Most in vitro electrophysiology studies extract information and draw conclusions from representative, temporally limited snapshot experiments. This approach bears the risk of missing decisive moments that may make a difference in our understanding of physiological events. This feasibility study presents a simple benchtop cell-culture perfusion system adapted to commercial microelectrode arrays (MEAs), multichannel electrophysiology equipment and common inverted microscopy stages for simultaneous and uninterrupted extracellular electrophysiology and time-lapse imaging at ambient CO2 levels. The concept relies on a transparent, replica-casted polydimethylsiloxane perfusion cap, gravity- or syringe-pump-driven perfusion and preconditioning of pH-buffered serum-free cell-culture medium to ambient CO2 levels at physiological temperatures. The low-cost microfluidic in vitro enabling platform, which allows us to image cultures immediately after cell plating, is easy to reproduce and is adaptable to the geometries of different cell-culture containers. It permits the continuous and simultaneous multimodal long-term acquisition or manipulation of optical and electrophysiological parameter sets, thereby considerably widening the range of experimental possibilities. Two exemplary proof-of-concept long-term MEA studies on hippocampal networks illustrate system performance. Continuous extracellular recordings over a period of up to 70 days revealed details on both sudden and gradual neural activity changes in maturing cell ensembles with large intra-day fluctuations. Correlated time-lapse imaging unveiled rather static macroscopic network architectures with previously unreported local morphological oscillations on the timescale of minutes.

The estimation of motion direction from time varying retinal images is a fundamental task of visual systems. Neurons that selectively respond to directional visual motion are found in almost all species. In many of them already in the retina direction selective neurons signal their preferred direction of movement. Scientific evidences suggest that direction selectivity is carried from the retina to higher brain areas. Here we adopt a simple integrate-and-fire neuron model, inspired by recent work of Casti et al. (2008), to investigate how directional selectivity changes in cells postsynaptic to directional selective retinal ganglion cells (DSRGC). Our model analysis shows that directional selectivity in the postsynaptic cells increases over a wide parameter range. The degree of directional selectivity positively correlates with the probability of burst-like firing of presynaptic DSRGCs. Postsynaptic potentials summation and spike threshold act together as a temporal filter upon the input spike train. Prior to the intricacy of neural circuitry between retina and higher brain areas, we suggest that sharpening is a straightforward result of the intrinsic spiking pattern of the DSRGCs combined with the summation of excitatory postsynaptic potentials and the spike threshold in postsynaptic neurons.

Invasions by alien pathogens are a major threat to forest conservation. The North American fungal pathogen of conifers Heterobasidion irregulare, inadvertently introduced in Central Italy in the 1940s, has been spreading causing high mortality of Italian stone pine (Pinus pinea). While invading newfound niches, H. irregulare has established itself in the current range of the native congener H. annosum. The aims of this study were to determine whether in time: (I) H. irregulare populations may be increasing in size; (II) H. irregulare may be replacing H. annosum, rather than simply coexisting with it; and, (III) H. annosum may disappear in forests infested by H. irregulare. The presence, abundance and distribution of H. annosum and H. irregulare were assessed through an aerobiological assay replicated ten years apart in a forest in which both species have been coexisting. Replacement index (RI), Markov chains and geometric progressions were used to model the interspecific interaction between the two species and to assess the invasiveness of H. irregulare. Results showed that, in 10 years, the incidence of H. annosum dropped from 39.4 to 6.1%, while that of H. irregulare increased from 57.6 to 81.8%, with the alien pathogen replacing the native species (RI = 84.6%) and spreading at a maximum rate of 139 ha/year. Although our models show that the extinction of H. annosum may be unlikely, the ability of H. irregulare to replace it suggests the alien pathogen may also readily colonize those parts of Europe where H. annosum is more abundant than in Central Italy.

No abstract

The online version of the original article can be found at http://dx.doi.