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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.


Tissue expression pattern of GLUT12 in the chicken.(A) Tissues from fed chickens were analysed for SLC12A12 expression by Real-Time PCR (N = 6). Values represent means and standard errors. Data were subjected to analysis of variance (ANOVA) to detect significant intergroup differences. Statistical model was tissue comparison. The means were compared by Fisher’s least significant difference test. P<0.05 was considered statistically significant. (B) Tissues from fed chickens were analysed by Western blotting using an anti-GLUT12 antibody for protein distribution analyses (N = 5). In grey: skeletal muscles (Pectoralis major and leg muscle). Vinculin was used as a protein loading control. (C) Protein distribution was analysed from Pectoralis major and the leg muscles, collected at 19 days during embryogenesis, hatch and 5 days post-hatch. (N = 7).
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pone.0139517.g006: Tissue expression pattern of GLUT12 in the chicken.(A) Tissues from fed chickens were analysed for SLC12A12 expression by Real-Time PCR (N = 6). Values represent means and standard errors. Data were subjected to analysis of variance (ANOVA) to detect significant intergroup differences. Statistical model was tissue comparison. The means were compared by Fisher’s least significant difference test. P<0.05 was considered statistically significant. (B) Tissues from fed chickens were analysed by Western blotting using an anti-GLUT12 antibody for protein distribution analyses (N = 5). In grey: skeletal muscles (Pectoralis major and leg muscle). Vinculin was used as a protein loading control. (C) Protein distribution was analysed from Pectoralis major and the leg muscles, collected at 19 days during embryogenesis, hatch and 5 days post-hatch. (N = 7).

Mentions: The tissue expression pattern of the SLC2A12 messenger RNA was analysed in various tissues from fed chickens by real-time PCR. SLC2A12 mRNA was detected in all tissue samples, but was the most abundant in skeletal (Pectoralis major and leg muscles) muscles, i.e. insulin-sensitive tissues (Fig 6A).


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)

Tissue expression pattern of GLUT12 in the chicken.(A) Tissues from fed chickens were analysed for SLC12A12 expression by Real-Time PCR (N = 6). Values represent means and standard errors. Data were subjected to analysis of variance (ANOVA) to detect significant intergroup differences. Statistical model was tissue comparison. The means were compared by Fisher’s least significant difference test. P<0.05 was considered statistically significant. (B) Tissues from fed chickens were analysed by Western blotting using an anti-GLUT12 antibody for protein distribution analyses (N = 5). In grey: skeletal muscles (Pectoralis major and leg muscle). Vinculin was used as a protein loading control. (C) Protein distribution was analysed from Pectoralis major and the leg muscles, collected at 19 days during embryogenesis, hatch and 5 days post-hatch. (N = 7).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139517.g006: Tissue expression pattern of GLUT12 in the chicken.(A) Tissues from fed chickens were analysed for SLC12A12 expression by Real-Time PCR (N = 6). Values represent means and standard errors. Data were subjected to analysis of variance (ANOVA) to detect significant intergroup differences. Statistical model was tissue comparison. The means were compared by Fisher’s least significant difference test. P<0.05 was considered statistically significant. (B) Tissues from fed chickens were analysed by Western blotting using an anti-GLUT12 antibody for protein distribution analyses (N = 5). In grey: skeletal muscles (Pectoralis major and leg muscle). Vinculin was used as a protein loading control. (C) Protein distribution was analysed from Pectoralis major and the leg muscles, collected at 19 days during embryogenesis, hatch and 5 days post-hatch. (N = 7).
Mentions: The tissue expression pattern of the SLC2A12 messenger RNA was analysed in various tissues from fed chickens by real-time PCR. SLC2A12 mRNA was detected in all tissue samples, but was the most abundant in skeletal (Pectoralis major and leg muscles) muscles, i.e. insulin-sensitive tissues (Fig 6A).

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.