Ursula C. Dräger

Restricted expression and retinoic acid-induced downregulation of the retinaldehyde dehydrogenase type 2 (RALDH-2) gene during mouse development

Retinaldehyde dehydrogenase type 2 (RALDH-2) was identified as a major retinoic acid generating enzyme in the early embryo. Here we report the expression domains of the RALDH-2 gene during mouse embryogenesis, which are likely to indicate regions of endogenous retinoic acid (RA) synthesis. During early gastrulation, RALDH-2 is expressed in the mesoderm adjacent to the node and primitive streak. At the headfold stage, mesodermal expression is restricted to posterior regions up to the base of the headfolds. Later, RALDH-2 is transiently expressed in the undifferentiated somites and the optic vesicles, and more persistently along the lateral walls of the intraembryonic coelom and around the hindgut diverticulum. The RALDH-2 expression domains in differentiating limbs, which include presumptive interdigital regions, coincide with, but slightly precede, those of the RA-inducible RAR beta gene. The RALDH-2 gene is also expressed in specific regions of the developing head, including the tooth buds, inner ear, meninges and pituitary gland, and in several viscera. Administration of a teratogenic dose of RA at embryonic day 8.5 results in downregulation of RALDH-2 transcript levels in caudal regions of the embryo, and may reflect a mechanism of negative feedback regulation of RA synthesis.

Birth Dates of Retinal Ganglion Cells Giving Rise to the Crossed and Uncrossed Optic Projections in the Mouse

In the mouse, as in most mammals, the crossed optic projections originate from the entire extent of the retina, whereas ganglion cells giving rise to the uncrossed (ipsilateral) projection are restricted to the temporal and ventral retina. The nasal border of this bilaterally projecting region in the retina corresponds to the midline of the visual field. Here the birth dates of ipsilaterally and contralaterally projecting ganglion cells were determined by combining tritiated thymidine labelling in the embryo with horseradish peroxidase tracings from the optic tract in the adult. Contralaterally projecting ganglion cells were found to be generated from embryonic day E11 to about E19 in a crude concentric fashion with the oldest cells in central and youngest ones in peripheral retina. Ipsilaterally projecting cells were born from E11 to E16, that is, during the earlier part of the period in which the contralateral projection was born. At the earliest time of ganglion cell generation (E11-12 ) ipsi- and contralaterally projecting cells were born within separate retinal regions, with the future midline representation forming the border between the two zones. This distinction became lost after E13, when both ipsi- and contralaterally projecting cells were born in the bilaterally projecting region. Hence at E11-12 the retina was found to have a bipartite organization that may allow the specification of the two maps of opposite topographical polarity in which the crossed and uncrossed projections are organized. Since in the adult retina this bipartite organization is preserved only in the large ganglion cells that project to the lateral geniculate nucleus, and since large ganglion cells are known to be the earliest ones formed in the mouse, these cells may be the ones that establish the early and bilateral projections of the retina. The conclusion that the bilateral projection system in the retina reflects an early developmental programme, and not the result of competition between the two eyes at later stages, was reinforced by observing a practically normal retinal origin of ipsilateral projections in mice which had only one normal eye from the earliest stages of eye development.

Regulation of retinoic acid signaling in the embryonic nervous system: a master differentiation factor

This review describes some of the properties of retinoic acid (RA) in its functions as a locally synthesized differentiation factor for the developing nervous system. The emphasis is on the characterization of the metabolic enzymes that synthesize and inactivate RA, and which determine local RA concentrations. These enzymes create regions of autocrine and paracrine RA signaling in the embryo. One mechanism by which RA can act as a differentiation agent is through the induction of growth factors and their receptors. Induction of growth factor receptors in neural progenitor cells can lead to growth factor dependency, and the consequent developmental fate of the cell will depend on the local availability of growth factors. Because RA activates the early events of cell differentiation, which then induce context-specific differentiation programs, RA may be called a master differentiation factor.

A retinoic acid synthesizing enzyme in ventral retina and telencephalon of the embryonic mouse

Most retinoic acid (RA) in the embryonic mouse is generated by three retinaldehyde dehydrogenases (RALDHs). RALDH1 (also called E1, AHD2 or ALDH1) is expressed in the dorsal retina, and RALDH2 (V2, ALDH11) generates most RA in the embryonic trunk. The third one, RALDH3 (V1), synthesizes the bulk of RA in the head of the early embryo. We show here that RALDH3 is a mouse homologue to ALDH6, an aldehyde dehydrogenase cloned from adult human salivary gland (Hsu, L.C., Chang, W.-C., Hiraoka, L., Hsien, C.-L., 1994. Molecular cloning, genomic organization, and chromosomal localization of an additional human aldehyde dehydrogenase gene, ALDH6. Genomics 24, 333-341), which was recently reported to act as a RALDH (Yoshida, A., Rzhetsky, A., Hsu, L.C., Chang, C., 1998. Human aldehyde dehydrogenase gene family. Eur. J. Biochem. 251, 549-557). RALDH3 expression begins in the surface ectoderm over the optic recess. In rapidly changing expression patterns it labels the appearance of several ectodermal structures: it marks the formation of the lens and the olfactory organ from ectodermal placodes, and it delineates the beginning eyelid field. Within the optic vesicle, RALDH3 is expressed in the ventral retina and the dorsal pigment epithelium. In the telencephalon, RALDH3 is expressed at high levels in the lateral part of the ganglionic eminence. From here it extends via the piriform cortex into the lower part of the septum. Of the three RALDHs, RALDH3 shows the strongest predilection for epithelia.

DORSAL AND VENTRAL RETINAL TERRITORIES DEFINED BY RETINOIC ACID SYNTHESIS,BREAK-DOWN AND NUCLEAR RECEPTOR EXPRESSION

Determination of the dorso-ventral dimension of the vertebrate retina is known to involve retinoic acid (RA), in that high RA activates expression of a ventral retinaldehyde dehydrogenase and low RA of a dorsal dehydrogenase. Here we show that in the early eye vesicle of the mouse embryo, expression of the dorsal dehydrogenase is preceded by, and transiently overlaps with, the RA-degrading oxidase CYP26. Subsequently in the embryonic retina, CYP26 forms a narrow horizontal boundary between the dorsal and ventral dehydrogenases, creating a trough between very high ventral and moderately high dorsal RA levels. Most of the RA receptors are expressed uniformly throughout the retina except for the RA-sensitive RARbeta, which is down-regulated in the CYP26 stripe. The orphan receptor COUP-TFII, which modulates RA responses, colocalizes with the dorsal dehydrogenase. The organization of the embryonic vertebrate retina into dorsal and ventral territories divided by a horizontal boundary has parallels to the division of the Drosophila eye disc into dorsal, equatorial and ventral zones, indicating that the similarities in eye morphogenesis extend beyond single molecules to topographical patterns.

Physiology of visual cells in mouse superior colliculus and correlation with somatosensory and auditory input

THE two main targets of the mammalian optic nerve fibres are the lateral geniculate body and the superior colliculus (optic tectum). From studies with various techniques, and in several mammalian species including the cat1–5, monkey6–9, rabbit10–12, rat13, and ground squirrel14 three major functions of the superior colliculus have been described. In the superficial layers the visual input is processed in a specific way; in deep layers several sense modalities, chiefly visual, auditory and somatosensory, are brought together; stimulation of the tectum results in an orienting of the animals eyes, head or body towards a location corresponding topographically to the part of the tectum stimulated.

Influence of the choroid plexus on cerebellar development: analysis of retinoic acid synthesis

The choroid plexus of the fourth ventricle is conspicuous both in location and size: it protrudes over the outer hindbrain, closely apposed to the caudal external surface of the cerebellum, and it is disproportionately large early on. While the developing cerebellum is known to respond to retinoic acid (RA), it does not express significant levels of RA synthesizing enzyme. Retinaldehyde dehydrogenase levels in the choroid plexus, however, are very high, with maxima during the pre- and postnatal periods of cerebellar morphogenesis. Explants assays demonstrate release of a neurite-outgrowth promoting activity from the choroid plexus, whose levels parallel the levels of RA synthesizing enzyme here, and which can be mimicked by RA. These observations characterize the choroid plexus as a paracrine, growth-promoting organ for the developing cerebellum, with the effects mediated through temporally regulated RA production.

Origins of uncrossed retinofugal projections in normal and hypopigmented mice

In albinos, the retinofugal projections to the ipsilateral side of the brain are reduced (e.g., see Guillery, 1969; La Vail et al., 1978; Lund, 1965). Although all ganglion cell types are affected, in mice the displaced ganglion cell population is the main target of the albino mutation (Dräger & Olsen, 1980). Here we tested whether this preferential effect on displaced ganglion cells is a general consequence of the melanin reduction or a pleiotropic effect unique to the albino locus, by retrogradely tracing retinal ganglion cells in normal C57BL/6J mice and in several non-allelic hypopigmentation mutants on the same background: albino (C57BL/6J-c2J), beige (C57BL/6J-bg), pale ear (C57BL/6J-ep), ruby-eye/haze (C57BL/6J-ru-2hz), and pearl (C57BL/6J-pe). All mutants have lower overall cell counts in the ipsilateral projection, but the displaced population is disproportionately affected: the albinos contain 42% of the normal number of displaced ganglion cells, and the other mutants have an average 57% of normal counts. The reduction in uncrossed retinofugal projections in albinos affects the inputs to the lateral geniculate nucleus and the superior colliculus, but not to the suprachiasmatic nucleus (Dräger, 1974). To address the question in which way the susceptible uncrossed projections differ from the nonsusceptible one, we compared ganglion cells backfilled from the suprachiasmatic nucleus to ganglion cells backfilled from the optic tract at geniculate level. Whereas the uncrossed optic tract projection originates from the binocular region in the ventro-temporal retina and contains a high fraction of large and displaced ganglion cells (Dräger & Olsen, 1980), both the crossed and uncrossed inputs to the suprachiasmatic nucleus originate from the entire retina with a relative preference for the lower nasal region that corresponds to part of the monocular visual field; all ganglion cells projecting to the suprachiasmatic nucleus are of medium size, and they are located in the ganglion cell layer. These observations allow the following conclusions: (1) All genetic mutants which cause a reduction in ocular melanin, regardless of the molecular or cell-biological mechanism underlying the pigment reduction, result in decreased uncrossed projections; this confirms previous reports (La Vail et al., 1978, Sanderson et al., 1974). (2) The decrease affects only projections involved in binocular vision. (3) In mice, the ganglion cells displaced to the inner nuclear layer, and hence located closer to the retinal pigment epithelium, are disproportionately affected by the melanin reductions. These observations may provide cues to the spatio-temporal mechanism of the

Retinoic Acid Synthesizing Enzymes in the Embryonic and Adult Vertebrate

The oxidation of retinaldehyde to retinoic acid (RA) provides the retinoid form of highest potency for a variety of cellular systems. RA has been implicated in many processes, such as growth and differentiation of epithelia in the adult organism (De Luca 1991), and determination of the antero-posterior axis for the limb bud (Eichele and Thaller 1987; Tickle et al. 1982) and the entire body of the vertebrate embryo (Durston et al. 1989; Hogan, Thaller, and Eichele 1992). In addition, RA is thought to promote neuronal survival, differentiation and neurite outgrowth (Haskell et al. 1987; Quinn and De Boni 1991; Wuarin, Sidell, and De Vellis 1990). RA exerts its effects by binding to specific nuclear receptors that regulate transcription. The diversity in RA actions is commonly attributed to differences in local expression patterns of different receptors and cytoplasmic binding proteins that modify the availability of intracellular RA (Giguere 1994). In addition, however, retinoid metabolism may contribute significantly to local diversity in RA actions. Retinoid metabolism includes the processes of precursor circulation and cellular uptake mediated by binding proteins, the reversible oxidation of retinol to retinaldehyde, the irreversible oxidation of retinaldehyde to RA, and RA degradation. Here we focus on the enzymes that mediate the oxidation of retinaldehyde to RA.

Retinoic acid signaling in the brain marks formation of optic projections, maturation of the dorsal telencephalon, and function of limbic sites.

As retinoic acid (RA) is known to regulate the expression of many neuronal proteins, it is likely to influence overall development and function of the brain; few particulars, however, are available about its role in neurobiological contexts due mainly to problems in RA detection. To ask whether the function of RA in the rostral brain is concentrated in particular neurobiological systems, we compared sites of RA synthesis and actions, as detected by RA signaling in reporter mice, for embryonic and adult ages. We found that most sites of RA actions in the forebrain do not colocalize with RA synthesis, consistent with a dominant RA supply by diffusion and the circulation. The changing RA patterns distinguish preferentially two complex functional schemes. (1) Within the visual system when the first optic axons grow toward their targets, RA signaling delineates the topographical adjustment of the retinal map, which is encoded in the coordinates of the visual world, to central visual maps, which are formed in the segmental brain coordinates. (2) The second scheme begins early in forebrain morphogenesis as a distinction of the dorsal telencephalon. With progressing development, and in the adult, the RA patterns then focus on widely distributed structures, most of which belong to the limbic system. These are sites in which emotional perception is combined with higher cognitive processes and in which normal function requires ongoing remodeling of synaptic connections, indicating that the developmental role of RA in promotion of neuronal differentiation programs continues in the adult brain for highly flexible neural circuits. J. Comp. Neurol. 470:297–316, 2004.