Limits...
Chromatin Modulatory Proteins and Olfactory Receptor Signaling in the Refinement and Maintenance of Fruitless Expression in Olfactory Receptor Neurons.

Hueston CE, Olsen D, Li Q, Okuwa S, Peng B, Wu J, Volkan PC - PLoS Biol. (2016)

Bottom Line: This regulation requires the chromatin modulatory protein Alhambra (Alh).Our results highlight two feed-forward regulatory mechanisms with both developmentally hardwired and olfactory receptor activity-dependent components that establish and maintain fru expression in ORNs.Such a dual mechanism of fru regulation in ORNs might be a trait of neurons driving plastic aspects of sex-specific behaviors.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Duke University, Durham, North Carolina, United States of America.

ABSTRACT
During development, sensory neurons must choose identities that allow them to detect specific signals and connect with appropriate target neurons. Ultimately, these sensory neurons will successfully integrate into appropriate neural circuits to generate defined motor outputs, or behavior. This integration requires a developmental coordination between the identity of the neuron and the identity of the circuit. The mechanisms that underlie this coordination are currently unknown. Here, we describe two modes of regulation that coordinate the sensory identities of Drosophila melanogaster olfactory receptor neurons (ORNs) involved in sex-specific behaviors with the sex-specific behavioral circuit identity marker fruitless (fru). The first mode involves a developmental program that coordinately restricts to appropriate ORNs the expression of fru and two olfactory receptors (Or47b and Ir84a) involved in sex-specific behaviors. This regulation requires the chromatin modulatory protein Alhambra (Alh). The second mode relies on the signaling from the olfactory receptors through CamK and histone acetyl transferase p300/CBP to maintain ORN-specific fru expression. Our results highlight two feed-forward regulatory mechanisms with both developmentally hardwired and olfactory receptor activity-dependent components that establish and maintain fru expression in ORNs. Such a dual mechanism of fru regulation in ORNs might be a trait of neurons driving plastic aspects of sex-specific behaviors.

Show MeSH
Maintenance of fru expression in adult ORNs requires CamK signaling and p300/CBP.(A) Wild-type antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta). (B–D) Antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta) as well as fruGal4 CamKI RNAi (B), UAS-creb(C), and UAS-p300 RNAi (D). (E) Quantification of antennal fru-positive ORN cell counts for experiments in Figs 5, 6 and 8. Data shown represents the fraction of Or47b-positive cells that are also fru-positive. For all graphs, asterisks indicate significant (p < .01) differences from fruGal4. Error bars represent SEM. A one-way ANOVA was performed and followed with Tukey’s HSD—see Materials and Methods. Cell count data also graphed in S10 Fig. All raw count data may be found in the Supporting Information as S1 Data.GENOTYPES:(A) Or47b-CD2 /+; fruGAL4UAS-40XCD8GFP(B) UAS-CamKI RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(C) UAS-CREB/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(D) UAS-p300RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4836687&req=5

pbio.1002443.g008: Maintenance of fru expression in adult ORNs requires CamK signaling and p300/CBP.(A) Wild-type antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta). (B–D) Antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta) as well as fruGal4 CamKI RNAi (B), UAS-creb(C), and UAS-p300 RNAi (D). (E) Quantification of antennal fru-positive ORN cell counts for experiments in Figs 5, 6 and 8. Data shown represents the fraction of Or47b-positive cells that are also fru-positive. For all graphs, asterisks indicate significant (p < .01) differences from fruGal4. Error bars represent SEM. A one-way ANOVA was performed and followed with Tukey’s HSD—see Materials and Methods. Cell count data also graphed in S10 Fig. All raw count data may be found in the Supporting Information as S1 Data.GENOTYPES:(A) Or47b-CD2 /+; fruGAL4UAS-40XCD8GFP(B) UAS-CamKI RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(C) UAS-CREB/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(D) UAS-p300RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP

Mentions: We also abolished Or47b function using mutants for the general Or coreceptor orco [28]. A loss of fru expression, similar to that seen in Or47b mutants, was observed in orco mutants (Figs 6A, 6B and 6H, 8E and S11 Fig) and was confirmed by qRT-PCR (S8 Fig). This phenotype could be rescued by expression of a UAS-orco transgene (Figs 6C and 6H, 8E and S11 Fig).


Chromatin Modulatory Proteins and Olfactory Receptor Signaling in the Refinement and Maintenance of Fruitless Expression in Olfactory Receptor Neurons.

Hueston CE, Olsen D, Li Q, Okuwa S, Peng B, Wu J, Volkan PC - PLoS Biol. (2016)

Maintenance of fru expression in adult ORNs requires CamK signaling and p300/CBP.(A) Wild-type antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta). (B–D) Antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta) as well as fruGal4 CamKI RNAi (B), UAS-creb(C), and UAS-p300 RNAi (D). (E) Quantification of antennal fru-positive ORN cell counts for experiments in Figs 5, 6 and 8. Data shown represents the fraction of Or47b-positive cells that are also fru-positive. For all graphs, asterisks indicate significant (p < .01) differences from fruGal4. Error bars represent SEM. A one-way ANOVA was performed and followed with Tukey’s HSD—see Materials and Methods. Cell count data also graphed in S10 Fig. All raw count data may be found in the Supporting Information as S1 Data.GENOTYPES:(A) Or47b-CD2 /+; fruGAL4UAS-40XCD8GFP(B) UAS-CamKI RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(C) UAS-CREB/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(D) UAS-p300RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836687&req=5

pbio.1002443.g008: Maintenance of fru expression in adult ORNs requires CamK signaling and p300/CBP.(A) Wild-type antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta). (B–D) Antennae expressing fruGal4 UAS-40XUASGFP (green) and Or47b-CD2 (magenta) as well as fruGal4 CamKI RNAi (B), UAS-creb(C), and UAS-p300 RNAi (D). (E) Quantification of antennal fru-positive ORN cell counts for experiments in Figs 5, 6 and 8. Data shown represents the fraction of Or47b-positive cells that are also fru-positive. For all graphs, asterisks indicate significant (p < .01) differences from fruGal4. Error bars represent SEM. A one-way ANOVA was performed and followed with Tukey’s HSD—see Materials and Methods. Cell count data also graphed in S10 Fig. All raw count data may be found in the Supporting Information as S1 Data.GENOTYPES:(A) Or47b-CD2 /+; fruGAL4UAS-40XCD8GFP(B) UAS-CamKI RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(C) UAS-CREB/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP(D) UAS-p300RNAi/+; Or47b-CD2/+; fruGAL4 UAS-40XCD8GFP
Mentions: We also abolished Or47b function using mutants for the general Or coreceptor orco [28]. A loss of fru expression, similar to that seen in Or47b mutants, was observed in orco mutants (Figs 6A, 6B and 6H, 8E and S11 Fig) and was confirmed by qRT-PCR (S8 Fig). This phenotype could be rescued by expression of a UAS-orco transgene (Figs 6C and 6H, 8E and S11 Fig).

Bottom Line: This regulation requires the chromatin modulatory protein Alhambra (Alh).Our results highlight two feed-forward regulatory mechanisms with both developmentally hardwired and olfactory receptor activity-dependent components that establish and maintain fru expression in ORNs.Such a dual mechanism of fru regulation in ORNs might be a trait of neurons driving plastic aspects of sex-specific behaviors.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Duke University, Durham, North Carolina, United States of America.

ABSTRACT
During development, sensory neurons must choose identities that allow them to detect specific signals and connect with appropriate target neurons. Ultimately, these sensory neurons will successfully integrate into appropriate neural circuits to generate defined motor outputs, or behavior. This integration requires a developmental coordination between the identity of the neuron and the identity of the circuit. The mechanisms that underlie this coordination are currently unknown. Here, we describe two modes of regulation that coordinate the sensory identities of Drosophila melanogaster olfactory receptor neurons (ORNs) involved in sex-specific behaviors with the sex-specific behavioral circuit identity marker fruitless (fru). The first mode involves a developmental program that coordinately restricts to appropriate ORNs the expression of fru and two olfactory receptors (Or47b and Ir84a) involved in sex-specific behaviors. This regulation requires the chromatin modulatory protein Alhambra (Alh). The second mode relies on the signaling from the olfactory receptors through CamK and histone acetyl transferase p300/CBP to maintain ORN-specific fru expression. Our results highlight two feed-forward regulatory mechanisms with both developmentally hardwired and olfactory receptor activity-dependent components that establish and maintain fru expression in ORNs. Such a dual mechanism of fru regulation in ORNs might be a trait of neurons driving plastic aspects of sex-specific behaviors.

Show MeSH