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Rainbow trout glucose transporter (OnmyGLUT1): functional assessment in Xenopus laevis oocytes and expression in fish embryos

Heli Teerijoki¹, Aleksei Krasnov^1,*, Yuri Gorodilov², Sanjeev Krishna³ and Hannu Mölsä¹

¹ Institute of Applied Biotechnology, University of Kuopio, PO Box 1627, Kuopio 70211, Finland,
² Biological Institute, University of Sanct Petersburg, Oranienbaum Chaussee 2, Stary Peterhof, Sanct Petersburg 198504, Russia and
³ Department of Infectious Diseases, St George’s Hospital Medical School, London SW17 0RE, UK

View larger version (11K): [in a new window]   Fig.1. The kinetics of D-glucose uptake by Xenopus leavis oocytes microinjected with OnmyGLUT1 mRNA was studied in three experiments. The results of a typical experiment are presented. Transport rates were determined under zero-trans conditions, as described in Materials and methods. Values are means ± S.E.M. (N=8). In this experiment, Km=9.3±2.3mmoll-1) and Vmax=8.8±1.0pmoloocyte-1h-1.

View larger version (1K): [in a new window]   Fig.2. Inhibition of D-glucose uptake by Xenopus laevis oocytes microinjected with OnmyGLUT1 mRNA. Results are expressed as a percentage of uptake of substrate into oocytes incubated without inhibitor (OmRNA). Values are means + S.E.M. (N=8). The results were corrected for uptake into oocytes injected with RNase-free water. *Significantly different from the control value (t-test; P<0.01).

View larger version (57K): [in a new window]   Fig.3. Expression of OnmyGLUT1 in rainbow trout embryos analysed by in situ hybridization. The developmental age of embryos was measured in ts units (see Materials and methods); the characteristic traits are as follows. (A) Early gastrula (36ts). The density of cells is highest in the germ ring (gr) and especially in the terminal node (tn). Scale bar, 1mm. (B) 5–7 somites (60–62ts). A multilayer embryo with the axial organs; h, head; t, tail. Scale bar, 1mm. (C) 14–15 somites (69–70ts). The length (L) of the embryo is 3mm. The most rostral part of the head (prospective forebrain, fb) expands as a result of the formation of two eye vesicles (ev). The neural crest (nc) is disposed along the borders separating the neural plate (np) from the lateral ectoderm (le). s, somite. Scale bar, 1mm. (D) 60–62 somites (115–117ts). In these embryos (L=5.8–6.0mm), the heart contracts and blood circulates in the dorsal aorta; the neural folds (nf) and anlagens of gill arches (ga) are visible. Scale bar, 10mm. (E) Early vitelline plexus (130ts). Somitogenesis has finished, and the development of the cerebral and caudal arteries is in progress; L=7.0–7.2mm. The upper jaw (uj), gill arches and pectoral fins (pf) are evident. Scale bar, 10mm. (F) Middle vitelline plexus (150ts). Bile fills the gut, and the semicircular canals in the otic vesicles have begun to develop; L=8.5–9.0mm.