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Decrease in retinal neuronal cells in streptozotocin-induced diabetic mice.

Yang Y, Mao D, Chen X, Zhao L, Tian Q, Liu C, Zhou BL - Mol. Vis. (2012)

Bottom Line: Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer.The temporal course of Brn3a+ RGC and Neuronal Nuclei+RGC (NeuN+ RGC) loss induced by intraperitoneal injection of streptozotocin followed a similar trend.It indicates that RGC loss may be an important component of diabetic retinopathy.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, the First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, Chongqing, China.

ABSTRACT

Purpose: Little is known about retinal neuronal loss in the retinas of diabetic mice. The purpose of this study was the quantitative assessment of retinal neural cell number in diabetic mice.

Methods: Five-week-old C57BL/6 mice were used as a diabetic model with streptozotocin. Mice were studied over the course of 6 and 12 weeks after the onset of diabetes. Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer. The retinal ganglion cells (RGCs) were counted at two different time points after the induction of diabetes and examined using the immunofluorescence technique and quantitative analysis.

Results: The diabetic mice had significantly elevated IOP levels at 6 and 12 weeks after the onset of diabetes compared with the age-matched control mice (p<0.01 and p<0.001, respectively). The temporal course of Brn3a+ RGC and Neuronal Nuclei+RGC (NeuN+ RGC) loss induced by intraperitoneal injection of streptozotocin followed a similar trend. At 6 and 12 weeks after the onset of diabetes, the number of Brn3a+ RGCs (p<0.05 at 6 weeks; p<0.001 at 12 weeks) and NeuN+ RGCs (p<0.05 at 6 weeks; p<0.001 at 12 weeks) was significantly lower in diabetic mice than age-matched control mice. In the retinal flatmounts, the number of Brn3a+ RGCs (p<0.05 at 6 weeks, p<0.01 at 12 weeks) was also significantly lower in diabetic mice than control mice. The IOP in diabetic mice was negatively related with RGCs in cross sections. The cut-off value of IOP was 14.2 mmHg for diabetes.

Conclusions: This is a specific quantitative study of neural cell loss in the retina during diabetes. These data suggest that retinal neural cell reduction occurs in diabetic mice. It indicates that RGC loss may be an important component of diabetic retinopathy.

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Quantification of retinal ganglion cell loss in the ganglion cell layer. Cell loss was assessed by counting the cells with Brn3a (A) and NeuN (C) immunoreactivity. The number of retinal ganglion cells (RGCs) exhibits a significantly difference in diabetic and control mice (n=10/group). In the percentage of RGCs for Brn3a (B) and NeuN (D), both markers were calculated in relation to the number of cells counted. Data are expressed as means±standard deviation (SD; * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001).
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f3: Quantification of retinal ganglion cell loss in the ganglion cell layer. Cell loss was assessed by counting the cells with Brn3a (A) and NeuN (C) immunoreactivity. The number of retinal ganglion cells (RGCs) exhibits a significantly difference in diabetic and control mice (n=10/group). In the percentage of RGCs for Brn3a (B) and NeuN (D), both markers were calculated in relation to the number of cells counted. Data are expressed as means±standard deviation (SD; * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001).

Mentions: Neurodegeneration of the retina is a critical component of DR and a significant loss of RGCs has been reported in STZ-induced diabetic mice [25,26]. We quantitatively analyzed RGCs in radial sections of the retina in diabetic mice using both the Brn3a and NeuN markers (Figure 2). Compared to control mice, the numbers of Brn3a+ RGCs (7.46±1.75 versus 8.93±2.14 cells/13.3 um at 6 weeks, p<0.05 and 5.96±1.37 versus 9.04±1.84 cells/13.3 um at 12 weeks, p<0.001) was significantly lower in diabetic mice (Figure 3A). Thus, the number of Brn3a+ RGCs exhibited a 7% loss over 6 weeks and a 15% loss over 12 weeks (Figure 3B). In contrast to the control mice, the numbers of NeuN+ RGCs (14.43±2.41 versus 12.3±2.39 cells/13.3 um at 6 weeks p<0.05 and 13.65 ± 2.22 versus 7.75±2.15 cells/13.3 um at 12 weeks, p<0.001) was also significantly decreased in diabetic mice (Figure 3C). Thus, the number of NeuN+ RGCs showed a 10% loss over 6 weeks and 22% loss over 12 weeks (Figure 3D).


Decrease in retinal neuronal cells in streptozotocin-induced diabetic mice.

Yang Y, Mao D, Chen X, Zhao L, Tian Q, Liu C, Zhou BL - Mol. Vis. (2012)

Quantification of retinal ganglion cell loss in the ganglion cell layer. Cell loss was assessed by counting the cells with Brn3a (A) and NeuN (C) immunoreactivity. The number of retinal ganglion cells (RGCs) exhibits a significantly difference in diabetic and control mice (n=10/group). In the percentage of RGCs for Brn3a (B) and NeuN (D), both markers were calculated in relation to the number of cells counted. Data are expressed as means±standard deviation (SD; * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3369894&req=5

f3: Quantification of retinal ganglion cell loss in the ganglion cell layer. Cell loss was assessed by counting the cells with Brn3a (A) and NeuN (C) immunoreactivity. The number of retinal ganglion cells (RGCs) exhibits a significantly difference in diabetic and control mice (n=10/group). In the percentage of RGCs for Brn3a (B) and NeuN (D), both markers were calculated in relation to the number of cells counted. Data are expressed as means±standard deviation (SD; * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001).
Mentions: Neurodegeneration of the retina is a critical component of DR and a significant loss of RGCs has been reported in STZ-induced diabetic mice [25,26]. We quantitatively analyzed RGCs in radial sections of the retina in diabetic mice using both the Brn3a and NeuN markers (Figure 2). Compared to control mice, the numbers of Brn3a+ RGCs (7.46±1.75 versus 8.93±2.14 cells/13.3 um at 6 weeks, p<0.05 and 5.96±1.37 versus 9.04±1.84 cells/13.3 um at 12 weeks, p<0.001) was significantly lower in diabetic mice (Figure 3A). Thus, the number of Brn3a+ RGCs exhibited a 7% loss over 6 weeks and a 15% loss over 12 weeks (Figure 3B). In contrast to the control mice, the numbers of NeuN+ RGCs (14.43±2.41 versus 12.3±2.39 cells/13.3 um at 6 weeks p<0.05 and 13.65 ± 2.22 versus 7.75±2.15 cells/13.3 um at 12 weeks, p<0.001) was also significantly decreased in diabetic mice (Figure 3C). Thus, the number of NeuN+ RGCs showed a 10% loss over 6 weeks and 22% loss over 12 weeks (Figure 3D).

Bottom Line: Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer.The temporal course of Brn3a+ RGC and Neuronal Nuclei+RGC (NeuN+ RGC) loss induced by intraperitoneal injection of streptozotocin followed a similar trend.It indicates that RGC loss may be an important component of diabetic retinopathy.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, the First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, Chongqing, China.

ABSTRACT

Purpose: Little is known about retinal neuronal loss in the retinas of diabetic mice. The purpose of this study was the quantitative assessment of retinal neural cell number in diabetic mice.

Methods: Five-week-old C57BL/6 mice were used as a diabetic model with streptozotocin. Mice were studied over the course of 6 and 12 weeks after the onset of diabetes. Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer. The retinal ganglion cells (RGCs) were counted at two different time points after the induction of diabetes and examined using the immunofluorescence technique and quantitative analysis.

Results: The diabetic mice had significantly elevated IOP levels at 6 and 12 weeks after the onset of diabetes compared with the age-matched control mice (p<0.01 and p<0.001, respectively). The temporal course of Brn3a+ RGC and Neuronal Nuclei+RGC (NeuN+ RGC) loss induced by intraperitoneal injection of streptozotocin followed a similar trend. At 6 and 12 weeks after the onset of diabetes, the number of Brn3a+ RGCs (p<0.05 at 6 weeks; p<0.001 at 12 weeks) and NeuN+ RGCs (p<0.05 at 6 weeks; p<0.001 at 12 weeks) was significantly lower in diabetic mice than age-matched control mice. In the retinal flatmounts, the number of Brn3a+ RGCs (p<0.05 at 6 weeks, p<0.01 at 12 weeks) was also significantly lower in diabetic mice than control mice. The IOP in diabetic mice was negatively related with RGCs in cross sections. The cut-off value of IOP was 14.2 mmHg for diabetes.

Conclusions: This is a specific quantitative study of neural cell loss in the retina during diabetes. These data suggest that retinal neural cell reduction occurs in diabetic mice. It indicates that RGC loss may be an important component of diabetic retinopathy.

Show MeSH
Related in: MedlinePlus