Production and specificity of monoclonal antibodies to retinal S antigen

An immunochemical approach was used to detect the expression of putative guanine nucleotide-binding proteins (C-proteins), arrestin, and nucleoside diphosphate kinases during wheat (Triticum aestivum) tissue culture initiated from immature embryos. Both the soluble and membrane extracts from the immature embryos revealed bands of 58, 40, and 16 kD with antibodies to C-protein ( a subunit), arrestin, and nucleoside diphosphate kinase, respectively. These proteins were overexpressed in vitro in both nonembryogenic callus and embryogenic cultures. An additional soluble protein (32 kD) was detected by anti-Gcu antibodies in cultured tissues but not in immature embryos, suggesting a possible function in cell multiplication. Moreover, somatic embryogenesis was associated with the appearance of a 29-kD protein reactive with anti-arrestin antibodies, both in soluble and membrane fractions. Tissue-cultured genetic stocks of Chinese Spring wheat, including the disomic, 36 ditelosomic, and 6 nullisomic-tetrasomic wheat lines, were used to ascertain the chromosomal location of the genes encoding the 29-kD arrestin-like protein. The lack of a signal with the nonembryogenic ditelosomic 3 D short chromosome arm line suggests that the 3 D long chromosome arm possesses at least one gene involved in the expression of the 29-kD protein. The putative role of the 29-kD protein in signal-transduction regulating embryogenesis is discussed.

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“…1). (b) Four mAbs against bovine retina1 arrestin (Faure et al, 1984;M. Mirshahi, unpublished data) were selected for this study.…”

Section: Antibodiesmentioning

confidence: 99%

Immunological Detection of Potential Signal-Transduction Proteins Expressed during Wheat Somatic Tissue Culture

Nato¹,

Mirshahi²,

Tichtinsky³

et al. 1997

Plant Physiology

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“…To define the cellular distribution of the prionimmunopositive structures in the OPL, photoreceptor terminals were immunolabeled with an antibody directed against arrestin, a protein expressed exclusively in photoreceptors showing their terminals in the OPL and their cell bodies with inner and outer segments in the outer nuclear layer. [22][23][24] In the OPL, all PrP c -immunopositive puncta were located in arrestin-immunolabeled structures, indicating that PrP c was located in photoreceptor terminals (Figure 2, A-C). To determine whether PrP c was present in cone pedicles or rod spherules, retinal sections were stained with the PNA, which labels cone pedicles in the OPL.…”

Section: Prp C Localization In the Rat Retinamentioning

confidence: 99%

The Toxicity of the PrP106-126 Prion Peptide on Cultured Photoreceptors Correlates with the Prion Protein Distribution in the Mammalian and Human Retina

Gong

Jellali

Forster

et al. 2007

The American Journal of Pathology

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In patients affected by Creutzfeldt-Jakob disease and in animals affected by transmissible spongiform encephalopathies, retinal functions are altered, and major spongiform changes are observed in the outer plexiform layer where photoreceptors have their synaptic terminals. In the present study, the prion protein PrP c was found to form aggregates in rod photoreceptor terminals from both rat and human retina, whereas no labeling was observed in cone photoreceptors. Discrete staining was also detected in the inner plexiform layer where the prion protein was located at human amacrine cell synapses. In mixed porcine retinal cell cultures, the PrP106-126 prion peptide triggered a 61% rod photoreceptor cell loss by apoptosis as indicated by terminal deoxynucleotidyl transferase dUTP nick-end labeling, whereas cone photoreceptors were not affected. Amacrine cells were also reduced by 47% in contrast to ganglion cells. Although this cell loss was associated with a 5.5-fold increase in microglial cells, the strict correlation between the PrP c prion protein expression and the peptide toxicity suggested that this toxicity did not rely on the release of a toxic compound by glial cells. These results provide new insights Transmissible spongiform encephalopathies in humans and in animals are linked to the transconformation of the prion protein PrP c into a proteinase-resistant form PrP res . 1 These neurological disorders exhibit common pathological symptoms like vacuolization of the neurophils, astrocytosis, and loss of neurons. 2 In patients with CreutzfeldtJakob disease, alteration of the retinal function was attested by the decrease in the electroretinogram b-wave amplitude, 3-5 which reflects the activity of bipolar cells postsynaptic to photoreceptors. These early electroretinogram changes were even proposed as a diagnostic measurement for the disease. 6 In histology, major spongiform changes were observed in the outer plexiform layer (OPL) where photoreceptors have their synaptic terminals, and only moderate changes were observed in the inner plexiform layer (IPL) and ganglion cell layer. 5 These localizations were consistent with the reported accumulation of the pathogenic form of the prion protein, PrP res , throughout the plexiform layers of the human retina. 7 The in vivo retina has been used on many occasions to study the progression of the disease in animal models because of its organized structure, its in vivo access for intraocular injection, and the possibility to correlate the

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“…Even FPLC-purified arrestin, which is essentially homogeneous on SDS-PAGE, is resolved into 2-4 major bands and 5-10 minor bands with isoelectric points between pH 5.5 and 6.1. All of these subspecies are stained by an antibody specific [13] for S-antigen (= arrestin; see fig.3, lanes 5-8). Qualitatively similar 1EF patterns are observed with arrestin purified by our method or that of Dorey et al [8], either crude or FPLC-purified ( fig.3, lanes 5-8).…”

Section: Resultsmentioning

confidence: 99%

“…For immunoblotting the proteins were transferred to nitrocellulose sheets [12] and stained using the anti-S-antigen monoclonal antibody $6H8 (kind gift of J.P. Faure, see [13]) and the biotin-streptavidin system (Amersham), with 1-chloro-4-naphthol as a substrate.…”

Section: Isoelectric Focusingmentioning

confidence: 99%

Rapid affinity purification of retinal arrestin (48 kDa protein) via its light‐dependent binding to phosphorylated rhodopsin

et al. 1986

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Arrestin (also named ‘48 kDa protein’ or ‘S‐antigen’) is a soluble protein involved in controlling light‐dependent cGMP phosphodiesterase activity in retinal rods, and is also known for its ability to induce autoimmune uveitis of the eye. We report a rapid and simple purification method based on the property of arrestin to bind specifically and reversibly to illuminated and phosphorylated rhodopsin [(1984) FEBS Lett. 176, 473–478]. This method does not require column chromatography and yields about 2–4 mg purified arrestin from 15 bovine retinas. Pure arrestin can be resolved by isoelectric focusing into at least 10 distinct bands, all of which stain with a monoclonal antibody specific for S‐antigen.

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Production and specificity of monoclonal antibodies to retinal S antigen

Cited by 50 publications

References 7 publications

Immunological Detection of Potential Signal-Transduction Proteins Expressed during Wheat Somatic Tissue Culture

Immunological Detection of Potential Signal-Transduction Proteins Expressed during Wheat Somatic Tissue Culture

The Toxicity of the PrP106-126 Prion Peptide on Cultured Photoreceptors Correlates with the Prion Protein Distribution in the Mammalian and Human Retina

Rapid affinity purification of retinal arrestin (48 kDa protein) via its light‐dependent binding to phosphorylated rhodopsin

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