Rainbow trout glucose transporter (OnmyGLUT1): functional assessment in Xenopus laevis oocytes and expression in fish embryos.
Recently, we reported the cloning of a putative glucose transporter (OnmyGLUT1) from rainbow trout embryos. In this paper, we describe the functional characteristics of OnmyGLUT1 and its expression during embryonic development of rainbow trout. Transport of D-glucose was analysed in Xenopus laevis oocytes following microinjection of mRNA transcribed in vitro. These experiments confirmed that OnmyGLUT1 is a facilitative Na(+)-independent transporter. Assessment of substrate selectivity, sensitivity to cytochalasin B and phloretin and kinetic parameters showed that the rainbow trout glucose transporter was similar to a carp transporter and to mammalian GLUT1. Embryonic expression of OnmyGLUT1 was studied using whole-mount in situ hybridization. Ubiquitous distribution of transcripts was observed until the early phase of somitogenesis. During the course of organogenesis, somitic expression decreased along the rostro-caudal axis, finally ceasing in the mature somites. The OnmyGLUT1 transcripts were detected in the neural crest during the whole study period. Transcripts were also found in structures that are likely to originate from the neural crest cells (gill arches, pectoral fins, upper jaw, olfactory organs and primordia of mouth lips). Hexose transport activity was detected at all developmental stages after blastulation. Cytochalasin B blocked the accumulation of phosphorylated 2-deoxy-D-glucose by dissociated embryonic cells, suggesting an important role for transport in glucose metabolism.
Teerijoki
H
H
Institute of Applied Biotechnology, University of Kuopio, PO Box 1627, Kuopio 70211, Finland.
Krasnov
A
A
Gorodilov
Y
Y
Krishna
S
S
Mölsä
H
H
D000818:Animals; D003847:Deoxyglucose / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D051272:Glucose Transporter Type 1; D017403:In Situ Hybridization; D007700:Kinetics; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014982:Xenopus laevis
D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; D003847:Deoxyglucose; D005947:Glucose