Microtubules in Xenopus oocytes are oriented with their minus-ends towards the cortex

Despite lacking centrosomes, stage VI Xenopus oocytes contain extensive networks of cytoplasmic microtubules (MTs). To gain additional insight into the factors regulating MT organization during oogenesis, we have used electron microscopy and "hook decoration" to examine the distribution and orientation of MTs in Xenopus oocytes. A limited survey of two "undecorated" stage VI oocytes revealed 218 MTs in images covering approximately 2,500 microm(2), and indicated that the MT number density of the animal cytoplasm was greater than that of the vegetal cytoplasm. Examination of five "decorated" stage VI oocytes (three animal and five vegetal hemispheres) revealed 653 MTs. Of these, 76% could be scored as having exclusively counterclockwise (CCW) or clockwise (CW) hooks. In the animal hemispheres, 93% of the scored MTs exhibited CCW hooks when viewed from the direction of the cortex, indicating that most MTs were oriented with their minus-ends out. MT orientation appeared relatively uniform throughout the animal cytoplasm: more than 90% of the scored MTs in the cortical (90%), subcortical (96%), or perinuclear (98%) cytoplasm were oriented with their minus-ends out. In the vegetal hemispheres, approximately 80% of the scored MTs exhibited CCW hooks, and thus were oriented with their minus-ends out; 96% of the scored MTs in stage III oocytes were oriented minus-end out. These observations support a model in which the cortex plays a significant role in MT nucleation and organization in Xenopus oocytes, and have significant implications for the MT-dependent transport and localization of cytoplasmic organelles and RNAs during oogenesis.