A quantitative analysis of frog optic nerve regeneration: is retrograde ganglion cell death or collateral axonal loss related to selective reinnervation?

The present study was designed to assess whether axon collateral formation and loss or retrograde cell death contribute to selective reinnervation during optic nerve regeneration in the frog, Rana pipiens. The right optic nerve was crushed in 18 frogs, and samples were taken near the optic disc (retinal segment) and near the optic chiasm (brain segment). These samples were studied quantitatively with the electron microscope at various postoperative survival times (1, 2, 3, 4, 6, 12 weeks, 6 months, 1 year; N = 2). The number and size of axons in each segment were estimated from a series of electron micrographs taken at intervals across the transverse extent of each nerve and compared with normal nerves (N = 4). Results show that there are 5.3 +/- 1.8 X 10(5) (S.D.) unmyelinated and 2.3 +/- .5 X 10(4) myelinated axons in the normal nerve. One week post-crush (p.c.) there is a 27% decrease in the number of axons in the retinal segment (4.1 +/- 1.4 X 10(5)), indicating early retrograde axonal loss. As expected, there is a greater loss of axons at this time in the brain segment (3.0 +/- 1.3 X 10(5)). Between 2 and 6 weeks p.c. the number of axons increases in the retinal segment to over twice the normal number of axons increases in the retinal segment to over twice the normal number (12.3 +/- 3.8 X 10(5)) and to over four times this number in the brain segment (20.0 +/- 3.0 X 10(5)), showing collateral axon formation results from this injury. A large loss in the number of axons occurs in both nerve segments between 6 and 12 weeks p.c. (4.3 +/- 1.5 X 10(5)) and an additional loss at 20 weeks p.c. (2.2 +/- .98 X 10(5)). Subsequently, the number remains constant, approximately 40% of normal. Visual recovery was seen in the two frogs tested one year after optic nerve crush that were used for optic axon counts. Autoradiography in these same animals showed the optic nerve projections normally seen after regeneration. Besides axonal loss, our results also indicate that the size of both myelinated and unmyelinated axons is significantly above normal at chronic postoperative periods. This increase in axonal size is interpreted to be related to the increased territory each remaining optic axon must fill to restore the optic projections.(ABSTRACT TRUNCATED AT 400 WORDS)