Expression of functional dopaminergic phenotype in purified cultured Müller cells from vertebrate retina

Kubrusly, Regina Célia Cussa; Panizzutti, Rogério; Gardino, P.F.; Stutz, Bernardo; Reis, Rodrigo S.; Ventura, Ana Lúcia Marques; Mello, Maria Christina F. de; Mello, Fernando G. de

doi:10.1016/j.neuint.2008.05.002

Cited by 29 publications

(27 citation statements)

References 50 publications

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“…A recent Western blot analysis performed in our laboratory using the AADC antiserum from the present study verified a band at the expected size ($55 kD) in the forebrain and midbrain of both prairie voles and laboratory rats, as well as a very heavy band in both species at $110 kD that is likely the dimerized AADC protein found in central nervous system tissue (Kubrusly et al, 2008;Siow and Dakshinamurti, 1990;Zhu and Juorio, 1995). Control blots that omitted either the primary or secondary antisera did not contain any immunoreactive bands.…”

Section: Tissue Collection and Immunohistochemistrysupporting

confidence: 75%

l-Amino acid decarboxylase- and tyrosine hydroxylase-immunoreactive cells in the extended olfactory amygdala and elsewhere in the adult prairie vole brain

Ahmed

Northcutt

Lonstein

2012

Journal of Chemical Neuroanatomy

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Section: Tissue Collection and Immunohistochemistrysupporting

confidence: 75%

l-Amino acid decarboxylase- and tyrosine hydroxylase-immunoreactive cells in the extended olfactory amygdala and elsewhere in the adult prairie vole brain

Ahmed

Northcutt

Lonstein

2012

Journal of Chemical Neuroanatomy

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“…On the other hand, treatment with 50 ng/ml BDNF not only reduces the response to muscimol, but also increases the number of cells responding to KCl or AMPA Cell Mol Neurobiol (2011) 31:835-846 843 Although in our experiments calcium shifts in glia cells were only observed after ATP stimulation, purified cultured Müller glia display several characteristics that are seen in retina neurons. For instance, these cells in culture accumulate cyclic AMP induced by catecholamines (Kubrusly et al 2008), display GABA uptake properties (De Sampaio Schitine et al 2007) and PACAP-induced activation of adenylyl cyclase (Kubrusly et al 2005). They also express glutamic acid decarboxylase, beta 2 nicotine acetylcholine receptors, enzymes of catecholamine metabolism, including tyrosine hydroxylase (Kubrusly et al 2005(Kubrusly et al , 2008Seki et al 2006), glutamate (Ramirez and Lamas 2009), and muscarinic acetylcholine receptors (Wakakura et al 1998).…”

Section: Discussionmentioning

confidence: 99%

“…For instance, these cells in culture accumulate cyclic AMP induced by catecholamines (Kubrusly et al 2008), display GABA uptake properties (De Sampaio Schitine et al 2007) and PACAP-induced activation of adenylyl cyclase (Kubrusly et al 2005). They also express glutamic acid decarboxylase, beta 2 nicotine acetylcholine receptors, enzymes of catecholamine metabolism, including tyrosine hydroxylase (Kubrusly et al 2005(Kubrusly et al , 2008Seki et al 2006), glutamate (Ramirez and Lamas 2009), and muscarinic acetylcholine receptors (Wakakura et al 1998). Chick Müller cells in culture express several excitatory amino acids (EAAs) receptors that are coupled to inositol triphosphate mobilization and 45 Ca 2?…”

Section: Discussionmentioning

confidence: 99%

Functional Identification of Cell Phenotypes Differentiating from Mice Retinal Neurospheres Using Single Cell Calcium Imaging

et al. 2011

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Degeneration of neural retina causes vision impairment and can lead to blindness. Neural stem and progenitor cells might be used as a tool directed to regenerative medicine of the retina. Here, we describe a novel platform for cell phenotype-specific drug discovery and screening of proneurogenic factors, able to boost differentiation of neural retinal progenitor cells. By using single cell calcium imaging (SCCI) and a rational-based stimulation protocol, a diversity of cells emerging from differentiated retinal neurosphere cultures were identified. Exposure of retinal progenitor cultures to KCl or to α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) stimulated Ca(2+) transients in microtubule-associated protein 2 (MAP-2) positive neurons. Doublecortin (DCX) and polysialated neural cell adhesion molecule (PSA-NCAM) positive neuroblasts were distinguished from differentiated neurons on the basis of their response to muscimol. Ca(2+) fluxes in glial fibrillary acidic protein (GFAP) or glutamine synthetase (GS) positive cells were induced by ATP. To validate the platform, neurospheres were treated with brain-derived neurotrophic factor (BDNF) (proneurogenic) or ciliary neurotrophic factor (CNTF) (gliogenic factor). BDNF increased the percentage of differentiated cells expressing Tuj-1 sensitive to KCl or AMPA and reduced the population of cells responding to muscimol. CNTF exposure resulted in a higher number of cells expressing GFAP responding to ATP. All together, our data may open new perspectives for cell type-specific discovery of drug targets and screening of novel proneurogenic factors to boost differentiation of neural retina cells to treat degenerative retinal diseases.

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“…After subculturing by conventional methods, the second generation of retinal Müller cells was used for experimentation. Retinal Müller cells were identified by immunocytochemistry for 3 specific markers: GFAP, vimentin and S-100 [14][15][16] .…”

Section: Retinal Müller Cell Culturementioning

confidence: 99%

Taurine Buffers Glutamate Homeostasis in Retinal Cells in vitro under Hypoxic Conditions

Chen

Zhang

et al. 2010

Ophthalmic Res

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Aims: We investigated whether taurine indirectly protects neurons under hypoxia by affecting retinal Müller cells, which are known to play important roles in the regulation of retinal glutamate content. Methods: Retinal cells isolated from rats were exposed to hypoxia for 24 h. We evaluated the retinal neuron survival, glutamate content in cultures with and without taurine under hypoxic conditions. The glutamate clearance function correlated with the expression of glutamine synthetase (GS) mRNA and L-glutamate/L-aspartate transporter (GLAST) mRNA. Immunohistochemical staining of glial fibrillary acidic protein (GFAP), vimentin and S-100 protein was performed to examine cytoskeletal changes in retinal Müller cells. Results: Retinal neurons treated with taurine exhibited significantly higher survival rates than those without taurine under hypoxia. Taurine inhibited the upregulation of GFAP and vimentin, and inhibited the downregulation of GLAST, GS and the nuclear-cytoplasmic ratio of S-100 under hypoxia. In addition, taurine inhibited the upregulation of the glutamate content in neurons and retinal Müller cells upon hypoxic exposure. Conclusion: These data suggest that hypoxic damage to cultured retinal cells is decreased by taurine. The neuroprotection by taurine may relate to buffering glutamate homeostasis via modulation of the glutamate clearance by retinal Müller cells.

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Expression of functional dopaminergic phenotype in purified cultured Müller cells from vertebrate retina

Cited by 29 publications

References 50 publications

l-Amino acid decarboxylase- and tyrosine hydroxylase-immunoreactive cells in the extended olfactory amygdala and elsewhere in the adult prairie vole brain

l-Amino acid decarboxylase- and tyrosine hydroxylase-immunoreactive cells in the extended olfactory amygdala and elsewhere in the adult prairie vole brain

Functional Identification of Cell Phenotypes Differentiating from Mice Retinal Neurospheres Using Single Cell Calcium Imaging

Taurine Buffers Glutamate Homeostasis in Retinal Cells in vitro under Hypoxic Conditions

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