Ribosomal targeting strategy and nuclear labeling to analyze photoreceptor phosphoinositide signatures

Rajala, Ammaji; Rajala, Rahul; Teel, Kenneth; Rajala, Raju V S

doi:10.1016/j.bbalip.2022.159161

Cited by 3 publications

(9 citation statements)

References 47 publications

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“…qRT-PCR was then performed to provide the accurate quantification of gene expression from various retinal cell types. We recently used the TRAP technique to isolate PI kinases and PI phosphatase signatures from rod photoreceptor cells using rhodopsin-Cre mice mated with HA-tagged Rpl22 mice ( 26 ). In this study, we have extended the scope of our study to isolate actively translating mRNAs from cone photoreceptors, Müller, RPE, and ganglion cells, and identified cell-type-specific PIP signatures.…”

Section: Resultsmentioning

confidence: 99%

“…We described the data in the context of enrichment and depletion, in the manner of previous studies ( 26 , 27 ), i.e. when a transcript is enriched, there is an increased level of transcript in our HA-TRAP samples when compared to input (total retina), suggesting that a particular cell type contributes a significant source of transcript in the retina.…”

Section: Resultsmentioning

confidence: 99%

“…A Venn diagram was generated (Fig. 5 C) comparing the rod-rich, cone-rich, rod-poor, and cone-poor transcripts according to the methodology we published previously ( 26 ). We also identified differentially expressed PI-converting enzymes between cones and rods and the total retina (Table S8 ).…”

Section: Resultsmentioning

confidence: 99%

“…In our previous study, we identified the PIP signatures in rods with TRAP technology taking the advantage of rhodopsin-Cre ( 26 ). In this study, we have identified PIP signatures using retina cell-specific Cre lines targeted to the cone, RPE, Müller glia, and ganglion cells and compared their expression profiles with each other.…”

Section: Discussionmentioning

confidence: 99%

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Atlas of phosphoinositide signatures in the retina identifies heterogeneity between cell types

Rajala

Nair

et al. 2023

PNAS Nexus

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Phosphoinositides (PIPs) are a family of minor acidic phospholipids in the cell membrane. Phosphoinositide (PI) kinases and phosphatases can rapidly convert one PIP product into another resulting in the generation of seven distinct PIPs. The retina is a heterogeneous tissue composed of several cell types. In the mammalian genome, around 50 genes encode PI kinases and PI phosphatases; however, there are no studies describing the distribution of these enzymes in the various retinal cell types. Using translating ribosome affinity purification, we have identified the in vivo distribution of PI-converting enzymes from the rod, cone, retinal pigment epithelium (RPE), Müller glia, and retinal ganglion cells, generating a physiological atlas for PI-converting enzyme expression in the retina. The retinal neurons: rods, cones, and RGCs are characterized by the enrichment of PI-converting enzymes whereas the Müller glia and RPE are characterized by the depletion of these enzymes. We also found distinct differences between the expression of PI kinases and PI phosphatases in each retinal cell type. Since mutations in PI-converting enzymes are linked to human diseases including retinal diseases, the results of this study will provide a guide for what cell types are likely to be affected by retinal degenerative diseases brought on by changes in PI metabolism.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Atlas of phosphoinositide signatures in the retina identifies heterogeneity between cell types

Rajala

Nair

et al. 2023

PNAS Nexus

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“…Rax-Cre ERT2 was induced with 1 mg tamoxifen every alternate day by gavage for 3 days, whereas VMD2-Cre was induced with doxycycline gavage at a dose of 0.4 mg/g (10 mg/mice) body weight for 2 days. We immunoprecipitated polyribosomes bound to mRNA ( 51 ) from those specific cell types and prepared RNA. The RNA is converted to cDNA and examined the expression of Ldha , Ldhb , Rpl38 , and β-actin by qRT-PCR (Table S6 .)…”

Section: Methodsmentioning

confidence: 99%

The function of lactate dehydrogenase A in retinal neurons: implications to retinal degenerative diseases

et al. 2023

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The postmitotic retina is highly metabolic and the photoreceptors depend on aerobic glycolysis for an energy source and cellular anabolic activities. Lactate dehydrogenase A (LDHA) is a key enzyme in aerobic glycolysis, which converts pyruvate to lactate. Here we show that cell-type-specific actively translating mRNA purification by translating ribosome affinity purification shows a predominant expression of LDHA in rods and cones and LDHB in the retinal pigment epithelium and Müller cells. We show that genetic ablation of LDHA in the retina resulted in diminished visual function, loss of structure, and a loss of dorsal-ventral patterning of the cone-opsin gradient. Loss of LDHA in the retina resulted in increased glucose availability, promoted oxidative phosphorylation, and upregulated the expression of glutamine synthetase, a neuron survival factor. However, lacking LDHA in Müller cells does not affect visual function in mice. Glucose shortage is associated with retinal diseases, such as age-related macular degeneration, and regulating the levels of LDHA may have therapeutic relevance. These data demonstrate the unique and unexplored roles of LDHA in the maintenance of a healthy retina.

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The Genetic Background of Abnormalities in Metabolic Pathways of Phosphoinositides and Their Linkage with the Myotubular Myopathies, Neurodegenerative Disorders, and Carcinogenesis

Derkaczew,

Martyniuk,

Hofman

et al. 2023

Biomolecules

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Myo-inositol belongs to one of the sugar alcohol groups known as cyclitols. Phosphatidylinositols are one of the derivatives of Myo-inositol, and constitute important mediators in many intracellular processes such as cell growth, cell differentiation, receptor recycling, cytoskeletal organization, and membrane fusion. They also have even more functions that are essential for cell survival. Mutations in genes encoding phosphatidylinositols and their derivatives can lead to many disorders. This review aims to perform an in-depth analysis of these connections. Many authors emphasize the significant influence of phosphatidylinositols and phosphatidylinositols’ phosphates in the pathogenesis of myotubular myopathies, neurodegenerative disorders, carcinogenesis, and other less frequently observed diseases. In our review, we have focused on three of the most often mentioned groups of disorders. Inositols are the topic of many studies, and yet, there are no clear results of successful clinical trials. Analysis of the available literature gives promising results and shows that further research is still needed.

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Ribosomal targeting strategy and nuclear labeling to analyze photoreceptor phosphoinositide signatures

Cited by 3 publications

References 47 publications

Atlas of phosphoinositide signatures in the retina identifies heterogeneity between cell types

Atlas of phosphoinositide signatures in the retina identifies heterogeneity between cell types

The function of lactate dehydrogenase A in retinal neurons: implications to retinal degenerative diseases

The Genetic Background of Abnormalities in Metabolic Pathways of Phosphoinositides and Their Linkage with the Myotubular Myopathies, Neurodegenerative Disorders, and Carcinogenesis

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