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.
The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.
The entire central nervous system develops from the neural plate, initially a single-cell layered epithelium. Anterior-posterior patterning is established by signals emanating from tissues outside the neural plate, while local signaling centers that are established within the neural plate itself will refine this initial patterning. One signaling center that plays an important role in the early development of midbrain and anterior hindbrain is the mid/hindbrain organizer (MHB). It is expresses Wnt1 on its rostral side and Fgf8 on its caudal side to exert its effects on future midand hindbrain.We have previously shown that Sip1/Zfhx1b is a multi-domain transcriptional repressor that can interact with several protein partners, thereby exerting several different functions depending on cellular context. We report here a new function of Sip1 in midbrain and hindbrain patterning.At E8.5, Sip1 is expressed in almost the entire neural plate. Sip1 represses Dkk1 expression in the early midbrain, leading to precocious Dkk1 expression at late headfold stage and increased expression at 5S stage in Sip1 knockout embryos. As Dkk1 is an inhibitor of Wnt signaling, the observed aberrant expression of Dkk1 leads to disturbed Wnt-b-catenin signaling in the future midbrain and anterior hindbrain. Hence, this aberrant Wnt signaling in Sip1 knockout embryos leads to early MHB patterning defects, as shown by the altered expression domain of MHB region genes, including En2 and Pax5.Sip1 is thus a regulator of Wnt-b-catenin signaling during early midbrain and anterior hindbrain development and thereby indirectly influences patterning of these tissues.We have identified a promoter element of chick Cerberus able to drive EGFP expression into the heart and the hemangioblast precursor cells. More importantly, these EGFP-positive H/HPC were able to be traced back to a population of cells that consistently exit from the anterior primitive streak region from as early as stage 4.In order to identify and study novel genes expressed and involved in the correct development and differentiation of the vertebrate H/HPC lineages, a differential screening using Affymetrix GeneChip system technologies was performed.Remarkably, this screening led to the identification of more than 200 new genes potentially expressed in these haematopoesis, angiogenesis or cardiogenesis precursor lineages.One of the novel genes identified, was initially denominated CHEP12 (Chick Heart/Haemangioblast Progenitor #12) due to its expression in anterior cardiac mesoderm precursor cells, both in chick and in mouse, and encodes for a novel secreted protein.Using anti-CHEP12 morpholino oligonucleotides resulted into several of heart tube malformations. We have also found that besides the canonical cCHEP12 transcript there are several alternativespliced form variants, which indicates that several isoform proteins may be synthesized in response to different signals. By performing developmental, genetic, biochemical and functional studies in chick, Xenopus and mouse models we aim to ...
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