Molecular determinants of retinal ganglion cell development, survival, and regeneration

Isenmann, Stefan; Kretz, Alexandra; Cellerino, Alessandro

doi:10.1016/s1350-9462(03)00027-2

Cited by 169 publications

(120 citation statements)

References 634 publications

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“…As expected, mRNA changes were relatively small because most RGCs are still present at 7 days after axotomy. 2,3,[6][7][8]24 It was not feasible to monitor the time course of channel protein loss by immunoblotting, because RGCs comprise B7% of cells in the healthy adult retina. 25 Kv1-selective blockers reduce RGC degeneration after axotomy.…”

Section: Resultsmentioning

confidence: 99%

“…Optic nerve transection is a popular model used to assess potential neuroprotective strategies in vivo. The highly coordinated signaling response in axotomized RGCs results in activation of cell death programs and loss of B90% of RGCs within 14 days [1][2][3][4] (reviewed by Watanabe et al, 5 Levin, 6 Isenmann et al, 7 Weishaupt and Bahr 8 ). RGC death occurs by apoptosis, as shown by the presence of fragmented nuclei and DNA, caspase 3 activation and apoptotic bodies.…”

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confidence: 99%

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Kv1.1 and Kv1.3 channels contribute to the degeneration of retinal ganglion cells after optic nerve transection in vivo

Koeberle

Wang²,

Schlichter

2009

Cell Death Differ

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Degeneration of retinal ganglion cells (RGCs) -an important cause of visual impairment -is often modeled by optic nerve transection, which leads to apoptotic death of these central nervous system neurons. With this model, we show that specific voltage-gated K þ channels (Kv1 family) contribute to the degeneration of rat RGCs and expression of apoptosis-related molecules in vivo. Retinal expression of Kv1.1, Kv1.2, Kv1.3 and Kv1.5 was examined by quantitative real-time reverse transcriptase-PCR and immunohistochemistry. Kv channel blockers and channel-specific short-interfering RNAs (siRNAs) were used to assess their roles in RGC degeneration. We found that (i) rat RGCs express Kv1.1, Kv1.2 and Kv1.3 (but not Kv1.5); (ii) intraocular injection of agitoxin-2 or margatoxin, potent blockers of Kv1.1, Kv1.2 and Kv1.3 channels, dose-dependently reduced the RGC degeneration; (iii) siRNAs applied to the cut optic nerve were rapidly transported throughout RGCs only, in which they reduced the expression of the cognate channel only. Our results show differential roles of the channels; siRNAs directed against Kv1.1 or Kv1.3 channels greatly reduced RGC death, whereas Kv1.2-targeted siRNAs had only a small effect, and siRNAs against Kv1.5 were without effect. (iv) Kv1.1 and Kv1.3 channels apparently contribute to cell-autonomous death of RGCs through different components of the apoptotic machinery. Kv1.1 depletion increased the antiapoptotic gene, Bcl-X L , whereas Kv1.3 depletion reduced the proapoptotic genes, caspase-3, caspase-9 and Bad.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Kv1.1 and Kv1.3 channels contribute to the degeneration of retinal ganglion cells after optic nerve transection in vivo

Koeberle

Wang²,

Schlichter

2009

Cell Death Differ

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“…It has been documented in rats, with the use of conventional tracing methods, that 96 -99% of RGC axons decussate at the optic chiasm and project to contralateral targets (1,24). In the present study MRI did not detect the 1-4% of axons that project ipsilaterally.…”

Section: Discussionmentioning

confidence: 99%

Manganese‐enhanced MRI of the optic visual pathway and optic nerve injury in adult rats

Thuen¹,

Singstad

Pedersen³

et al. 2005

Magnetic Resonance Imaging

106

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Purpose: To evaluate manganese (Mn 2ϩ )-enhanced MRI in a longitudinal study of normal and injured rat visual projections. Materials and Methods:MRI was performed 24 hours after unilateral intravitreal injection of MnCl 2 (150 nmol) into adult Fischer rats that were divided into four groups: 1) controls (N ϭ 5), 2) dose-response (N ϭ 10, 0.2-200 nmol), 3) time-response with repeated MRI during 24 -168 hours post injection (N ϭ 4), and 4) optic nerve crush (ONC) immediately preceding the MnCl 2 injection (N ϭ 7). Control and ONC animals were reinjected with MnCl 2 20 days after the first injection, and MRI was performed 24 hours later. Results:In the control group, the optic projection was visualized from the retina to the superior colliculus, with indications of transsynaptic transport to the cortex. There was a semilogarithmic relationship between the Mn 2ϩ dose and Mn 2ϩ enhancement from 4 to 200 nmol, and the enhancement decayed gradually to 0 by 168 hours. No Mn 2ϩ -enhanced signal was detected distal to the ON crush site. In the control group, similar enhancement was obtained after the first and second MnCl 2 injections, while in the ONC group the enhancement proximal to the crush site was reduced 20 days post lesion (20dpl). Conclusion:Mn 2ϩ -enhanced MRI is a viable method for temporospatial visualization of normal and injured ON in the adult rat. The observed reduction in the Mn 2ϩ signal proximal to the ONC is probably a result of retrograde damage to the retinal ganglion cells, and not of Mn 2ϩ toxicity.

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“…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.…”

Section: Introductionmentioning

confidence: 99%

Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

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. Viral vectors, plasmids or short interfering RNAs (siRNAs) can also be delivered to the vitreous chamber in order to infect or transfect retinal cells [5][6][7][8][9][10][11][12] . The high tropism of Adeno-Associated Virus (AAV) vectors is beneficial to target RGCs, with an infection rate approaching 90% of cells near the injection site 6,7,[13][14][15] . Moreover, RGCs can be selectively transfected by applying siRNAs, plasmids, or viral vectors to the cut end of the optic nerve [16][17][18][19] or injecting vectors into their target the superior colliculus 10 . This allows researchers to study apoptotic mechanisms in the injured neuronal population without confounding effects on other bystander neurons or surrounding glia. 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 window for experimental manipulations directed against pathways involved in apoptosis. Manipulations that directly target RGCs from the transected optic nerve stump are performed at the time of axotomy, immediately after cutting the nerve. In contrast, when substances are delivered via an intraocular route, they can be injected prior to surgery or within the first 3 days after surgery, preceding the initiation of apoptosis in axotomized RGCs. In the present article, we demonstrate several methods for experimental manipulations after optic nerve transection. 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 ...

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Molecular determinants of retinal ganglion cell development, survival, and regeneration

Cited by 169 publications

References 634 publications

Kv1.1 and Kv1.3 channels contribute to the degeneration of retinal ganglion cells after optic nerve transection in vivo

Kv1.1 and Kv1.3 channels contribute to the degeneration of retinal ganglion cells after optic nerve transection in vivo

Manganese‐enhanced MRI of the optic visual pathway and optic nerve injury in adult rats

Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

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