Romero-Carvajal A et al. (2009), Embryogenesis and laboratory maintenance of the...

XB-ART-39609

Dev Dyn 2009 Jun 01;2386:1444-54. doi: 10.1002/dvdy.21952.

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Embryogenesis and laboratory maintenance of the foam-nesting túngara frogs, genus Engystomops (= Physalaemus).

Romero-Carvajal A , Sáenz-Ponce N , Venegas-Ferrín M , Almeida-Reinoso D , Lee C , Bond J , Ryan MJ , Wallingford JB , Del Pino EM .

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The vast majority of embryological research on amphibians focuses on just a single genus of frogs, Xenopus. To attain a more comprehensive understanding of amphibian development, experimentation on non-model frogs will be essential. Here, we report on the early development, rearing, and embryological analysis of túngara frogs (genus Engystomops, also called Physalaemus). The frogs Engystomops pustulosus, Engystomops coloradorum, and Engystomops randi construct floating foam-nests with small eggs. We define a table of 23 stages for the developmental period in the foam-nest. Embryos were immunostained against Lim1, neural, and somite-specific proteins and the expression pattern of RetinoBlastoma Binding Protein 6 (RBBP6) was analyzed by in situ hybridization. Due to their brief life-cycle, frogs belonging to the genus Engystomops are attractive for comparative and genetic studies of development. Developmental Dynamics 238:1444-1454, 2009. (c) 2009 Wiley-Liss, Inc.

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Species referenced: Xenopus
Genes referenced: lhx1 lsamp ncam1 rbbp6 uqcc6

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	Figure 3. Morphology of early embryos. A: Blastocoel of a 16-cell embryo. B: Blastocoel of a large-cell blastula. The arrow signals a darkly pigmented nucleus. C: Blastocoel of a medium-cell blastula. D: RBBP6 in situ hybridization signal is in the animal hemisphere. E: Lim1-positive nuclei occurred in the dorsal blastopore lip. F: Sagittal section of an early gastrula. G: Higher magnification from the embryo in F. The arrow signals a bottle cell. H: Three cell layers in the blastocoel roof of a mid gastrula. I: Two cell layers in the blastocoel roof of a late gastrula. J: Blastopore of a late gastrula. K: Sagittal section of a late gastrula. L: Sagittal section of a stage-13 embryo. M: Lim1-positive notochord of a late gastrula. N: Transverse section through the caudal region of a late gastrula. Arrows indicate the lateral endodermal crests at the limits of the gastrocoel roof plate. O: Transversal section through the rostral region of an early neurula. The endoderm covered the archenteron roof. a, archenteron; bl, blastocoel; dl, dorsal lip; n, notochord; np, neural plate; yp, yolk plug. Scale bars = 50 μm in G,N,O; 100 μm in H,I; 200 μm in A–D,E,M; 250 μm in F,J,K,L.Download figure to PowerPoint
	Figure 4. Morphology of the neurula. A: Neurula stage, anterior view, dorsal to the top. In situ hybridization signal of RBBP6 was detected in the presumptive eye region. B: Para-sagittal section through the somites. C: Higher magnification, of the embryo in B. D: The antigen 2G9–positive neural folds. E: The Lim1-positive notochord, cns cells, and pronephros anlage in a stage-15 embryo. F: Higher magnification of the embryo in E. The arrows in E,F signal a cns Lim 1-positive nucleus. G: Transversal section through the rostral region. The archenteron roof is covered by endoderm. H: Transversal section through the caudal region. The notochord is exposed in the gastrocoel roof plate. Arrows signal the lateral endodermal crests. I: A late neurula in dorsal view. J: The antigen 2G9–positive neural tube and cranial neural crest cell streams. K: Embryo immunostained against sarcomeric meromyosin (indicated as Myosin in the image). L: Horizontal section through the somites. a, archenteron; cnc, cranial neural crest; n, notochord; nt, neural tube; p, pronephros anlage; s, somite. Scale bars = 100 μm in C,F–H,J,L; 200 μm in D,E; 250 μm in A,B,K; 400 μm in I.Download figure to PowerPoint
	Figure 5. Morphology of tailbud embryos. A: RBBP6 in situ hybridization signal was detected in the neural tube. B: Dorsal view of an embryo immunostained against antigen 2G9. Rhombomere 5 is 2G9-negative. C: Lateral view of the embryo in B that shows the streams of cranial neural crest. D: Dorsal view of an embryo immunostained against NCAM. The neural tube and optic vesicles are NCAM-positive. E: Somites are sarcomeric meromyosin–positive. F: The Lim 1–positive pronephros. G: Stage-18 embryo. The somites and heart were positive for sarcomeric meromyosin (sarcomeric meromyosin is indicated as Myosin in the images E,G). H: Stage-18.5 embryo. The Lim 1–positive pronephros and pronephric ducts are Lim 1 positive. The Lim 1–positive cns cells are not shown. ba, branchial anterior stream of cranial neural crest cells; bp, branchial posterior stream of cranial neural crest cells; br, branchial arch; h, heart; hy, hyoid stream of cranial neural crest cells; m, mandibular stream of cranial neural crest cells; nt, neural tube; o, optic vesicle; p, pronephros; pd, pronephric duct; r5, rhombomere 5; s, somite. Scale bars = 200 μm in B,C,F,H; 300 μm in A; 400 μm in D,E; 500 μm in G.Download figure to PowerPoint

References [+] :

Bader, Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. 1982, Pubmed