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Conserved RNA-binding proteins required for dendrite morphogenesis in Caenorhabditis elegans sensory neurons.

Antonacci S, Forand D, Wolf M, Tyus C, Barney J, Kellogg L, Simon MA, Kerr G, Wells KL, Younes S, Mortimer NT, Olesnicky EC, Killian DJ - G3 (Bethesda) (2015)

Bottom Line: Homologs of each of these genes were previously identified as important in the Drosophila melanogaster dendritic arborization sensory neurons.Our results suggest that RNA processing, mRNA localization, mRNA stability, and translational control are all important mechanisms that contribute to dendrite morphogenesis, and we present a conserved set of RNA-binding proteins that regulate these processes in diverse animal species.Furthermore, homologs of these genes are expressed in the human brain, suggesting that these RNA-binding proteins are candidate regulators of dendrite development in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Colorado College, Colorado Springs, Colorado 80903.

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PVD-specific expression of RNA-binding protein::green fluorescent protein (RBP::GFP) fusion proteins rescues dendrite defects in most of the RBP mutants. Control (ctl) animals (gray), mutant animals (red), and mutant animals bearing an extrachromosomal array (Ex) that expresses the corresponding RBP::GFP fusion protein in the PVD neuron (blue) were scored for the number of dendritic termini using the ser-2prom3::myr-mCherry PVD marker. Points within each scatter column represent counts of dendritic termini from the PVD cell body to the tail on the dorsal or ventral side of the worm. Lines represent the means and the 95% confidence interval of the mean. All of the mutants are significantly lower than the control. *Mutants expressing RBP::GFP are significantly different from mutants and significantly different from controls indicating a partial rescue. **Mutants with RBP::GFP are significantly different from mutants and not significantly different from controls indicating a complete rescue. ***GFP::LARP-5 causes a significant reduction in dendritic termini compared to the larp-5 mutant alone. All statistics are based on a one-way ANOVA test with a Fisher’s Least Significant Difference multiple comparisons test with a 95% confidence interval.
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fig7: PVD-specific expression of RNA-binding protein::green fluorescent protein (RBP::GFP) fusion proteins rescues dendrite defects in most of the RBP mutants. Control (ctl) animals (gray), mutant animals (red), and mutant animals bearing an extrachromosomal array (Ex) that expresses the corresponding RBP::GFP fusion protein in the PVD neuron (blue) were scored for the number of dendritic termini using the ser-2prom3::myr-mCherry PVD marker. Points within each scatter column represent counts of dendritic termini from the PVD cell body to the tail on the dorsal or ventral side of the worm. Lines represent the means and the 95% confidence interval of the mean. All of the mutants are significantly lower than the control. *Mutants expressing RBP::GFP are significantly different from mutants and significantly different from controls indicating a partial rescue. **Mutants with RBP::GFP are significantly different from mutants and not significantly different from controls indicating a complete rescue. ***GFP::LARP-5 causes a significant reduction in dendritic termini compared to the larp-5 mutant alone. All statistics are based on a one-way ANOVA test with a Fisher’s Least Significant Difference multiple comparisons test with a 95% confidence interval.

Mentions: To test whether these RBPs function cell-autonomously in the PVD neuron and to determine if RBP::GFP fusion protein subcellular localization is biologically relevant, we expressed each RBP::GFP transgene specifically in the PVD neuron and tested for its ability to rescue PVD dendrite defects in RBP mutant animals (see the section Materials and Methods). We found that PVD-specific expression of CGH-1, MBL-1, and Y55F3AM.3 fusion proteins to GFP confer a statistically significant complete rescue of the PVD dendritic termini defects in the respective mutants (Figure 7). This strongly suggests that CGH-1, MBL-1, and Y55F3AM.3 each function cell-autonomously in the PVD to control dendrite development. Furthermore, the subcellular localizations inferred from these RBP::GFP fusions proteins are likely accurate because these proteins are functional.


Conserved RNA-binding proteins required for dendrite morphogenesis in Caenorhabditis elegans sensory neurons.

Antonacci S, Forand D, Wolf M, Tyus C, Barney J, Kellogg L, Simon MA, Kerr G, Wells KL, Younes S, Mortimer NT, Olesnicky EC, Killian DJ - G3 (Bethesda) (2015)

PVD-specific expression of RNA-binding protein::green fluorescent protein (RBP::GFP) fusion proteins rescues dendrite defects in most of the RBP mutants. Control (ctl) animals (gray), mutant animals (red), and mutant animals bearing an extrachromosomal array (Ex) that expresses the corresponding RBP::GFP fusion protein in the PVD neuron (blue) were scored for the number of dendritic termini using the ser-2prom3::myr-mCherry PVD marker. Points within each scatter column represent counts of dendritic termini from the PVD cell body to the tail on the dorsal or ventral side of the worm. Lines represent the means and the 95% confidence interval of the mean. All of the mutants are significantly lower than the control. *Mutants expressing RBP::GFP are significantly different from mutants and significantly different from controls indicating a partial rescue. **Mutants with RBP::GFP are significantly different from mutants and not significantly different from controls indicating a complete rescue. ***GFP::LARP-5 causes a significant reduction in dendritic termini compared to the larp-5 mutant alone. All statistics are based on a one-way ANOVA test with a Fisher’s Least Significant Difference multiple comparisons test with a 95% confidence interval.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390579&req=5

fig7: PVD-specific expression of RNA-binding protein::green fluorescent protein (RBP::GFP) fusion proteins rescues dendrite defects in most of the RBP mutants. Control (ctl) animals (gray), mutant animals (red), and mutant animals bearing an extrachromosomal array (Ex) that expresses the corresponding RBP::GFP fusion protein in the PVD neuron (blue) were scored for the number of dendritic termini using the ser-2prom3::myr-mCherry PVD marker. Points within each scatter column represent counts of dendritic termini from the PVD cell body to the tail on the dorsal or ventral side of the worm. Lines represent the means and the 95% confidence interval of the mean. All of the mutants are significantly lower than the control. *Mutants expressing RBP::GFP are significantly different from mutants and significantly different from controls indicating a partial rescue. **Mutants with RBP::GFP are significantly different from mutants and not significantly different from controls indicating a complete rescue. ***GFP::LARP-5 causes a significant reduction in dendritic termini compared to the larp-5 mutant alone. All statistics are based on a one-way ANOVA test with a Fisher’s Least Significant Difference multiple comparisons test with a 95% confidence interval.
Mentions: To test whether these RBPs function cell-autonomously in the PVD neuron and to determine if RBP::GFP fusion protein subcellular localization is biologically relevant, we expressed each RBP::GFP transgene specifically in the PVD neuron and tested for its ability to rescue PVD dendrite defects in RBP mutant animals (see the section Materials and Methods). We found that PVD-specific expression of CGH-1, MBL-1, and Y55F3AM.3 fusion proteins to GFP confer a statistically significant complete rescue of the PVD dendritic termini defects in the respective mutants (Figure 7). This strongly suggests that CGH-1, MBL-1, and Y55F3AM.3 each function cell-autonomously in the PVD to control dendrite development. Furthermore, the subcellular localizations inferred from these RBP::GFP fusions proteins are likely accurate because these proteins are functional.

Bottom Line: Homologs of each of these genes were previously identified as important in the Drosophila melanogaster dendritic arborization sensory neurons.Our results suggest that RNA processing, mRNA localization, mRNA stability, and translational control are all important mechanisms that contribute to dendrite morphogenesis, and we present a conserved set of RNA-binding proteins that regulate these processes in diverse animal species.Furthermore, homologs of these genes are expressed in the human brain, suggesting that these RNA-binding proteins are candidate regulators of dendrite development in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Colorado College, Colorado Springs, Colorado 80903.

Show MeSH
Related in: MedlinePlus