Project description:The face is one of the three regions most frequently affected by congenital defects in humans. In order to understand the molecular mechanisms involved it is necessary to have a more complete picture of gene expression in the embryo. Here we use microarrays to profile expression in chicken facial prominences, post neural crest migration and prior to differentiation of mesenchymal cells. Chip-wide analysis revealed that maxillary and mandibular prominences had similar expression profiles while the frontonasal mass chips were distinct. Of the 3094 genes that were differentially expressed in one or more regions of the face, a group of 56 genes was subsequently validated with quantitative PCR and a subset examined with in situ hybridization. Microarrays trends were consistent with the QPCR data for the majority of genes (81%). On the basis of QPCR and microarray data, groups of genes that characterize each of the facial prominences can be determined. We used microarrays to detail the global programme of gene expression underlying facial morphogensis Experiment Overall Design: Chicken embryos were selected at a stage after neural crest cells have ceased migration, when facial prominences have formed and prior to ctyodifferentiation of cartilage, bone, muscle. Microdissection was used to isolate facial prominences. Embryos were dissected on ice in Hanks Buffered Salt Solution and batches of 26-32 pieces were snap frozen in liquid Nitrogen. These pooled facial prominences comprised one biological replicate. A total of three biological replicates were generated for the frontonasal mass, maxillary and mandibular prominences.
Project description:The face is one of the three regions most frequently affected by congenital defects in humans. In order to understand the molecular mechanisms involved it is necessary to have a more complete picture of gene expression in the embryo. Here we use microarrays to profile expression in chicken facial prominences, post neural crest migration and prior to differentiation of mesenchymal cells. Chip-wide analysis revealed that maxillary and mandibular prominences had similar expression profiles while the frontonasal mass chips were distinct. Of the 3094 genes that were differentially expressed in one or more regions of the face, a group of 56 genes was subsequently validated with quantitative PCR and a subset examined with in situ hybridization. Microarrays trends were consistent with the QPCR data for the majority of genes (81%). On the basis of QPCR and microarray data, groups of genes that characterize each of the facial prominences can be determined. We used microarrays to detail the global programme of gene expression underlying facial morphogensis keyword: craniofacial
Project description:The patterning of the facial midline involves early specification of neural crest cells to form skeletal tissues that support the upper jaw . In order to understand the molecular mechanisms involved we have taken advantage of a beak duplication model developed in the chicken embryo. Here we can induce the transformation of the side of the beak into a second midline that is easily identifiable by the formation of a supernumerary egg tooth. The phenotype is induced by implanting two microscopic beads, one soaked in retinoic acid and the other soaked in Noggin into the side of the head of the chicken embryo. Here we use microarrays to profile expression of maxillary mesenchyme 16h after placing the beads. A subset of genes were validated using in situ hybridization and QPCR. The aims of the study are to test the function of these genes using retroviral transgenesis, knockdown with morpholinos or expression of secreted proteins and their application to the embryo. Embryos were incubated at 38 degrees C until they reached Hamburger Hamilton Stage 15 or 25 somites. Four different treatments were carried out consisting of two beads simultaneously implanted into the side of the head between the eye and mandibular arch. The beads were either Noggin+retinoic acid, Tris+retinoic acid, Noggin+DMSO or the control, Tris+DMSO. Embryos were reincubated for another 16h until they reached stage 18. Embryos were dissected on ice in Hanks Buffered Salt Solution and batches of 12-15 maxillary pieces were snap frozen in liquid Nitrogen. These pooled facial prominences comprised one biological replicate. A total of three biological replicates were generated for each treatment.
Project description:We report the RNA profiles of both control and talpid2 frontonasal, maxillary, and mandibular prominences of the chick face at Hamburger and Hamilton (HH) stage 25. For more details please see: "The cellular and molecular etiology of the craniofacial defects in the avian ciliopathic mutant, talpid2." Facial prominences (frontonasal, maxillary, and mandibular) from 8 control and 8 talpid2 HH 25 embryos were harvested, pooled, and RNA-seq was preformed on samples.
Project description:The aim of the present study was to investigated the difference of Nrf2-regulated genes in livers between normal and heat-stressed chickens. The CUT&Tag and high-throughput sequencing technologies were used in this experiment. Results showed that 13171838- 15417444 clean reads were obtained in this study. These data suggested that there were many Nrf2- regulated genes in the liver of heat-stressed chicken.