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Short-term molecular polarization of cells on symmetric and asymmetric micropatterns.

ABSTRACT: The ability of cells to sense geometrical/physical constraints of local environment is important for cell movements during development, immune surveillance, and in cancer invasion. In this paper, we quantify "front-rear" polarization - the crucial step in initiating cell migration - based on cytoskeleton and substrate adhesion anisotropy in micropatterned cells of well-defined shapes. We then show that the general viewpoint that asymmetric cell shape is one of the defining characteristics of polarized cells is incomplete. Specifically, we demonstrate that cells on circular micropatterned islands can exhibit asymmetric distribution of both filamentous actin (f-actin) and focal adhesions (FAs) as well as directional, lamellipodial-like ruffling activity. This asymmetry, however, is transient and persists only for the period of several hours during which actin filaments and adhesion structures reorganize into symmetric peripheral arrangement. Cells on asymmetric tear-drop shape islands also display polarized f-actin and FAs, but polarization axes are oriented towards the wide end of the islands. Polarization of actin filaments on tear-drop islands is short-term, while focal adhesions remain asymmetrically distributed for long times. From a practical perspective, circular cells constitute a convenient experimental system, in which phenomena related to cell polarization are "decoupled" from the effects of cells' local curvature (constant along circular cell's perimeter), while asymmetric (tear-drop) micropatterned cells standardize the organization of motility machinery of polarized/ moving cells. Both systems may prove useful for the design of diagnostic tools with which to probe and quantify ex vivo the motility/invasiveness status of cells from cancer patients.

SUBMITTER: Kandere-Grzybowska K

PROVIDER: S-EPMC3697907 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature

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Short-term molecular polarization of cells on symmetric and asymmetric micropatterns.

Kandere-Grzybowska Kristiana K Soh Siowling S Mahmud Goher G Komarova Yulia Y Pilans Didzis D Grzybowski Bartosz A BA

Soft matter 20100701 14

The ability of cells to sense geometrical/physical constraints of local environment is important for cell movements during development, immune surveillance, and in cancer invasion. In this paper, we quantify "front-rear" polarization - the crucial step in initiating cell migration - based on cytoskeleton and substrate adhesion anisotropy in micropatterned cells of well-defined shapes. We then show that the general viewpoint that asymmetric cell shape is one of the defining characteristics of pol ...[more]

PMID: 23826026

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Project description:While the epithelial cell cortex displays profound asymmetries in protein distribution and morphology along the apico-basal axis, the extent to which the cytoplasm is similarly polarized within epithelial cells remains relatively unexplored. We show that cytoplasmic organelles within C. elegans embryonic intestinal cells develop extensive apico-basal polarity at the time they establish cortical asymmetry. Nuclei and conventional endosomes, including early endosomes, late endosomes, and lysosomes, become polarized apically. Lysosome-related gut granules, yolk platelets, and lipid droplets become basally enriched. Removal of par-3 activity does not disrupt organelle positioning, indicating that cytoplasmic apico-basal asymmetry is independent of the PAR polarity pathway. Blocking the apical migration of nuclei leads to the apical positioning of gut granules and yolk platelets, whereas the asymmetric localization of conventional endosomes and lipid droplets is unaltered. This suggests that nuclear positioning organizes some, but not all, cytoplasmic asymmetries in this cell type. We show that gut granules become apically enriched when WHT-2 and WHT-7 function is disrupted, identifying a novel role for ABCG transporters in gut granule positioning during epithelial polarization. Analysis of WHT-2 and WHT-7 ATPase mutants is consistent with a WHT-2/WHT-7 heterodimer acting as a transporter in gut granule positioning. In wht-2(-) mutants, the polarized distribution of other organelles is not altered and gut granules do not take on characteristics of conventional endosomes that could have explained their apical mispositioning. During epithelial polarization wht-2(-) gut granules exhibit a loss of the Rab32/38 family member GLO-1 and ectopic expression of GLO-1 is sufficient to rescue the basal positioning of wht-2(-) and wht-7(-) gut granules. Furthermore, depletion of GLO-1 causes the mislocalization of the endolysosomal RAB-7 to gut granules and RAB-7 drives the apical mispositioning of gut granules when GLO-1, WHT-2, or WHT-7 function is disrupted. We suggest that ABC transporters residing on gut granules can regulate Rab dynamics to control organelle positioning during epithelial polarization.

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Short-term molecular polarization of cells on symmetric and asymmetric micropatterns.

Publications

Short-term molecular polarization of cells on symmetric and asymmetric micropatterns.

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