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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.


Comparative maps of SLC2A12 genomic region.(A) The genomic location of SLC2A12. synteny blocks between human Chromosome 6 (Chr 6) and chicken chromosomes 1, 2, 3 and 26 (Chr 1, Chr 2, Chr3 and Chr 26) according to Ensembl (http://www.ensembl.org). The arrows indicate SLC2A12 regions. (B) Gene arrangement of the genomic region encompassing SLC2A12 according to the genome browser Genomicus (http://www.genomicus.biologie.ens.fr, Genomicus—database version: 79.01 / Web-code version: 2014-09-19). Schematic gene maps of the conserved syntenic regions of SLC2A12 in Gallus gallus and in other vertebrates.The gene initially placed in the centre of the display and aligned over a vertical black line is the gene that was used as query (reference gene = SLC2A12). Coloured genes over a light blue background corresponds to extant or ancestral genes that are orthologous to genes from the species used in the query that show the same colour. The coding direction of the genes is indicated by the pointed end. Chr: denotes the chromosome. Neighbouring genes upstream and downstream in the chicken and other species: SGK1, serum/glucocorticoid regulated kinase 1; ALDH8A1, aldehyde dehydrogenase 8 family, member A1; HBS1L, uncharacterized protein; TBPL1, TBP-like 1; TCF21, transcription factor 21; EYA4, eyes absent homolog 4; RPS12, ribosomal protein S12.
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pone.0139517.g004: Comparative maps of SLC2A12 genomic region.(A) The genomic location of SLC2A12. synteny blocks between human Chromosome 6 (Chr 6) and chicken chromosomes 1, 2, 3 and 26 (Chr 1, Chr 2, Chr3 and Chr 26) according to Ensembl (http://www.ensembl.org). The arrows indicate SLC2A12 regions. (B) Gene arrangement of the genomic region encompassing SLC2A12 according to the genome browser Genomicus (http://www.genomicus.biologie.ens.fr, Genomicus—database version: 79.01 / Web-code version: 2014-09-19). Schematic gene maps of the conserved syntenic regions of SLC2A12 in Gallus gallus and in other vertebrates.The gene initially placed in the centre of the display and aligned over a vertical black line is the gene that was used as query (reference gene = SLC2A12). Coloured genes over a light blue background corresponds to extant or ancestral genes that are orthologous to genes from the species used in the query that show the same colour. The coding direction of the genes is indicated by the pointed end. Chr: denotes the chromosome. Neighbouring genes upstream and downstream in the chicken and other species: SGK1, serum/glucocorticoid regulated kinase 1; ALDH8A1, aldehyde dehydrogenase 8 family, member A1; HBS1L, uncharacterized protein; TBPL1, TBP-like 1; TCF21, transcription factor 21; EYA4, eyes absent homolog 4; RPS12, ribosomal protein S12.

Mentions: The analysis of chicken GLUTs (Table 1) showed the existence of a GLUT12 ortholog (corresponding to the SLC2A12 gene) in the chicken. The SLC2A12 gene is located on chromosome 3, and comprises 5 exons and encodes a putative 596 amino acid protein. The phylogenetic analysis of GLUT12 amino acid sequences in different species (Table 2) is presented in Fig 3. We decided to determine whether selection pressures acted on the avian GLUT12 sequence. No positive selection pressure was evidenced in these 19 species. Synteny analysis and comprehensive genome location searches (Fig 4A and 4B) showed a similar arrangement of the SLC2A12 gene and its neighbouring genes upstream and downstream in the chicken and other species.


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)

Comparative maps of SLC2A12 genomic region.(A) The genomic location of SLC2A12. synteny blocks between human Chromosome 6 (Chr 6) and chicken chromosomes 1, 2, 3 and 26 (Chr 1, Chr 2, Chr3 and Chr 26) according to Ensembl (http://www.ensembl.org). The arrows indicate SLC2A12 regions. (B) Gene arrangement of the genomic region encompassing SLC2A12 according to the genome browser Genomicus (http://www.genomicus.biologie.ens.fr, Genomicus—database version: 79.01 / Web-code version: 2014-09-19). Schematic gene maps of the conserved syntenic regions of SLC2A12 in Gallus gallus and in other vertebrates.The gene initially placed in the centre of the display and aligned over a vertical black line is the gene that was used as query (reference gene = SLC2A12). Coloured genes over a light blue background corresponds to extant or ancestral genes that are orthologous to genes from the species used in the query that show the same colour. The coding direction of the genes is indicated by the pointed end. Chr: denotes the chromosome. Neighbouring genes upstream and downstream in the chicken and other species: SGK1, serum/glucocorticoid regulated kinase 1; ALDH8A1, aldehyde dehydrogenase 8 family, member A1; HBS1L, uncharacterized protein; TBPL1, TBP-like 1; TCF21, transcription factor 21; EYA4, eyes absent homolog 4; RPS12, ribosomal protein S12.
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pone.0139517.g004: Comparative maps of SLC2A12 genomic region.(A) The genomic location of SLC2A12. synteny blocks between human Chromosome 6 (Chr 6) and chicken chromosomes 1, 2, 3 and 26 (Chr 1, Chr 2, Chr3 and Chr 26) according to Ensembl (http://www.ensembl.org). The arrows indicate SLC2A12 regions. (B) Gene arrangement of the genomic region encompassing SLC2A12 according to the genome browser Genomicus (http://www.genomicus.biologie.ens.fr, Genomicus—database version: 79.01 / Web-code version: 2014-09-19). Schematic gene maps of the conserved syntenic regions of SLC2A12 in Gallus gallus and in other vertebrates.The gene initially placed in the centre of the display and aligned over a vertical black line is the gene that was used as query (reference gene = SLC2A12). Coloured genes over a light blue background corresponds to extant or ancestral genes that are orthologous to genes from the species used in the query that show the same colour. The coding direction of the genes is indicated by the pointed end. Chr: denotes the chromosome. Neighbouring genes upstream and downstream in the chicken and other species: SGK1, serum/glucocorticoid regulated kinase 1; ALDH8A1, aldehyde dehydrogenase 8 family, member A1; HBS1L, uncharacterized protein; TBPL1, TBP-like 1; TCF21, transcription factor 21; EYA4, eyes absent homolog 4; RPS12, ribosomal protein S12.
Mentions: The analysis of chicken GLUTs (Table 1) showed the existence of a GLUT12 ortholog (corresponding to the SLC2A12 gene) in the chicken. The SLC2A12 gene is located on chromosome 3, and comprises 5 exons and encodes a putative 596 amino acid protein. The phylogenetic analysis of GLUT12 amino acid sequences in different species (Table 2) is presented in Fig 3. We decided to determine whether selection pressures acted on the avian GLUT12 sequence. No positive selection pressure was evidenced in these 19 species. Synteny analysis and comprehensive genome location searches (Fig 4A and 4B) showed a similar arrangement of the SLC2A12 gene and its neighbouring genes upstream and downstream in the chicken and other species.

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.