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The Full Globin Repertoire of Turtles Provides Insights into Vertebrate Globin Evolution and Functions.

Schwarze K, Singh A, Burmester T - Genome Biol Evol (2015)

Bottom Line: Quantitative real-time reverse transcription polymerase chain reaction experiments revealed that myoglobin, neuroglobin, and globin E are highly expressed with tissue-specific patterns, which are in line with their roles in the oxidative metabolism of the striated muscles, the brain, and the retina, respectively.Histochemical analyses showed high levels of globin E in the pigment epithelium of the eye.Globin E probably has a myoglobin-like role in transporting O2 across the pigment epithelium to supply in the metabolically highly active retina.

View Article: PubMed Central - PubMed

Affiliation: Institute of Zoology, Department of Biology, University of Hamburg, Germany.

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Localization of GbE in the retina of softshell turtle and chicken. ISH was carried out with a species-specific antisense probe to detect the GbE mRNA (A, C); for IHC, a specific GbE-peptide antibody was used to detect the GbE protein (B, D). Both GbE mRNA and protein were detected in retinal PE, indicated by red arrows. In ISH, the nuclei were stained with Hoechst dye 33258, shown in yellow. Negative controls with sense probes (ISH) and omitted first antibody (IHC) are shown in supplementary fig. S4, Supplementary Material online. Scale bar = 100 µm. PE: pigment epithelium, OS: outer segments of the photoreceptor cells, IS: inner segments of the photoreceptor cells, ONL: outer nuclear layer, INL: inner nuclear layer, IPL: inner plexiform layer, GC: ganglion cells.
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evv114-F5: Localization of GbE in the retina of softshell turtle and chicken. ISH was carried out with a species-specific antisense probe to detect the GbE mRNA (A, C); for IHC, a specific GbE-peptide antibody was used to detect the GbE protein (B, D). Both GbE mRNA and protein were detected in retinal PE, indicated by red arrows. In ISH, the nuclei were stained with Hoechst dye 33258, shown in yellow. Negative controls with sense probes (ISH) and omitted first antibody (IHC) are shown in supplementary fig. S4, Supplementary Material online. Scale bar = 100 µm. PE: pigment epithelium, OS: outer segments of the photoreceptor cells, IS: inner segments of the photoreceptor cells, ONL: outer nuclear layer, INL: inner nuclear layer, IPL: inner plexiform layer, GC: ganglion cells.

Mentions: The distribution of GbE protein and mRNA was characterized in cryosections of the eyes of Chinese softshell turtle and chicken (fig. 5). For immunostaining, we used affinity-purified polyclonal anti-GbE antibodies (Blank, Kiger, et al. 2011). The specificity of the antibodies was checked by SDS-PAGE and western blotting employing whole eye protein extracts along with recombinantly expressed chicken GbE as control. The antibody showed high specificity for GbE in western blot (fig. 6). A second band at ∼30 kDa is likely due to the dimerization of the GbE, which has frequently observed for globins in SDS-PAGE (Schmidt et al. 2003). The predominant band at 15 kDa in the turtle eye was partially sequenced de novo with mass spectrometric methods. The identified peptide (ADAEDNGTTVLVR) corresponds to positions 20–32 of the GbE protein; an exchange of an asparagine to an aspartic acid at position 21 is probably due to deamination during processing.Fig. 5.—


The Full Globin Repertoire of Turtles Provides Insights into Vertebrate Globin Evolution and Functions.

Schwarze K, Singh A, Burmester T - Genome Biol Evol (2015)

Localization of GbE in the retina of softshell turtle and chicken. ISH was carried out with a species-specific antisense probe to detect the GbE mRNA (A, C); for IHC, a specific GbE-peptide antibody was used to detect the GbE protein (B, D). Both GbE mRNA and protein were detected in retinal PE, indicated by red arrows. In ISH, the nuclei were stained with Hoechst dye 33258, shown in yellow. Negative controls with sense probes (ISH) and omitted first antibody (IHC) are shown in supplementary fig. S4, Supplementary Material online. Scale bar = 100 µm. PE: pigment epithelium, OS: outer segments of the photoreceptor cells, IS: inner segments of the photoreceptor cells, ONL: outer nuclear layer, INL: inner nuclear layer, IPL: inner plexiform layer, GC: ganglion cells.
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Related In: Results  -  Collection

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evv114-F5: Localization of GbE in the retina of softshell turtle and chicken. ISH was carried out with a species-specific antisense probe to detect the GbE mRNA (A, C); for IHC, a specific GbE-peptide antibody was used to detect the GbE protein (B, D). Both GbE mRNA and protein were detected in retinal PE, indicated by red arrows. In ISH, the nuclei were stained with Hoechst dye 33258, shown in yellow. Negative controls with sense probes (ISH) and omitted first antibody (IHC) are shown in supplementary fig. S4, Supplementary Material online. Scale bar = 100 µm. PE: pigment epithelium, OS: outer segments of the photoreceptor cells, IS: inner segments of the photoreceptor cells, ONL: outer nuclear layer, INL: inner nuclear layer, IPL: inner plexiform layer, GC: ganglion cells.
Mentions: The distribution of GbE protein and mRNA was characterized in cryosections of the eyes of Chinese softshell turtle and chicken (fig. 5). For immunostaining, we used affinity-purified polyclonal anti-GbE antibodies (Blank, Kiger, et al. 2011). The specificity of the antibodies was checked by SDS-PAGE and western blotting employing whole eye protein extracts along with recombinantly expressed chicken GbE as control. The antibody showed high specificity for GbE in western blot (fig. 6). A second band at ∼30 kDa is likely due to the dimerization of the GbE, which has frequently observed for globins in SDS-PAGE (Schmidt et al. 2003). The predominant band at 15 kDa in the turtle eye was partially sequenced de novo with mass spectrometric methods. The identified peptide (ADAEDNGTTVLVR) corresponds to positions 20–32 of the GbE protein; an exchange of an asparagine to an aspartic acid at position 21 is probably due to deamination during processing.Fig. 5.—

Bottom Line: Quantitative real-time reverse transcription polymerase chain reaction experiments revealed that myoglobin, neuroglobin, and globin E are highly expressed with tissue-specific patterns, which are in line with their roles in the oxidative metabolism of the striated muscles, the brain, and the retina, respectively.Histochemical analyses showed high levels of globin E in the pigment epithelium of the eye.Globin E probably has a myoglobin-like role in transporting O2 across the pigment epithelium to supply in the metabolically highly active retina.

View Article: PubMed Central - PubMed

Affiliation: Institute of Zoology, Department of Biology, University of Hamburg, Germany.

Show MeSH
Related in: MedlinePlus