Immunoassay for glial fibrillary acidic protein: Antigen recognition is affected by its phosphorylation state

Tramontina, Francine; Leite, Marina Concli; Ceresér, Keila M.; Souza, Daniela Fraga de; Tramontina, Ana Carolina; Nardin, Patrícia; Andreazza, Ana Cristina; Gottfried, Carmem; Kapczinski, Flávio; Gonçalves, Carlos-Alberto

doi:10.1016/j.jneumeth.2007.01.001

Cited by 65 publications

(21 citation statements)

References 26 publications

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“…H-1152P might have directly interfered with the Müller cell hypertrophy by suppressing the Rho-kinase dependent phosphorylation of GFAP at the head domain and thereby preventing the reorganization of these filaments [56]. Indeed, a novel study demonstrates that several commercially available antibodies against GFAP have a higher affinity towards the phosphorylated protein compared to the unphosphorylated GFAP-isoforms, suggesting that the changes in the levels of GFAP immunoreactivity might in fact reflect the changes in the degree of phosphorylation rather than the total level of this protein [57]. The Rho-kinase may also regulate the cytoskeletal contraction in the microglial cells, resulting in the activation of these cells [31] whereas the inhibitor prevented the transition into the reactive, ameboid morphology.…”

Section: Discussionmentioning

confidence: 99%

The Neuroprotective Potential of Rho-Kinase Inhibition in Promoting Cell Survival and Reducing Reactive Gliosis in Response to Hypoxia in Isolated Bovine Retina

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Hilgers

Tura

et al. 2013

Cell Physiol Biochem

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Aims: To investigate the outcomes of Rho-kinase inhibition in the electrophysiological ex vivo model of the isolated perfused vertebrate retina under hypoxia. Methods: Bovine retinas were perfused with an oxygen saturated nutrient solution with or without the Rho-kinase inhibitor H-1152P. The retinas were stimulated repeatedly until stable amplitudes were reached and the electroretinogram was recorded at five minute intervals. Hypoxia was induced for 15, 30, and 45 minutes, after which the oxygen saturation was restored. The extent of the cell damage and glial reactivity was determined by Ethidium homodimer-1 staining, immunohistochemistry, and Western blot. Results: Hypoxia caused a time-dependent reduction of the b-wave amplitudes, which could not be prevented by the H-1152P. Although the Rho-kinase inhibitor maintained higher b-wave amplitudes, these effects did not reach statistical significance. Hypoxia also resulted in an increase in cell damage and the activation of the glial cells in the untreated retinas whereas the administration of H-1152P significantly reduced the extent of these events. Conclusion: H-1152P exerted a neuroprotective effect against necrosis on the isolated bovine retina under hypoxia together with a reduction in glial cell reactivity. However, the inhibitor could not prevent the hypoxia induced retinal dysfunction possibly due to the interference with synaptic modulation.

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Section: Discussionmentioning

confidence: 99%

The Neuroprotective Potential of Rho-Kinase Inhibition in Promoting Cell Survival and Reducing Reactive Gliosis in Response to Hypoxia in Isolated Bovine Retina

Alt

Hilgers

Tura

et al. 2013

Cell Physiol Biochem

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show abstract

“…Post-translational modifications and/or non-covalent ligants are elements that interfere in the antibody-antigen interaction, however these modifications are often not fully considered during the elaboration and interpretation of immunoassays (Tramontina et al, 2007). S100B is a calcium-binding protein and its calcium-binding status greatly affects its antibody recognition (Gonçalves et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

A simple, sensitive and widely applicable ELISA for S100B: Methodological features of the measurement of this glial protein

Leite

Galland

Brolese

et al. 2008

Journal of Neuroscience Methods

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116

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“…Post-translational modifications and/or non-covalent ligands are elements that interfere in the antibody-antigen interaction, however these modifications are often not fully considered during the elaboration and interpretation of immunoassays [105]. The calcium-binding status of the S100B greatly affects its antibody recognition [106].…”

Section: Methodological Procedures For S100b Measurementmentioning

confidence: 99%

Biological and methodological features of the measurement of S100B, a putative marker of brain injury

Gonçalves

Leite

Nardin

2008

Clinical Biochemistry

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224

166

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Immunoassay for glial fibrillary acidic protein: Antigen recognition is affected by its phosphorylation state

Cited by 65 publications

References 26 publications

The Neuroprotective Potential of Rho-Kinase Inhibition in Promoting Cell Survival and Reducing Reactive Gliosis in Response to Hypoxia in Isolated Bovine Retina

The Neuroprotective Potential of Rho-Kinase Inhibition in Promoting Cell Survival and Reducing Reactive Gliosis in Response to Hypoxia in Isolated Bovine Retina

A simple, sensitive and widely applicable ELISA for S100B: Methodological features of the measurement of this glial protein

Biological and methodological features of the measurement of S100B, a putative marker of brain injury

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