quantifications of western blots with imagej ncbi

#QUANTIFICATIONS OF WESTERN BLOTS WITH IMAGEJ NCBI SOFTWARE#

In addition, it was shown in experiments performed in anesthetized rats that whisker stimulation caused a significant increase of glucose uptake in astrocytes, but not in neurons, in the activated somatosensory cortex ( Chuquet et al., 2010). First, it was demonstrated in cerebellar slices using fluorescent glucose analogs that both glucose transport and metabolism occur preferentially in Bergmann glia over Purkinje cells while the same observation was made for astrocytes over neurons in hippocampal slices ( Barros et al., 2009 Jakoby et al., 2014). Such a view is reinforced by recent observations made ex vivo and in vivo. As a consequence, a major part of glucose uptake taking place in an activated brain region and visualized by functional brain imaging is due to astrocytic utilization ( Pellerin and Magistretti, 1996b, 2012 Figley and Stroman, 2011). Glutamate enhances both glucose transport ( Loaiza et al., 2003) as well as uptake and utilization ( Pellerin and Magistretti, 1994, 1996a) in cultured astrocytes. Glutamatergic synapses are by far the predominant type of synapses in the central nervous system and constitute the main source of excitatory neurotransmission. Despite these important developments, the specific fate of glucose at the cellular and molecular levels remains elusive. This tight relationship has formed the basis for the development of some functional brain imaging techniques (e.g., FDG-PET) to visualize changes in brain activity during cognitive tasks. A linear correlation between glucose utilization and brain activity has been reported for various brain regions in different conditions ( Sokoloff, 1993). Under physiological conditions, glucose is considered the major energy substrate taken up and used by the adult brain to supply its energy needs. The human brain represents only 2% of the total body weight but it accounts for a much larger proportion of the total energy expenditures of the organism ( Magistretti, 1999). These data suggest that glutamate exerts an influence on neuronal energy metabolism likely through a regulation of the expression of some key mitochondrial proteins. In parallel, a similar increase in the protein expression of LDHA was evidenced without an effect on the mRNA level. Exposure of cultured hippocampal neurons to glutamate 100 μM for 1 h led to enhanced expression of both AQP9 and MCT2 at the protein level without any significant change at the mRNA level. Moreover, they not only co-localize but they interact with each other as they were found to co-immunoprecipitate from hippocampal neuron homogenates. The aquaglyceroporin AQP9 and the monocarboxylate transporter MCT2, two transporters for oxidative energy substrates, appear to be present in mitochondria of these neurons. Using primary cultures of rat hippocampal neurons, we observed that glutamate reduces glucose utilization in this cell type, suggesting alteration in mitochondrial oxidative metabolism. The excitatory neurotransmitter glutamate has been reported to have a major impact on brain energy metabolism. 2Laboratory of Neuroenergetics, Department of Physiology, University of Lausanne, Lausanne, Switzerland.1Departamento de Fisiologia, Universidade Federal de São Paulo, São Paulo, Brazil.All rights reserved.Fabio Tescarollo 1,2 Luciene Covolan 1 Luc Pellerin 2 * The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability.īackground subtraction Densitometry ImageJ Quantification Western blot.Ĭopyright © 2018 Elsevier B.V.

#QUANTIFICATIONS OF WESTERN BLOTS WITH IMAGEJ NCBI SOFTWARE#

In this work, we describe the use of office scanner coupled with the ImageJ software together with a new image background subtraction method for accurate Western blot quantification. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. Due to the technical knowledge required for densitometry analysis together with the resources availability, standard office scanners are often used for the imaging acquisition of developed Western blot films. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Since its first description, Western blot has been widely used in molecular labs.

Quantifications of western blots with imagej ncbi