates on the west coast of Japan.