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Non-classical gluconeogenesis-dependent glucose metabolism in Rhipicephalus microplus embryonic cell line BME26.

da Silva RM, Noce BD, Waltero CF, Costa EP, de Abreu LA, Githaka NW, Moraes J, Gomes HF, Konnai S, Vaz Ida S, Ohashi K, Logullo C - Int J Mol Sci (2015)

Bottom Line: In this work we evaluated several genes involved in gluconeogenesis, glycolysis and glycogen metabolism, the major pathways for carbohydrate catabolism and anabolism, in the BME26 Rhipicephalus microplus embryonic cell line.Surprisingly, the transcription of gluconeogenic enzymes was found to increase alongside that of glycolytic enzymes, especially pyruvate kinase, with high glucose treatment.In addition, RNAi data from this study revealed that the transcription of gluconeogenic genes in BME26 cells is controlled by GSK-3.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Chemistry and Function of Proteins and Peptides, Animal Experimentation Unit, UENF, Av. Alberto Lamego, 2000, Horto, CEP 28013-602 Campos dos Goytacazes, RJ, Brazil. rjrenato@ig.com.br.

ABSTRACT
In this work we evaluated several genes involved in gluconeogenesis, glycolysis and glycogen metabolism, the major pathways for carbohydrate catabolism and anabolism, in the BME26 Rhipicephalus microplus embryonic cell line. Genetic and catalytic control of the genes and enzymes associated with these pathways are modulated by alterations in energy resource availability (primarily glucose). BME26 cells in media were investigated using three different glucose concentrations, and changes in the transcription levels of target genes in response to carbohydrate utilization were assessed. The results indicate that several genes, such as glycogen synthase (GS), glycogen synthase kinase 3 (GSK3), phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6 phosphatase (GP) displayed mutual regulation in response to glucose treatment. Surprisingly, the transcription of gluconeogenic enzymes was found to increase alongside that of glycolytic enzymes, especially pyruvate kinase, with high glucose treatment. In addition, RNAi data from this study revealed that the transcription of gluconeogenic genes in BME26 cells is controlled by GSK-3. Collectively, these results improve our understanding of how glucose metabolism is regulated at the genetic level in tick cells.

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Glycogen degradation has a transcriptional control in BME26 cells. Transcriptional analysis of glycogen debranching enzyme (A) and phosphoglucomutase (B) in embryonic Rhipicephalus microplus cells (BME26) in response to glucose treatment. Control: cells maintained with 50 mM of glucose; Low: cell maintained without glucose addition; and High: cells maintained with 100 mM of glucose. The experiment was performed with three independent biological samples in three experimental replicates each (*p < 0.05; **p < 0.001, ANOVA).
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ijms-16-01821-f006: Glycogen degradation has a transcriptional control in BME26 cells. Transcriptional analysis of glycogen debranching enzyme (A) and phosphoglucomutase (B) in embryonic Rhipicephalus microplus cells (BME26) in response to glucose treatment. Control: cells maintained with 50 mM of glucose; Low: cell maintained without glucose addition; and High: cells maintained with 100 mM of glucose. The experiment was performed with three independent biological samples in three experimental replicates each (*p < 0.05; **p < 0.001, ANOVA).

Mentions: The regulation of glycogen mobilization was assessed by analyzing PGM and GDE transcription. Both enzymes are involved in glycogen polymer degradation. GDE cleaves the α-1,6-glycosidic bond between adjacent glucose molecules in the glycogen polymer, assisting glycogen phosphorylase and PGM [35,36]. PGM transcription is elevated in low-glucose cells (Figure 6B). This result suggests a type of genetic regulation that increases the transcript level of PGM when glucose availability is low and glycogen mobilization is necessary. On the other hand, the transcription of GDE does not change when glucose amounts are low (Figure 6A). However, when the cells were treated under high-glucose conditions, GDE transcription increased significantly, indicating a differential genetic regulation in relation to PGM.


Non-classical gluconeogenesis-dependent glucose metabolism in Rhipicephalus microplus embryonic cell line BME26.

da Silva RM, Noce BD, Waltero CF, Costa EP, de Abreu LA, Githaka NW, Moraes J, Gomes HF, Konnai S, Vaz Ida S, Ohashi K, Logullo C - Int J Mol Sci (2015)

Glycogen degradation has a transcriptional control in BME26 cells. Transcriptional analysis of glycogen debranching enzyme (A) and phosphoglucomutase (B) in embryonic Rhipicephalus microplus cells (BME26) in response to glucose treatment. Control: cells maintained with 50 mM of glucose; Low: cell maintained without glucose addition; and High: cells maintained with 100 mM of glucose. The experiment was performed with three independent biological samples in three experimental replicates each (*p < 0.05; **p < 0.001, ANOVA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307336&req=5

ijms-16-01821-f006: Glycogen degradation has a transcriptional control in BME26 cells. Transcriptional analysis of glycogen debranching enzyme (A) and phosphoglucomutase (B) in embryonic Rhipicephalus microplus cells (BME26) in response to glucose treatment. Control: cells maintained with 50 mM of glucose; Low: cell maintained without glucose addition; and High: cells maintained with 100 mM of glucose. The experiment was performed with three independent biological samples in three experimental replicates each (*p < 0.05; **p < 0.001, ANOVA).
Mentions: The regulation of glycogen mobilization was assessed by analyzing PGM and GDE transcription. Both enzymes are involved in glycogen polymer degradation. GDE cleaves the α-1,6-glycosidic bond between adjacent glucose molecules in the glycogen polymer, assisting glycogen phosphorylase and PGM [35,36]. PGM transcription is elevated in low-glucose cells (Figure 6B). This result suggests a type of genetic regulation that increases the transcript level of PGM when glucose availability is low and glycogen mobilization is necessary. On the other hand, the transcription of GDE does not change when glucose amounts are low (Figure 6A). However, when the cells were treated under high-glucose conditions, GDE transcription increased significantly, indicating a differential genetic regulation in relation to PGM.

Bottom Line: In this work we evaluated several genes involved in gluconeogenesis, glycolysis and glycogen metabolism, the major pathways for carbohydrate catabolism and anabolism, in the BME26 Rhipicephalus microplus embryonic cell line.Surprisingly, the transcription of gluconeogenic enzymes was found to increase alongside that of glycolytic enzymes, especially pyruvate kinase, with high glucose treatment.In addition, RNAi data from this study revealed that the transcription of gluconeogenic genes in BME26 cells is controlled by GSK-3.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Chemistry and Function of Proteins and Peptides, Animal Experimentation Unit, UENF, Av. Alberto Lamego, 2000, Horto, CEP 28013-602 Campos dos Goytacazes, RJ, Brazil. rjrenato@ig.com.br.

ABSTRACT
In this work we evaluated several genes involved in gluconeogenesis, glycolysis and glycogen metabolism, the major pathways for carbohydrate catabolism and anabolism, in the BME26 Rhipicephalus microplus embryonic cell line. Genetic and catalytic control of the genes and enzymes associated with these pathways are modulated by alterations in energy resource availability (primarily glucose). BME26 cells in media were investigated using three different glucose concentrations, and changes in the transcription levels of target genes in response to carbohydrate utilization were assessed. The results indicate that several genes, such as glycogen synthase (GS), glycogen synthase kinase 3 (GSK3), phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6 phosphatase (GP) displayed mutual regulation in response to glucose treatment. Surprisingly, the transcription of gluconeogenic enzymes was found to increase alongside that of glycolytic enzymes, especially pyruvate kinase, with high glucose treatment. In addition, RNAi data from this study revealed that the transcription of gluconeogenic genes in BME26 cells is controlled by GSK-3. Collectively, these results improve our understanding of how glucose metabolism is regulated at the genetic level in tick cells.

Show MeSH