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The RHO-1 RhoGTPase modulates fertility and multiple behaviors in adult C. elegans.

McMullan R, Nurrish SJ - PLoS ONE (2011)

Bottom Line: Changes in RHO-1 signaling in cholinergic neurons affected locomotion, pharyngeal pumping and fecundity.Changes in RHO-1 signaling outside the cholinergic neurons resulted in defective defecation, ovulation, and changes in C. elegans body morphology.Finally both increased and decreased RHO-1 signaling in adults resulted in death within hours.

View Article: PubMed Central - PubMed

Affiliation: MRC Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Neuroscience, Physiology and Pharmacology, University College, London, United Kingdom.

ABSTRACT
The Rho family of small GTPases are essential during early embryonic development making it difficult to study their functions in adult animals. Using inducible transgenes expressing either a constitutively active version of the single C. elegans Rho ortholog, RHO-1, or an inhibitor of endogenous Rho (C3 transferase), we demonstrate multiple defects caused by altering Rho signaling in adult C. elegans. Changes in RHO-1 signaling in cholinergic neurons affected locomotion, pharyngeal pumping and fecundity. Changes in RHO-1 signaling outside the cholinergic neurons resulted in defective defecation, ovulation, and changes in C. elegans body morphology. Finally both increased and decreased RHO-1 signaling in adults resulted in death within hours. The multiple post-developmental roles for Rho in C. elegans demonstrate that RhoA signaling pathways continue to be used post-developmentally and the resulting phenotypes provide an opportunity to further study post-developmental Rho signaling pathways using genetic screens.

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RHO-1 regulates fecundity.(A) The number of unlaid eggs remaining in animals was determined as a steady-state measure of egg laying. Expression of hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) led to a significant decrease in the number of eggs remaining in animals when compared to wild type controls. Inhibition of endogenous RHO-1 using hsC3 transferase (nzEx4) did not have any significant effect on the number of eggs remaining in animals. (B) The stage of eggs laid was determined by visual inspection. Experiments were repeated four times and the data presented is the sum of these experiments. Animals expressing activated hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) laid some early stage eggs containing fewer than eight cells and hsRHO-1(G14V) (nzIs1) the total number of eggs laid was decreased. This decrease was also observed in animals expressing hsC3 transferase (nzEx4).
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pone-0017265-g003: RHO-1 regulates fecundity.(A) The number of unlaid eggs remaining in animals was determined as a steady-state measure of egg laying. Expression of hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) led to a significant decrease in the number of eggs remaining in animals when compared to wild type controls. Inhibition of endogenous RHO-1 using hsC3 transferase (nzEx4) did not have any significant effect on the number of eggs remaining in animals. (B) The stage of eggs laid was determined by visual inspection. Experiments were repeated four times and the data presented is the sum of these experiments. Animals expressing activated hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) laid some early stage eggs containing fewer than eight cells and hsRHO-1(G14V) (nzIs1) the total number of eggs laid was decreased. This decrease was also observed in animals expressing hsC3 transferase (nzEx4).

Mentions: Two sets of neurons (VC and HSN) control egg laying in C. elegans [42]. Both release ACh and serotonin; serotonin stimulates egg laying [32] whereas ACh has both positive and negative effects [40], [43]. To investigate the role of RHO-1 in this neuronally regulated behavior we tested the ability of RHO-1 to alter egg laying using two methods. We determined the number of unlaid eggs remaining in animals as a steady state measure of egg laying; animals with increased egg laying rates should retain fewer eggs than wild type while those with decreased egg laying should retain more. Even in the absence of heat shock hsRHO-1(G14V) animals showed a decrease in the number of eggs remaining in the animals suggesting that our heat shock-inducible transgenes were leaky in some cells under certain conditions (Figure 3A [44]). Expression of hsRHO-1(G14V) in adults decreased the number of eggs remaining in animals relative to heat shocked wild type and non-heat shocked hsRHO-1(G14V) animals (Figure 3A).


The RHO-1 RhoGTPase modulates fertility and multiple behaviors in adult C. elegans.

McMullan R, Nurrish SJ - PLoS ONE (2011)

RHO-1 regulates fecundity.(A) The number of unlaid eggs remaining in animals was determined as a steady-state measure of egg laying. Expression of hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) led to a significant decrease in the number of eggs remaining in animals when compared to wild type controls. Inhibition of endogenous RHO-1 using hsC3 transferase (nzEx4) did not have any significant effect on the number of eggs remaining in animals. (B) The stage of eggs laid was determined by visual inspection. Experiments were repeated four times and the data presented is the sum of these experiments. Animals expressing activated hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) laid some early stage eggs containing fewer than eight cells and hsRHO-1(G14V) (nzIs1) the total number of eggs laid was decreased. This decrease was also observed in animals expressing hsC3 transferase (nzEx4).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3046162&req=5

pone-0017265-g003: RHO-1 regulates fecundity.(A) The number of unlaid eggs remaining in animals was determined as a steady-state measure of egg laying. Expression of hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) led to a significant decrease in the number of eggs remaining in animals when compared to wild type controls. Inhibition of endogenous RHO-1 using hsC3 transferase (nzEx4) did not have any significant effect on the number of eggs remaining in animals. (B) The stage of eggs laid was determined by visual inspection. Experiments were repeated four times and the data presented is the sum of these experiments. Animals expressing activated hsRHO-1(G14V) (nzIs1) or nRHO-1(G14V) (nzIs29) laid some early stage eggs containing fewer than eight cells and hsRHO-1(G14V) (nzIs1) the total number of eggs laid was decreased. This decrease was also observed in animals expressing hsC3 transferase (nzEx4).
Mentions: Two sets of neurons (VC and HSN) control egg laying in C. elegans [42]. Both release ACh and serotonin; serotonin stimulates egg laying [32] whereas ACh has both positive and negative effects [40], [43]. To investigate the role of RHO-1 in this neuronally regulated behavior we tested the ability of RHO-1 to alter egg laying using two methods. We determined the number of unlaid eggs remaining in animals as a steady state measure of egg laying; animals with increased egg laying rates should retain fewer eggs than wild type while those with decreased egg laying should retain more. Even in the absence of heat shock hsRHO-1(G14V) animals showed a decrease in the number of eggs remaining in the animals suggesting that our heat shock-inducible transgenes were leaky in some cells under certain conditions (Figure 3A [44]). Expression of hsRHO-1(G14V) in adults decreased the number of eggs remaining in animals relative to heat shocked wild type and non-heat shocked hsRHO-1(G14V) animals (Figure 3A).

Bottom Line: Changes in RHO-1 signaling in cholinergic neurons affected locomotion, pharyngeal pumping and fecundity.Changes in RHO-1 signaling outside the cholinergic neurons resulted in defective defecation, ovulation, and changes in C. elegans body morphology.Finally both increased and decreased RHO-1 signaling in adults resulted in death within hours.

View Article: PubMed Central - PubMed

Affiliation: MRC Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Neuroscience, Physiology and Pharmacology, University College, London, United Kingdom.

ABSTRACT
The Rho family of small GTPases are essential during early embryonic development making it difficult to study their functions in adult animals. Using inducible transgenes expressing either a constitutively active version of the single C. elegans Rho ortholog, RHO-1, or an inhibitor of endogenous Rho (C3 transferase), we demonstrate multiple defects caused by altering Rho signaling in adult C. elegans. Changes in RHO-1 signaling in cholinergic neurons affected locomotion, pharyngeal pumping and fecundity. Changes in RHO-1 signaling outside the cholinergic neurons resulted in defective defecation, ovulation, and changes in C. elegans body morphology. Finally both increased and decreased RHO-1 signaling in adults resulted in death within hours. The multiple post-developmental roles for Rho in C. elegans demonstrate that RhoA signaling pathways continue to be used post-developmentally and the resulting phenotypes provide an opportunity to further study post-developmental Rho signaling pathways using genetic screens.

Show MeSH