Retinoids, eye development, and maturation of visual function.

Vitamin A is known to be critical for the beginning of eye development as well as for photoreception in the functional retina. Hardly anything, however, is known about whether retinoic acid (RA)-regulated gene expression also plays a role in the long intervening period, during which the neurobiological retinal structure takes shape. The eye contains a highly intricate architecture of RA-synthesizing (RALDH) and degrading (CYP26) enzymes.

A retinoic-acid critical period in the early postnatal mouse brain.

Background: A normal supply of vitamin A, which regulates gene expression through its active form retinoic acid, is required by many organs; both excess and deficiency can be teratogenic. Very little is known about the role of retinoic acid in maturation of the mammalian forebrain.

Methods: As retinoic acid cannot be visualized directly, we mapped its actions in the forebrain with indirect morphologic methods and by applying retinoic acid overdoses to early postnatal mice.

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 meninges is a source of retinoic acid for the late-developing hindbrain.

One general function for retinoic acid (RA) is pattern organization in the CNS. This regulatory factor has an essential role in spinal cord motor neuron and early posterior hindbrain development. In the anterior CNS, however, there is only a limited number of foci of RA synthesis, and less attention has been placed on regions such as the anterior hindbrain where RA synthesizing enzymes are absent.

Retinoic acid synthesis in the postnatal mouse brain marks distinct developmental stages and functional systems.

Retinoic acid (RA) affects development and function of the brain, but little is known about how much is made locally and where it is distributed. To identify RA-sensitive neural processes, we mapped the RA-synthesizing retinaldehyde dehydrogenases (RALDHs) during postnatal brain formation of the mouse. High and stable RALDH expressions mark the basal ganglia, olfactory bulbs, hippocampus and auditory afferents as major sites of RA actions in the functional brain.

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.

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.

Dorsal and ventral rentinoic 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 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.

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.

Postnatal effects of retinoic acid on cerebellar development.

Retinoic acid(RA) is a potent teratogen to which the early CNS is known to be highly sensitive. However, very little is know about the postnatal effects of RA. The cerebellum is a candidate for postnatal RA toxicity, as it develops late and exhibits temporal patterns of RA synthesis that are synchronized with developmental stages.

Retinoid-binding proteins in the cerebellum and choroid plexus and their relationship to regionalized retinoic acid synthesis and degradation.

The expression of cellular retinoic-acid-binding protein (CRABP) and cellular retinol-binding protein (CRBP), as well as their relationship to retinoic acid (RA) synthesis and degradation were examined in the developing mouse cerebellum and choroid plexus of the fourth ventricle. The choroid plexus, which expresses the RA-synthesizing retinaldehyde dehydrogenase RALDH-2, is likely to represent a diffusion source of RA for the closely apposed cerebellum, regulating its development. We found CRBP to be expressed in the choroid plexus and, in an in-vitro assay, addition of recombinant CRBP to RALDH-2 increased RA synthesis from retinaldehyde, with the amount of increase depending on the CRBP/retinaldehyde ratio.

A novel assay for retinoic acid catabolic enzymes shows high expression in the developing hindbrain.

We have employed a novel technique that determines the relative capacity of tissues to catabolize all-trans retinoic acid (RA) to a metabolite incapable of activating a RA reporter cell line. This assay uses the microsomal fraction of tissues from the developing mouse and detects a pathway which requires NADPH and is inhibitable by ketoconazole, suggesting that a cytochrome P450-dependent enzyme may be required. High catabolic activity was detected transiently in the developing cerebellum which peaked at postnatal day 2.

Aldehyde dehydrogenases in the generation of retinoic acid in the developing vertebrate: a central role of the eye.

In the developing vertebrate, retinoic acid is distributed in patterns that are highly regulated, both in the spatial and temporal domains. These patterns are generated by the localized expression of retinoic acid-synthesizing aldehyde dehydrogenases, which form the origins of retinoic acid-diffusion gradients in the surrounding tissues. The developing eye, known to be exceptionally vulnerable to vitamin A deficiency, is one of the retinoic acid-richest regions in the embryo.

Retinoic acid synthesis in the developing chick retina.

The transcriptional activator retinoic acid (RA) has been shown to influence the early patterning of the vertebrate eye. Models for the establishment of the retinofugal projection postulate gradients of cell-surface markers across the retinal surface that are expressed by ganglion cells and mediate the correct connection of fibers within central target fields. Spatial asymmetries of RA and RA-producing enzymes, as have been found in the eyes of mice and zebrafish, could induce the required asymmetry in gene expression.

A sensitive bioassay for enzymes that synthesize retinoic acid.

Retinoic acid (RA) is a potent regulator of gene transcription and it plays a pivotal role in neural development. As the compound is active at nanomolar concentrations, standard RA detection methods based on high-pressure liquid chromatography (HPLC) are poorly suited for neurodevelopmental questions and single RA measurements require pooling of tissues from very large numbers of embryos. An alternative approach is to determine the potential for RA synthesis by assaying for the enzymes that catalyze the last step of RA synthesis, the irreversible oxidation of retinaldehyde to RA.

Retinoic aid increases arrestin mRNA levels in the mouse retina.

Arrestin, which plays a role in the termination of the visual transduction cascade, is one of several photoreceptor proteins whose mRNA levels are increased by light. Retinoic acid, a by-product of photoreceptor signaling and a potent modulator of hormonal transcription control, is one candidate for regulating the arrestin mRNA levels. Here we show that retinoic acid, injected intraperitoneally into dark-adapted mice, increases the arrestin mRNA levels and mimics the effect of light.

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.

Retinoic acid in the anteroposterior patterning of the zebrafish trunk.

Retinoic acid has been linked to pattern formation in the vertebrate anteroposterior axis. This report describes the spatial and temporal distributions of both endogenous retinoic acid and retinoic acid synthase activity along the anteroposterior axis of neurulating zebrafish embryos, as detected by a transient transgenic assay and by a zymography bioassay. Both retinoic acid levels and synthase activity were found to be highest in anterior regions of the trunk at all of the stages which were analysed.