Two distinct filopodia populations at the growth cone allow to sense nanotopographical extracellular matrix cues to guide neurite outgrowth

BACKGROUND The process of neurite outgrowth is the initial step in producing the neuronal processes that wire the brain. Current models about neurite outgrowth have been derived from classic two-dimensional (2D) cell culture systems, which do not recapitulate the topographical cues that are present in the extracellular matrix (ECM) in vivo. Here, we explore how ECM nanotopography influences neurite outgrowth. METHODOLOGY/PRINCIPAL FINDINGS We show that, when the ECM protein laminin is presented on a line pattern with nanometric size features, it leads to orientation of neurite outgrowth along the line pattern. This is also coupled with a robust increase in neurite length. The sensing mechanism that allows neurite orientation occurs through a highly stereotypical growth cone behavior involving two filopodia populations. Non-aligned filopodia on the distal part of the growth cone scan the pattern in a lateral back and forth motion and are highly unstable. Filopodia at the growth cone tip align with the line substrate, are stabilized by an F-actin rich cytoskeleton and enable steady neurite extension. This stabilization event most likely occurs by integration of signals emanating from non-aligned and aligned filopodia which sense different extent of adhesion surface on the line pattern. In contrast, on the 2D substrate only unstable filopodia are observed at the growth cone, leading to frequent neurite collapse events and less efficient outgrowth. CONCLUSIONS/SIGNIFICANCE We propose that a constant crosstalk between both filopodia populations allows stochastic sensing of nanotopographical ECM cues, leading to oriented and steady neurite outgrowth. Our work provides insight in how neuronal growth cones can sense geometric ECM cues. This has not been accessible previously using routine 2D culture systems.

Abundance and biomass of zooplankton from 1986-1994 collected in front of Constanta city, Black Sea

The Est Constanta 1986-1994 dataset contains zooplankton data collected allong a 5 station transect in front of the city Constanta (44°10'N, 28°41.5'E - EC1; 44°10'N, 28°47'E - EC2; 44°10'N, 28°54'E - EC3; 44°10'N, 29°08'E - EC4; 44°10'N, 29°22'E - EC5). Zooplankton sampling was undertaken at 5 stations where samples were collected using a Juday closing net in the 0-10, 10-25, 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance of mesozooplankton in the western part of the Black Sea in summer 1998 during the National Monitoring Programme

The dataset is based on samples collected in the summer of 1998 in the Western Black Sea in front of Bulgaria coast. The whole dataset is composed of 69 samples (from 22 stations of National Monitoring Grid) with data of mesozooplankton species composition abundance and biomass. Samples were collected in discrete layers 0-10, 0-20, 0-50, 10-25, 25-50, 50-100 and from bottom up to the surface at depths depending on water column stratification and the thermocline depth. Zooplankton samples were collected with vertical closing Juday net,diameter - 36cm, mesh size 150 µm. Tows were performed from surface down to bottom meters depths in discrete layers. Samples were preserved by a 4% formaldehyde sea water buffered solution. Sampling volume was estimated by multiplying the mouth area with the wire length. Mesozooplankton abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Lyudmila Kamburska using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972). Taxon-specific abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Copepods and Cladoceras were identified and enumerated; the other mesozooplankters were identified and enumerated at higher taxonomic level (commonly named as mesozooplankton groups). Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Lyudmila Kamburska using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972).

Abundance and biomass of mesozooplankton from the Romanian littoral during DANUBS_2002 in spring and autumn 2002

The Danubs 2002 dataset contains zooplankton data collected in April, June,September and October 2002 in 11 station allong 5 transect in front of the Romanian littoral. Zooplankton sampling was undertaken at 11 stations where samples were collected using a Juday closing net in the 0-10, 10-25, and 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance of mesozooplankton in the western part of the Black Sea in summer 2001 during the National Monitoring Programme of the Bulgarian Institute of Oceanography

The dataset is based on samples collected in the summer of 2001 in the Western Black Sea in front of Bulgaria coast (transects at c. Kaliakra and c. Galata). The whole dataset is composed of 26 samples (from 10 stations of National Monitoring Grid) with data of mesozooplankton species composition abundance and biomass. Samples were collected in discrete layers 0-10, 10-20, 10-25, 25-50, 50-75, 75-90. Zooplankton samples were collected with vertical closing Juday net,diameter - 36cm, mesh size 150 µm. Tows were performed from surface down to bottom meters depths in discrete layers. Samples were preserved by a 4% formaldehyde sea water buffered solution. Sampling volume was estimated by multiplying the mouth area with the wire length. Mesozooplankton abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Lyudmila Kamburska and Kremena Stefanova using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972). Taxon-specific abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Copepods and Cladoceras were identified and enumerated; the other mesozooplankters were identified and enumerated at higher taxonomic level (commonly named as mesozooplankton groups). Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Lyudmila Kamburska and Kremena Stefanova using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972).