Retinol dehydrogenase and rpe-retinal G protein-coupled receptor expression patterns in mice after monocular deprivation

doi:10.3877/cma.j.issn.2095-2007.2022.01.002

Abstract

Abstract:

Objective

The aim of this study was to examine the expression pattern differences of retinol dehydrogenase (RDH) and rpe-retinal G protein-coupled receptor (RGR) in deprivation myopic mice.

Methods

100 four-week-old C57BL6/J mice (200 eyes) were randomly distributed into myopia group and control group, 50 for each group (100 eyes). Monocular deprivation of was induced by covering the right eyes with handmade, translucent occluders, while the left eyes served as contralateral eyes. Mean-while, the control group received no treatment. After deprivation for four weeks, refraction errors were measured via an eccentric infrared photorefractor, and axial lengths were estimated using a hand-held digital microscope. The messenger ribonucleic acid (mRNA) transcript levels of Rdh5, Rdh10 and Rgr genes were detected using real-time reverse transcription polymerase chain reaction. The corresponding protein concentrations of these genes were detected by Western blot analysis. Refractive errors, axial length, mRNA levels and the relative gray value of protein were described as ±s. The refraction interocular difference was used to compare by paired t-test. One-way ANOVA was used to determine statistic differences in axial length, mRNA levels and relative gray value of protein among multiple samples, and the LSD test was used for further pairwise comparisons.

Results

The refractive error of the covered myopic eyes and contralateral eyes in myopia group were (-5.631±4.052) D, (4.231±2.828) D, resepctively. There was a significant difference between them (t=-10.914, P<0.05). The control group was found to have refractive error for right and left eyes, which were (3.774±4.079) D and (4.171±4.425) D, respectively. There was not a significant difference between them (t=-1.344, P>0.05). The axial length of covered myopic eyes, contralateral eyes and control group were (3305.0±86.4) μm, (3221.0±90.0) μm and (3232.0±68.6) μm, respectively. After one-way ANOVA, there was a statistically significant difference among them (F=6.248, P<0.05). In the retinas of covered myopic eyes, the relative mRNA levels of Rdh5, Rdh10 and Rgr genes was (2.032±0.162), (2.611±0.258) and (3.876±0.576), respectively, which were all lower than that in the contralateral retinas. Compared with control group, the mRNA levels of gene increased significantly in myopia group. After one-way ANOVA, there was a significant difference between them (F=38.671, 88.510, 78.557; P<0.05). The results of Western blot analysis showed that relative gray value of protein expression pattern of these genes in myopic group were in accordance with the mRNA expression trend. The relative gray value of proteins of RDH5, RDH10 and RGR in covered myopic eyes were lower than that in the contralateral uncovered eyes. After one-way ANOVA, there was a significant difference between them (F=12.189, 56.956, 6.620; P<0.05).

Conclusions

Early monocular deprivation induces a robust increase in mRNA levels of Rdh5, Rdh10 and Rgr genes, while the relative gray value of protein is inconsistent with that. The relative gray value of protein of RDH5 and RDH10 are high, and RGR low in covered myopic eyes. Abnormal visual stimulus is helpful to impede the expression of retinol dehydrogenases and promote the myopia progression through the potential effects of visual cycle.

Key words: Form deprivation myopia, Retinol dehydrogenase 5, Retinol dehydrogenase 10, Rpe-retinal G protein-coupled receptor, Retina

Tiantian Liu, Bingjie Chen, Longqian Liu. Retinol dehydrogenase and rpe-retinal G protein-coupled receptor expression patterns in mice after monocular deprivation[J]. Chinese Journal of Ophthalmologic Medicine(Electronic Edition), 2022, 12(01): 6-11.

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References 41

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引物名称	引物序列(5′-3′)
GAPDH-F	GGTTGTCTCCTGCGACTTCA
GAPDH-R	TGGTCCAGGGTTTCTTACTCC
RGR-F	GTTCATGGTTTCCAGGGATTTG
RGR-R	CAGATGACATCCACACAAACAG
RDH5-F	GGAGAAACTGGACTTTTTGGTC
RDH5-R	TGTGTTCACACTCAGTACTCTC
RDH10-F	TGGGGAAGAGAGAGAATGTCTA
RDH10-R	AAACCACACCAGCATTATTGAC

引物名称	引物序列(5′-3′)
GAPDH-F	GGTTGTCTCCTGCGACTTCA
GAPDH-R	TGGTCCAGGGTTTCTTACTCC
RGR-F	GTTCATGGTTTCCAGGGATTTG
RGR-R	CAGATGACATCCACACAAACAG
RDH5-F	GGAGAAACTGGACTTTTTGGTC
RDH5-R	TGTGTTCACACTCAGTACTCTC
RDH10-F	TGGGGAAGAGAGAGAATGTCTA
RDH10-R	AAACCACACCAGCATTATTGAC

组别	只数	右眼	左眼	眼间差值
近视组	50	-5.631±4.052	4.231±2.828	-9.863±4.781
对照组	50	3.774±4.079	4.171±4.425	-0.398±2.001
t值		-9.644	0.064
P值		<0.05	>0.05

组别	只数	右眼	左眼	眼间差值
近视组	50	-5.631±4.052	4.231±2.828	-9.863±4.781
对照组	50	3.774±4.079	4.171±4.425	-0.398±2.001
t值		-9.644	0.064
P值		<0.05	>0.05

组别	只数	右眼	左眼
近视组	50	3305.0±86.4	3221.0±90.0
对照组	50		3232.0±68.6
F值		6.248
P值		<0.05

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