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bis_104_pq_3_ans_spring_08 - 2 Apply fluorescently labeled...

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Bis 104 Practice Question #3 In their initial set of experiments (the results depicted in Figure 1 A – N, page 694) Mukouyama’s group obtained results that led them to conclude that: “Peripheral nerves associate preferentially with arterial vessels but not with veins.” These experiments were done using 15.5 day old (E15.5) mouse embryo limb skin. a. Briefly describe the design of the EXPERIMENT that led them to this conclusion. List the procedural steps in chronological order. EXPERIMENT: Experimental strategy: Use whole mount immune-histochemistry of mouse embryo limb skin to examine the branching pattern of arteries, veins, and peripheral nerves in the same tissue. Experimental procedure: 1. Remove and fix limb skin samples from 15.5 day old mouse embryos. Use two different strains of transgenic mice. One whose arterial endothelial cells express a marker (ephrinB2 lacZ) and One whose venous endothelial cells express a marker (EphB4) lac Z)
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Unformatted text preview: 2. Apply fluorescently labeled antibodies to skin specimens to detect presence and pattern of cell-specific markers. (2H3 for nerves = green; PECAM-1 for undifferentiated endothelial cells = blue). 3. Wash away any unbound antibody. 4. Examine labeled skin whole mount samples by confocal fluorescence microscopy. 5. Compare staining patterns of nerves and blood vessels . b. Describe the key RESULTS of this experiment. The pattern of arterial cell staining mimics that of peripheral nerves (Fig. 1abc). The pattern of venous cell staining does not mimic peripheral nerve pattern (Fig. 1ghi) c. Do these results establish plausibility, necessity, or sufficiency? Explain the basis for your choice. It is plausible that arteries and neurons influence each other. A correlation in space has been established, but no proof of cause and effect....
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