RESUMEN
Placodes are discrete thickenings of the vertebrate cranial ectoderm that generate morpho-functionally distinct structures, such as the adenohypophysis, olfactory epithelium and lens. All placodes arise from a horseshoe-shaped preplacodal ectoderm in which the precursors of individual placodes are intermingled. However, fate-map studies indicated that cells positioned at the preplacodal midline give rise to only the adenohypophyseal placode, suggesting a unique organization of these precursors within the preplacode. To test this possibility, we combined embryological and molecular approaches in chick embryos to show that, at gastrula stage, adenohypophyseal precursors are clustered in the median preplacodal ectoderm, largely segregated from those of the adjacent olfactory placode. Median precursors are elongated, densely packed and, at neurula stage, express a molecular signature that distinguishes them from the remaining preplacodal cells. Olfactory placode precursors and midline neural cells can replace ablated adenohypophyseal precursors up to head-fold stage, although with a more plastic organization. We thus propose that adenohypophyseal placode precursors are unique within the preplacodal ectoderm possibly because they originate the only single placode and the only one with an endocrine character.
Asunto(s)
Ectodermo/embriología , Adenohipófisis/citología , Adenohipófisis/embriología , Células Madre/citología , Animales , Tipificación del Cuerpo , Forma de la Célula , Embrión de Pollo , Ectodermo/citología , Cristalino/citología , Cristalino/embriología , Mucosa Olfatoria/citología , Mucosa Olfatoria/embriologíaRESUMEN
All vertebrate brains develop following a common Bauplan defined by anteroposterior (AP) and dorsoventral (DV) subdivisions, characterized by largely conserved differential expression of gene markers. However, it is still unclear how this Bauplan originated during evolution. We studied the relative expression of 48 genes with key roles in vertebrate neural patterning in a representative amphioxus embryonic stage. Unlike nonchordates, amphioxus develops its central nervous system (CNS) from a neural plate that is homologous to that of vertebrates, allowing direct topological comparisons. The resulting genoarchitectonic model revealed that the amphioxus incipient neural tube is unexpectedly complex, consisting of several AP and DV molecular partitions. Strikingly, comparison with vertebrates indicates that the vertebrate thalamus, pretectum, and midbrain domains jointly correspond to a single amphioxus region, which we termed Di-Mesencephalic primordium (DiMes). This suggests that these domains have a common developmental and evolutionary origin, as supported by functional experiments manipulating secondary organizers in zebrafish and mice.
Asunto(s)
Encéfalo/embriología , Embrión no Mamífero/embriología , Anfioxos/embriología , Tubo Neural/embriología , Vertebrados/embriología , Animales , Evolución Biológica , Tipificación del Cuerpo/genética , Encéfalo/metabolismo , Embrión de Pollo , Embrión no Mamífero/metabolismo , Regulación del Desarrollo de la Expresión Génica , Hibridación Fluorescente in Situ , Anfioxos/metabolismo , Masculino , Ratones Noqueados , Modelos Biológicos , Modelos Genéticos , Tubo Neural/metabolismo , Vertebrados/metabolismo , Pez CebraRESUMEN
The anterior border of the neural plate, presumed to contain the prospective peripheral portion (roof) of the prospective telencephalon, emerges within a vaguely defined proneural ectodermal region. Fate maps carried out at HH4 in the chick reveal that this region still produces indistinctly neural, placodal and non-neural derivatives; it does not express neural markers. We examined how the definitive anterior border domain of the rostral forebrain becomes established and comes to display a neural molecular profile, whereas local non-neural derivatives become separated. The process, interpreted as a border sharpening mechanism via intercalatory cell movements, was studied using fate mapping, time-lapse microscopy and in situ hybridization. Separation of neural and non-neural domains proceeds along stages HH4-HH4+, is well advanced at HH5, and is accompanied by a novel dorsoventral intercalation, oriented orthogonal to the border, that distributes transitional cells into molecularly distinct neural and non-neural fields. Meanwhile, neuroectodermal Sox2 expression spreads peripherally from the neighbourhood of the node, reaching the nascent anterior border domain at HH5. We also show that concurrent signals from the endodermal layer are necessary to position and sharpen the neural border, and suggest that FGF8 might be a component of this signalling.
Asunto(s)
Embrión de Pollo , Endodermo/citología , Endodermo/fisiología , Morfogénesis/fisiología , Placa Neural/anatomía & histología , Placa Neural/fisiología , Transducción de Señal/fisiología , Animales , Movimiento Celular , Embrión de Pollo/anatomía & histología , Embrión de Pollo/fisiología , Factor 8 de Crecimiento de Fibroblastos/genética , Factor 8 de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Sistema Nervioso , Factores de Transcripción SOXB1/genética , Factores de Transcripción SOXB1/metabolismoRESUMEN
Sonic Hedgehog (Shh) signaling is an important determinant of vertebrate retinal ganglion cell (RGC) development. In mice, there are two major RGC populations: (1) the Islet2-expressing contralateral projecting (c)RGCs, which both produce and respond to Shh; and (2) the Zic2-expressing ipsilateral projecting RGCs (iRGCs), which lack Shh expression. In contrast to cRGCs, iRGCs, which are generated in the ventrotemporal crescent (VTC) of the retina, specifically express Boc, a cell adhesion molecule that acts as a high-affinity receptor for Shh. In Boc(-/-) mutant mice, the ipsilateral projection is significantly decreased. Here, we demonstrate that this phenotype results, at least in part, from the misspecification of a proportion of iRGCs. In Boc(-/-) VTC, the number of Zic2-positive RGCs is reduced, whereas more Islet2/Shh-positive RGCs are observed, a phenotype also detected in Zic2 and Foxd1 null embryos. Consistent with this observation, organization of retinal projections at the dorsal lateral geniculate nucleus is altered in Boc(-/-) mice. Analyses of the molecular and cellular consequences of introducing Shh into the developing VTC and Zic2 and Boc into the central retina indicate that Boc expression alone is insufficient to fully activate the ipsilateral program and that Zic2 regulates Shh expression. Taking these data together, we propose that expression of Boc in cells from the VTC is required to sustain Zic2 expression, likely by regulating the levels of Shh signaling from the nearby cRGCs. Zic2, in turn, directly or indirectly, counteracts Shh and Islet2 expression in the VTC and activates the ipsilateral program.
Asunto(s)
Lateralidad Funcional/fisiología , Regulación del Desarrollo de la Expresión Génica/genética , Proteínas Hedgehog/metabolismo , Inmunoglobulina G/metabolismo , Receptores de Superficie Celular/metabolismo , Células Ganglionares de la Retina/fisiología , Transducción de Señal/fisiología , Animales , Electroporación , Retroalimentación Fisiológica/fisiología , Factores de Transcripción Forkhead/deficiencia , Lateralidad Funcional/genética , Cuerpos Geniculados/fisiología , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Inmunoglobulina G/genética , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptores de Superficie Celular/genética , Retina/citología , Transducción de Señal/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vías Visuales/fisiologíaRESUMEN
Members of the Meis family of TALE homeobox transcription factors are involved in many processes of vertebrate development and morphogenesis, showing extremely complex transcriptional and spatiotemporal expression patterns. In this work, we performed a comprehensive study of chicken Meis genes using multiple approaches. First, we assessed whether the chicken genome contains a Meis3 ortholog or harbors only two Meis genes; we gathered several lines of evidence pointing to a specific loss of the Meis3 ortholog in an early ancestor of birds. Next, we studied the transcriptional diversity generated from chicken Meis genes through alternative splicing during development. Finally, we performed a detailed analysis of chick Meis1/2 expression patterns during early embryogenesis and organogenesis. We show that the expression of both Meis genes begins at the gastrulation stage in the three embryonic layers, presenting highly dynamic patterns with overlapping as well as distinct expression domains throughout development.
Asunto(s)
Pollos/genética , Desarrollo Embrionario/genética , Regulación del Desarrollo de la Expresión Génica , Variación Genética/genética , Proteínas de Homeodominio/genética , Proteínas de Neoplasias/genética , Animales , Animales Modificados Genéticamente , Aves/embriología , Aves/genética , Aves/metabolismo , Embrión de Pollo , Pollos/crecimiento & desarrollo , Pollos/metabolismo , Desarrollo Embrionario/fisiología , Dosificación de Gen/fisiología , Proteínas de Homeodominio/metabolismo , Proteínas de Homeodominio/fisiología , Modelos Biológicos , Proteína 1 del Sitio de Integración Viral Ecotrópica Mieloide , Proteínas de Neoplasias/metabolismo , Organogénesis/genética , Organogénesis/fisiología , Homología de Secuencia , Factores de Transcripción/genética , Transcripción Genética/fisiologíaRESUMEN
We correlated available fate maps for the avian neural plate at stages HH4 and HH8 with the progress of local molecular specification, aiming to determine when the molecular specification maps of the primary longitudinal and transversal domains of the anterior forebrain agree with the fate mapped data. To this end, we examined selected gene expression patterns as they normally evolved in whole mounts and sections between HH4 and HH8 (or HH10/11 in some cases), performed novel fate-mapping experiments within the anterior forebrain at HH4 and examined the results at HH8, and correlated grafts with expression of selected gene markers. The data provided new details to the HH4 fate map, and disclosed some genes (e.g., Six3 and Ganf) whose expression domains initially are very extensive and subsequently retract rostralwards. Apart from anteroposterior dynamics, some genes soon became downregulated at the prospective forebrain floor plate, or allowed to identify an early roof plate domain (dorsoventral pattern). Peculiarities of the telencephalon (initial specification and differentiation of pallium versus subpallium) are contemplated. The basic anterior forebrain subdivisions seem to acquire correlated specification and fate mapping patterns around stage HH8.
Asunto(s)
Tipificación del Cuerpo/fisiología , Regulación del Desarrollo de la Expresión Génica , Placa Neural/citología , Prosencéfalo , Animales , Mapeo Encefálico , Trasplante de Tejido Encefálico , Linaje de la Célula , Embrión de Pollo , Hibridación in Situ , Datos de Secuencia Molecular , Prosencéfalo/anatomía & histología , Prosencéfalo/embriología , Prosencéfalo/fisiologíaRESUMEN
Visual system development involves the formation of neuronal projections connecting the retina to the thalamic dorso-lateral geniculate nucleus (dLGN) and the thalamus to the visual cerebral cortex. Patients carrying mutations in the SOX2 transcription factor gene present severe visual defects, thought to be linked to SOX2 functions in the retina. We show that Sox2 is strongly expressed in mouse postmitotic thalamic projection neurons. Cre-mediated deletion of Sox2 in these neurons causes reduction of the dLGN, abnormal distribution of retino-thalamic and thalamo-cortical projections, and secondary defects in cortical patterning. Reduced expression, in mutants, of Sox2 target genes encoding ephrin-A5 and the serotonin transport molecules SERT and vMAT2 (important for establishment of thalamic connectivity) likely provides a molecular contribution to these defects. These findings unveil thalamic SOX2 function as a novel regulator of visual system development and a plausible additional cause of brain-linked genetic blindness in humans.
RESUMEN
We quantitatively analyzed the developing prospective neural and non-neural ectoderm during chicken gastrulation on semithin transverse sections. At stage PS8 (primitive streak stage 8 of Lopez-Sanchez et al. [C. Lopez-Sanchez, L. Puelles, V. Garcia-Martinez, L. Rodriguez-Gallardo, Morphological and molecular analysis of the early developing chick requires an expanded series of primitive streak stages, J. Morphol. 264 (2005) 105-116.], equivalent to stage HH4), the thickest area of the ectoderm agrees in extent with the fate-mapped neural plate we had reported previously. The thickness of the median ectoderm is constantly higher up to a distance of 250mum from Hensen's node, and thickness decreases along a mediolateral gradient with a further drop at the prospective lateral border of the neural plate. A higher cell density of the developing ectoderm also coincided with the prospective neural plate. We observed that cell death does not play an important role in the spatial definition of the neural plate.
Asunto(s)
Tipificación del Cuerpo/fisiología , Embrión de Pollo/citología , Gastrulación/fisiología , Placa Neural/citología , Animales , Recuento de Células , Embrión de Pollo/crecimiento & desarrollo , Proteínas de Unión al ADN/metabolismo , Proteínas del Grupo de Alta Movilidad/metabolismo , Etiquetado Corte-Fin in Situ , Neuronas , Factores de Transcripción SOXB1 , Factores de Transcripción/metabolismoRESUMEN
A changing network of gene activity settles the molecular basis of regionalization in the nervous system. As a consequence, analysis of combined gene expressions patterns represents a powerful initial approach to decode the complex process that drives neurohistogenesis and generates distinct morphological features. We have started to do a comparative screening of molecular regionalization in the mouse and chicken pretectal region at selected developmental stages. The pretectal region is composed of alar and roof plate derivatives of prosomere 1. This is a poorly understood region, best characterized in avian embryos and adults because nuclear cytoarchitectonic delimitation is clearer in these animals. During the early regionalization process the main pretectal boundaries and histogenetic/progenitor domains are established. We explore here Pax3, Pax6 and Six3 mRNA expression (and PAX3 immunoreactivity) in both chicken and mice, with the aim to compare their respective patterns. Our focus is centered on stages HH22-HH24 in chicken and embryonic days E11.5-E12.5 in mice. We found that, in both vertebrates, the same three main anteroposterior subdivisions are distinguished by these markers. They were defined as precommissural, juxtacommissural and commissural pretectal domains. These preliminary data represent an initial scaffold to explore more detailed pretectal regionalization processes and provide an important new key to approach unresolved pretectal homologies between vertebrates.
Asunto(s)
Tipificación del Cuerpo/genética , Diencéfalo/embriología , Desarrollo Embrionario/genética , Regulación del Desarrollo de la Expresión Génica , Animales , Embrión de Pollo , Embrión de Mamíferos , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Ratones , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Factores de Transcripción Paired Box/genética , Factores de Transcripción Paired Box/metabolismo , Proteína Homeobox SIX3RESUMEN
The adenohypophysis (ADH) is an important endocrine organ involved in the regulation of many physiological processes. The late morphogenesis of this organ at neural tube stages is well known: the epithelial ADH primordium is recognized as an invagination of the stomodeal roof (Rathke's pouch), whose walls later thicken and differentiate as the primordium becomes pediculated, and then fully separated from the stomodeum. The primordium attaches to the pial surface of the basal hypothalamus, next to the neurohypophyseal field (NH; future posterior pituitary), from which it was previously separated by migrating prechordal plate (pp) cells. Once the NH evaginates, the ADH surrounds it and jointly forms with it the pituitary gland. In contrast, little is known about the precise origin of the ADH precursors at neural plate stages and how the primordium reaches the stomodeum. For that reason, we produced in the chicken a specific ADH fate map at early neural plate stages, which was amplified with gene markers. By means of experiments labeling the mapped presumptive ADH, we were able to follow the initial anlage into its transformation into Rathke's pouch. The ADH origin was corroborated to be strictly extraneural, i.e., to lie at stage HH4/5 outside of the anterior neural plate (anp) within the pre-placodal field. The ADH primordium is fully segregated from the anterior neural border cells and the neighboring olfactory placodes both in terms of precursor cells and molecular profile from head fold stages onwards. The placode becomes visible as a molecularly characteristic ectodermal thickening from stage HH10 onwards. The onset of ADH genoarchitectonic regionalization into intermediate and anterior lobes occurs at closed neural tube stages.
RESUMEN
Patterning of the vertebrate optic vesicle into proximal/optic stalk and distal/neural retina involves midline-derived Hedgehog (Hh) signalling, which promotes stalk specification. In the absence of Hh signalling, the stalks are not specified, causing cyclopia. Recent studies showed that the cell adhesion molecule Cdon forms a heteromeric complex with the Hh receptor Patched 1 (Ptc1). This receptor complex binds Hh and enhances signalling activation, indicating that Cdon positively regulates the pathway. Here we show that in the developing zebrafish and chick optic vesicle, in which cdon and ptc1 are expressed with a complementary pattern, Cdon acts as a negative Hh signalling regulator. Cdon predominantly localizes to the basolateral side of neuroepithelial cells, promotes the enlargement of the neuroepithelial basal end-foot and traps Hh protein, thereby limiting its dispersion. This Ptc-independent function protects the retinal primordium from Hh activity, defines the stalk/retina boundary and thus the correct proximo-distal patterning of the eye.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Ojo/embriología , Proteínas Hedgehog/metabolismo , Proteínas de Pez Cebra/metabolismo , Animales , Animales Modificados Genéticamente , Sitios de Unión , Tipificación del Cuerpo , Embrión de Pollo , Células HEK293 , Humanos , Proteínas de la Membrana , Moléculas de Adhesión de Célula Nerviosa/metabolismo , Receptores Patched , Receptor Patched-1 , Receptores de Superficie Celular/metabolismo , Pez CebraRESUMEN
The raphe nuclei represent the origin of central serotonergic projections. The literature distinguishes seven nuclei grouped into rostral and caudal clusters relative to the pons. The boundaries of these nuclei have not been defined precisely enough, particularly with regard to developmental units, notably hindbrain rhombomeres. We hold that a developmental point of view considering rhombomeres may explain observed differences in connectivity and function. There are twelve rhombomeres characterized by particular genetic profiles, and each develops between one and four distinct serotonergic populations. We have studied the distribution of the conventional seven raphe nuclei among these twelve units. To this aim, we correlated 5-HT-immunoreacted neurons with rhombomeric boundary landmarks in sagittal mouse brain sections at different developmental stages. Furthermore, we performed a partial genoarchitectonic analysis of the developing raphe nuclei, mapping all known serotonergic differentiation markers, and compared these results, jointly with others found in the literature, with our map of serotonin-containing populations, in order to examine regional variations in correspondence. Examples of regionally selective gene patterns were identified. As a result, we produced a rhombomeric classification of some 45 serotonergic populations, and suggested a corresponding modified terminology. Only a minor rostral part of the dorsal raphe nucleus lies in the midbrain. Some serotonergic neurons were found in rhombomere 4, contrary to the conventional assumption that it lacks such neurons. We expect that our reclassification of raphe nuclei may be useful for causal analysis of their differential molecular specification, as well as for studies of differential connectivity and function.
Asunto(s)
Núcleos del Rafe/citología , Núcleos del Rafe/crecimiento & desarrollo , Rombencéfalo/citología , Neuronas Serotoninérgicas/fisiología , Serotonina/metabolismo , Animales , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Hibridación in Situ , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neuronas Serotoninérgicas/clasificación , Terminología como AsuntoRESUMEN
Cdon and Boc, and their Drosophila homologues Ihog and Boi, are evolutionary conserved transmembrane glycoproteins belonging to a subgroup of the Immunoglobulin superfamily of cell adhesion molecules (CAMs). Initially isolated in vertebrates as CAMs that link cadherin function with MAPK signaling in myoblast differentiation, they have thereafter been shown to act as essential receptors for the Hedgehog (Hh) family of secreted proteins. They associate with both ligand and other Hh receptor components, including Ptch and Gas1, thus forming homo- and heteromeric complexes. In Drosophila, they are also involved in ligand processing and release from Hh producing cells. Cdon/Boc and Ihog/Boi can substitute one another and play redundant functions is some contexts. In addition, Boc, but not Cdon, mediates axon guidance information provided by Hh in specific neuronal populations, whereas mutations in the CDON cause holoprosencephaly, a human congenital anomaly defined by forebrain midline defects prominently associated with diminished Hh pathway activity.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Comunicación Celular , Inmunoglobulina G/metabolismo , Neuronas/metabolismo , Receptores de Superficie Celular/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Animales , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Moléculas de Adhesión Celular/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Regulación del Desarrollo de la Expresión Génica , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Holoprosencefalia/genética , Humanos , Inmunoglobulina G/genética , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Mutación , Neuronas/citología , Unión Proteica , Receptores de Superficie Celular/genética , Transducción de Señal , Proteínas Supresoras de Tumor/genéticaRESUMEN
Sonic hedgehog (SHH) is a secreted signaling factor that is implicated in the molecular patterning of the central nervous system (CNS), somites, and limbs in vertebrates. SHH has a crucial role in the generation of ventral cell types along the entire rostrocaudal axis of the neural tube. It is secreted early in development by the axial mesoderm (prechordal plate and notochord) and the overlying ventral neural tube. Recent studies clarified the impact of SHH signaling mechanisms on dorsoventral patterning of the spinal cord, but the corresponding phenomena in the rostral forebrain are slightly different and more complex. This notably involves separate Shh expression in the preoptic part of the forebrain alar plate, as well as in the hypothalamic floor and basal plates. The present work includes a detailed spatiotemporal description of the singular alar Shh expression pattern in the rostral preoptic forebrain of chick embryos, comparing it with FoxG1, Dlx5, Nkx2.1, and Nkx2.2 mRNA expression at diverse stages of development. As a result of this mapping, we report a subdivision of the preoptic region in dorsal and ventral zones; only the dorsal part shows Shh expression. The positive area impinges as well upon a median septocommissural preoptic domain. Our study strongly suggests tangential migration of Shh-positive cells from the preoptic region into other subpallial domains, particularly into the pallidal mantle and the intermediate septum.
RESUMEN
A number of gene markers are currently claimed to allow positive or negative visualization of the early chick neural plate at stages 3d/4, when its fate becomes determined. Some markers labeled by various authors as either "neural" or "non-neural" indeed show ectodermal expression patterns roughly correlative with widespread yet vague ideas on the shape and size of the early neural plate, based on previous fate maps. However, for technical reasons, it is not clear how precisely these expression patterns correlate with any experimentally determined fate boundaries. An eventual mismatch between fate and marker interpretation might bear importantly on ideas about gene functions and causal hypotheses in issues such as the establishment of the neural/non-neural border or the earliest mechanisms of neural regionalization. In this review, we correlated a set of epiblastic and mesendodermal gene expression patterns with the novel neuroectoderm proportions suggested by our recent fate map of the chick neural plate at stages HH 3d/4 [P. Fernández-Garre, L. Rodriguez-Gallardo, V. Gallego-Diaz, I.S. Alvarez, L. Puelles, Fate map of the chicken neural plate at stage 4, Development 129 (2002) 2807-2822.]. This analysis suggests the existence of various nested subregions of the epiblast with boundaries codefined by given sets of gene patterns. No gene expression studied reproduces exactly or even approximately the entire neural plate shape, leading to a combinatorial hypothesis on its specification. This kind of analysis (fate and molecular maps), jointly with competence maps, provides the basis for understanding gene functions and the mechanisms of neural induction, specification and regionalization. Several gene patterns observed are consistent with precocious incipient regionalization of the neural plate along the dorsoventral and anteroposterior axes.
Asunto(s)
Tipificación del Cuerpo/fisiología , Embrión de Pollo/metabolismo , Ectodermo/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , Animales , Biomarcadores , Inducción Embrionaria/fisiología , Proteínas de Homeodominio/genética , Modelos Biológicos , Transactivadores/genéticaRESUMEN
Fate maps are essential to understand embryonic development; they provide a background for deducing maps of differential cellular specification in the context of other experimental data and molecular expression patterns. Due to its accessibility, the chick neural plate has been fate-mapped many times, albeit without complete agreement with respect to its shape, extent and fated subdivisions. In this review, we first comment about avian neural plate fate maps reported since the early period of experimental embryology, referring to the different methods followed. We next review a perfected fate-mapping methodology, which recently allowed us rather precise delimitation of the chick neural plate at stages 3d/4. This leads to a general discussion about the apparent border of the neural plate and the prospective main rostrocaudal and longitudinal divisions of the neural tube.