0000000000139497
AUTHOR
Rosa M. Guasch
Neuronal polarization is impaired in mice lacking RhoE expression
J. Neurochem. (2012) 121, 903–914. Abstract Proper development of neuronal networks relies on the polarization of the neurons, thus the establishment of two compartments, axons and dendrites, whose formation depends on cytoskeletal rearrangements. Rnd proteins are regulators of actin organization and they are important players in several aspects of brain development as neurite formation, axon guidance and neuron migration. We have recently demonstrated that mice lacking RhoE/Rnd3 expression die shortly after birth and have neuromotor impairment and neuromuscular alterations, indicating an abnormal development of the nervous system. In this study, we have further investigated the specific ro…
Study of surface carbohydrates on isolated Golgi subfractions by fluorescent-lectin binding and flow cytometry
The Golgi complex is a functionally heterogeneous subcellular structure that plays a key role in the synthesis, maturation, and sorting of newly synthesized glycoproteins. Fluorescent lectins have been used extensively to analyze surface glycoproteins by flow cytometry in whole cells and more recently in isolated subcellular organelles, such as mitochondria and chloroplasts. We report here the use of several fluorescein-isothiocyanate-conjugated lectins to detect and quantify specific surface sugars by flow cytometry on isolated elements from purified cis and trans-Golgi fractions from rat liver. Our results show that this approach may be useful to study Golgi composition and function, sinc…
Flow cytometric analysis of concanavalin A binding to isolated Golgi fractions from rat liver.
Flow cytometry (FCM) has been used repeatedly to study lectin binding to whole cells. However, there are very few attempts to analyze glycoconjugates in isolated subcellular organelles. We have applied FCM to quantitate the specific binding of fluorescein-conjugated concanavalin A (FITC-Con A) to isolated cis and trans fractions of rat liver Golgi complex, the cell compartment involved in glycoprotein maturation and sorting. Our results show similar intensities of Con A-specific binding in the two fractions. Using this method we show a decreased FITC-Con A binding to both Golgi fractions in ethanol-treated rats, which is consistent to previous work on alcoholic effects on galactosyltransfer…
Analysis of Subcellular Components by Fluorescent-Lectin Binding and Flow Cytometry
Because of their extensive availability and the wide spectrum of carbohydrates that may be specifically bound, lectins have become essential reagents for detection and quantitation of glycoconjugates in solution and in cell surfaces, identification and separation of cells, and functional studies based on membrane properties (1,2).
Flow-cytometric enumeration of reticulocytes with the new fluorochrome 1′,3′-diethyl-4,2′-quinolylthiacyanine
Several flow-cytometric methods for reticulocyte enumeration in whole blood have been developed, with different degrees of practical use. Recently, a new fluorochrome, 1′,3′-diethyl-4,2′-quinolylthyacianine (DEQTC) was proposed in a brief report, as an alternative to thiazole orange for reticulocyte counting. We have evaluated the usefulness of this fluorescent stain by assessing the optimal conditions for the flow-cytometric analysis, and by comparing in double-blind assays the quantitative results of this technique with those obtained by manual counting with brilliant cresyl blue. Our results show that flow cytometry with DEQTC is highly correlated to the manual method (r=0.95–0.99), supp…