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Phylogenesis and Biological Characterization of a New Glucose Transporter in the Chicken (Gallus gallus), GLUT12.

Coudert E, Pascal G, Dupont J, Simon J, Cailleau-Audouin E, Crochet S, Duclos MJ, Tesseraud S, Métayer-Coustard S - PLoS ONE (2015)

Bottom Line: Third a physiological characterization was performed: SLC2A12 mRNA levels were significantly lowered in fed chickens subjected to insulin immuno-neutralization.Finally, recruitment of immuno-reactive GLUT12 to the muscle plasma membrane was increased following 1h of intraperitoneal insulin administration (compared to a control fasted state).In conclusion, these results suggest that the facilitative glucose transporter protein GLUT12 could act in chicken muscle as an insulin-sensitive transporter that is qualitatively similar to GLUT4 in mammals.

View Article: PubMed Central - PubMed

Affiliation: UR83 Recherches Avicoles, Institut National de la Recherche Agronomique, Nouzilly, France.

ABSTRACT
In mammals, insulin-sensitive GLUTs, including GLUT4, are recruited to the plasma membrane of adipose and muscle tissues in response to insulin. The GLUT4 gene is absent from the chicken genome, and no functional insulin-sensitive GLUTs have been characterized in chicken tissues to date. A nucleotide sequence is predicted to encode a chicken GLUT12 ortholog and, interestingly, GLUT12 has been described to act as an insulin-sensitive GLUT in mammals. It encodes a 596 amino acid protein exhibiting 71% identity with human GLUT12. First, we present the results of a phylogenetic study showing the stability of this gene during evolution of vertebrates. Second, tissue distribution of chicken SLC2A12 mRNA was characterized by RT-PCR. It was predominantly expressed in skeletal muscle and heart. Protein distribution was analysed by Western blotting using an anti-human GLUT12 antibody directed against a highly conserved region (87% of identity). An immuno-reactive band of the expected size (75kDa) was detected in the same tissues. Third a physiological characterization was performed: SLC2A12 mRNA levels were significantly lowered in fed chickens subjected to insulin immuno-neutralization. Finally, recruitment of immuno-reactive GLUT12 to the muscle plasma membrane was increased following 1h of intraperitoneal insulin administration (compared to a control fasted state). Thus insulin administration elicited membrane GLUT12 recruitment. In conclusion, these results suggest that the facilitative glucose transporter protein GLUT12 could act in chicken muscle as an insulin-sensitive transporter that is qualitatively similar to GLUT4 in mammals.

No MeSH data available.


GLUT12 translocation after insulin stimulation.In the upper part, a representative Western blot shows GLUT12 content in membrane fractions prepared from leg muscle of fasted and fed animals without insulin injection or 1hr after insulin injection (1U/kg). In the lower part, a representative Western blot shows GLUT12 content in the corresponding cytosolic proteins used as a control of the cytosolic GLUT12 content. Vinculin was used as a loading control for the two fractions.
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pone.0139517.g008: GLUT12 translocation after insulin stimulation.In the upper part, a representative Western blot shows GLUT12 content in membrane fractions prepared from leg muscle of fasted and fed animals without insulin injection or 1hr after insulin injection (1U/kg). In the lower part, a representative Western blot shows GLUT12 content in the corresponding cytosolic proteins used as a control of the cytosolic GLUT12 content. Vinculin was used as a loading control for the two fractions.

Mentions: Membrane and cytosolic proteins were prepared from leg muscles and analysed by Western blotting (Fig 8). The 75 kDa immuno-reactive band was observed in membrane fractions (Fig 8, upper part) from fed animals in the absence of insulin injection, but not in fasted animals. Membrane GLUT12 protein significantly increased in fasted and fed conditions in response to insulin injection. In the cytosolic fraction (Fig 8, lower part), the immuno-reactive GLUT12 band was detected at identical levels, whatever the experimental treatment. These observations demonstrate that GLUT12 translocates to the plasma membrane in response to insulin in chicken skeletal muscle.


Phylogenesis and Biological Characterization of a New Glucose Transporter in the Chicken (Gallus gallus), GLUT12.

Coudert E, Pascal G, Dupont J, Simon J, Cailleau-Audouin E, Crochet S, Duclos MJ, Tesseraud S, Métayer-Coustard S - PLoS ONE (2015)

GLUT12 translocation after insulin stimulation.In the upper part, a representative Western blot shows GLUT12 content in membrane fractions prepared from leg muscle of fasted and fed animals without insulin injection or 1hr after insulin injection (1U/kg). In the lower part, a representative Western blot shows GLUT12 content in the corresponding cytosolic proteins used as a control of the cytosolic GLUT12 content. Vinculin was used as a loading control for the two fractions.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4592010&req=5

pone.0139517.g008: GLUT12 translocation after insulin stimulation.In the upper part, a representative Western blot shows GLUT12 content in membrane fractions prepared from leg muscle of fasted and fed animals without insulin injection or 1hr after insulin injection (1U/kg). In the lower part, a representative Western blot shows GLUT12 content in the corresponding cytosolic proteins used as a control of the cytosolic GLUT12 content. Vinculin was used as a loading control for the two fractions.
Mentions: Membrane and cytosolic proteins were prepared from leg muscles and analysed by Western blotting (Fig 8). The 75 kDa immuno-reactive band was observed in membrane fractions (Fig 8, upper part) from fed animals in the absence of insulin injection, but not in fasted animals. Membrane GLUT12 protein significantly increased in fasted and fed conditions in response to insulin injection. In the cytosolic fraction (Fig 8, lower part), the immuno-reactive GLUT12 band was detected at identical levels, whatever the experimental treatment. These observations demonstrate that GLUT12 translocates to the plasma membrane in response to insulin in chicken skeletal muscle.

Bottom Line: Third a physiological characterization was performed: SLC2A12 mRNA levels were significantly lowered in fed chickens subjected to insulin immuno-neutralization.Finally, recruitment of immuno-reactive GLUT12 to the muscle plasma membrane was increased following 1h of intraperitoneal insulin administration (compared to a control fasted state).In conclusion, these results suggest that the facilitative glucose transporter protein GLUT12 could act in chicken muscle as an insulin-sensitive transporter that is qualitatively similar to GLUT4 in mammals.

View Article: PubMed Central - PubMed

Affiliation: UR83 Recherches Avicoles, Institut National de la Recherche Agronomique, Nouzilly, France.

ABSTRACT
In mammals, insulin-sensitive GLUTs, including GLUT4, are recruited to the plasma membrane of adipose and muscle tissues in response to insulin. The GLUT4 gene is absent from the chicken genome, and no functional insulin-sensitive GLUTs have been characterized in chicken tissues to date. A nucleotide sequence is predicted to encode a chicken GLUT12 ortholog and, interestingly, GLUT12 has been described to act as an insulin-sensitive GLUT in mammals. It encodes a 596 amino acid protein exhibiting 71% identity with human GLUT12. First, we present the results of a phylogenetic study showing the stability of this gene during evolution of vertebrates. Second, tissue distribution of chicken SLC2A12 mRNA was characterized by RT-PCR. It was predominantly expressed in skeletal muscle and heart. Protein distribution was analysed by Western blotting using an anti-human GLUT12 antibody directed against a highly conserved region (87% of identity). An immuno-reactive band of the expected size (75kDa) was detected in the same tissues. Third a physiological characterization was performed: SLC2A12 mRNA levels were significantly lowered in fed chickens subjected to insulin immuno-neutralization. Finally, recruitment of immuno-reactive GLUT12 to the muscle plasma membrane was increased following 1h of intraperitoneal insulin administration (compared to a control fasted state). Thus insulin administration elicited membrane GLUT12 recruitment. In conclusion, these results suggest that the facilitative glucose transporter protein GLUT12 could act in chicken muscle as an insulin-sensitive transporter that is qualitatively similar to GLUT4 in mammals.

No MeSH data available.