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To further characterize Trak1 depletion-induced mitochondrial morphological phenotype, we performed super-resolution imaging analyses using three-dimensional structured illumination microscopy (Huang et al., 2009; Fallaize et al., 2015).
Because CK19CreERT has a high rate of recombination in the small intestine but no morphological phenotype, we also immunolabeled this tissue for markers of proliferation, phospho-histone H3 and Ki67.
To interrogate this morphological phenotype we analyzed gene expression of patterns through growth in the tbpD non-native expression strain.
Consistent with the observed enhanced skeletal muscle morphological phenotype, we observed a concomitant up-regulation of skeletal muscle genes including Myh1-8, Myf5, and Pax3.
Consistent with its morphological phenotype, we identified genes encoding several mucins including MUC2, MUC3A (MUC3) and MUC17 but not MUC16 (CA125).
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Intrigued by the consistent and somewhat unexpected appearance of morphological phenotypes, we continued by examining the effects of phosphomimetic mutations at the serine-5 position.
We next wanted to characterize the gross morphological phenotypes we observed at higher resolution to glean better insight into whether exposure to Macondo oil affected specific developmental processes during zebrafish embryogenesis.
To determine when these morphological phenotypes occurred, we tracked individual 4 dpf Tg(fabp10 dsRed) larvae exposed to 350 mM ethanol over 32 hours (Fig. 1C) and verified these findings in larger cohorts (Fig. 1D,E).
Given the robust morphological phenotype induced by ApLRRTK, we sought to determine whether this structural remodeling was also accompanied by functional electrical changes in the synaptic strength of the sensory-motor neuron synapses.
Therefore, we speculated that the morphological phenotype of motor mutants and drug-treated cells was due to defects in CHS secretion.
We confirmed that cvy1 morphological phenotype was indeed caused by the described T-DNA insertion by constitutively overexpressing CVY1 gene in cvy1 mutant background.
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