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Tamm S.L.,Bell Center
Biological Bulletin | Year: 2015

Ctenophores, or comb jellies, are geotactic with a statocyst that controls the activity of the eight ciliary comb rows. If a ctenophore is tilted or displaced from a position of vertical balance, it rights itself by asymmetric frequencies of beating on the uppermost and lowermost comb rows, turning to swim up or down depending on its mood. I recently discovered that the statocyst of ctenophores has an asymmetric architecture related to the sagittal and tentacular planes along the oral-aboral axis. The four groups of pacemaker balancer cilia are arranged in a rectangle along the tentacular plane, and support a superellipsoidal statolith elongated in the tentacular plane. By controlled tilting of immobilized ctenophores in either body plane with video recording of activated comb rows, I found that higher beat frequencies occurred in the sagittal than in the tentacular plane at orthogonal orientations. Similar tilting experiments on isolated statocyst slices showed that statolith displacement due to gravity and the resulting deflection of the mechanoresponsive balancers are greater in the sagittal plane. Finally, tilting experiments on a mechanical model gave results similar to those of real statocysts, indicating that the geometric asymmetries of statolith design are sufficient to account for my findings. The asymmetric architecture of the ctenophore statocyst thus has functional consequences, but a possible adaptive value is not known. © 2015 Marine Biological Laboratory.

Dundon S.E.R.,Dartmouth College | Chang S.-S.,University of California at Los Angeles | Kumar A.,U.S. National Institutes of Health | Occhipinti P.,Dartmouth College | And 4 more authors.
Molecular Biology of the Cell | Year: 2016

Nuclei in syncytia found in fungi, muscles, and tumors can behave independently despite cytoplasmic translation and the homogenizing potential of diffusion. We use a dynaction mutant strain of the multinucleate fungus Ashbya gossypii with highly clustered nuclei to assess the relative contributions of nucleus and cytoplasm to nuclear autonomy. Remarkably, clustered nuclei maintain cell cycle and transcriptional autonomy; therefore some sources of nuclear independence function even with minimal cytosol insulating nuclei. In both nuclear clusters and among evenly spaced nuclei, a nucleus' transcriptional activity dictates local cytoplasmic contents, as assessed by the localization of several cyclin mRNAs. Thus nuclear activity is a central determinant of the local cytoplasm in syncytia. Of note, we found that the number of nuclei per unit cytoplasm was identical in the mutant to that in wild-type cells, despite clustered nuclei. This work demonstrates that nuclei maintain autonomy at a submicrometer scale and simultaneously maintain a normal nucleocytoplasmic ratio across a syncytium up to the centimeter scale. © 2016 Dundon et al.

Peinado G.,National University of Colombia | Osorno T.,National University of Colombia | Del Pilar Gomez M.,National University of Colombia | Del Pilar Gomez M.,Bell Center | And 2 more authors.
Proceedings of the National Academy of Sciences of the United States of America | Year: 2015

Melanopsin, the photopigment of the "circadian" receptors that regulate the biological clock and the pupillary reflex in mammals, is homologous to invertebrate rhodopsins. Evidence supporting the involvement of phosphoinositides in light-signaling has been garnered, but the downstream effectors that control the light-dependent conductance remain unknown. Microvillar photoreceptors of the primitive chordate amphioxus also express melanopsin and transduce light via phospholipase-C, apparently not acting through diacylglycerol. We therefore examined the role of calcium in activating the photoconductance, using simultaneous, high time-resolution measurements of membrane current and Ca2+ fluorescence. The light-induced calcium rise precedes the onset of the photocurrent, making it a candidate in the activation chain. Moreover, photolysis of caged Ca elicits an inward current of similar size, time course and pharmacology as the physiological photoresponse, but with a much shorter latency. Internally released calcium thus emerges as a key messenger to trigger the opening of light-dependent channels in melanopsin-expressing microvillar photoreceptors of early chordates. © 2015, National Academy of Sciences. All rights reserved.

Stevenson J.W.,Providence College | Stevenson J.W.,Bell Center | Conaty E.A.,Providence College | Conaty E.A.,Bell Center | And 10 more authors.
PLoS ONE | Year: 2016

The amyloid precursor protein (APP) is a causal agent in the pathogenesis of Alzheimer's disease and is a transmembrane protein that associates with membrane-limited organelles. APP has been shown to co-purify through immunoprecipitation with a kinesin light chain suggesting that APP may act as a trailer hitch linking kinesin to its intercellular cargo, however this hypothesis has been challenged. Previously, we identified an mRNA transcript that encodes a squid homolog of human APP770. The human and squid isoforms share 60% sequence identity and 76% sequence similarity within the cytoplasmic domain and share 15 of the final 19 amino acids at the C-terminus establishing this highly conserved domain as a functionally import segment of the APP molecule. Here, we study the distribution of squid APP in extruded axoplasm as well as in a well-characterized reconstituted organelle/microtubule preparation from the squid giant axon in which organelles bind microtubules and move towards the microtubule plus-ends. We find that APP associates with microtubules by confocal microscopy and co-purifies with KI-washed axoplasmic organelles by sucrose density gradient fractionation. By electron microscopy, APP clusters at a single focal point on the surfaces of organelles and localizes to the organelle/microtubule interface. In addition, the association of APP-organelles with microtubules is an ATP dependent process suggesting that the APP-organelles contain a microtubule-based motor protein. Although a direct kinesin/APP association remains controversial, the distribution of APP at the organelle/microtubule interface strongly suggests that APP-organelles have an orientation and that APP like the Alzheimer's protein tau has a microtubule-based function. © 2016 Stevenson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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