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Gene Expr Patterns
2003 Aug 01;34:503-7. doi: 10.1016/s1567-133x(03)00057-7.
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The vesicular glutamate transporter 1 (xVGlut1) is expressed in discrete regions of the developing Xenopus laevis nervous system.
Gleason KK
,
Dondeti VR
,
Hsia HL
,
Cochran ER
,
Gumulak-Smith J
,
Saha MS
.
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As the major excitatory neurotransmitter in the vertebrate nervous system, glutamate not only plays an essential role in adult neural signaling, but has also been implicated as a trophic factor in neuronal cell maturation, differentiation, and survival. An essential component of the glutamatergic neurotransmission system is the family of glutamate transporters, a multigene family that codes for plasma membrane-bound as well as vesicle-bound proteins responsible for the removal of glutamate from the cleft and its re-uptake into the synaptic vesicle. Here we describe the spatial and temporal expression of the vesicular glutamate transporter (xVGlut1) during the early developmental stages of the amphibian Xenopus laevis. RNAse protection analysis and in situ hybridization reveal that xVGlut1 is first expressed at late neurula stages in the developing spinal cord and trigeminal nerve. By tailbud stages xVGlut1 transcripts are detected in several of the cranial nerves, the pineal gland, and medialforebrain. By hatching stages xVGlut1 expression reappears in localized tracts within the spinal cord. Expression levels increase throughout development into adulthood.
Fig. 2. RNAse protection assay showing the expression of xVGlut1 at various embryonic stages of development and in the adult brain and eye. Corresponding bands for EF1a, which was used a loading control, are shown below each experimental xVGlut1 band.
Fig. 3. Analysis of xVGlut1 spatial and temporal expression pattern throughout development using whole-mount in situ hybridization. In all cases, embryos are viewed laterally, with anterior to the right. (A) Stage 21 (late neurula). (B) Stage 25 (tailbud). (C) Stage 27 (late tailbud). (D) Stage 30 (hatching). (E) Stage 33 (hatching). (F) Stage 35 (swimming tadpole). (G) Stage 38 (swimming tadpole). sc, spinal cord; pg, pineal gland; e, eye; ol, olfactory placodes; fb, forebrain; ot, otic vesicle; hb, hindbrain. V, cranial nerve V, trigeminal nerve; VII, cranial nerve VII, geniculate (facialis) nerve; VIII, cranial nerve VIII, acousticus nerve; IX, cranial nerve IX, glossopharyngeal nerve; X, cranial nerve X, vagal nerve. Scale bars are equivalent to 0.5 mm for A–E, and 1.0 mm for F,G.
Fig. 4. Histological analysis of xVGlut1 mRNA expression. Following whole-mount in situ hybridization, embryos were embedded and serially sectioned. All sections are transverse, 10-mm sections. (A) Staining is present in the neural tube of an early tailbud stage (stage 23) embryo. Panels (B–I) represent serial, anterior-to-posterior sections of a stage 35 (swimming tadpole) stage embryo. (B) Most anteriorly, xVGlut1 expression is present in the olfactory placodes (designated by arrow) and the forebrain region of the neural tube. (C) Slightly more posterior, signal is present in the pineal gland (designated by arrow). (D) At the level of the anterior aspect of the eye, xVGlut1 mRNA is present in the medialforebrain. (E,F) At the level of the posterior aspect of the eye, signal is prominent in cranial nerves V and VII (designated by arrows). (G,H) In sections at the level of the otic vesicle, xVGlut1 mRNA is expressed in cranial nerves VIII and IX (designated by arrows) and in the medialhindbrain. (I) Further posterior, signal is restricted to two bilateral stripes in the developing spinal cord. np, neural plate; nt, neural tube; cg, cement glands, ol, olfactory placodes; pg, pineal gland; e, eye; no, notochord; ot, otic vesicle; so, somites. Cranial nerves: V, trigeminal; VII, facialis; VIII, acousticus; IX, glossopharyngeal; X, vagus. Scale bars represent 0.5 mm for A–C and 0.75 mm for D–I.
