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Techniques in Dev. B
Key Techniques in Developmental Biology (Fate Mapping & Experimental Approaches)
| Question | Answer |
|---|---|
| What are four techniques used in an Anatomical Approach to Embryology? | 1. Comparative embryology 2. Evolutionary embryology 3. Teratology 4. Mathematical modeling |
| What is comparative embryology? | Studies how anatomy changes during development in different organisms |
| What is evolutionary embryology? | Studies how development changes may cause evolutionary changes |
| What is teratology? | The study of birth defects |
| What is mathematical modeling? | A way of making predictions on how structures form during development |
| What are two techniques used in an Experimental Approach to Embryology? | 1. Ablation 2. Transplantation |
| Define ablation | Ablation is the destruction of or the removal of cells in an embryo |
| Define transplantation | Transplantation is where we take cells from one embryo and transplant them into another |
| Why do we perform transplantation in embryos? | To find out if those cells have the potential to take on a different fate (Beat Differentiation) |
| What are two techniques used in a Genetic Approach to Embryology? | 1. Mutagenesis 2. Transgenics |
| Define Mutagenesis | To cause a gene mutation and observe what problems arise |
| Define Transgenics | To insert a gene that isn't normally expressed in a cell |
| What is the purpose of using transgenics? | To artificially express genes and observe differentiation changes |
| What is Fate Mapping? | Fate Mapping is the study of embryonic origin of various adult tissues and structures |
| What is a "Fate"? | The "Fate" of each cell or cell group is mapped onto the embryo, showing which parts of the embryo will develop into which tissue |
| What is a Fate Map? | A Fate Map labels groups of cells to see what that area of the embryo will become |
| What is the purpose of a Fate Map? | To provide researchers with information on which portions of the embryo normally become which larval or adult structures |
| What are five techniques used in Fate Mapping? | 1. Living embryos 2. Vital dye marking 3. Radioactive labeling 4. Fluorescent dyes 5. Genetic marking |
| Out of those five techniques, what three are a form of "tagging"? | 1. Vital dye marking 2. Radioactive labeling 3. Fluorescent dyes |
| What are some techniques used in Genetic Embryology? | -RNA & Protein Localization Techniques -DNA Insertion -Transgenic and Knockout Mice -DNA & Antisense RNA Inhibition |
| Describe common RNA & Protein Localization Techniques | -Northern Blotting -RT-PCR -Microarrays -In situ Hybridization -Immunohistochemistry |
| Define Northern Blotting | A way of determining where RNA is outside of the cell |
| Define RT-PCR | Taking RNA and cloning it |
| Define Microarrays | Gives you hundreds of RNA at a time; similar to northern blots |
| Define In situ Hybridization | Looks at where RNA is expressed inside the embryo; antibodies are introduced and causes staining--stains where the RNA is |
| Define Immunohistochemistry | Similar to northern blots but labels proteins and RNA; looks at protein detection through florescent antibodies |
| Describe common DNA Insertion Techniques | -Microinjection -Lipid transfection -Electroporation |
| Define Microinjection | DNA is injected directly into the nucleus of a cell |
| Define Lipid transfection | DNA enters the cell encapsulated within a lipid bilayer |
| Define Electroporation | A high voltage pulse creates transient pores in the cell membrane which allows the DNA to enter the cell |
| What are Knockout Mice? | Replace/insert DNA into embryonic stem cell of a mouse to genetically modify it. They knockout expression of genes. |
| Which Genetic Development question does Knockout mice answer? | Is this gene necessary? |
| Describe Antisense RNA Inhibition | Targets the RNA itself and inhibits proteins that inhibit RNA translation and observe the downstream effects |
| What are the 3 Genetic Development Questions? | 1. Time and Place -- Is a gene expressed at the right time and place to play a particular role in development? 2. Necessity -- Is the gene required for the appropriate activity? 3. Sufficiency -- Is expression of the gene enough by itself? |
| Describe a Defect Experiment | Embryo portion is destroyed, and the impaired embryo development is observed |
| Describe an Isolation Experiment? | A portion of the embryo is removed; partial embryo and isolated portion is observed (FATE) |
| Describe a Recombination Experiment? | Replacing an original part of an embryo with another part from the same embryo (DIFFERENTIATION) |
| Describe a Transplantation Experiment? | Replacing a portion of an embryo with a portion from a different embryo |
| What is the main purpose of Time and Place? | Localization, natural processes of development Objective --> visualize wild-type expression patterns |
| What is the main purpose of Necessity? | Is it necessary for the cells fate? Objective --> Knockout or block a gene from being expressed |
| What is the main purpose of Sufficiency? | Activate a gene in a different cell type; is the gene by itself sufficient enough to do what it needs to do Objective --> Misexpress the gene in a region where not normally expressed |
Created by:
IsabellaHixson
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