Zebrafish Von Willebrand Factor Page: 55
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Discussion
The above experiments were able to use VMO to examine whether or not
creating a dysfunctional protein or a knockdown of VWF will have an affect on thrombus
formation upon injury. This is the second hemostatic gene knockdown by VMO in
zebrafish which adds credence to the VMO technology and its application to zebrafish
hemostasis. The use of two different types of morpholino provides different modes of
investigation while both providing the same efficacy and ability to target the same
protein at any stage of development and also providing confirmation of the mutual
results.
Here the employment of antisense morpholino was used to examine the effects
of alternate splicing on VWF function; however, upon examination of the RT-PCR
results it appears that an unstable VWF mRNA was created by the VWF-MO based
alternative splicing. A knockdown of VWF is similar to the human vWD type 1, where
individuals have very little VWF [139]. Because the resulting effect was a knockdown,
this provided a convenient way to observe whether or not VWF is functioning similarly to
human VWF, using a variety of bleeding assays. The assays include determining
whether or not VWF is essential for a firm clot formation as well as determining whether
or not VWF might be stored, and whether FVIII is being affected by a loss of VWF.
Creating a model for type 1 vWD on zebrafish is significant despite the fact other type 1
models exist. This is significant because zebrafish provide advantages that no other
animal models posses. A high fecundity allows for a researcher to investigate possible
drugs that might initiate transcription of new VWF on a large scale with a variety of
treatments all at the same time. The ease with which zebrafish adults and larvae can55
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Carrillo, Maira M. Zebrafish Von Willebrand Factor, dissertation, August 2012; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc177228/m1/63/: accessed April 25, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .