Microtubules of axons of crayfish nerve cord normally have 12 wall protofilaments and microtubules of surrounding glial cells have 13 protofilaments. Tubulin was isolated from such nerve cords and polymerized in vitro; tannic acid staining of sedimented microtubules showed that microtubules with 12 and 13 wall protofilaments were present, suggesting that the ability to form a microtubule with a given number of protofilaments is inherent in the tubulin or its associated proteins. The temperature of polymerization was found to influence the number of protofilaments in resultant microtubules. With assembly at 20 degrees C, for example, most of the complete microtubules had 13 protofilaments, while at 40 degrees C most showed 10 protofilaments. It is suggested that the temperature of in vitro polymerization, among other factors, can influence the angle of binding between adjacent protofilaments and thus alter the number of protofilaments required to complete the circumference of the tubule.