Threshold-Free Population Analysis Identifies Larger DRG Neurons to Respond Stronger to NGF StimulationReportar como inadecuado




Threshold-Free Population Analysis Identifies Larger DRG Neurons to Respond Stronger to NGF Stimulation - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Sensory neurons in dorsal root ganglia DRG are highly heterogeneous in terms of cell size, protein expression, and signaling activity. To analyze their heterogeneity, threshold-based methods are commonly used, which often yield highly variable results due to the subjectivity of the individual investigator. In this work, we introduce a threshold-free analysis approach for sparse and highly heterogeneous datasets obtained from cultures of sensory neurons. This approach is based on population estimates and completely free of investigator-set parameters. With a quantitative automated microscope we measured the signaling state of single DRG neurons by immunofluorescently labeling phosphorylated, i.e., activated Erk1-2. The population density of sensory neurons with and without pain-sensitizing nerve growth factor NGF treatment was estimated using a kernel density estimator KDE. By subtraction of both densities and integration of the positive part, a robust estimate for the size of the responsive subpopulations was obtained. To assure sufficiently large datasets, we determined the number of cells required for reliable estimates using a bootstrapping approach. The proposed methods were employed to analyze response kinetics and response amplitude of DRG neurons after NGF stimulation. We thereby determined the portion of NGF responsive cells on a true population basis. The analysis of the dose dependent NGF response unraveled a biphasic behavior, while the study of its time dependence showed a rapid response, which approached a steady state after less than five minutes. Analyzing two parameter correlations, we found that not only the number of responsive small-sized neurons exceeds the number of responsive large-sized neurons—which is commonly reported and could be explained by the excess of small-sized cells—but also the probability that small-sized cells respond to NGF is higher. In contrast, medium-sized and large-sized neurons showed a larger response amplitude in their mean Erk1-2 activity.



Autor: Christine Andres , Jan Hasenauer , Frank Allgower, Tim Hucho

Fuente: http://plos.srce.hr/



DESCARGAR PDF




Documentos relacionados