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 57 - Number 11/12 (2013) 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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CONTENTS + ABSTRACTS
Meeting Report
Mammalian Embryology Conference: celebrating the pioneering work of Andrzej K. Tarkowski (Warsaw, Poland, 25-26 October, 2013)
Magdalena Krupa, Aneta Suwińska and Marek Maleszewski
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 799-808
Review
The problem of the origin of primordial germ cells (PGCs) in vertebrates: historical review and a possible solution
Giovanni Pilato, Vera D’Urso, Fabio Viglianisi, Francesca Sammartano, Giorgio Sabella and Oscar Lisi
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 809-819
Original Articles
Dual embryonic origin of the hyobranchial apparatus in the Mexican axolotl (Ambystoma mexicanum)
Asya Davidian and Yegor Malashichev
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 821-828
Essential role of AWP1 in neural crest specification in Xenopus
Jeong-Han Seo, Dong-Seok Park, Mina Hong, Eun-Ju Chang and Sun-Cheol Choi
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 829-836
Insulin-like growth factor 1 acts as an autocrine factor to improve early embryogenesis in vitro
Charmaine J. Green and Margot L. Day
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 837-844
The Oct4 promoter-EGFP transgenic rabbit: a new model for monitoring the pluripotency of rabbit stem cells
Mingru Yin, Zhenfu Fang, Weihua Jiang, Fengying Xing, Manxi Jiang, Pengcheng Kong, Yao Li, Xiaomei Zhou, Lan Tang, Shangang Li and Xuejin Chen
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 845-852
Remodeling of the myocardium in early trabeculation and cardiac valve formation; a role for TGFβ2
Boudewijn P.T. Kruithof, Marianna Kruithof-De-Julio, Robert E. Poelmann, Adriana C. Gittenberger-De-Groot, Vinciane Gaussin and Marie-José Goumans
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 853-863
AP-1c-Jun/FosB mediates xFoxD5b expression in Xenopus early developmental neurogenesis
Jaeho Yoon, Jung-Ho Kim, Ok-Joo Lee, Sung-Young Lee, Seung-Hwan Lee, Jae-Bong Park, Jae-Yong Lee, Sung-Chan Kim and Jaebong Kim
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 865-872
The expression and function of thymosin beta 10 in tooth germ development
Maho Shiotsuka, Hiroko Wada, Tamotsu Kiyoshima, Kengo Nagata, Hiroaki Fujiwara, Makiko Kihara, Kana Hasegawa, Hirotaka Someya, Ichiro Takahashi and Hidetaka Sakai
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 873-883
Technical Article
Matrigel supports neural, melanocytic and chondrogenic differentiation of trunk neural crest cells
Ana B. Ramos-Hryb, Meline C. Da-Costa, Andréa G. Trentin and Giordano W. Calloni
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 885-890
Short Communication
Sexual dimorphism of AMH, DMRT1 and RSPO1 localization in the developing gonads of six anuran species
Rafal P. Piprek, Anna Pecio, Katarzyna Laskowska-Kaszub,Jacek Z. Kubiak and Jacek M. Szymura
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 891-895
Developmental Expression Patterns
Identification and characterization of VEGF and FGF from Hydra
Lakshmi-Surekha Krishnapati and Surendra Ghaskadbi
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 897-906
Differential expression of angiogenic and anti-angiogenic molecules in the chick embryo chorioallantoic membrane and selected organs during embryonic development
Christian Marinaccio, Beatrice Nico and Domenico Ribatti
EHU/UPV/UBC - The International Journal of Developmental Biology (2013) 57: 907-916
The International Journal of Developmental Biology
ISSN 1696-3547 (online) and 0214-6282 (print)
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ABSTRACTS
Meeting Report
EHU/UPV/UBC - The International Journal of Developmental Biology 57: 799 - 808 (2013)
doi: 10.1387/ijdb.130321mm / © UBC Press (www.a360grados.net)
Mammalian Embryology Conference: celebrating the pioneering work of Andrzej K. Tarkowski (Warsaw, Poland, 25-26 October, 2013)
Magdalena Krupa, Aneta Suwińska and Marek Maleszewski
Department of Embryology, Faculty of Biology, University of Warsaw, Poland
ABSTRACT: During this Conference, a distinguished group of scientists from all over the world presented their latest studies on early mammalian development. Among them there were former students and colleagues of Professor Tarkowski. The Conference was opened with an address from the Dean of the Faculty of Biology, Professor Agnieszka Mostowska, who talked about the significance of the scientific and organizational activities of Professor Tarkowski, which, for the last 50 years, have been very important for Warsaw University. Sixteen lectures were presented over the two-days of the Conference. The topics covered studies of the regulation of the cell cycle during gametogenesis, activation of the oocyte at fertilization, control of cleavage, formation of the earliest cell lines of the embryo, cell totipotency, pluripotency, differentiation during early development, primordial germ cells, developmental epigenetics and evolutionary aspects of the study of development. Many of the topics presented had a beginning in the seminal works of Professor Tarkowski, who pioneered studies on the developmental potency of isolated blastomeres, on the formation of experimental chimaeras, on the artificial activation of oocytes and the development of parthenogenetic embryos. A detailed program of the Conference can be found at the following link:http://embryconf.biol.uw.edu.pl
Keywords: mammalian experimental embryology, embryo, blastomere potency, chimaera, parthenogenesis, stem cell, germ cell, gamete
Review -------------------------------------------------------
EHU/UPV/UBC - The International Journal of Developmental Biology 57: 809 - 819 (2013)
doi: 10.1387/ijdb.120261gp / © UBC Press (www.a360grados.net)
The problem of the origin of primordial germ cells (PGCs) in vertebrates: historical review and a possible solution
Giovanni Pilato, Vera D’Urso, Fabio Viglianisi, Francesca Sammartano, Giorgio Sabella and Oscar Lisi
Department of Biological, Geological and Environmental Sciences, Section of Animal Biology, University of Catania, Italy
ABSTRACT: A concise review of the articles about the origin of primordial germ cells (PGCs) in vertebrates is provided. Differences among various taxa concerning the origin of PGCs, not easily understandable on the base of traditional knowledge, are pointed out. All those differences can be explained taking into consideration the recent “theory of the endoderm as secondary layer”. That theory allows us to understand that those differences are only apparent, being related to modifications of stages of the consequent embryogeny, overall, to a different amount of yolk in the egg. Eggs very rich in yolk became meroblastic, and the portion of primordial ectomesenchyme destined to give rise to a part of the mesoderm and the PGCs separates early from the part destined to give rise to the rest of the mesoderm and to the digestive endoderm in order to form the vitelline hypoblast lamina. To this lamina, in contrast to the traditional interpretation, a mesodermal, not endodermal, origin must be attributed. With the misunderstanding regarding the origin of this lamina clarified, all the differences about the origin of PGCs disappears. Furthermore, in taxa where PGCs were considered to be of endodermal origin, they too have a mesodermal origin. Considering that a mesodermal origin of PGCs has been demonstrated in all sponges and cnidarians, as well, a unique, mesodermal origin of germinal cells in all pluricellular animals results.
Keywords: vertebrate, primordial germ cell origin
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 821 - 828 (2013)
doi: 10.1387/ijdb.130213ym / © UBC Press (www.a360grados.net)
Dual embryonic origin of the hyobranchial apparatus in the Mexican axolotl (Ambystoma mexicanum)
Asya Davidian1 and Yegor Malashichev,1,2
1. Department of Embryology and 2. Department of Vertebrate Zoology, Faculty of Biology and Soil Sciences, Saint-Petersburg State University, St. Petersburg, Russia.
ABSTRACT: Traditionally, the cartilaginous viscerocranium of vertebrates is considered as neural crest (NC)-derived. Morphological work carried out on amphibian embryos in the first half of the XX century suggested potentially mesodermal origin for some hyobranchial elements. Since then, the embryonic sources of the hyobranchial apparatus in amphibians has not been investigated due to lack of an appropriate long-term labelling system. We performed homotopic transplantations of neural folds along with the majority of cells of the presumptive NC, and/or fragments of the head lateral plate mesoderm (LPM) from transgenic GFP+ into white embryos. In these experiments, the NC-derived GFP+ cells contributed to all hyobranchial elements, except for basibranchial 2, whereas the grafting of GFP+ head mesoderm led to a reverse labelling result. The grafting of only the most ventral part of the head LPM resulted in marking of the basibranchial 2 and the heart myocardium, implying their origin from a common mesodermal region. This is the first evidence of contribution of LPM of the head to cranial elements in any vertebrate. If compared to fish, birds, and mammals, in which all branchial skeletal elements are NC-derived, the axolotl (probably this is true for all amphibians) demonstrates an evolutionary deviation, in which the head LPM replaces NC cells in a hyobranchial element. This implies that cells of different embryonic origin may have the same developmental program, leading to the formation of identical (homologous) elements of the skeleton.
Keywords: neural crest, head lateral plate mesoderm, branchial arch, viscerocranium, basibranchial 2
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 829 - 836 (2013)
doi: 10.1387/ijdb.130109sc / © UBC Press (www.a360grados.net)
Essential role of AWP1 in neural crest specification in Xenopus
Jeong-Han Seo 1, Dong-Seok Park 1, Mina Hong 1, Eun-Ju Chang 1 and Sun-Cheol Choi 1,2
1. Department of Biomedical Sciences, Cell Dysfunction Research Center (CDRC)
2. Department of Biochemistry and Molecular Biology. University of Ulsan College of Medicine, Pungnap-Dong, Songpa-Gu, Seoul, Republic of Korea
ABSTRACT: The neural crest (NC) comprises a transient and multipotent embryonic cell population, which gives rise to a wide variety of cell types, including craniofacial cartilage, melanocytes, and neurons and glia of the peripheral nervous system. The NC is induced by the integrated action of Wnt, FGF, and BMP signaling, and its cell fates are subsequently specified by a genetic cascade of specific transcription factors. Here we describe a critical role of AWP1 in NC induction during Xenopus early development. Xenopus AWP1 (XAWP1) was found to be expressed in the presumptive preplacodal ectoderm, neural tissue, and posterior dorsal mesoderm, but was absent in the neural fold along the anterior-posterior axis of the neurulae. Notably, XAWP1 was induced by FGF8a in naïve ectodermal tissue. XAWP1-depleted embryos exhibited defects in pigmentation, craniofacial cartilage, and in the dorsal fin. A knockdown of XAWP1 impaired both endogenous and the FGF8a or Wnt8-induced expression of NC markers without affecting mesoderm formation. Furthermore, NC induction inhibited by XAWP1 depletion was rescued by co-expression of activating forms of beta-catenin or TCF3. In addition, overexpression of XAWP1, in concert with BMP inhibition, induced the expression of neural plate border specifiers, Pax3 and Msx1, and these regulatory factors recovered NC induction in the XAWP1-depleted embryos. Beta-catenin stability and Wnt-responsive reporter activity were also impaired in AWP1-depleted cells. Taken together, these results suggest that XAWP1 functions as a mediator of Wnt signaling to regulate NC specification.
Keywords: AWP1, neural crest induction, Wnt, Xenopus
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 837 - 844 (2013)
doi: 10.1387/ijdb.130044md / © UBC Press (www.a360grados.net)
Insulin-like growth factor 1 acts as an autocrine factor to improve early embryogenesis in vitro
Charmaine J. Green and Margot L. Day
Department of Physiology, Bosch Institute, University of Sydney, Sydney, Australia
ABSTRACT: Consideration of embryo-derived growth factors, such as IGF1, is important when culturing an embryo in an in vitro fertilization (IVF) setting, or when studying the effect of growth factors on embryo development in vitro. Addition of IGF1 to the culture medium has been reported to cause a range of developmental responses in preimplantation mouse embryos. This variability may be due to culture of embryos in suboptimal culture media and at different culture densities/volumes. This study examined the role of exogenous and autocrine IGF1 on mouse preimplantation development in vitro, by treatment of embryos with an IGF1R neutralising antibody (IGF1R nAb) under low density (1 embryo/100 microl) or high density (1 embryo/1 microl) culture conditions. At low density, IGF1R nAb reduced development to the blastocyst stage, hatching, and total cell numbers in blastocysts and increased the number of apoptotic cells in blastocysts, suggesting that autocrine IGF1 signalling is occurring, even at low density. This signalling is independent of IGF1 present in the zona pellucida, since culturing embryos in the absence of their zona pellucida had no effect on blastocyst development. Addition of 10 ng/ml IGF1 increased blastocyst development at low density, but decreased hatching at high density. Similarly, high levels of exogenous IGF1 at low density decreased hatching. IGF1 appears to play a role in cell survival and treatment of blastocysts with IGF1 increased Akt phosphorylation. The IGF1R antagonist picropodophyllin was also used in this study, but was found to have non-specific effects on the mitotic spindle. In conclusion, IGF1 is an important growth factor for the improvement of preimplantation development; however, for optimal development the total amount of IGF1 present must be tightly controlled.
Keywords: IGF1, embryogenesis, autocrine growth factors
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 845 - 852 (2013)
doi: 10.1387/ijdb.130128sl / © UBC Press (www.a360grados.net)
The Oct4 promoter-EGFP transgenic rabbit: a new model for monitoring the pluripotency of rabbit stem cells
Mingru Yin, Zhenfu Fang, Weihua Jiang, Fengying Xing, Manxi Jiang, Pengcheng Kong, Yao Li, Xiaomei Zhou, Lan Tang, Shangang Li and Xuejin Chen
Department of Laboratory Animal Sciences, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
ABSTRACT: The rabbit has long been used as a laboratory animal model for developing reproductive and stem cell-related technologies, as well as for studying human disease. The Oct4 transcription factor plays a crucial role in the maintenance and regulation of pluripotency in embryos and stem cells. We constructed a reporter plasmid containing the gene encoding the enhanced green fluorescent protein (EGFP) under the control of the rabbit Oct4 promoter (prOG) and transfected it into E14 mouse stem cells and rabbit ESCs. In addition, prOG transgenic fibroblasts were derived and prOG transgenic rabbits were produced by somatic cell nuclear transfer (SCNT). The pattern of expression of ectopic EGFP was similar in E14 mouse stem cells whether under the control of the rabbit (prOG) or mouse Oct4 promoter (pmOG). EGFP expression was observed in rabbit ESCs following prOG transfection. Both prOG transgenic SCNT embryos and F1 prOG transgenic embryos derived from adult transgenic rabbits expressed green fluorescence at the morula and blastocyst stages. EGFP was clearly detected in gonads isolated from fetuses at 27 dpc. The prOG transgenic rabbit represents a new model for studying the derivation and maintenance of rabbit pluripotent cells, and for investigating rabbit embryo development.
Keywords: Oct4, rabbit, stem cell, EGFP, development
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 853 - 863 (2013)
doi: 10.1387/ijdb.130302bk / © UBC Press (www.a360grados.net)
Remodeling of the myocardium in early trabeculation and cardiac valve formation; a role for TGFβ2
Boudewijn P.T. Kruithof 1,2, Marianna Kruithof-De-Julio 2, Robert E. Poelmann 3, Adriana C. Gittenberger-De-Groot 4, Vinciane Gaussin 1 and Marie-José Goumans 2
1. Cardiovascular Research Institute, Department of Cell Biology and Molecular Medicine, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey, USA
2. Department of Molecular Cell Biology
3. Department of Anatomy and Embryology
4. Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands
ABSTRACT: Trabeculation and the formation of the leaflets of the mitral and tricuspid valves both involve remodeling of the embryonic myocardium. The nature and possible connection of these myocardial remodeling processes, however, are unclear. Therefore, we examined the morphogenesis of the early ventricular and atrioventricular (AV) myocardium and report for the first time that the formation of the early trabeculae and the positioning of the valve primordia (endocardial cushions) into the ventricular lumen are part of one continuous myocardial remodeling process, which involves the dissociation of the myocardial layers. For the endocardial cushions, this process results in delamination from the AV myocardium. The AV myocardium that will harbor the right lateral cushion is the exception and becomes positioned in the ventricular lumen by folding of the right ventricle. As a consequence, remodeling of the left and right AV myocardium occurs differently with implications for the formation of the mural leaflets and annulus fibrosis. At both the right and left side, the valvular myocardium harbors a distinct molecular phenotype and its removal from the cardiac leaflets involves a second wave of delamination. Interestingly, in the TGFβ2-KO mouse, which is a known model for cushion and valve defects, remodeling of the early myocardium is disturbed as indicated by defective trabeculae formation, persistence of valvular myocardium, disturbed myocardial phenotypes and differential defects at left and right side of the AV canal. Based on these results we propose a new model clarifying early trabeculae formation and AV valve formation and provide new inroads for an enhanced understanding of congenital heart defects.
Keywords: valve development, trabeculae, atrioventricular canal, Tbx3, TGFβ2
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 865 - 872 (2013)
doi: 10.1387/ijdb.130163jk / © UBC Press (www.a360grados.net)
AP-1c-Jun/FosB mediates xFoxD5b expression in Xenopus early developmental neurogenesis
Jaeho Yoon 1, Jung-Ho Kim 1, Ok-Joo Lee 1, Sung-Young Lee 1,2, Seung-Hwan Lee 1, Jae-Bong Park 1, Jae-Yong Lee 1, Sung-Chan Kim 1 and Jaebong Kim 1
1. Department of Biochemistry, Institute of Cell Differenciation and Aging, College of Medicine, Hallym University, ChunCheon, Kangwon-Do, Republic of Korea
2. WCU, Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Sciences and Technology, Seoul National University, Seoul, Republic of Korea
ABSTRACT: AP-1 (activator protein-1) is composed of Jun and Fos proteins and functions in cell proliferation, apoptosis and differentiation. Previous studies have demonstrated that different AP-1 complexes participate in the determination of various cell fates. However, the role of different AP-1 complexes during early vertebrate development is not yet fully understood. In the present study, we demonstrate that AP-1c-Jun/FosB regulates neurogenesis via FoxD5b expression. We show that FoxD5 was induced by the inhibition of BMP and that FoxD5b plays roles in neurogenesis. Additionally, we show that the FoxD5b promoter region within -1336 and -1316 contains an AP-1 binding site, which is required for the transcriptional regulation of FoxD5b and is induced by the inhibition of BMP signaling in animal cap explants. Moreover, c-Jun, a component of AP-1, directly binds to the AP-1 binding site of the FoxD5b promoter and induces FoxD5b expression cooperatively with FosB, but not with c-Fos or Fra-1. Altogether, these results suggest that AP-1c-Jun/FosB may play a role in neurogenesis via the induction of FoxD5b expression during early vertebrate development.
Keywords: AP-1c-Jun/FosB, FoxD5b, BMP, Neurogenesis, Xenopus
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 873 - 883 (2013)
doi: 10.1387/ijdb.120240hs / © UBC Press (www.a360grados.net)
The expression and function of thymosin beta 10 in tooth germ development
Maho Shiotsuka 1,2, Hiroko Wada 1, Tamotsu Kiyoshima 1, Kengo Nagata 1, Hiroaki Fujiwara 1, Makiko Kihara 1,2, Kana Hasegawa 1,3, Hirotaka Someya 1,4, Ichiro Takahashi 2 and Hidetaka Sakai 1
1. Laboratory of Oral Pathology
2. Department of Orthodontics
3. Department of Endodontology and Operative Dentistry
4. Department of Removable Prosthodontics, Faculty of Dental Science, Kyushu University, Fukuoka, Japan
ABSTRACT: This study presents the expression pattern and functions of thymosin beta 10 (Tbeta10), a Tbeta4 homologue during the development of mouse lower first molars. An in situ signal of Tbeta10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tbeta4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tbeta10 expression and preameloblasts with strong Tbeta4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tbeta10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig’s epithelial root sheath, thus suggesting that Tbeta10 participates in the formation of the outline of the tooth root. An inhibition assay using Tbeta10-siRNA in E11.0 mandibles showed significant growth inhibition in the tooth germ. The Tbeta10-siRNA-treated E15.0 tooth germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tbeta10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the tooth germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tbeta10 in the developing tooth germ is indicative of multiple functions of Tbeta10 in the developmental course from initiation to root formation of the tooth germ.
Keywords: thymosin beta 10, thymosin beta 4, tooth germ, development, knockdown assay
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 885 - 890 (2013)
doi: 10.1387/ijdb.130206gw / © UBC Press (www.a360grados.net)
Matrigel supports neural, melanocytic and chondrogenic differentiation of trunk neural crest cells
Ana B. Ramos-Hryb 1, Meline C. Da-Costa 1, Andréa G. Trentin 1,2 and Giordano W. Calloni 1,2
1. Programa de Pós-graduação em Biologia Celular e do Desenvolvimento and 2. Departamento de Biologia Celular, Embriologia e Genética, CCB, UFSC, Florianópolis, Brazil
ABSTRACT: The neural crest (NC) is composed of highly multipotent precursor cells able to differentiate into both neural and mesenchymal phenotypes. Until now, most studies focusing on NC cell differentiation have been performed with traditional two-dimensional (2D) cell culture systems. However, such culture systems do not reflect the complex three-dimensional (3D) microenvironments of in vivo NC cells. To address this limitation, we have developed a method of Matrigel™ coating to create 2D and 3D microenvironments in the same culture well. When we performed cultures of trunk neural crest cells (TNCCs) on three different lots of basement membrane matrix (Matrigel™), we observed that all analyzed Matrigel™ lots were equally efficient in allowing the appearance of glial cells, neurons, melanocytes, smooth muscle cells and chondrocytes. We further observed that chondrocytes were found predominantly in the 3D microenvironment, whereas smooth muscle cells were almost exclusively located in the 2D microenvironment. Glial cells were present in both environments, but with broader quantities on the 2D surface. Melanocytes and neurons were equally distributed in both 2D and 3D microenvironments, but with distinct morphologies. It is worth noting the higher frequency of chondrocytes detected in this study using the 3D Matrigel™ microenvironment compared to previous reports of chondrogenesis obtained from TNCCs on traditional 2D cultures. In conclusion, Matrigel™ represents an attractive scaffold to study NC multipotentiality and differentiation, since it permits the appearance of the major NC phenotypes.
Keywords: Matrigel, trunk neural crest, cell differentiation, chondrogenesis, 3D microenvironment
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 891 - 895 (2013)
doi: 10.1387/ijdb.130192rp / © UBC Press (www.a360grados.net)
Sexual dimorphism of AMH, DMRT1 and RSPO1 localization in the developing gonads of six anuran species
Rafal P. Piprek 1, Anna Pecio 1, Katarzyna Laskowska-Kaszub 2,3, Jacek Z. Kubiak 2,3 and Jacek M. Szymura 1
1. Department of Comparative Anatomy, Institute of Zoology, Jagiellonian University, Krakow, Poland
2. CNRS, UMR 6290, Institute of Genetics and Development of Rennes, Cell Cycle Group, France
3. Université Rennes 1, UEB, UMS Biosit, Faculty of Medicine, Rennes, France
ABSTRACT: In vertebrates, several genes which are differentially expressed in various species, have been implicated in sex determination and gonadal differentiation. We used immunolocalization to study the expression pattern of three proteins AMH, DMRT1, RSPO1 involved in the sexual differentiation of gonads. The pattern of AMH, DMRT1 and RSPO1 expression was analyzed in X. laevis and in five other divergent anuran species: Bombina bombina, Bufo viridis, Hyla arborea, Rana arvalis and Rana temporaria during gonadal development. The pattern of expression of AMH in the developing testes of six studied anuran species was similar to that described for other vertebrates. AMH was strongly expressed in differentiating Sertoli cells. Interestingly, in B. viridis, R. arvalis and R. temporaria, AMH was also expressed in ovaries. In all studied species, DMRT1 was highly expressed in the developing testes, in both the somatic and germ cells. It was also expressed at low level in ovaries in all studied species, with the exception of H. arborea. RSPO1 was expressed in the developing ovaries, especially in the somatic cells, and was almost undetectable in developing testes in all examined anurans. These developmental expression patterns strongly suggest an involvement of AMH and DMRT1 in the development of male gonads and of RSPO1 in the female gonads. The differences in the expression patterns of these proteins in the gonads of different species might reflect the diversity of gonadal development patterns in anurans resulting from long lasting and diverged paths of their evolution.
Keywords: AMH, DMRT1, RSPO1, sex differentiation, amphibian
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 897 - 906 (2013)
doi: 10.1387/ijdb.130077sg / © UBC Press (www.a360grados.net)
Identification and characterization of VEGF and FGF from Hydra
Lakshmi-Surekha Krishnapati and Surendra Ghaskadbi
Division of Animal Sciences, Agharkar Research Institute, Pune, India
ABSTRACT: Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) play important roles in the formation of the blood vascular system and in axon guidance, nervous system development and function. Here, we report isolation and characterization of VEGF and FGF homologues from Hydra vulgaris Ind-Pune, a Cnidarian which exhibits an organized nervous system and primitive epithelio-muscular cells. VEGF expression was prominent in the endoderm of the peduncle region and tentacles, as evident from in situ hybridization of whole polyps and its transverse sections. High levels of FGF were detected in the ectoderm of the budding region. The expression of VEGF in endodermal and FGF in interstitial cells was confirmed using sf-1 hydra, a temperature-sensitive mutant strain of Hydra magnipapillata. Tissue-specific expression of VEGF and FGF was confirmed by semi quantitative RT-PCR for ectodermal and endodermal tissues in H. vulgaris Ind-Pune. Treatment with SU5416, a specific inhibitor of the VEGF receptor, did not affect the whole polyp, but did delay both budding and head regeneration, suggesting a possible role of VEGF in nerve cell development, tube formation and/or in branching. FGF expression in the ectoderm of budding region, where the majority of interstitial stem cells reside suggests its role in interstitial stem cell maintenance. Further, activation of canonical Wnt signalling with the glycogen synthase kinase-3β (GSK-3β) inhibitor alsterpaullone caused down-regulation of VEGF and FGF, suggesting an antagonistic relationship between the Wnt and VEGF/FGF pathways. Our results indicate that VEGF and FGF evolved early in evolution, before the development of the blood vascular system, and open up the possibility of elucidating the evolutionarily ancient functions of VEGF and FGF.
Keywords: Evolution, FGF, hydra, nervous system, VEGF
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EHU/UPV/UBC - The International Journal of Developmental Biology 57: 907 - 916 (2013)
doi: 10.1387/ijdb.130317dr / © UBC Press (www.a360grados.net)
Differential expression of angiogenic and anti-angiogenic molecules in the chick embryo chorioallantoic membrane and selected organs during embryonic development
Christian Marinaccio, Beatrice Nico and Domenico Ribatti
Department of Basic Medical Sciences, Neurosciences, and Sensory Organs, University of Bari Medical School, Bari, Italy
ABSTRACT: In this study we have investigated by real time polymerase chain reaction (RT-PCR) the expression of cytokines, which are well known angiogenic and anti-angionenic molecules, during chick embryo development in four organs, namely the chorioallantoic membrane (CAM), heart, liver and optic tectum at four different stages. We have studied the expression of vascular endothelial growth factor-A (VEGF-A), fibroblast growth factor-2 (FGF-2), angiopoietin-1 (ANG-1), hypoxia inducible factor-1 and -2 alpha (HIF-1α and HIF-2α), hepatocyte growth factor (HGF), and of endostatin. All the pro-angiogenic cytokines examined showed a progressive increase in their expression in brain, heart, and liver, through all the stages of development, and parallel endostatin reduces its expression. In contrast, in the CAM, all the pro-angiogenic factors examined, with the exception of ANG-1, showed a stably-decreased expression during development, whereas endostatin progressively increased its expression. The CAM as an extraembryonic membrane which mediates gas and nutrient exchange until hatching, may be considered an outer organ and not an intrinsic organ of the developing embryo in which the mechanisms regulating the development of the vascular tree are different.
Keywords: angiogenesis, chorioallantoic membrane, chick embryo, heart, liver, optic tectum
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