Uniform apicobasal polarity of microtubules and apical location of gamma-tubulin in polarized intestinal epithelium in situ

Polarized differentiation of the intestinal epithelium has been previously shown to depend on an intact microtubular system that is essential for vectorial delivery of apical membrane proteins to the apical cell surfaces. Uniform alignment and polarization of microtubules have been suggested to provide the ultrastructural basis for vectorial transport between the Golgi apparatus and the apical cell surface. In the present study we applied the hook decoration technique to analyse the polarity of microtubules in the rat jejunal epithelium. By immunocytochemistry we studied the subcellular location of gamma-tubulin, an essential component of microtubule-organizing centers. Microtubules were found to be mainly aligned parallel to the apicobasal axis of the cells and to extend from the subterminal space underneath the apical terminal web down to the cellular basis. We found that 98% out of 1122 decorated microtubules displayed uniform apicobasal polarity with the growing ends (plus ends) pointing basally and the non-growing ends (minus ends) pointing towards the cellular apex. No differences were observed with respect to microtubular polarity between the apical, perinuclear and infranuclear cellular portions. Immunostaining specific for gamma-tubulin was restricted to the apical subterminal space underneath the rootlets of microvilli. These findings indicate that the apical subterminal space of enterocytes serves as the predominant if not exclusive microtubule-organizing compartment from which uniformly polarized microtubules grow out with their plus ends towards the cellular basis.