An Immunohistochemical Study of Protein Kinase C in the Bovine Retina

Shin, Taekyun; Kim, Seungjoon; Ahn, Meejung; Kim, Heechul

doi:10.1292/jvms.68.71

Cited by 9 publications

(5 citation statements)

References 11 publications

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“…GFP-expressing cells in the outer border of the INL, with round cell bodies, showed Lim1+2 staining at each of the developmental stages. Pkcα has been shown to specifically label bipolar cells in the developing retina [ 48 - 50 ]. Pkcα labeling showed no staining at E10 (J).…”

Section: Resultsmentioning

confidence: 99%

Analysis of retinal cell development in chick embryo by immunohistochemistry and in ovo electroporation techniques

et al. 2010

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BackgroundRetinal cell development has been extensively investigated; however, the current knowledge of dynamic morphological and molecular changes is not yet complete.ResultsThis study was aimed at revealing the dynamic morphological and molecular changes in retinal cell development during the embryonic stages using a new method of targeted retinal injection, in ovo electroporation, and immunohistochemistry techniques. A plasmid DNA that expresses the green fluorescent protein (GFP) as a marker was delivered into the sub-retinal space to transfect the chick retinal stem/progenitor cells at embryonic day 3 (E3) or E4 with the aid of pulses of electric current. The transfected retinal tissues were analyzed at various stages during chick development from near the start of neurogenesis at E4 to near the end of neurogenesis at E18. The expression of GFP allowed for clear visualization of cell morphologies and retinal laminar locations for the indication of retinal cell identity. Immunohistochemistry using cell type-specific markers (e.g., Visinin, Xap-1, Lim1+2, Pkcα, NeuN, Pax6, Brn3a, Vimentin, etc.) allowed further confirmation of retinal cell types. The composition of retinal cell types was then determined over time by counting the number of GFP-expressing cells observed with morphological characteristics specific to the various retinal cell types.ConclusionThe new method of retinal injection and electroporation at E3 - E4 allows the visualization of all retinal cell types, including the late-born neurons, e.g., bipolar cells at a level of single cells, which has been difficult with a conventional method with injection and electroporation at E1.5. Based on data collected from analyses of cell morphology, laminar locations in the retina, immunohistochemistry, and cell counts of GFP-expressing cells, the time-line and dynamic morphological and molecular changes of retinal cell development were determined. These data provide more complete information on retinal cell development, and they can serve as a reference for the investigations in normal retinal development and diseases.

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Section: Resultsmentioning

confidence: 99%

Analysis of retinal cell development in chick embryo by immunohistochemistry and in ovo electroporation techniques

et al. 2010

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“…The monoclonal anti-PKC␣ antibody (raised against amino acids 650 -690 at the C-terminus of human PKC␣) reveals only a major band at Ϸ80 kDa. In addition to this product, the polyclonal anti-PKC␣ antibody (raised against the same epitope) detects lighter bands likely corresponding to ␤I, ␤II (Ϸ74 kDa; as notified by the manufacturer) and ␦ isoforms (Ϸ40 -51 kDa; Shin et al, 2006). The labeling pattern was identical to that seen in mouse retina with different monoclonal antibodies (Haverkamp and Wä ssle, 2000).…”

Section: Primary Antibodiesmentioning

confidence: 86%

Retinal anatomy and visual performance in a diurnal cone‐rich laboratory rodent, the Nile grass rat (Arvicanthis niloticus)

Gaillard

Bonfield

Gilmour

et al. 2008

J of Comparative Neurology

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Unlike laboratory rats and mice, muridae of the Arvicanthis family (A. ansorgei and A. niloticus) are adapted to functioning best in daylight. To date, they have been used as experimental models mainly in studies of circadian rhythms. However, recent work aimed at optimizing photoreceptor-directed gene delivery vectors (Khani et al. [2007] Invest Ophthalmol Vis Sci 48:3954-3961) suggests their potential usefulness for studying retinal pathologies and therapies. In the present study we analyzed the retinal anatomy and visual performance of the Nile grass rat (A. niloticus) using immunohistofluorescence and the optokinetic response (OKR). We found that approximately 35-40% of photoreceptors are cones; that many neural features of the inner retina are similar to those in other diurnal mammals; and that spatial acuity, measured by the OKR, is more than two times that of the usual laboratory rodents. These observations are consistent with the known diurnal habits of this animal, and further support its pertinence as a complementary model for studies of structure, function, and pathology in cone-rich mammalian retinae.

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“…In contrast, light causes the retraction of the spinules in the depolarizing bipolar cells concurrent with the formation of filamentous actin in the contracted synaptic pedicles and an increase in protein kinase C (PKC) activity [59]. PKC is indeed a frequently used marker for the bipolar cells, which is localized mainly to the bipolar cell dendrites, axons, and photoreceptor presynaptic terminals, suggesting a key role for PKC in the establishment of the retinal circuitry [60,61]. …”

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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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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An Immunohistochemical Study of Protein Kinase C in the Bovine Retina

Cited by 9 publications

References 11 publications

Analysis of retinal cell development in chick embryo by immunohistochemistry and in ovo electroporation techniques

Analysis of retinal cell development in chick embryo by immunohistochemistry and in ovo electroporation techniques

Retinal anatomy and visual performance in a diurnal cone‐rich laboratory rodent, the Nile grass rat (Arvicanthis niloticus)

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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