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Planar Spindle Orientation and Asymmetric Cytokinesis in the Mouse Small Intestine

Elizabeth S. Fleming, Mark Zajac, Darcy M. Moschenross, David C. Montrose, Daniel W. Rosenberg, Ann E. Cowan and Jennifer S. Tirnauer

Center for Molecular Medicine (ESF,DMM,DCM,DWR,JST), Colon Cancer Prevention Program of the Neag Comprehensive Cancer Center (DWR,JST), and Center for Cell Analysis and Modeling (MZ,AEC), University of Connecticut Health Center, Farmington, Connecticut

Correspondence to: Jennifer S. Tirnauer, Center for Molecular Medicine, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030-3101. E-mail: tirnauer{at}uchc.edu

A major feature of epithelial cell polarity is regulated positioning of the mitotic spindle within the cell. Spindles in cells of symmetrically expanding tissues are predicted to align parallel to the tissue plane. Direct measurement of this alignment has been difficult in mammalian tissues. Here, we analyzed the position of spindles in intact mouse intestinal epithelium using microtubule immunofluorescence and three-dimensional confocal imaging. Mitotic cells were identified in the proliferative zone of intestinal crypts. Spindle angle relative to the apical cell surface was determined either by direct measurement from confocal images or with a computational algorithm. Angles averaged within 10° of parallel to the apical surface in metaphase and anaphase cells, consistent with robust planar spindle positioning, whereas spindles in prometaphase cells showed much greater angle variability. Interestingly, cytokinetic furrows appeared to extend from the basal cell surface toward the apical surface. This type of image analysis may be useful for studying the regulation of spindle position during tissue remodeling and tumor formation. (J Histochem Cytochem 55:1173–1180, 2007)

Key Words: spindle orientation • planar cell polarity • cytokinesis • three-dimensional tissue • mitosis

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