RESUMEN
The human gene AGTRL1 is an angiotensin II receptor-like gene expressed in vasculature, which acts as the receptor for the small peptide APELIN, and a co-receptor for Human Immunodeficiency Virus. Mammalian AGTRL1 has been shown to modulate cardiac contractility, venous and arterial dilation, and endothelial cell migration in vitro, but no role in the development of the vasculature, or other tissues, has been described. We report the identification and expression of the zebrafish ortholog of the human gene AGTRL1. Zebrafish agtrl1a is first expressed before epiboly in dorsal precursors. During epiboly it is expressed in the enveloping layer, yolk syncytial layer and migrating mesendoderm. During segmentation stages, expression is observed in epithelial structures such as adaxial cells, border cells of the newly formed somites, developing lens, otic vesicles and venous vasculature.
Asunto(s)
Epitelio/embriología , Regulación del Desarrollo de la Expresión Génica , Receptor de Angiotensina Tipo 1/genética , Proteínas de Pez Cebra/genética , Pez Cebra/embriología , Secuencia de Aminoácidos , Animales , Blástula/metabolismo , Vasos Sanguíneos/embriología , Vasos Sanguíneos/metabolismo , Fase de Segmentación del Huevo/metabolismo , Clonación Molecular , Embrión no Mamífero/metabolismo , Epitelio/metabolismo , Gástrula/metabolismo , Humanos , Hibridación in Situ , Mesodermo/metabolismo , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia , Somitos/citología , Somitos/metabolismo , Pez Cebra/anatomía & histología , Pez Cebra/genéticaRESUMEN
Notch encodes a transmembrane protein that functions in intercellular signaling. Although there is one Notch gene in Drosophila, vertebrates have three or more with overlapping patterns of embryonic expression. We cloned the entire 7575-bp coding region of an amphioxus Notch gene (AmphiNotch), encoding 2524 amino acids, and obtained the exon/intron organization from a genomic cosmid clone. Southern blot and PCR data indicate that AmphiNotch is the only Notch gene in amphioxus. AmphiNotch, like Drosophila Notch and vertebrate Notch1 and Notch2, has 36 EGF repeats, 3 Notch/lin-12 repeats, a transmembrane region, and 6 ankyrin repeats. Phylogenetic analysis places it at the base of all the vertebrate genes, suggesting it is similar to the ancestral gene from which the vertebrate Notch family genes evolved. AmphiNotch is expressed in all three embryonic germ layers in spatiotemporal patterns strikingly similar to those of all the vertebrate homologs combined. In the developing nerve cord, AmphiNotch is first expressed in the posteriormost part of the neural plate, then it becomes more broadly expressed and later is localized dorsally in the anteriormost part of the nerve cord corresponding to the diencephalon. In late embryos and larvae, AmphiNotch is also expressed in parts of the pharyngeal endoderm, in the anterior gut diverticulum, and, like AmphiPax2/5/8, in the rudiment of Hatschek's kidney. A comparison with Notch1 and Pax5 and Pax8 expression in the embryonic mouse kidney helps support homology of the amphioxus and vertebrate kidneys. AmphiNotch is also an early marker for presumptive mesoderm, transcripts first being detectable at the gastrula stage in a ring of mesendoderm just inside the blastopore and subsequently in the posterior mesoderm, notochord, and somites. As in sea urchins and vertebrates, these domains of AmphiNotch expression overlap with those of several Wnt genes and brachyury. These relationships suggest that amphioxus shares with other deuterostomes a common mechanism for patterning along the anterior/posterior axis involving a posterior signaling center in which the Notch and Wnt pathways and brachyury interact.