Detecting Glycogen in Peripheral Blood Mononuclear Cells with Periodic Acid Schiff Staining

Shafiei, Mahdieh Tabatabaei; Gonczi, Catalina M. Carvajal; Rahman, Mohammed Samiur; East, Ashley; François, Jonathan; Darlington, Peter J.

doi:10.3791/52199-v

Cited by 5 publications

(7 citation statements)

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“…In our results, we found periodic acid to be the best oxidizing agent when compared to the other evaluated oxidizers in the current study. This nding agrees with what is found in literature where periodic acid appears to be the most commonly used oxidizing agent to be used with Schiff's reagent for glycogen demonstration [12,13,14].…”

Section: Declarations Fundingsupporting

confidence: 92%

Optimization of PAS reaction and similar Schiff`s based methods for glycogen demonstration in liver tissue.

Abdellah

Abdallah

Abdalrheem

et al. 2023

Preprint

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Purpose Demonstration of glycogen in tissue has valuable diagnostic significance in several lesions, including certain tumors. Histochemical staining of glycogen with Schiff's dependent methods is affected by the type of fixative, the temperature of fixation, and oxidizing agents. This study aimed to evaluate different fixatives, temperatures of fixation, and oxidizing agents with variable durations of treatment to be used with Schiff's reagent for glycogen demonstration. Methods Archived paraffin blocks prepared from one rabbit's liver were used and 340 paraffin sections were stained with different oxidizers-Schiff methods. Results For tissues fixed at 4°C: 10% NBF, 80% alcohol, and Bouins's solution provided good staining results with PAS reaction. For tissues fixed at RT: both 10% NBF and 80% alcohol provided good staining results with PAS reaction. The other oxidizing agents provided poor results with all fixatives and temperature of fixation, with two exceptions of satisfactory staining results for 5% chromic acid for 10 minutes at both room temperature (RT) and 60°C. Conclusion Both 10% NBF and 80% alcohol seem to be the fixatives of choice for glycogen when the PAS reaction is used. However, Bouin's solution could provide good results at 4° C.

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Section: Declarations Fundingsupporting

confidence: 92%

Optimization of PAS reaction and similar Schiff`s based methods for glycogen demonstration in liver tissue.

Abdellah

Abdallah

Abdalrheem

et al. 2023

Preprint

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“…Glucose conversion into glucose-1-P, the first step in glycogen synthesis, also appeared to be severely impacted by HK2-deficiency based on low glucose-1-P/glucose ratios. While little is known about the role of glycogen storage in lymphocytes, they do express glycogen synthase, contain measurable stores of glycogen and functional insulin receptor 60,61 . Together our results indicate that HK2-deficiency substantially impairs glucose utilization, particularly for anabolic pathways, and may impact glucose storage.…”

Section: Discussionmentioning

confidence: 99%

PI3K-dependant reprogramming of hexokinase isoforms controls glucose metabolism and functional responses of B lymphocytes

Paradoski,

Hou,

Mejia

et al. 2024

Preprint

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B lymphocyte metabolic reprogramming is essential for B cell differentiation and mounting a healthy immune response. The PI3K signaling pathway regulates B cell metabolism, but the mechanisms involved are not well understood. Here we report that signaling via PI3Kδ can impact B cell glucose metabolism and immune functions via selective upregulation of hexokinase 2 (HK2). Three HK enzymes can catalyze the critical first step for glucose utilization and may selectively direct glucose into specific catabolic and anabolic pathways. While HK1 is constitutively expressed in B cells, HK2 is strikingly upregulated during B cell activation in a PI3Kδ-dependent manner. HK2 shows a unique distribution between mitochondrial and cytoplasmic pools that is also regulated by PI3K. Genetic deletion of HK2 significantly impairs extracellular acidification rate and glycolytic ATP production despite strong expression of HK1. B cell-specific deletion of HK2 in mice caused mild perturbations in B cell development but did not prevent generation of mature B cell subsets. HK2-deficient B cells show altered functional responses in vitro and evidence of metabolic adaptation to become less dependent on glucose and more dependent on glutamine. HK2-deficient B cells exhibit impaired glycolysis, altered metabolite profiles and altered flux of labeled glucose carbons into the pentose phosphate pathway. Upon immunization, HK2-deficient mice exhibit impaired generation of germinal centre B cells, plasmablasts and antibody responses. We further found that HK2 expression in primary human chronic lymphocytic leukemia (CLL) cells was associated with recent proliferation and could be reduced by PI3K inhibition. Our study identifies hexokinase 2 upregulation as a functionally important component of B cell metabolic reprogramming dependent on the PI3K pathway.

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“…Finally, the cells were washed with deionized water, and the cell nuclei were differentiated using acidic hematoxylin. The prepared slides were examined under a fluorescence microscope (Axioskop, Carl Zeiss, Oberkochen, Germany) with the aid of filter 09 (Filter set 09-487909-0000; Carl Zeiss, Oberkochen, Germany) following the protocol described by Tabatabaei Shafiei et al in 2014 [83].…”

Section: Fluorescent Microscopy: Imaging Of Glycogen Accumulation Usi...mentioning

confidence: 99%

Fluoride as a Potential Repressor of Glycogen Metabolism in Skeletal Muscle Cell Line CCL136

et al. 2023

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The exposure of humans to fluorine is connected with its presence in the air, food and water. It is well known that fluorides even at a low concentration but with long time exposure accumulate in the body and lead to numerous metabolic disorders. Fluoride is recognised as a factor modulating the energy metabolism of cells. This interaction is of particular importance in muscle cells, which are cells with high metabolic activity related to the metabolism of glucose and glycogen. In someone suffering from chronic fluoride poisoning, frequent symptoms are chronic fatigue not relieved by extra sleep or rest, muscular weakness, muscle spasms, involuntary twitching. The aim of this study was to examine the effect of fluorine at concentrations determined in blood of people environmentally exposed to fluorides on activity and expression of enzymes taking part in metabolism of muscle glycogen. CCL136 cells were cultured under standard conditions with the addition of NaF. The amount of ATP produced by the cells was determined using the HPLC method, the amount and expression of genes responsible for glycogen metabolism using WB and RT PCR methods and the amount of glycogen in cells using the fluorimetric and PAS methods. It has been shown that in CCL136 cells exposed to 1, 3 and 10 μM NaF there is a change in the energy state and expression pattern of enzymes involved in the synthesis and breakdown of glycogen. It was observed that NaF caused a decrease in ATP content in CCL136 cells. Fluoride exposure also increased glycogen deposition. These changes were accompanied by a decrease in gene expression and the level of enzymatic proteins related to glycogen metabolism: glycogen synthase, glycogen synthase kinase and glycogen phosphorylase. The results obtained shed new light on the molecular mechanisms by which fluoride acts as an environmental toxin.

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Detecting Glycogen in Peripheral Blood Mononuclear Cells with Periodic Acid Schiff Staining

Cited by 5 publications

References 0 publications

Optimization of PAS reaction and similar Schiff`s based methods for glycogen demonstration in liver tissue.

Optimization of PAS reaction and similar Schiff`s based methods for glycogen demonstration in liver tissue.

PI3K-dependant reprogramming of hexokinase isoforms controls glucose metabolism and functional responses of B lymphocytes

Fluoride as a Potential Repressor of Glycogen Metabolism in Skeletal Muscle Cell Line CCL136

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