Culture of rat retinal ganglion cells

Xu, Zhirong; Jiang, Fagang; Yancai, Zeng; Alkhodari, Hamed Tm; Chen, Fei

doi:10.1007/s11596-011-0389-0

Cited by 4 publications

(2 citation statements)

References 11 publications

(12 reference statements)

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“…As such, for the in vitro experiments, it was important for the mixed retinal cell culture to contain RGCs. To test for the presence of RGCs, cells were stained for Class III β-tubulin (TUJ-1) which is a constituent of neuronal microtubules and frequently expressed on RGCs, as well as Brn3a and Thy 1 which are also RGC-specific markers.23 Thy 1 is a glycophosphatidylinositol-anchored cell surface protein of the immunoglobulin superfamily which has been shown to be specifically expressed on RGCs 24–26. The expression of Thy 1 in the mixed retinal cell culture was at a level similar to past studies 24,27,28.…”

Section: Discussionmentioning

confidence: 99%

<p>Safety of anti-VEGF treatments in a diabetic rat model and retinal cell culture</p>

Filek¹,

Hooper²,

Sheidow³

et al. 2019

OPTH

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Purpose To analyze the safety of different concentrations of anti-VEGF on retinal cells. Methods Non-diabetic and streptozotocin (STZ)-induced diabetic rats received intravitreal rat anti-VEGF injections that had final vitreous concentrations of 0, 0.0625, 0.125 (clinical dose), and 0.25 mg/mL. Rats were also injected with the clinical dose of ranibizumab. TUNEL assay was performed on sectioned eyes to evaluate apoptotic cells. In vitro, rat retinal cell cultures were exposed to 0, 0.0625, 0.125 (clinical dose), and 0.25 mg/mL of ranibizumab for 48 and 72 hrs. Cellular metabolic activity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, necrosis by lactate dehydrogenase (LDH), and apoptosis by cell death enzyme-linked immunosorbent assay (ELISA). Results Diabetic rats had a significant increase ( p <0.03) in apoptotic cell death at half the clinical dose, at the clinical dose, and at double the clinical dose. In vitro, MTT showed a significant decrease ( p <0.04) in cellular metabolic activity at the clinical dose and double the clinical dose compared to control at 48 and 72 hrs. LDH showed a significant increase ( p <0.04) in necrosis at the clinical dose and double the clinical dose compared to control at 48 and 72 hrs. ELISA showed a significant increase ( p <0.04) in apoptosis at half the clinical dose, at the clinical dose, and double the clinical dose, compared to control at 48 and 72 hrs. Conclusions Anti-VEGF treatment may be potentially detrimental to the retina by decreasing cellular metabolic activity and increasing cytotoxicity of retinal cells. The results provide a cautionary note to monitor both the retina and optic nerve status in patients undergoing frequent injections.

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Section: Discussionmentioning

confidence: 99%

<p>Safety of anti-VEGF treatments in a diabetic rat model and retinal cell culture</p>

Filek¹,

Hooper²,

Sheidow³

et al. 2019

OPTH

View full text Add to dashboard Cite

show abstract

“…An important advantage of these mixed cultures is the improved survival time and viability of the cells. However, RGCs may constitute only a limited percentage of the total retinal cells in these cultures, compromising the study of specific effects on RGCs (Zhang et al 2002;Xu et al 2011).…”

Section: In Vitro Cellular Studies In Rodentsmentioning

confidence: 99%

Complementary research models and methods to study axonal regeneration in the vertebrate retinofugal system

et al. 2017

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Due to the lack of axonal regeneration, age-related deterioration in the central nervous system (CNS) poses a significant burden on the wellbeing of a growing number of elderly. To overcome this regenerative failure and to improve the patient's life quality, the search for novel regenerative treatment strategies requires valuable (animal) models and techniques. As an extension of the CNS, the retinofugal system, consisting of retinal ganglion cells that send their axons along the optic nerve to the visual brain areas, has importantly contributed to the current knowledge on mechanisms underlying the restricted regenerative capacities and to the development of novel strategies to enhance axonal regeneration. It provides an extensively used research tool, not only in amniote vertebrates including rodents, but also in anamniote vertebrates, such as zebrafish. Indeed, the latter show robust regeneration capacities, thereby providing insights into the factors that contribute to axonal regrowth and proper guidance, complementing studies in mammals. This review provides an integrative and critical overview of the classical and state-of-the-art models and methods that have been employed in the retinofugal system to advance our knowledge on the signaling pathways underlying the restricted versus robust axonal regeneration in rodents and zebrafish, respectively. In vitro, ex vivo and in vivo models and techniques to improve the visualization and analysis of regenerating axons are summarized. As such, the retinofugal system is presented as a valuable model to further facilitate research on axonal regeneration and to open novel therapeutic avenues for CNS pathologies.

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Secretion of Down Syndrome Critical Region 1 Isoform 4 in Ischemic Retinal Ganglion Cells Displays Anti-Angiogenic Properties Via NFATc1-Dependent Pathway

Yang

et al. 2016

Mol Neurobiol

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Down syndrome candidate region 1 (DSCR1) has two differentially regulated isoforms (DSCR1-1 and DSCR1-4) and is reported to play a role in a number of physiological processes, such as the inhibition of cardiac hypertrophy, attenuation of angiogenesis and carcinogenesis, and protection against neuronal death. However, the function of DSCR1 in the retina is still not clear. Therefore, we analyzed the expression and location of DSCR1 in the retina of neonatal mice with oxygen-induced retinopathy (OIR), and studied its effects on angiogenesis. The neonatal C57BL/6J mice were exposed to 75 % O for 5 days from postnatal day 7 (P7) to P12. At P12, the mice were returned to 21 % O at room air. The primary retinal ganglion cells (RGCs) were exposed to hypoxia (93 % N, 5 % CO, and 2 % O) at 37 °C for 36 h. And then the mouse retinal microvascular endothelial cells (mRMECs) were treated with 25 ng/mL vascular endothelial growth factor (VEGF) or culture medium conditioned by hypoxic RGCs alone, hypoxic RGCs treated with DSCR1-4-siRNA (siDSCR1-4) or hypoxic RGCs treated with siDSCR1-4 and 200 ng/mL cyclosporin A (CsA), and then primed with VEGF (25 ng/mL). The expression of DSCR1-4 increased strongly at P16 after OIR. There was no change in messenger RNA (mRNA) expression of DSCR1-1 at P16 after OIR. The increased DSCR1 was mainly located in the RGCs of avascular retina. In addition, DSCR1-4 expression was increased in primary RGCs after hypoxia exposure. There was no change in mRNA expression of DSCR1-1 in primary RGCs after hypoxia exposure. Moreover, DSCR1-4 produced by hypoxic RGCs showed anti-angiogenic properties, with decreased cell proliferation, migration, tube formation, and inflammatory cytokines production. These properties were due to inhibited nuclear factor of activated T cell (NFATc) 1 dephosphorylation and translocation into nuclear in VEGF-treated mRMECs. Using siRNA-mediated knockdown of DSCR1-4 and NFATc1 inhibitor (Cs A) further demonstrated the inhibitory effect of DSCR1-4 on angiogenic properties in VEGF-induced mRMECs, and this effect was NFATc1-dependent. This report describes a novel effect of DSCR1-4 in the aspect of anti-angiogenesis, suggesting potential therapeutic strategies for proliferative retinopathies.

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Culture of rat retinal ganglion cells

Cited by 4 publications

References 11 publications

<p>Safety of anti-VEGF treatments in a diabetic rat model and retinal cell culture</p>

<p>Safety of anti-VEGF treatments in a diabetic rat model and retinal cell culture</p>

Complementary research models and methods to study axonal regeneration in the vertebrate retinofugal system

Secretion of Down Syndrome Critical Region 1 Isoform 4 in Ischemic Retinal Ganglion Cells Displays Anti-Angiogenic Properties Via NFATc1-Dependent Pathway

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