This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator.
This aim i s 90% complete. We have generated transgenic animals for RNAi studies. Since our previous data shows that cof-2 expression is eliminated when animals are stressed, we are assaying for changes in behavior and development when cof-2 expression is reduced in vivo. We will also continue our attempts to generate animals carrying cof-2::GFP by (1) generating a new transgene under using serial overlap extension and (2) altering the dosage of cof-2::GFP DNA. Once trangenic lines are generated we plan to assay for changes in protein accumulation and animal development when cof-2 is ectopically expressed under induced, stressful conditions. Likewise, expression of cof-2 under the ribosomal protein promoter will allow continuous expression of cof-2, rather than the expression seen by developmental profile.
Aim 3 is completed.
Specific Aim 2 & 4:
These aims are 50% complete. We plan to continue isolating total RNA from brain sections at each developmental stage in stressed animals and to finalize qPCR measurements. We are currently growing and treating animals for similar developmental profile in protein expression. We also plan to continue with immunolocalization studies in unstressed animals for a baseline of myocilin protein accumulation.
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