May 1, 1993;
Expression of a Xenopus Distal-less homeobox gene involved in forebrain and cranio-facial development.
Homeobox-containing genes are thought to perform essential functions in the process of pattern formation in vertebrates and invertebrates. They provide cells with positional information critical for normal embryonic development. Since most of the identified homeobox genes in Xenopus seem to provide positional information for the development of the trunk
, we have concentrated on genes that may be specifically involved in the formation of the head
region. Using a polymerase chain reaction strategy we have searched for Xenopus homeobox-containing genes that might provide positional cues for correct development of the brain
. In this paper we report the identification and cloning of a novel gene that by homology appears to be a member of the Distal
-less homeobox gene family. We show that its temporal expression patterns in the cement gland, neural crest derived visceral arches, retina
, while quite diverse, does suggest shared developmental features which may be required for correct craniofacial development and the regionalization of the Xenopus brain
. Furthermore, expression of this gene at later stages is primarily restricted to the tadpole forebrain
suggesting that the Distal
-less gene product continues to play a role after the initial brain
patterning is complete.
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Fig. 3. (A) Wholemount in situ hybridization of XDLL-1 cRNA to stage 18 embryo visualizes expression of this gene in the cement gland (arrow), This is a lateral view of the embryo. (B) Anterior view of the same embryo. (C) Lateral view of stage 24 embryo demonstrates the expression of XDLL-1 in the neural crest derived visceral arches. 1, cement gland; 2, mandibular arch; 3, hyoid arch; 4, anterior branchial arch; 5, posterior branchial arch. Triangle points to the otic vesicle. (D) Anterior view of the same embryo as in (C), In this view the cement gland and the mandibular arch show strong hybridization. Arrow points to the hybridisation signal in the lateral diencephalon. (E) In situ hybridization to stage 30 embryo (lateral view), At this stage in addition to the cement gland and visceral arches, the diencephalon (arrow) shows strong hybridization. (F) A dorsal view of the same embryo as in (E) shows the segment-like hybridization of XDLL-1 to the visceral arches in the head region. The posterior part of the embryo in (E) and (F) has been removed.
Fig. 4. (A) Whole-mount in situ hybridization to a stage 32 embryo (anterior view) showing the most intense area of XDLL-1 expression in the forebrain - the lateral diencephalic region (arrow) (B) In situ hybridization of XDLL-1 to a stage 33/34 embryo (anterior view), At this stage the entire ventral forebrain is uniformly labeled. (C) A lateral view of the embryo in (B) showing the boundaries of XDLL-1 hybridization within the brain. Upper arrow (e) points at the epiphysis which delineates the dorsal limit of the telencephalon. The lower arrow (d) points at the dorsal limit of XDLL-1 expression boundary in the telencephalon which is ventral to the epiphysis. O, otic vesicle, upper triangle points to the plane of section in (H), lower triangle points to the plane of section in (G). (D) In situ hybridization of XDLL-1 to stage 42 brain. Lateral view. Forebrain is to the left. t, telencephalon; d, diencephalon; i, infundibulum. Apparent signal in the hindbrain area is due to pigment granules and not to hybridization. (E) Wholemount hybridization to stage 43 brain. Arrows indicate the plane of section in (F). (F) Cross section of the telencephalic region of the brain in (E) Notice the sharp boundaries of labeling. (G) Coronal section of the embryo in (c) at the level of pharyngeal arches (lower triangle in (C)). 1, ectoderm; 2, neural crest; 3, mesoderm; 4, endoderm. (H) Coronal section of the embryo in (C) at the level of the eyes (upper triangle in (C)). d, diencephalon; 1, lens; o, otic vesicle; r, retina; t, telencephalon. Notice the presence of hybridization in the retina and the absence of it in the lens.
Fig. 5. (A) sagital section of a stage 45 brain. Arrow, area olfactoria primitiva; triangle, ventromedial thalamus; a, b, c, d indicate the planes of sections in Fig. 6A, B, C and D respectively. (B) A ventral view of the brain in (A) visualizing an increased level of expression of XDLL-1 in the neuroepithelium of medial preoptic nucleus (1), arcuate nucleus (2) and hypothalamus, pars ventralis (3).
Fig. 6. (A) Transverse section of the telencephalic region in Fig. 5A. Plane of section is indicated by (a). (B) Transverse section of the telencephalic region of the brain in Fig. 5A. Level of the section indicated by letter (b). (C) Transverse section through the mesencephalon and dorsal diencephalon of the brain in Fig. 5A. Plane of section indicated by and letter (c). (D) Coronal section of the brain in 5A. Plane of section is indicated by a letter (d). Arrow points to the strongly labeled area olfactoria primitiva.