mRNA expression analysis of the SUMO pathway genes in the adult mouse retina

Abstract

BSTRACTSumoylation is a reversible post-translational modification that regulates different cellular processes by conjugation/deconjugation of SUMO moieties to target proteins. Most work on the functional relevance of SUMO has focused on cell cycle, DNA repair and cancer in cultured cells, but data on the inter-dependence of separate components of the SUMO pathway in highly specialized tissues, such as the retina, is still scanty. Nonetheless, several retinal transcription factors (TFs) relevant for cone and rod fate, as well as some circadian rhythm regulators, are regulated by sumoylation. Here we present a comprehensive survey of SUMO pathway gene expression in the murine retina by quantitative RT-PCR and in situ hybridization (ISH). The mRNA expression levels were quantified in retinas obtained under four different light/dark conditions, revealing distinct levels of gene expression. In addition, a SUMO pathway retinal gene atlas based on the mRNA expression pattern was drawn. Although most genes are ubiquitously expressed, some patterns could be defined in a first step to determine its biological significance and interdependence. The wide expression of the SUMO pathway genes, the transcriptional response under several light/dark conditions, and the diversity of expression patterns in different cell layers clearly support sumoylation as a relevant post-translational modification in the retina. This expression atlas intends to be a reference framework for retinal researchers and to depict a more comprehensive view of the SUMO-regulated processes in the retina.

Retinal ganglion cell loss in an ex vivo mouse model of optic nerve cut is prevented by curcumin treatment

“…SUMOylation is known to play an important role in retinal function [ 22 , 34 ] and in oxidative stress [ 21 ]. Interestingly, SUMO-1ylation was highly increased in retinas at time 24 (2.25 ± 0.16, P < 0.0001 vs t 0; Fig.…”

“…Recently, we reported that SUMO-1 plays an important role in the hyperactivation of JNK and Tau in a cellular model of oxidative stress and that curcumin, a strong antioxidant and anti-inflammatory natural molecule, was able to re-equilibrate the JNK–SUMO-1–Tau axis [ 21 ]. Since the involvement of SUMO in retinal function has been already shown [ 22 ], SUMO-1ylation levels in our ex vivo model has been investigated. In addition, photoreceptors and RGCs are known to be highly vulnerable to oxidative stress as that reactive oxidative species (ROS) imbalance is involved in many retinal diseases [ 23 , 24 ].…”

The tumor-associated YB-1 protein: new player in the circadian control of cell proliferation

“…How does the clock regulate zfYB-1 nuclear localization? One mechanism that has been implicated in clock-directed shuttling of proteins between the nucleus and cytoplasm is SUMOylation [ 5 , 35 ]. SUMOylation has been documented to time the nuclear entry of the BMAL1 clock protein.…”

Expression Atlas of the Deubiquitinating Enzymes in the Adult Mouse Retina, Their Evolutionary Diversification and Phenotypic Roles

“…A RT-qPCR was performed on mouse neuroretinas to assess the expression levels of the whole set of 87 mouse genes that encode the deubiquitinating enzymes belonging to the five aforementioned families (11 JAMM, 4 MJD, 15 OTU, 4 UCH, and 53 USP genes). Two reference genes, Rhodopsin and Cerkl , were included in the analysis due to their previously reported high and low levels of expression in the mouse retina, respectively [ 29 ]. The relative expression levels have been normalized to the expression of Gapdh , and the Z-score was calculated for the whole set of genes per each sample, so that they could be directly compared among them and between different samples.…”