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Gene profiling of postnatal Mfrprd6 mutant eyes reveals differential accumulation of Prss56, visual cycle and phototransduction mRNAs.

Soundararajan R, Won J, Stearns TM, Charette JR, Hicks WL, Collin GB, Naggert JK, Krebs MP, Nishina PM - PLoS ONE (2014)

Bottom Line: In Mfrprd6 eyes, a significant 1.5- to 2.0-fold decrease was observed among transcripts of genes linked to retinal degeneration, including those involved in visual cycle (Rpe65, Lrat, Rgr), phototransduction (Pde6a, Guca1b, Rgs9), and photoreceptor disc morphogenesis (Rpgrip1 and Fscn2).Levels of RPE65 were significantly decreased by 2.0-fold.In summary, the Mfrprd6 allele causes significant postnatal changes in transcript and protein levels in the retina and RPE.

View Article: PubMed Central - PubMed

Affiliation: The Jackson Laboratory, Bar Harbor, Maine, United States of America.

ABSTRACT
Mutations in the membrane frizzled-related protein (MFRP/Mfrp) gene, specifically expressed in the retinal pigment epithelium (RPE) and ciliary body, cause nanophthalmia or posterior microphthalmia with retinitis pigmentosa in humans, and photoreceptor degeneration in mice. To better understand MFRP function, microarray analysis was performed on eyes of homozygous Mfrprd6 and C57BL/6J mice at postnatal days (P) 0 and P14, prior to photoreceptor loss. Data analysis revealed no changes at P0 but significant differences in RPE and retina-specific transcripts at P14, suggesting a postnatal influence of the Mfrprd6 allele. A subset of these transcripts was validated by quantitative real-time PCR (qRT-PCR). In Mfrprd6 eyes, a significant 1.5- to 2.0-fold decrease was observed among transcripts of genes linked to retinal degeneration, including those involved in visual cycle (Rpe65, Lrat, Rgr), phototransduction (Pde6a, Guca1b, Rgs9), and photoreceptor disc morphogenesis (Rpgrip1 and Fscn2). Levels of RPE65 were significantly decreased by 2.0-fold. Transcripts of Prss56, a gene associated with angle-closure glaucoma, posterior microphthalmia and myopia, were increased in Mfrprd6 eyes by 17-fold. Validation by qRT-PCR indicated a 3.5-, 14- and 70-fold accumulation of Prss56 transcripts relative to controls at P7, P14 and P21, respectively. This trend was not observed in other RPE or photoreceptor mutant mouse models with similar disease progression, suggesting that Prss56 upregulation is a specific attribute of the disruption of Mfrp. Prss56 and Glul in situ hybridization directly identified Müller glia in the inner nuclear layer as the cell type expressing Prss56. In summary, the Mfrprd6 allele causes significant postnatal changes in transcript and protein levels in the retina and RPE. The link between Mfrp deficiency and Prss56 up-regulation, together with the genetic association of human MFRP or PRSS56 variants and ocular size, raises the possibility that these genes are part of a regulatory network influencing postnatal posterior eye development.

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Volcano plots showing the relationship between fold change (represented as mean A – mean B) and the level of significance (represented by the Fs permutated p-value).Differentially expressed probe sets (q<0.05 shown in red across all fold change levels) at and fold change greater than 2 are depicted in volcano plots in three pairwise comparisons. (A) rd6/rd6 (Mfrprd6/Mfrprd6) P0 vs B6 (C57BL/6J) P0, (B) rd6/rd6 P14 vs B6 P14 and (C) rd6/rd6 P14 vs rd6/rd6 P0.
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pone-0110299-g001: Volcano plots showing the relationship between fold change (represented as mean A – mean B) and the level of significance (represented by the Fs permutated p-value).Differentially expressed probe sets (q<0.05 shown in red across all fold change levels) at and fold change greater than 2 are depicted in volcano plots in three pairwise comparisons. (A) rd6/rd6 (Mfrprd6/Mfrprd6) P0 vs B6 (C57BL/6J) P0, (B) rd6/rd6 P14 vs B6 P14 and (C) rd6/rd6 P14 vs rd6/rd6 P0.

Mentions: Gene expression analysis was performed using the Affymetrix Mouse 430v2 microarray. Volcano plots were generated to graphically represent differentially accumulated gene transcripts at a significance level of q<0.05 using six pairwise analyses. Comparison of Mfrprd6 and C57BL/6J samples at the P0 time point did not yield any significant differences in transcript levels, indicating that the effects of Mfrp mutation occur postnatally (Fig. 1A). In contrast, comparison at the P14 time point resulted in 2,454 differentially expressed probe sets (Fig. 1B). Not surprisingly, comparison of the P0 and P14 time points in Mfrprd6 mice yielded 28,452 significant differentially accumulated probe sets (Fig. 1C), and a similar result was obtained in C57BL/6J mice (Fig. S1A). Further analysis of differential accumulated gene transcripts between the two strains (Mfrprd6 and C57BL/6J) irrespective of the time point yielded a relatively small set of significantly accumulated probe sets (Fig. S1B). By contrast, the comparison of the difference in time point (P0 vs P14) irrespective of the strain difference yielded a relatively large set of significant differentially accumulated probe sets (Fig. S1C). Taken together, this analysis suggests that these probe sets likely change as a consequence of ocular development between P0 and P14.


Gene profiling of postnatal Mfrprd6 mutant eyes reveals differential accumulation of Prss56, visual cycle and phototransduction mRNAs.

Soundararajan R, Won J, Stearns TM, Charette JR, Hicks WL, Collin GB, Naggert JK, Krebs MP, Nishina PM - PLoS ONE (2014)

Volcano plots showing the relationship between fold change (represented as mean A – mean B) and the level of significance (represented by the Fs permutated p-value).Differentially expressed probe sets (q<0.05 shown in red across all fold change levels) at and fold change greater than 2 are depicted in volcano plots in three pairwise comparisons. (A) rd6/rd6 (Mfrprd6/Mfrprd6) P0 vs B6 (C57BL/6J) P0, (B) rd6/rd6 P14 vs B6 P14 and (C) rd6/rd6 P14 vs rd6/rd6 P0.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4214712&req=5

pone-0110299-g001: Volcano plots showing the relationship between fold change (represented as mean A – mean B) and the level of significance (represented by the Fs permutated p-value).Differentially expressed probe sets (q<0.05 shown in red across all fold change levels) at and fold change greater than 2 are depicted in volcano plots in three pairwise comparisons. (A) rd6/rd6 (Mfrprd6/Mfrprd6) P0 vs B6 (C57BL/6J) P0, (B) rd6/rd6 P14 vs B6 P14 and (C) rd6/rd6 P14 vs rd6/rd6 P0.
Mentions: Gene expression analysis was performed using the Affymetrix Mouse 430v2 microarray. Volcano plots were generated to graphically represent differentially accumulated gene transcripts at a significance level of q<0.05 using six pairwise analyses. Comparison of Mfrprd6 and C57BL/6J samples at the P0 time point did not yield any significant differences in transcript levels, indicating that the effects of Mfrp mutation occur postnatally (Fig. 1A). In contrast, comparison at the P14 time point resulted in 2,454 differentially expressed probe sets (Fig. 1B). Not surprisingly, comparison of the P0 and P14 time points in Mfrprd6 mice yielded 28,452 significant differentially accumulated probe sets (Fig. 1C), and a similar result was obtained in C57BL/6J mice (Fig. S1A). Further analysis of differential accumulated gene transcripts between the two strains (Mfrprd6 and C57BL/6J) irrespective of the time point yielded a relatively small set of significantly accumulated probe sets (Fig. S1B). By contrast, the comparison of the difference in time point (P0 vs P14) irrespective of the strain difference yielded a relatively large set of significant differentially accumulated probe sets (Fig. S1C). Taken together, this analysis suggests that these probe sets likely change as a consequence of ocular development between P0 and P14.

Bottom Line: In Mfrprd6 eyes, a significant 1.5- to 2.0-fold decrease was observed among transcripts of genes linked to retinal degeneration, including those involved in visual cycle (Rpe65, Lrat, Rgr), phototransduction (Pde6a, Guca1b, Rgs9), and photoreceptor disc morphogenesis (Rpgrip1 and Fscn2).Levels of RPE65 were significantly decreased by 2.0-fold.In summary, the Mfrprd6 allele causes significant postnatal changes in transcript and protein levels in the retina and RPE.

View Article: PubMed Central - PubMed

Affiliation: The Jackson Laboratory, Bar Harbor, Maine, United States of America.

ABSTRACT
Mutations in the membrane frizzled-related protein (MFRP/Mfrp) gene, specifically expressed in the retinal pigment epithelium (RPE) and ciliary body, cause nanophthalmia or posterior microphthalmia with retinitis pigmentosa in humans, and photoreceptor degeneration in mice. To better understand MFRP function, microarray analysis was performed on eyes of homozygous Mfrprd6 and C57BL/6J mice at postnatal days (P) 0 and P14, prior to photoreceptor loss. Data analysis revealed no changes at P0 but significant differences in RPE and retina-specific transcripts at P14, suggesting a postnatal influence of the Mfrprd6 allele. A subset of these transcripts was validated by quantitative real-time PCR (qRT-PCR). In Mfrprd6 eyes, a significant 1.5- to 2.0-fold decrease was observed among transcripts of genes linked to retinal degeneration, including those involved in visual cycle (Rpe65, Lrat, Rgr), phototransduction (Pde6a, Guca1b, Rgs9), and photoreceptor disc morphogenesis (Rpgrip1 and Fscn2). Levels of RPE65 were significantly decreased by 2.0-fold. Transcripts of Prss56, a gene associated with angle-closure glaucoma, posterior microphthalmia and myopia, were increased in Mfrprd6 eyes by 17-fold. Validation by qRT-PCR indicated a 3.5-, 14- and 70-fold accumulation of Prss56 transcripts relative to controls at P7, P14 and P21, respectively. This trend was not observed in other RPE or photoreceptor mutant mouse models with similar disease progression, suggesting that Prss56 upregulation is a specific attribute of the disruption of Mfrp. Prss56 and Glul in situ hybridization directly identified Müller glia in the inner nuclear layer as the cell type expressing Prss56. In summary, the Mfrprd6 allele causes significant postnatal changes in transcript and protein levels in the retina and RPE. The link between Mfrp deficiency and Prss56 up-regulation, together with the genetic association of human MFRP or PRSS56 variants and ocular size, raises the possibility that these genes are part of a regulatory network influencing postnatal posterior eye development.

Show MeSH
Related in: MedlinePlus