TY - JOUR
T1 - Microtubules are organized independently of the centrosome in Drosophila neurons
AU - Nguyen, Michelle M.
AU - Stone, Michelle C.
AU - Rolls, Melissa M.
N1 - Funding Information:
We are very thankful to everyone who sent us fly stocks and antibodies, including Renata Basto and Jordan Raff, Miki Fujioka and Jim Jaynes, Li Cheng and Yuh Nung Jan, Tim Megraw, and Greg Rogers. We are also very grateful to the Bloomington Drosophila Stock Center and Developmental Studies Hybridoma Bank for serving as important resources to the scientific community. We thank Wendy Hanna-Rose for allowing use of her UV laser for experiments. We are grateful to Floyd Mattie for comments on the manuscript and other members of the Rolls lab for helpful discussions throughout. Funding for this work was provided by the National Institutes of Health, grant R01 GM085115.
PY - 2011/12/6
Y1 - 2011/12/6
N2 - Background: The best-studied arrangement of microtubules is that organized by the centrosome, a cloud of microtubule nucleating and anchoring proteins is clustered around centrioles. However, noncentrosomal microtubule arrays are common in many differentiated cells, including neurons. Although microtubules are not anchored at neuronal centrosomes, it remains unclear whether the centrosome plays a role in organizing neuronal microtubules. We use Drosophila as a model system to determine whether centrosomal microtubule nucleation is important in mature neurons.Results: In developing and mature neurons, centrioles were not surrounded by the core nucleation protein γ-tubulin. This suggests that the centrioles do not organize functional centrosomes in Drosophila neurons in vivo. Consistent with this idea, centriole position was not correlated with a specific region of the cell body in neurons, and growing microtubules did not cluster around the centriole, even after axon severing when the number of growing plus ends is dramatically increased. To determine whether the centrosome was required for microtubule organization in mature neurons, we used two approaches. First, we used DSas-4 centriole duplication mutants. In these mutants, centrioles were present in many larval sensory neurons, but they were not fully functional. Despite reduced centriole function, microtubule orientation was normal in axons and dendrites. Second, we used laser ablation to eliminate the centriole, and again found that microtubule polarity in axons and dendrites was normal, even 3 days after treatment.Conclusion: We conclude that the centrosome is not a major site of microtubule nucleation in Drosophila neurons, and is not required for maintenance of neuronal microtubule organization in these cells.
AB - Background: The best-studied arrangement of microtubules is that organized by the centrosome, a cloud of microtubule nucleating and anchoring proteins is clustered around centrioles. However, noncentrosomal microtubule arrays are common in many differentiated cells, including neurons. Although microtubules are not anchored at neuronal centrosomes, it remains unclear whether the centrosome plays a role in organizing neuronal microtubules. We use Drosophila as a model system to determine whether centrosomal microtubule nucleation is important in mature neurons.Results: In developing and mature neurons, centrioles were not surrounded by the core nucleation protein γ-tubulin. This suggests that the centrioles do not organize functional centrosomes in Drosophila neurons in vivo. Consistent with this idea, centriole position was not correlated with a specific region of the cell body in neurons, and growing microtubules did not cluster around the centriole, even after axon severing when the number of growing plus ends is dramatically increased. To determine whether the centrosome was required for microtubule organization in mature neurons, we used two approaches. First, we used DSas-4 centriole duplication mutants. In these mutants, centrioles were present in many larval sensory neurons, but they were not fully functional. Despite reduced centriole function, microtubule orientation was normal in axons and dendrites. Second, we used laser ablation to eliminate the centriole, and again found that microtubule polarity in axons and dendrites was normal, even 3 days after treatment.Conclusion: We conclude that the centrosome is not a major site of microtubule nucleation in Drosophila neurons, and is not required for maintenance of neuronal microtubule organization in these cells.
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U2 - 10.1186/1749-8104-6-38
DO - 10.1186/1749-8104-6-38
M3 - Article
C2 - 22145670
AN - SCOPUS:82655186566
SN - 1749-8104
VL - 6
JO - Neural Development
JF - Neural Development
IS - 1
M1 - 38
ER -