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 59 - Numbers 4-5-6 (2015) / Pages 159-256 Editor-in-Chief: Juan Aréchaga
MORE INFORMATION [Abstract - FullText / Full Text Open Access]
ISSN: 0214-6282 / ISSN-e: 1696-3547 www.intjdevbiol.com
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ABSTRACTS
Reviews
EHU/UPV/UBC - The International Journal of Developmental Biology 59: 159-170(2015)
doi: 10.1387/ijdb.150223sg / © UBC Press ( www.a360grados.net )
The significance of Hox gene collinearity
Stephen J. Gaunt
Department of Zoology, University of Cambridge, Downing Street, Cambridge, UK
Abstract: Arthropods and vertebrates inherited their Hox clusters from an ancestral cluster of at least six genes already present in their last common ancestor, Urbilateria. Clustering and a common transcriptional direction are both likely features of the way that the gene complex first arose in a process of tandem gene duplication. Spatial collinearity (correspondence between ordering of Hox genes along the chromosome and their expression patterns along the head-tail axis) has been conserved in many animal groups and is likely to have been already present in Urbilateria. It is not known why the Hox cluster evolved with spatial collinearity. Four models are discussed. These vary in the significance they place upon Hox chromatin structure, and also on whether they propose that collinearity is primarily concerned with establishment or maintenance of Hox expression. Published proposals to explain spatial collinearity, which invoke enhancer sharing, chromatin closing or chromatin opening, are either problematic or can offer only partial explanations. In an alternative proposal it is suggested here that spatial collinearity evolved principally to maximise physical segregation, and thereby minimise incidence of boundaries, between active and inactive genes within the Hox cluster. This is to minimise erroneous transfer of transcriptional activity, or inactivity, between adjacent Hox genes.
Keywords: evolution, embryo, homology, chromatin, polycomb
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 171-177(2015)
doi: 10.1387/ijdb.150071tz / © UBC Press ( www.a360grados.net )
Histone deacetylases 1 and 2 are required for brain development
Joanna Jaworska, Malgorzata Ziemka-Nalecz and Teresa Zalewska
NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Abstract: Epigenetic modifications of histones have been implicated in the regulation of cell specific expression of genes required for neuronal development. The best studied post-translational (epigenetic) modification of histones is the process of reversible acetylation. Two types of enzymes - histone acetyltransferases (HATs) and histone deacetylases (HDACs) establish and maintain specific patterns of histone acetylation in balance, thereby contributing to both transcriptional activation and repression of specific sets of genes. Histone deacetylases catalyze the removal of acetyl groups from selected lysine residues in the conserved tails of core histone proteins and are considered as transcriptional corepressors. A significant amount of data implicates HDACs in diverse biological processes, including tissue specific developmental program by silencing specific growth-inhibitory genes. In line with this, gene disruption studies have shown that the class I deacetylases, HDAC1 and HDAC2 play an essential role in nervous system development. In the present review, we briefly highlight current insights supporting the function of histone deacetylases in rodent brain development and discuss present knowledge referring to their role in neurogenesis, taking into consideration results obtained in culture systems and in in vivo studies.
Keywords: histone, deacetylase, brain, development
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: (2015)
doi: 10.1387/ijdb.150042fh / © UBC Press ( www.a360grados.net )
SCNT versus iPSCs: proteins and small molecules in reprogramming
Fei Han, Xia Li, Dandan Song, Shaoshuai Jiang, Qun Xu and Yunhai Zhang
Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei, China
Abstract: Somatic cell nuclear transplantation (SCNT) and induced pluripotent stem cell (iPSC) technologies can be employed to change cell fate by reprogramming. The discoveries of SCNT and iPSCs were awarded the Nobel Prize for Physiology and Medicine in 2012, which reaffirmed the importance of cell fate plasticity. However, the low cloning efficiency of SCNT and differences between iPSCs and embryonic stem cells (ESCs) are great barriers and may be caused by incomplete or aberrant reprogramming. Additionally, the well characterized reprogramming factors Oct4, Sox2, Klf4 and c-Myc (OSKM) are not simultaneously expressed at high levels in enucleated or early embryonic oocytes, suggesting reprogramming may be different in the above two methods. Recent studies have demonstrated that small molecules and specific proteins expressed in oocytes and in early embryonic development play important roles in reprogramming by replacing transcription factors, erasing reprogramming memory and accelerating the speed and extent of reprogramming. In this review, we summarize the current state of SCNT and iPSCs technologies and discuss the latest advances in the research of proteins and small molecules affecting SCNT and iPSCs. This is an area of research in which chemical biology and proteomics are combining to facilitate improving cellular reprogramming and production of clinical grade iPSCs.
Keywords: SCNT, iPSC, early embryo, oocyte, differentiation, protein, small molecule
Original Articles --------------------
EHU/UPV/UBC - The International Journal of Developmental Biology 59: 187-194(2015)
doi: 10.1387/ijdb.140190LS / © UBC Press ( www.a360grados.net )
Gliolectin positively regulates Notch signalling during wing-vein specification in Drosophila
Naveen Prasad and Lingadahalli S. Shashidhara
Indian Institute of Science Education and Research (IISER), Pune, India
Abstract: Notch signalling is essential for animal development. It integrates multiple pathways controlling cell fate and specification. Here we report the genetic characterization of Gliolectin, presumably a lectin, a cytoplasmic protein, significantly enriched in Golgi bodies. Its expression overlaps with regions where Notch is activated. Loss of gliolectin function results in ectopic veins, while gain of its function causes loss of wing veins. It positively regulates Enhancer of split mβ, a target of Notch signalling. These observations suggest that it is a positive regulator of Notch signalling during wing development in Drosophila.
Keywords: Glec, Notch, wing vein development, D/V boundary
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 195-203(2015)
doi: 10.1387/ijdb.140200md / © UBC Press ( www.a360grados.net )
Down-regulation of msrb3 and destruction of normal auditory system development through hair cell apoptosis in zebrafish
Xiaofang Shen1, Fei Liu 1, Yingzhi Wang 1, Huijun Wang 2, Jing Ma 1, Wenjun Xia 3, Jin Zhang 1, Nan Jiang 1, Shaoyang Sun 1, Xu Wang 1 and Duan Ma 1,2,3
1. Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Institute of Medical Sciences, School of Basic Medical Sciences
2. Children’s Hospital of Fudan University
3. Institute of Biomedical Science, School of Basic Medical Sciences, Fudan University, Shanghai, China
Abstract: Hearing defects can significantly influence quality of life for those who experience them. At this time, 177 deafness genes have been cloned, including 134 non-syndromic hearing-loss genes. The methionine sulfoxide reductase B3 (Ahmed et al., 2011) gene (also called DFNB74) is one such newly discovered hearing-loss gene. Within this gene c.265 T>G and c.55 T>C mutations are associated with autosomal recessive hearing loss. However, the biological role and mechanism underlying how it contributes to deafness is unclear. Thus, to better understand this mutation, we designed splicing morpholinos for the purpose of down-regulating msrb3 in zebrafish. Morphants exhibited small, tiny, fused, or misplaced otoliths and abnormal numbers of otoliths. Down-regulation of msrb3 also caused shorter, thinner, and more crowded cilia. Furthermore, L1-8 neuromasts were reduced and disordered in the lateral line system; hair cells in each neuromast underwent apoptosis. Co-injection with human MSRB3 mRNA partially rescued auditory system defects, but mutant MSRB3 mRNA could not. Thus, msrb3 is instrumental for auditory system development in zebrafish and MSRB3-related deafness may be caused by promotion of hair cell apoptosis.
Keywords: deafness, zebrafish, otolith, hair cell, neuromast
Short Communication --------------------
EHU/UPV/UBC - The International Journal of Developmental Biology 59: 205-209(2015)
doi: 10.1387/ijdb.140348mj / © UBC Press ( www.a360grados.net )
Generation and characterization of mice harboring a conditional CXCL12 allele
Ryan E. Hillmer 1, Joanne P. Boisvert 2, Monica J. Cucciare 1, Michael B. Dwinell 2 and Milan Joksimovic 1
1. Department of Cell Biology, Neurobiology and Anatomy
2. Department of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, WI, USA
Abstract: The chemokine CXCL12 has important functions in immune and central nervous systems. Moreover, a global disruption of CXCL12 in mice results in perinatal lethality. To circumvent this impediment and provide a tool for analyzing CXCL12 functions in specific organ systems, we have generated a mouse line harboring a loxP-site flanked exon 2 of CXCL12. A germ line deleter, β???-actin::cre was used to remove a CXCL12 exon 2 and subsequently systemic CXCL12 exon 2 deficient embryos were generated. These mutant embryos showed a marked depletion of CXCL12 transcript. As expected from the global mutant phenotype, our mutants were also characterized by highly irregular cerebellar cytoarchitecture of the external granule layer as well as altered radial migration of midbrain dopaminergic neurons. Importantly, migration of the pontine grey nucleus (PGN) was derailed and remarkably resembled the global mutant phenotype of the CXCL12 receptor – CXCR4 in this system. Despite the fact that CXCL12 signaling can be mediated through receptors other than CXCR4, our results indicate a monogamous relationship between the CXCL12 ligand and CXCR4 receptor in controlling PGN migration. Our findings further expand on the understanding of CXCL12 function in PGN development. Moreover, phenotypic similarities between our mutants and mice harboring a global CXCL12 disruption support the validity of our line. Importantly, these results strongly suggest that our conditional CXCL12 line can be used as a powerful tool to manipulate CXCL12 signaling and function in vivo.
Keywords: SDF1 floxed, pontine grey nucleus, external granule layer, midbrain dopaminergic neuron
Technical Articles -------------------
EHU/UPV/UBC - The International Journal of Developmental Biology 59: 211-216(2015)
doi: 10.1387/ijdb.150172mz / © UBC Press ( www.a360grados.net )
3D culture of ovarian follicles: a system towards their engineering?
Maurizio Zuccotti 1, Valeria Merico 2, Paola Rebuzzini 2, Martina Belli 2, Giulia Vigone 2, Francesca Mulas 3, Lorenzo Fassina 3,4, Wasco Wruck 5, James Adjaye 5,6, Riccardo Bellazzi 3 and Silvia Garagna 2,4
1. Sezione di Anatomia, Istologia ed Embriologia, Dipartimento di Scienze Biomediche, Biotecnologiche e Traslazionali (S.Bi.Bi.T.), Università degli Studi di Parma, Italy
2. Laboratorio di Biologia dello Sviluppo, Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Università degli Studi di Pavia, Italy
3. Dipartimento di Ingegneria Industriale e dell’Informazione, Università degli Studi di Pavia, Pavia, Italy
4. Centro di Ingegneria Tissutale, Università degli Studi di Pavia, Italy
5. Institute for Stem Cell Research and Regenerative Medicine, Medical Faculty, Heinrich Heine University, Düsseldorf, Germany
6. Molecular Embryology and Aging Group, Department of Vertebrate Genomics. Max-Planck Institute for Molecular Genetics, Berlin, Germany.
Abstract: Infertility in women is a health priority. Designing a robust culture protocol capable of attaining complete follicle growth is an exciting challenge, for its potential clinical applications, but also as a model to observe and closely study the sequence of molecular events that lie behind the intricate relationship existing between the oocyte and surrounding follicle cells. Here, we describe the procedures used to maintain the ovarian follicle 3D architecture employing a variety of in vitro systems and several types of matrices. Collagen and alginate are the matrices that led to better results, including proof-of-concept of full-term development. Pioneer in its kind, these studies underlie the drawbacks encountered and the need for a culture system that allows more quantitative analyses and predictions, projecting the culture of the ovarian follicle into the realm of tissue engineering.
Keywords: Ovarian follicle, 3D culture, matrix, collagen, alginate, tissue engineering
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 217-219(2015)
doi: 10.1387/ijdb.150025dr / © UBC Press ( www.a360grados.net )
A simple method of image analysis to estimate CAM vascularization by APERIO ImageScope software
Christian Marinaccio 1 and Domenico Ribatti 1,2
1. Department of Basic Medical Sciences, Neurosciences, and Sensory Organs, University of Bari Medical School
2. National Cancer Institute “Giovanni Paolo II”, Bari, Italy
Abstract: The chick chorioallantoic membrane (CAM) assay is a well-established method to test the angiogenic stimulation or inhibition induced by molecules and cells administered onto the CAM. The quantification of blood vessels in the CAM assay relies on a semi-manual image analysis approach which can be time consuming when considering large experimental groups. Therefore we present here a simple and fast volumetric method to inspect differences in vascularization between experimental conditions related to the stimulation and inhibition of CAM angiogenesis based on the Positive Pixel Count algorithm embedded in the APERIO ImageScope software.
Keywords: chick chorioallantoic membrane assay, image analysis, angiogenesis
Developmental Expression Patterns ------------------
EHU/UPV/UBC - The International Journal of Developmental Biology 59: 221-228(2015)
doi: 10.1387/ijdb.140195LB / © UBC Press ( www.a360grados.net )
Analysis of AtGUS1 and AtGUS2 in Arabidopsis root apex by a highly sensitive TSA-MISH method
Leonardo Bruno 1, Matteo Ronchini 2, Olimpia Gagliardi 1, Tamara Corinti 2, Adriana Chiappetta 1, Paolo Gerola 2 and Maria B. Bitonti 1
1. Department of Biology, Ecology and Earth Science, University of Calabria, Laboratory of Plant Biology, Cosenza
2. Department of Teorethical and Applied Sciences, University of Insubria, Varese, Italy
Abstract: A new highly sensitive whole-mount in situ hybridization method, based on tyramide signal amplification (TSA-MISH) was developed and a combined GFP detection and TSA-MISH procedure was applied for the first time in plants, to precisely define the spatial pattern of AtGUS1 and AtGUS2 expression in the root apex. ?-glucuronidases (GUSs) belonging to the glycosyl hydrolases (GHs) 79 family, are widely distributed in plants, but their functional role has not yet been fully investigated. In the model system Arabidopsis Thaliana, three different AtGUS genes have been identified which encode proteins with putative different fates. Endogenous GUS expression has been detected in different organs and tissues, but the cyto-histological domains of gene expression remain unclear. The results here reported show co-expression of AtGUS1 and AtGUS2 in different functional zones of the root apex (the cap central zone, the root cap meristem, the staminal cell niche and the cortical cell layers of the proximal meristem), while AtGUS2 is exclusively expressed in the cap peripheral layer and in the epidermis in the elongation zone. Interestingly, both genes are not expressed in the stelar portion of the proximal meristem. A spatial (cortex vs. stele) and temporal (proximal meristem vs. transition zone) regulation of AtGUS1 and AtGUS2 expression is therefore active in the root apex. This expression pattern, although globally consistent with the involvement of GUS activity in both cell proliferation and elongation, clearly indicates that AtGUS1 and AtGUS2 could control distinct downstream process depending on the developmental context and the interaction with other players of root growth control. In the future, the newly developed approaches may well be very useful to dissect such interactions.
Keywords: Arabidopsis thaliana, multiprobe in situ hybridization, GUS, root, tyramide signal amplification (TSA)
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 229-234(2015)
doi: 10.1387/ijdb.140197LV / © UBC Press ( www.a360grados.net )
Expression patterns of CREB binding protein (CREBBP) and its methylated species during zebrafish development
Julie Batut 1,2, Carine Duboé 1,2 and Laurence Vandel 1,2
1. Université de Toulouse, UPS, CBD (Centre de Biologie du Développement)
2. CNRS, CBD UMR 5547, Toulouse, France
Abstract: Proper embryonic development requires a fine-tuned control of gene expression, which is achieved in part through the activity of transcription coactivators or corepressors. The nuclear coactivator cAMP-response element-binding protein (CREB) binding protein (CREBBP or CBP) interacts with numerous transcription factors and thereby plays a key role in various signaling pathways. Interestingly, in cell-based studies CREBBP activity is modulated by post-translational modifications such as methylation on arginine residues which is catalyzed by coactivator-associated arginine methyltransferase 1 (CARM1). However, whether and where CREBBP, and in particular its methylated forms, are expressed during development in vertebrates has not been addressed so far. Here, we analyzed the expression of the two crebbp genes (crebbpa & crebbpb) during zebrafish development using both RT-qPCR and in situ hybridization. We found that while crebbpa expression is higher in posterior, caudal nascent somites during somitogenesis, crebbpb accumulates in anterior, rostral, and more mature somites. In addition, crebbpa mRNA is enriched in the central myotome at 24 hpf indicating that its expression is spatially and temporally controlled. We next characterized the expression of CREBBP protein from blastula to gastrula stages by immunohistochemistry. We found that while CREBBP is clearly cytoplasmic in the early blastula, it becomes both cytoplasmic and nuclear at 30% epiboly before turning mainly nuclear during gastrulation. Of interest, CREBBP methylated species appear to be mainly nuclear from 30% epiboly to 6-somite stage. This suggests that methylation may regulate CREBBP import to the nucleus during zebrafish development and could therefore participate in the control of early developmental processes.
Keywords: CREB binding protein, arginine methylation, expression, embryo, zebrafish
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 235-240(2015)
doi: 10.1387/ijdb.140275ma / © UBC Press ( www.a360grados.net )
Comparative expression analysis of pfdn6a and tcp1α during Xenopus development
Silvia Marracci 1, Davide Martini 1, Martina Giannaccini 1,2, Guido Giudetti 1, Luciana Dente 1 and Massimiliano Andreazzoli 1
1. Unità di Biologia Cellulare e dello Sviluppo, Dipartimento di Biologia, Università di Pisa
2. Istituto di Scienze della Vita, Scuola Superiore Sant'Anna, Pisa, Italy
Abstract: We recently identified pfdn6a and tcp1α (also known as cct-α) as genes coregulated by the transcription factor Rx1. The proteins encoded by these genes belong to two interacting complexes (Prefoldin and “chaperonin containing t-complex polypeptide 1”), which promote the folding of actin and tubulin and have more recently been reported to be involved in a variety of additional functions including cell cycle control and transcription regulation. However, little is known about the expression and function of these two genes during vertebrate development. To assess whether pfdn6a and tcp1α display a general coordinated expression during Xenopus development, we determined, by RT-PCR and in situ hybridization, the spatio-temporal expression pattern of pfnd6a, which was not previously described, and compared it to that of tcp1α, extending the analysis to stages not previously investigated for this gene. We detected maternal transcripts of pfnd6a in the animal hemisphere at early blastula stage. During gastrulation, pfdn6a was expressed in the involuting mesoderm and subsequently in the anterior and dorsal neural plate. At tailbud and tadpole stages, pfdn6a RNA was mainly detected in the forebrain, midbrain, eye vesicle, otic vesicle, branchial arches, and developing pronephros. The pfnd6a expression pattern largely overlaps with that of tcp1α indicating a spatio-temporal transcriptional coregulation of these genes in the majority of their expression sites, which is suggestive of a possible involvement in the same developmental events.
Keywords: pfdn6, hke2, tcp1, cct, Xenopus embryogenesis
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 241-246(2015)
doi: 10.1387/ijdb.140263ho / © UBC Press ( www.a360grados.net )
Identification of distal enhancers for Six2 expression in pronephros
Nanoka Suzuki 1, Kodai Hirano 1, Hajime Ogino 2,3 and Haruki Ochi 1
1. Faculty of Medicine, Yamagata University, Yamagata-shi, Yamagata
2. Department of Animal Bioscience, Nagahama Institute of Bio-Science and Technology, Nagahama-shi, Shiga
3. JST, CREST, 5, Sanbancho, Chiyoda-ku, Tokyo, Japan
Abstract: The embryonic nephric mesenchyme contains pluripotent progenitor cells. Six2, a homeodomain transcription factor, is expressed in a subset of the nephric mesenchyme, and it functions to maintain a progenitor state by suppressing nephrogenesis. Despite the functional significance of Six2 in nephric development, its regulatory mechanisms remain unclear. To identify the cis-regulatory elements for Six2, we focused on the evolutionarily conserved sequences known as conserved noncoding sequences (CNSs) associated with the Six2 locus. Transgenic experiments using Xenopus laevis embryos revealed that three of the eight CNSs located within a 317-kb segment of the Six2 genomic locus were nephric enhancers. Motif analysis of transcription factors combined with phylogenetic footprinting revealed the enrichment of putative T-cell factor (Tcf)-, Hox-, and SWI/SNF complex helicase-like transcription factor (Hltf)- and AT-rich interactive domain 3A (Arid3a)-binding motif sequences in these enhancers.
Keywords: Six2, conserved noncoding sequence, nephric enhancer, Xenopus laevis, transgenic system
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EHU/UPV/UBC - The International Journal of Developmental Biology 59: 247-254(2015)
doi: 10.1387/ijdb.140224fh / © UBC Press ( www.a360grados.net )
Expression patterns and immunohistochemical localization of PITX2B transcription factor in the developing mouse heart
Francisco Hernandez-Torres, Diego Franco, Amelia E. Aranega and Francisco Navarro
Department of Experimental Biology, Faculty of Experimental Sciences, University of Jaen, Jaen, Spain
Abstract: The Pitx2 gene is involved in the establishment of vertebrate left-right axis with an important role in subsequent heart organogenesis. Mutations in the Pitx2 gene have been associated with Axenfeld-Rieger syndrome, which is characterized by ocular, craniofacial, and umbilical anomalies, as well as cardiac defects. In addition, recent data have unravelled a molecular link between PITX2 loss of function and atrial fibrillation (AF), supporting an important role of Pitx2 not only in development but also in heart homeostasis. Three PITX2 isoforms have been described in mice: PITX2A, PITX2B, and PITX2C. During heart organogenesis, PITX2C seems to play a determinant role in left–right signalling from early somitogenesis onwards. However the participation of the PITX2A and/or PITX2B isoforms during cardiogenesis is controversial. Here we report for the first time that the Pitx2a and Pitx2b isoforms are jointly expressed with the Pitx2c isoform during heart development. Interestingly, in terms of relative quantification of mRNA, the Pitx2b and Pitx2c isoforms display similar expression profiles during cardiogenesis, decreasing with further development but maintaining their expression until adult stages. Moreover, a detailed analysis of PITX2B protein during cardiac development shows that PITX2B is dynamically expressed in the developing ventricular septum and asymmetrically expressed in the tricuspid valve primordia, suggesting a putative role of the PITX2B isoform during ventricular septation as well as in the maturation of the right portion of the atrioventricular canal.
Keywords: Pitx2 isoforms, heart development, mouse and ventricular septation
The International Journal of Developmental Biology
ISSN 1696-3547 (online) and 0214-6282 (print)
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