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- Expression and Function of Ccbe1 in the Chick Early Cardiogenic Regions Are Required for Correct Heart DevelopmentPublication . Furtado, João; Bento, Margaret; Correia, Elizabeth; Inacio, Jose Manuel; Belo, José A.During the course of a differential screen to identify transcripts specific for chick heart/hemangioblast precursor cells, we have identified Ccbe1 (Collagen and calcium-binding EGF-like domain 1). While the importance of Ccbe1 for the development of the lymphatic system is now well demonstrated, its role in cardiac formation remained unknown. Here we show by whole-mount in situ hybridization analysis that cCcbe1 mRNA is initially detected in early cardiac progenitors of the two bilateral cardiogenic fields (HH4), and at later stages on the second heart field (HH9-18). Furthermore, cCcbe1 is expressed in multipotent and highly proliferative cardiac progenitors. We characterized the role of cCcbe1 during early cardiogenesis by performing functional studies. Upon morpholino-induced cCcbe1 knockdown, the chick embryos displayed heart malformations, which include aberrant fusion of the heart fields, leading to incomplete terminal differentiation of the cardiomyocytes. cCcbe1 overexpression also resulted in severe heart defects, including cardia bifida. Altogether, our data demonstrate that although cardiac progenitors cells are specified in cCcbe1 morphants, the migration and proliferation of cardiac precursors cells are impaired, suggesting that cCcbe1 is a key gene during early heart development.
- Study of the role of cCcbe1, a novel gene coding for an EGF-like domain protein in the process of induction and organogenesis of the heartPublication . Furtado, João Francisco Venturinha; Belo, José AntónioThe vertebrate heart is a complex organ composed of several cell types, being developed through cardiogenic regions that have different expressing specific genes involved in heart specification. Understanding heart development on a molecular level is a requirement for unravel the causes of congenital heart diseases since specific cardiac lineages have been associated with cardiovascular malformations. During the course of a differential screen to identify transcripts specific for chick heart/hemangioblast precursor cells, we have identified Ccbe1 (Collagen and calcium-binding EGF-like domain 1). The current study intends to accomplish a detailed characterization of the expression pattern and functional analyses, by overexpression and knockdown approaches, of chick (c)Ccbe1. Whole-mount in situ hybridization analysis demonstrate that cCcbe1 is expressed in the early cardiac precursors of the heart forming regions at stage HH4 and at later stages is highly specific for the second heart field. Furthermore, functional analyses of cCcbe1 revealed an important role of cCcbe1 in early heart tube formation. In addition, the results presented in this thesis suggested that cCcbe1 is an important gene during heart development, is required for proper proliferation and migration of the heart precursors, and might be limited to multipotent and highly proliferative progenitors and downregulated upon cellular commitment into more specific cardiac phenotypes.
- Aulas da unidade curricular de Métodos Citohistoquímica 2016/2017Publication . Furtado, João1. Teoria da coloração 2. Hematoxilina e Eosina (princípios e exemplos de aplicação) 3. Hidratos Carbono (princípios e exemplos de aplicação) 4. Tecido Conjuntivo – Colagénio, Elastina e Reticulina (princípios e exemplos de aplicação) 5. Deteção de Lípidos (princípios e exemplos de aplicação) 6. Deteção de Pigmentos e Minerais (princípios e exemplos de aplicação) 7. Deteção Amiloide (princípios e exemplos de aplicação) 8. Deteção de Microorganismos (princípios e exemplos de aplicação)
- Aulas da unidade curricular de Citopatologia I 2016/2017Publication . Furtado, JoãoPermite ao estudante conhecer os principais constituintes/células do sistema Ginecológico estudado, para conhecimento da sua organização, função. Estes conteúdos possibilitam ainda o estudo da patologia associada a citologia Ginecológica.
- Chick heart/hemangioblast progenitor 12 (chep 12: study of a novel gene expressed in the heart/hemangioblast percursors cellsPublication . Furtado, João Francisco Venturinha; Belo, José AntónioEstablishment of the biochemical and molecular nature of cardiac development is essential to understand the relationship between genetic and morphological aspects of heart formation. The need to identify genes that are differentially expressed between cell populations within the embryo and within the embryonic heart is critical to elucidate the complex processes of heart development. With this in mind, our laboratory focused on the identification and study of novel genes expressed and involved in the correct development of the vertebrate heart/hemangioblast precursor cell (HPC) lineages. A differential screening using Affymetrix GeneChip Chick system was performed, leading to the identification of several new genes expressed in the haematopoiesis, angiogenesis or cardiogenesis precursor lineages. One of these novel genes, initially denominated CHEP12 (Chick Heart/Hemangioblast Progenitor 12), encodes for a novel secreted protein that contains a Calcium-binding EGF-like domain. Additionally, it was found that besides the canonical CHEP12 transcript, there are several alternative-spliced form variants, indicating that several isoform proteins may be synthesized in response to different signals. The current study intends to accomplish a detailed characterization of the expression pattern of the canonical CHEP12 gene and one of its alternative-spliced form variant (CHEP12-A). WISH and histological sections allowed us to suggest that CHEP12 may be involved in cardiogenic processes due to its expression in the Secondary Heart Field. Moreover, CHEP12 might also be implicated in the development of the venous system as its expression was also detected in the anterior, posterior and common cardinal veins. In addition, comparison between the expression pattern of the full transcript CHEP12 and its isoform CHEP12-A showed no significant differences.