Sumario:
The International Journal of Developmental Biology
Linking Development, Stem Cells and Cancer Research
Euskal Herriko Unibertsitateko Argitalpen Zerbitzua / Servicio Editorial de la Universidad del País Vasco / University of the Basque Country Press
Volume 58 - Number 5 (2014) pp.299-384 Editor-in-Chief: Juan Aréchaga
MORE INFORMATION [Abstract - FullText / FullText Open Access]
ISSN: 0214-6282 / ISSN-e: 1696-3547 www.intjdevbiol.com
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C O N T E N T S
In Memoriam
EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 299-302 (2014)
doi: 10.1387/ijdb.140117ed / © UBC Press (www.a360grados.net)
Herbert Steinbeisser: a life with the Xenopus embryo
Eddy M. De Robertis and Christof Niehrs
Los Angeles, USA and Mainz, Germany
Abstract: Herbert Steinbeisser was a developmental biologist completely immersed in science. Of a cheerful disposition and constant good humor, he was the best collaborator one could hope for. When such a nice, kind colleague is taken by cancer at age 56 it seems so unjust. Yet, his life is an illustration of how wonderful a life in science can be and we would like to relate it here.
Keywords: Herbert Steinbeisser
Essay ----------------------------------
EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 303-306 (2014)
doi: 10.1387/ijdb.140143dr / © UBC Press (www.a360grados.net)
Epithelial-mesenchymal interactions: a fundamental Developmental Biology mechanism
Domenico Ribatti and Marcello Santoiemma
Department of Basic Medical Sciences, Neurosciences and Sensory Organs, University of Bari Medical School, National Cancer Institute “Giovanni Paolo II”, Bari, Italy
Abstract: Interactions between epithelium and mesenchyme are common features of early stages of morphogenesis in different organs. In this historical review article, we retrospectively analyze the most important contribution to the definition and characterization of these interactions in three different organogenetic systems, including kidney, lung and limb bud. Tubule formation in the kidney is an example of an organogenetic event which involves interaction between the ureteric epithelium and the underlying mesenchyme that, in turn, induces the branching of the ureteric epithelium. In contrast, in lung organogenesis, interactive signaling occurs between the endodermal epithelium and the mesenchyme, leading to an alveolar structure. Finally, limb bud development results from a series of epithelial-mesenchymal interactions between the mesenchymal cells of the lateral plate mesoderm and the overlying ectodermal cells.
Keywords: embryology, epithelia, kidney, limb bud, lung, mesenchyme
Original Articles ---------------------------------
EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 307-314 (2014)
doi: 10.1387/ijdb.140003sg / © UBC Press (www.a360grados.net)
Synergistic action in P19 pluripotential cells of retinoic acid and Wnt3a on Cdx1 enhancer elements
Stephen J. Gaunt 1,2, and Yu-Lee Paul 2
1. Department of Zoology, University of Cambridge, Cambridge, U.K.
2. The Babraham Institute, Babraham, Cambridge, U.K.
Abstract: Cdx1 encodes a homeodomain protein that regulates expression of some Hox genes. Cdx1 itself is known to be regulated in the primitive streak/tailbud by both retinoic acid (RA) and Wnt3a. Cdx1 in eutherian mammals has two retinoic acid response elements (RAREs), located upstream and in the first intron, and each is adjacent to structural Lef/Tcf motifs. Upstream Lef/Tcf motifs respond to canonical Wnt signalling to activate Cdx1 synergistically with RA. By combined use of reporter assays, immunofluorescence and flow cytometry in mouse P19 embryonal carcinoma cells we show that the Cdx1 intron Lef/Tcf motif also responds to Wnt3a signalling. Synergy between individual Cdx1 RARE and Lef/Tcf motifs can occur whether they are adjacent or distant in the gene. Part, though not all, of the Cdx1 stimulation by RA (in absence of added Wnt3a) likely depends upon Wnt protein produced by the cells themselves, since it is inhibited by mutation of Lef/Tcf motifs, or by IWP-2, an inhibitor of Wnt production. RA and Wnt3a stimulate Cdx1 by increasing both the proportion of P19 cells that are expressing and also their mean level of expression. The expressing/non-expressing sub-populations do not simply correspond with those that express a marker of pluripotentiality, Nanog. We conclude that RA and Wnt3a activate Cdx1 synergistically by overlapping use of both upstream and intron enhancers, and that mouse embryonal carcinoma cell populations display heterogeneity in their response to these activators.
Keywords: mouse Cdx1, chicken Cdx1, Lef/Tcf, retinoic acid response element, stochastic
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 315-323 (2014)
doi: 10.1387/ijdb.130333yi / © UBC Press (www.a360grados.net)
Egg activation in physiologically polyspermic newt eggs: involvement of IP3 receptor, PLCγ, and microtubules in calcium wave induction
Tomoyo Ueno, Takehiro Ohgami, Yuichirou Harada, Shuichi Ueno and Yasuhiro Iwao
Laboratory of Molecular Developmental Biology, Department of Applied Molecular Biosciences, Graduate School of Medicine, Yamaguchi University, 753-8512 Yamaguchi, Japan
Abstract: The egg of the polyspermic newt is activated by Ca2+ waves induced by several sperm at fertilization. A major component of the sperm factor for egg activation is the sperm-specific citrate synthase (CS), which is introduced into the egg cytoplasm after sperm-egg fusion. We tried to clarify the mechanism for sperm-specific CS to induce [Ca2+]i increase in egg cytoplasm. The injection of the sperm factor into the unfertilized egg induces a [Ca2+]i increase that propagates over the whole egg surface as a Ca2+ wave. The propagation of the Ca2+ wave is inhibited by depolymerization of microtubules in the egg cytoplasm. The sperm-specific CS is highly phosphorylated and binds the component containing microtubules and the IP3 receptor. The sperm CS localized in the midpiece region was dispersed in the egg cytoplasm, but most of the CS accumulates at the sperm entry site and is distributed in association with the microtubules around the midpiece region and the nucleus. Phospholipase C (PLC) γ in egg cytoplasm also accumulates around the midpiece region in association with the sperm CS. Thus, CS at the initiation site of the Ca2+ wave forms a complex of microtubules and endoplasmic reticulum (ER) with the IP3 receptor, in addition to PLCγ, indicating close involvement of those complexes in Ca2+ releases from the ER by the sperm factor.
Keywords: Ca2+ wave, microtubule, citrate synthase, IP3 receptor, PLCγ
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 325-333 (2014)
doi: 10.1387/ijdb.130327cm / © UBC Press (www.a360grados.net)
α integrin cytoplasmic tails have tissue-specific roles during C. elegans development
Christopher M. Meighan 1, and Jean E. Schwarzbauer 2
1. Department of Molecular Biology and Chemistry, Christopher Newport University, Newport News, VA, USA
2. Department of Molecular Biology, Princeton University, Princeton, NJ, USA
Abstract: Integrin signaling impacts many developmental processes. The complexity of these signals increases when multiple, unique integrin heterodimers are expressed during a single developmental event. Since integrin heterodimers have different signaling capabilities, the signals originating at each integrin type must be separated in the cell. C. elegans have two integrin heterodimers, α INA-1/β PAT-3 and α PAT-2/β PAT-3, which are expressed individually or simultaneously, based on tissue type. We used chimeric α integrins to assess the role of α integrin cytoplasmic tails during development. Chimeric integrin ina-1 with the pat-2 cytoplasmic tail rescued lethality and maintained neuron fasciculation in an ina-1 mutant. Interestingly, the pat-2 tail was unable to completely restore distal tip cell migration and vulva morphogenesis. Chimeric integrin pat-2 with the ina-1 cytoplasmic tail had a limited ability to rescue a lethal mutation in pat-2, with survivors showing aberrant muscle organization, yet normal distal tip cell migration. In a wild type background, α integrin pat-2 with the ina-1 cytoplasmic tail had a dominant negative effect which induced muscle disorganization, cell migration defects and lethality. These results show the α integrin cytoplasmic tails impact unique cellular behaviors that vary by tissue type during development.
Keywords: integrin, morphogenesis, distal tip cell, muscle
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 335-341 (2014)
doi: 10.1387/ijdb.130289yk / © UBC Press (www.a360grados.net)
Palatal adhesion is dependent on Src family kinases and p38MAPK
Yukiko Kitase and Charles F. Shuler
Oral Biological and Medical Sciences, Faculty of Dentistry, University of British Columbia, Vancouver BC, Canada
Abstract: During secondary palate development, palatal shelves adhere to each other in the midline to form a midline epithelial seam leading to palatal closure. Cell-cell and cell-extracellular matrix adhesions, which are mediated by cell adhesion receptors, E-cadherin and integrins, are implicated in the process of adhesion of the palatal shelves. Src family kinases (SFK) function downstream of both receptors. In this study, we focused on the role of SFK in the process of palatal adhesion. During palatal adhesion, the expression of SFK mRNA, as well as localization and quantitation of the protein in the activated form, were examined by real-time qPCR and immunofluorescence. Palatal organ cultures were performed to identify the effect of pharmacological inhibition of SFK on palatal adhesion. Activated SFKs were found to be co-localized with adhesion receptors, E-cadherin and integrins in the palatal medial edge epithelium. Src, Fyn and Yes subfamily members were expressed in the palatal tissue. The expression of SFK mRNA and the quantity of the activated form of the protein were upregulated during palatal adhesion. An SFK inhibitor, PP2, blocked palatal adhesion, but another SFK inhibitor, SU6656 was not inhibitory. However, the combination of SU6656 and either of the p38MAPK inhibitors, SB203580 or BIRB0796, showed similar inhibitory effects on palatal adhesion compared to PP2 alone. The p38MAPK inhibitors alone did not alter palatal adhesion. Real-time qPCR revealed that p38MAPK alpha and delta were elevated during palatal adhesion. This study indicates that palatal cell adhesion is dependent on signaling from integrin receptors and E-cadherin through SFK and p38MAPK.
Keywords: Src family kinases (SFK), p38MAPK, palatal adhesion
Short Communications ---------------------------------
EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 343-347 (2014)
doi: 10.1387/ijdb.140058rj / © UBC Press (www.a360grados.net)
Gene expression suggests double-segmental and single-segmental patterning mechanisms during posterior segment addition in the beetle Tribolium castaneum
Ralf Janssen
Uppsala University, Department of Earth Sciences, Palaeobiology, Uppsala, Sweden
Abstract: In the model arthropod Drosophila, all segments are patterned simultaneously in the blastoderm. In most other arthropods, however, posterior segments are added sequentially from a posterior segment addition zone. Posterior addition of single segments likely represents the ancestral mode of arthropod segmentation, although in Drosophila, segments are patterned in pairs by the pair-rule genes. It has been shown that in the new model insect, the beetle Tribolium, a segmentation clock operates that apparently patterns all segments in pairs as well. Here, I report on the expression of the segment polarity gene H15/midline in Tribolium. In the anterior embryo, segmental stripes of H15 appear in pairs, but in the posterior of the embryo stripes appear in a single-segmental periodicity. This implies that either two completely different segmentation-mechanisms may act in the germ band of Tribolium, that the segmentation clock changes its periodicity during development, or that the speed in which posterior segments are patterned changes. In any case, the data suggest the presence of another (or modified), yet undiscovered, mechanism of posterior segment addition in one of the best-understood arthropod models. The finding of a hitherto unrecognized segmentation mechanism in Tribolium may have major implications for the understanding of the origin of segmentation mechanisms, including the origin of pair rule patterning. It also calls for (re)-investigation of posterior segment addition in Tribolium and other previously studied arthropod models.
Keywords: segmentation, arthropod development, arthropod evolution, segment polarity
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 349-353 (2014)
doi: 10.1387/ijdb.140016yl / © UBC Press (www.a360grados.net)
Dipeptidyl peptidase IV reduces trophoblast invasion by inhibiting the activity of MMPs
Youfei Li 1, Zhen Li1 and Jian Zhang 2
1. Department of Obstetrics and Gynecology, Xinqiao Hospital
2. Department of Pathobiology, Third Military Medical University, Chongqing, China
Abstract: Preeclampsia is a severe pregnancy complication in part due to insufficient implantation. This study aimed at elucidating the mechanism of action of dipeptidyl peptidase IV (DPPIV) in preeclampsia. Small activating RNAs (saRNA) were used to upregulate DPPIV expression in human trophoblast JAR cells. The DPPIV expression level was analyzed by real-time quantitative PCR and western blot and its activity was measured by luminescent protease assay. MMP-9 activity was analyzed by zymography and cell invasion by matrigel invasion assay. DPPIV expression level and activity was significantly increased by saRNA in JAR cells. DDPIV specific inhibitor diprotin A inhibited its activity at both basal and activated levels. DPPIV did not regulate MMP-9 expression but did repress MMP-9 activity. The invading ability of JAR cells was reduced by saRNA but increased by diprotin A. DPPIV might be responsible for the shallow implantation of the placenta due to its inhibition of the invading ability of extravillous trophoblasts, causing preeclampsia at later stage of pregnancy.
Keywords: preeclampsia, trophoblast, dipeptidyl peptidase IV, small activating RNA, cell invasion, diprotin A
Developmental Expression Pattern -------------------------------
EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 355-362 (2014)
doi: 10.1387/ijdb.140106ja / © UBC Press (www.a360grados.net)
Expression and evolution of the Tiki1 and Tiki2 genes in vertebrates
Alice H. Reis 1, Bryan T. Macdonald 2, Kerstin Feistel 3, Jose M. Brito 1, Nathalia G. Amado 1, Chiwei Xu 2, Jose G. Abreu 1 and Xi He 2
1. Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
2 F. M. Kirby Center, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA
3. Institute of Zoology, University of Hohenheim, Stuttgart, Germany
Abstract: Tiki1 is a Wnt protease and antagonist specifically expressed in the Spemann-Mangold Organizer and is required for head formation in Xenopus embryos. Here we report neighbor-joining phylogenetic analysis of vertebrate Tiki genes and their mRNA expression patterns in chick, mouse, and rabbit embryos. Tiki1 and Tiki2 orthologues are highly conserved, and exhibit similar but also different developmental expression patterns among the vertebrate/mammalian species analyzed. The Tiki1 gene is noticeably absent in the rodent lineage, but is present in lagomorphs and all other vertebrate/mammalian species examined. Expression in Hensen's node, the equivalent of the Xenopus Organizer, was observed for Chick Tiki2 and Rabbit Tiki1 and Tiki2. Mouse Tiki2 was detected at low levels at gastrulation and head fold stages, but not in the node. Mouse Tiki2 and chick Tiki1 display similar expression in the dorsal spinal cord. Chick Tiki1 expression was also detected in the surface ectoderm and maxillary bud, while chick Tiki2 was found in the anterior intestinal portal, head mesenchyme and primitive atrium. Our expression analyses provide evidence that Tiki1 and Tiki2 are evolutionarily conserved among vertebrate species and their expression in the Organizer and other regions suggests contributions of these Wnt inhibitors to embryonic patterning, as well as organogenesis. Our analyses further reveal mis-regulation of TIKI1 and TIKI2 in human cancer and diseases.
Keywords: Tiki1, Tiki2, Wnt, organizer, head induction, organogenesis
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 363-368 (2014)
doi: 10.1387/ijdb.140029mu / © UBC Press (www.a360grados.net)
Differential expression of arid5b isoforms in Xenopus laevis pronephros
Ronan Le Bouffant, Anne-Claire Cunin, Isabelle Buisson, Jérôme Cartry, Jean-François Riou and Muriel Umbhauer
Sorbonne Universités, UPMC Univ Paris 06 and CNRS, UMR7622 Developmental Biology, Paris, France
Abstract: Arid5b belongs to the ARID family of transcription factors characterised by a helix-turn-helix motif- based DNA-binding domain called ARID (A-T Rich Interaction Domain). In human, alternative splicing leads to long and short isoforms (isoform1 and 2, respectively) which differ in their N-terminal part. In this study, we report the cloning and expression pattern of Xenopus laevis arid5b. We have isolated a full length cDNA that shows homology with the human arid5b isoform1. Furthermore, 5’RACE experiments revealed the presence of a shorter isoform equivalent to the human isoform2. Temporal expression analysis by RT-qPCR indicated that X. laevis arid5b isoform1 and isoform2 are differentially expressed during development. Isoform1 is strongly expressed maternally, while isoform2 expression is essentially restricted to tailbud stages. Spatial expression analysis by whole mount in situ showed that arid5b is predominantly expressed in the developing pronephros. Arid5b mRNAs are detected in the antero-dorsal part of the pronephros anlage at the early tailbud stage and later on, in the proximal part of the pronephric tubule. RT-qPCR analyses with primers that allow to discriminate isoform1 from isoform2 showed that the latter is enriched in the pronephros anlage. In agreement with a specific pronephric signature of the isoform2, we also observed that isoform2 but not isoform1 is upregulated in animal caps induced to form pronephric tissue in response to activin A and retinoic acid. These results indicate that the two arid5b isoforms are differentially expressed and likely play different roles during early Xenopus development.
Keywords: arid5b, Xenopus, pronephros
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 369-377 (2014)
doi: 10.1387/ijdb.130353ct / © UBC Press (www.a360grados.net)
Two different vestigial like 4 genes are differentially expressed during Xenopus laevis development
María-Guadalupe Barrionuevo 1, Manuel J. Aybar 1, 2 and Celeste Tríbulo 1,2
1. Instituto Superior de Investigaciones Biológicas (INSIBIO, CONICET-UNT)
2. Instituto de Biología “Dr. Francisco D. Barbieri”, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, San Miguel de Tucumán, Tucumán, Argentina
Abstract: The vestigial gene (vg) was first characterized in Drosophila and several homologues were identified in vertebrates and called vestigial like 1-4 (vgll1-4). Vgll proteins interact with the transcription factors TEF-1 and MEF-2 through a conserved region called TONDU (TDU). Vgll4s are characterized by two tandem TDU domains which differentiate them from other members of the vestigial family. In Xenopus two genes were identified as vgll4. Our bioinformatic analysis demonstrated that these two genes are paralogues and must be named differently. We designated them as vgll4 and vgll4l. In situ hybridization analysis revealed that the expression of these two genes is rather different. At gastrula stage, both were expressed in the animal pole. However, at neurula stage, vgll4 was mainly expressed in the neural plate and neural folds, while vgll4l prevailed in the neural folds and epidermis. From the advanced neurula stage onward, expression of both genes was strongly enhanced in neural tissues, anterior neural plate, migrating neural crest, optic and otic vesicles. Nevertheless, there were some differences: vgll4 presented somite expression and vgll4l was localized at the skin and notochord. Our results demonstrate that Xenopus has two orthologues of the vgll4 gene, vgll4 and vgll4l with differential expression in Xenopus embryos and they may well have different roles during development.
Keywords: ectoderm, Vgll4, Vgll4l, TONDU
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EHU/UPV/UBC - The International Journal of Developmental Biology, Vol 58-5: 379-384 (2014)
doi: 10.1387/ijdb.140024jt / © UBC Press (www.a360grados.net)
Analysis of NUAK1 and NUAK2 expression during early chick development reveals specific patterns in the developing head
Abdelhamid Bekri, Marc Billaud and Jacques Thélu
Univ. Grenoble Alpes, INSERM, CNRS, France
Abstract: Several human diseases are associated with the NUAK1 and NUAK2 genes. These genes encode kinases, members of the AMPK-related kinases (ARK) gene family. Both NUAK1 and NUAK2 are known targets of the serine threonine kinase LKB1, a tumor suppressor involved in regulating cell polarity. While much is known about their functions in disease, their expression pattern in normal development has not been extensively studied. Here, we present the expression patterns for NUAK1 and NUAK2 in the chick during early-stage embryogenesis, until day 3 (Hamburger and Hamilton stage HH20). Several embryonic structures, in particular the nascent head, showed distinct expression levels. NUAK1 expression was first detected at stage HH6 in the rostral neural folds. It was then expressed (HH7-11) throughout the encephalalon, predominantly in the telencephalon and mesencephalon. NUAK1 expression was also detected in the splanchnic endoderm area at HH8-10, and in the vitellin vein derived from this area, but not in the heart. NUAK2 expression was first detected at stage HH6 in the neural folds. It was then found throughout the encephalon at stage HH20. Particular attention was paid in this study to the dorsal ectoderm at stages HH7 and HH8, where a local deficit or accumulation of NUAK2 mRNA were found to correlate with the direction of curvature of the neural plate. This is the first description of NUAK1 and NUAK2 expression patterns in the chick during early development; it reveals non-identical expression profiles for both genes in neural development.
Keywords: NUAK, chick embryo, in situ hybridization
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