The synapses in the cerebral cortex can be classified into two main types, Gray's type I and type II, which correspond to asymmetric (mostly glutamatergic excitatory) and symmetric (inhibitory GABAergic) synapses, respectively. Hence, the quantification and identification of their different types and the proportions in which they are found, is extraordinarily important in terms of brain function. The ideal approach to calculate the number of synapses per unit volume is to analyze 3D samples reconstructed from serial sections. However, obtaining serial sections by transmission electron microscopy is an extremely time consuming and technically demanding task. Using focused ion beam/scanning electron microscope microscopy, we recently showed that virtually all synapses can be accurately identified as asymmetric or symmetric synapses when they are visualized, reconstructed, and quantified from large 3D tissue samples obtained in an automated manner. Nevertheless, the analysis, segmentation, and quantification of synapses is still a labor intensive procedure. Thus, novel solutions are currently necessary to deal with the large volume of data that is being generated by automated 3D electron microscopy. Accordingly, we have developed ESPINA, a software tool that performs the automated segmentation and counting of synapses in a reconstructed 3D volume of the cerebral cortex, and that greatly facilitates and accelerates these processes.
Geometrical features of chemical synapses are relevant to their function. Two critical components of the synaptic junction are the active zone (AZ) and the postsynaptic density (PSD), as they are related to the probability of synaptic release and the number of postsynaptic receptors, respectively. Morphological studies of these structures are greatly facilitated by the use of recent electron microscopy techniques, such as combined focused ion beam milling and scanning electron microscopy (FIB/SEM), and software tools that permit reconstruction of large numbers of synapses in three dimensions. Since the AZ and the PSD are in close apposition and have a similar surface area, they can be represented by a single surface—the synaptic apposition surface (SAS). We have developed an efficient computational technique to automatically extract this surface from synaptic junctions that have previously been three-dimensionally reconstructed from actual tissue samples imaged by automated FIB/SEM. Given its relationship with the release probability and the number of postsynaptic receptors, the surface area of the SAS is a functionally relevant measure of the size of a synapse that can complement other geometrical features like the volume of the reconstructed synaptic junction, the equivalent ellipsoid size and the Feret's diameter.
The possibilities for improving the control of greenhouse fogging systems, were studied by comparing several combinations of ventilation cooling techniques, shade screening and low-pressure fogging. The study was divided into three parts: experiments, modelling and simulations.In the first part of the paper, ten combinations of five cooling techniques were tested during the summers of 2002 and 2003 in a 132 m 2 greenhouse with a steel structure and a single-layer methacrylate cover located in Madrid, Spain. An analysis of variance of the climatic parameters was carried out to determine which combinations produced significant differences in inside temperature or relative humidity. Comparing the values for the inside to outside temperature difference, the combination of a shade screen and above-screen fogging achieved a difference in temperature almost the same as that for under-screen fogging, but the relative humidity was significantly lower.In the second part of the study a dynamic model was developed (2002) and validated (2003). The mean absolute error obtained for inside temperature was similar in the fit and the validation and it was less than 1.5 °C in both cases.The model was used to simulate the inside air temperature for a fog system working without shading, and above and under a shade screen. Control algorithms were developed for reducing system water consumption. In the three cases a simple on/off control with a fixed fogging cycle was compared with a pulse width modulation (PWM) strategy, in which the duration of the fogging pulse was increased as a function of inside temperature. The strategies with PWM applied to the fog system were able to reduce water consumption by 8-15% with respect to the strategies with a fixed fogging cycle.
Dendritic spines establish most excitatory synapses in the brain and are located in Purkinje cell's dendrites along helical paths, perhaps maximizing the probability to contact different axons. To test whether spine helixes also occur in neocortex, we reconstructed Ͼ500 dendritic segments from adult human cortex obtained from autopsies. With Fourier analysis and spatial statistics, we analyzed spine position along apical and basal dendrites of layer 3 pyramidal neurons from frontal, temporal, and cingulate cortex. Although we occasionally detected helical positioning, for the great majority of dendrites we could not reject the null hypothesis of spatial randomness in spine locations, either in apical or basal dendrites, in neurons of different cortical areas or among spines of different volumes and lengths. We conclude that in adult human neocortex spine positions are mostly random. We discuss the relevance of these results for spine formation and plasticity and their functional impact for cortical circuits.
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