Use mice as the experiment's test subject. Make sure the animals are kept in a pathogen-free environment to prevent compromising the intended investigations. Hysterectomies should be done to remove the embryos. Before performing the experiment, make sure the animals are given isofluorane to make them unconscious (Ranajeet S 2427). To ensure that the medicine does not negatively affect the embryos to be researched, the pregnant females should be put to sleep at specified gestational stages. The intended embryos must be preserved for an hour in paraformaldehyde that is 4% concentrated. Label the cryosections using various antibodies that would be provided. Visualize the nuclei and the F-actin using Phalloidin. Determine the cells that have been identified using the Photoshop. Analyze your results suing the SPSS soft wares.
Question three
To test for the functionality of the Shroom3 and Pitx1 in the development of the gut. A mouse should be stored in a sterile environment under injection of gonadotropin hormone to enhance their ovulation (Mei-I Chung1 p 1340). The animal should be checked whether ovulation actually occurred then sperms are pumped in. after fertilization, the embryo I extracted through hysterectomies. The embryo is evaluated during its growth process and the period in which the effects of Shroom3 and Pitx1 are seen are noted. The mouse should be monitored through the use of ultra sound machines to ensure that the development of the embryo is monitored to determine the time that is essential for study is reached (Mei-I Chung1 p 1340). The extract is fixed using the four percent concentrated formalin at room temperature. The extract is then waxed using increasing concentrations of alcohol. The resultant is then microtome in many small segments. Imaging of the resultant microtomes should be done and the various effects of Shroom3 and Pitx1 noted and recorded.
Question four
Mouse are the main source of the embryos during the experiments. Ensure that they are injected with human gonadotropic hormone to allow for their ovulation. Extract the embryo using the female animal then fertilize and analyze the various developmental stages of the animal. The mouse and the embryo should be kept and maintained under specific environmental conditions to ensure that they are not infected by pathogens. Before the harvest, the embryos should be monitored under ultra sounds to help determine the best time to harvest the embryo for studies (Rachel A. Reed 2311). In should be noted that the mouse need to be under anesthesia to ensure that it does not feel too much pain. The embryo should be extracted, persevered under four percent formalin, and then analyzed using the image processing techniques. The imaging process should involve the combination of two related sections to ensure that their reactions are effectively viewed. The imaging should be done under eighty percent glycerol and 10-micrometer sections. The sectioning should be done following the paraffin wax procedures. The fixation should be done for one hour at room temperatures. Fixation should be followed by temperatures of about -20 under the influence of methane.
Question five
I expect to find out a great relationship between mammals and Xenopus conservation. The developmental stages of mammals are the same. Therefore, there must be similarity in their conservation patterns and effects. In case there was no conversion, there could also be no conversion among the mammals.
Works Cited
Mei-I Chung1, Nanette M. Nascone-Yoder2, Stephanie A. Grover3, Thomas A. Drysdale3,4,5 and John B. Wallingford1,6,. “Direct activation of Shroom3 transcription by Pitx proteins drives epithelial morphogenesis in the developing gut.” Development 137.8 (2010): 1339-1349.
Rachel A. Reed,1 Mandy A. Womble,1 Michel K. Dush,1 Rhesa R. Tull,1 Stephanie K. Bloom,1 Allison R. Morckel,1 Edward W. Devlin,2 and Nanette M. Nascone-Yoder1*. “Morphogenesis of the primitive gut tube is generated by Rho/ROCK/myosin II–mediated endoderm rearrangements.” Developmental Dynamics 238.12 (2009): 3111-3125.
Ranajeet S. Saund1, Masami Kanai-Azuma2, Yoshiakira Kanai3, Injune Kim4, Mary T. Lucero5,6 and Yukio Saijoh1. “Gut endoderm is involved in the transfer of left-right asymmetry from the node to the lateral plate mesoderm in the mouse embryo.” Development 139.13 (2012): 2426-2435.
