Specific transcellular staining of microglia in the adult rat after traumatic degeneration of carbocyanine-filled retinal ganglion cells

Thanos, Solon; Pavlidis, Christos; Mey, Jörg; Thiel, Holger

doi:10.1016/0014-4835(92)90098-d

Cited by 69 publications

(41 citation statements)

References 40 publications

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“…Therefore there may be differences in glial activation mechanisms between retina and cortex. Note that we also failed to find aggregations of microglia around ganglion cell somas at early stages as might be expected from the work suggesting that microglia may actively kill ganglion cells (Thanos, 1992(Thanos, , 1993 but it is possible that these are not obvious without double labelling.…”

Section: Correlation With Muller Cell Reactioncontrasting

confidence: 41%

“…The microglia were labelled by histochemistry for the enzyme nucleoside di-phosphatase (NDP-ase) because this provides excellent morphological detail (Murabe and Sano, 1982;Vorbrodt and Wisnieswski, 1982j, is known to be regulated by the injury response (Miyamoto et al, 1992) and makes our data directly comparable to studies of retinal microglia following optic nerve section (Schnitzer and Scherer, 1990). It has recently been suggested that microglia can actively kill retinal ganglion cells (Thanos et al, 1992(Thanos et al, , 1993 and therefore the microglial reaction was also compared to the neuronal reaction as shown by phosphorylated neurofilament immunocytochemistry.…”

Section: Introductionmentioning

confidence: 60%

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Microglial responses to focal lesions of the rabbit retina: Correlation with neural and macroglial reactions

Humphrey

Moore

1996

Glia

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Section: Correlation With Muller Cell Reactioncontrasting

confidence: 41%

Section: Introductionmentioning

confidence: 60%

Microglial responses to focal lesions of the rabbit retina: Correlation with neural and macroglial reactions

Humphrey

Moore

1996

Glia

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“…However, the proportion of dying cells in this layer remained high until P7 (Young, 1984;Pequignot et al, 2003). Therefore, phagocytic cells labeled in the developing optic nerve may be participating in the clearance of axonal debris of degenerating ganglion cells, as has been proposed for microglia in the postnatal rabbit retina (Schnitzer, 1989) or for activated microglia after optic nerve cut (Thanos et al, 1992;Lawson et al, 1994). Furthermore, during development of other regions of the murine central nervous system, apoptotic neurons and their connections are actively and rapidly removed by microglia (Frade and Barde, 1998;Marin-Teva et al, 2004).…”

Section: Spatiotemporal Distribution Of Cathepsin B/d-expressing Macrmentioning

confidence: 88%

Macrophage and microglia ontogeny in the mouse visual system can be traced by the expression of Cathepsins B and D

Bejarano-Escobar

Holguín-Arévalo

Montero

et al. 2011

Developmental Dynamics

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Here, we show a detailed chronotopographical analysis of cathepsin B and D expression during development of the mouse visual system. Both proteases were detected in large rounded/ameboid cells usually located in close relationship with prominent sites of extensive physiological cell death. In concordance with their morphological features and topographical distribution, we demonstrate that expressing cells corresponded with macrophages and microglial precursors. We found that as microglial precursors differentiated the expression of both cathepsins was down-regulated. Of interest, cathepsin B and D transcripts were never observed in degenerating cells. Our findings point to a role for cathepsin D and B in cell debris degradation after apoptotic processes rather than promoting cell death, as proposed for other developmental models. Additionally their pattern of expression suggests a role in the maturation of the microglial precursors.

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“…By 14 days after axotomy, the majority of RGCs have died, and a few remaining RGCs are interspersed between retinal microglia (Fig 1b). When RGCs undergo apoptosis, microglia phagocytose the dead cells and as a result become transcellularly labeled with the fluorescent tracer that was used to label the RGCs 13,14 . The appearance of the tracer in the phagosomes of microglia is different from that in surviving RGCs.…”

Section: Journal Of Visualized Experiments Wwwjovecommentioning

confidence: 99%

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

Magharious

D'Onofrio

Koeberle

2011

JoVE

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Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. The optic nerve can be accessed within the orbit of the eye and completely transected (axotomized), cutting the axons of the entire RGC population. Optic nerve transection is a reproducible model of apoptotic neuronal cell death in the adult CNS [1][2][3][4] . This model is particularly attractive because the vitreous chamber of the eye acts as a capsule for drug delivery to the retina, permitting experimental manipulations via intraocular injections. The diffusion of chemicals through the vitreous fluid ensures that they act upon the entire RGC population. Moreover, RGCs can be selectively transfected by applying short interfering RNAs (siRNAs), plasmids, or viral vectors to the cut end of the optic nerve [5][6][7] or injecting vectors into their target, the superior colliculus 8 . This allows researchers to study apoptotic mechanisms in the desired neuronal population without confounding effects on other bystander neurons or surrounding glia. An additional benefit is the ease and accuracy with which cell survival can be quantified after injury. The retina is a flat, layered tissue and RGCs are localized in the innermost layer, the ganglion cell layer. The survival of RGCs can be tracked over time by applying a fluorescent tracer (3% Fluorogold) to the cut end of the optic nerve at the time of axotomy, or by injecting the tracer into the superior colliculus (RGC target) one week prior to axotomy. The tracer is retrogradely transported, labeling the entire RGC population. Because the ganglion cell layer is a monolayer (one cell thick), RGC densities can be quantified in flat-mounted tissue, without the need for stereology. Optic nerve transection leads to the apoptotic death of 90% of injured RGCs within 14 days postaxotomy [9][10][11] . RGC apoptosis has a characteristic time-course whereby cell death is delayed 3-4 days postaxotomy, after which the cells rapidly degenerate. This provides a time window for experimental manipulations directed against pathways involved in apoptosis. Experiments should be carried out using aseptic technique and following the animal use protocols of your specific institution. Instruments and materials (solutions, test substances, tracers, needles, etc.) coming into contact with living tissue must be sterile to prevent infection and adverse impacts on animal welfare and potential negative impacts on the study.1. Rats will be anaesthetized using a veterinary isoflurane vaporizer system. Use medical grade oxygen at a rate of 0.8 L/min to vaporize the isoflurane gas. Place the animal in the attached anesthesia box and dial in an isoflurane concentration of 4% until the breathing has slowed and the animal is sedate. 2. Next, switch the gas flow to the gas mask attachment for the stereotaxic frame and place the animal in the stereotaxic apparatus. Turn the isoflurane concentration down to 2% and monitor anesthesia. Larger animals (>300g) may require a hi...

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Specific transcellular staining of microglia in the adult rat after traumatic degeneration of carbocyanine-filled retinal ganglion cells

Cited by 69 publications

References 40 publications

Microglial responses to focal lesions of the rabbit retina: Correlation with neural and macroglial reactions

Microglial responses to focal lesions of the rabbit retina: Correlation with neural and macroglial reactions

Macrophage and microglia ontogeny in the mouse visual system can be traced by the expression of Cathepsins B and D

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

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