90 likes | 187 Views
Gene Function in Vulval Morphogenesis of C. elegans. Erin Weston, Jimmy Goncalves , and Karina Gonzalez . Introduction . Tubulogenesis in early development is an important process in the function of many organisms. Vulval development of C. elegans is the working model
E N D
Gene Function in VulvalMorphogenesis of C. elegans Erin Weston, Jimmy Goncalves, and Karina Gonzalez
Introduction • Tubulogenesis in early development is an important process in the function of many organisms. • Vulval development of C. elegansis the working model • Specifically the role of egl-26 • Egl-26 is active in the maintenance of the shape of the vulva towards the end of the development of the vulva • This is the only gene that has been found to control vulF in this way • Why haven’t more genes been found to have this role? • Forward genetics • RNAishould give better insight. Why?
Vulval Morphogenesis • Made up of 7 multi-nucleated toroids stack on top of each other • Form the connection between the vulval and uterine lumens to the outside environment • Formation begins when anchor cell (AC) sends out an EGF-like signal to 3 precursor cells • P6.p adopts the primary fate while P5.p and P7.p both adopt a secondary fate
Precursor cells go through division creating 22 cells • Cells begin to move proximally which causes the invagination of the epithelial lining • AC punches hole through top of vulva to create the connection to the uterine lumen then retracts
egl-26 • Encodes a protein that is essential in the morphology of the dorsally located vulF cell • In mutants, the vulF cell collapses after the retraction of the AC • Expressed non-cell autonomously in the vulE cell and is evident in L4 • Required in the primary lineage of the vulva Normal L4 vulva egl-26 vulva
Hypothesis • Of a set of genes found to affect vulval development, through the use of RNAi, some of those genes act closely with EGL-26 • To test this hypothesis, RNAi was used to knockdown the expression of a desired gene to observe the loss of function phenotype • Special attention was given to genes in which vulval morphogenesis had been compromised
Results • Around 50 genes have been screened • dcp-66, F27C1.6, ncbp-2, egl-30 are of interest because phenotypes expressed in the vulva are similar to egl-26 dcp-66 F27C1.6 ncbp-2
What’s next? • Continue screening • Rescreen genes of interest • More careful phenotypical analysis • Transcriptional and translational GFP reporters • Protein localization • Gene expression