Growth of segmental nerves to the developing rat diaphragm: absence of pioneer axons

A study has been made of the growth of cervical nerves C3-C6 to the rat diaphragm. At 11 days of embryonic age these cervical nerves first project out of the spinal cord toward the cardinal veins and later form the left and right phrenic nerve trunks. During the next 2 days, the phrenic nerves grow caudally in close association with the cardinal veins toward the diaphragm. At the growing tips of these nerve trunks the growth cones of axons were observed every 1-2 micrometers. The last axon did not project more than 2 micrometers ahead of any neighbouring axons. At 14 days the phrenic nerves reach the level of the developing diaphragm and converge into pools of premuscle cells. Previous studies have suggested that the phrenic nerve enters the premuscle masses of the diaphragm at an early developmental stage when the premuscle masses are at approximately the segmental levels C3-C6. This study shows that the phrenic nerves must grow to more caudal levels in order to reach the premuscle cells of the diaphragm. Furthermore, the leading axons of the phrenic nerve trunk do not project in a pioneering fashion, far in advance of the trailing axons.