Ultrastructural and histochemical observations in the developing iris musculature in the chick.

Irises from chick embryos and from 2- and 3-week-old chicks were studied using ultrastructural and histochemical methods in order to clarify the relationship between cell loss in the ciliary ganglion and the establishment of permanent peripheral connections between the ciliary neurons and the iris muscle. The iris muscle undergoes morphological and biochemical differentiation between 11 and 13 days of incubation. This period coincides with the critical period in the development of the ciliary ganglion when massive cell degeneration occurs. During this period, the iris develops typical sarcomeric structure, with AChE activity in the nuclear envelope, Golgi, and the "T' system. At 15 days of incubation AChE activity is found localized in discrete areas on the muscle fiber, forming specific neuromuscular junctions. Between 13 and 15 days of incubation, there is a shift in the localization of AChE activity in the iris muscle, from the sarcoplasmic structures to the junctional membranes. Few synaptic terminals are observed in the iris musculature prior to 11 days of incubation. There is a marked increase in the number of synaptic terminals between 11 and 13 days of incubation which also coincides with the period of cell loss in the ciliary ganglion. The establishment of neuromuscular junctions at 15 days of incubation corresponds with the period when the number of neurons in the ciliary ganglion has attained the adult level. The time table of the events described above, leads us to conclude that during development only those neurons in the ciliary ganglion which make peripheral contacts survive, and only such contacts differentiate into mature neuromuscular junctions on the iris muscle. This will imply that neurons which are doomed to die, although they may send out fibers to the periphery, do not make peripheral contacts before death.