PVR is a novel way to present volumetric data and may enable noninvasive diagnostic endoscopy and provide an alternate method to analyze volumetric imaging data for primary diagnosis.

Neurites of superior cervical ganglion neurons from embryonic, perinatal, and adult rats extended at different rates when placed in tissue culture on similar collagen substrata. Using high resolution cinematography and a time-lapse video recording system, we concluded that these differences arise from variations in individual growth cone behavior. Growth cones of embryonic and perinatal neuronal origin exhibited high peak rates of advance and filopodial and lamellipodial excresences. Perinatal cones differed from embryonic ones in that they were somewhat larger, advanced in straighter paths, and retracted less, consequently translocating at 14 to 29 microns/hr compared with 8 to 22 microns/hr for embryonic cones (ranges of 4-hr means). The growth cones of neurons obtained from adult rats had scant cytoplasm and short branched filopodia, lacked definitive lamellipodia, and traversed the terrain at 4 to 13 microns/hr due to lack of high peak rates of advance and more time spent in stationary or minimal advance phases. Periodic pauses lasting 10 to 20 min, occurring every 20 to 90 min, interrupted the forward advance of growth cones of all ages. During pauses or slow forward movement, the growth cone displayed numerous filopodia whereas, during brief episodes when embryonic and perinatal growth cones moved at peak rates of 200 microns/hr or more, the cone periphery was predominantly lamellipodial. We conclude that the predominance of a lamellipodial or filopodial conformation correlates with the rate of growth cone advance and that age-dependent variations in neurite extension rates are related to differences in growth cone form and pattern of translocation. This is the first documentation of differing behavior of single growth cones of neurons of varying developmental ages in culture.

No abstract

The extension of filopodia from growth cones of regenerating neurites from rat superior cervical ganglion neurons in tissue culture was studied. Cultures were grown on a thin layer of fibrous collagen and maintained in a medium containing serum and nerve growth factor. Time-lapse cinematography and computer-assisted morphometry were used to observe and measure the kinetics of extension of individual filopodia. Filopodia extended from the growth cone margin, trailing neurite, or from each other. Frequently, extension was preceded by the appearance at the cone margin of a nodule of cytoplasm which appeared dense in phase-contrast optics. Branch points between adjacent extending filopodia remained fixed with respect to the growth cone while the filopodia lengthened. The rate of extension was maximum just after initiation (0.12 +/- 0.4 micron/sec; mean +/- SD; n = 36) and declined thereafter until the filopodium collapsed. This initial rate of extension was directly correlated with the eventual length of the filopodium (r = 0.67). Filopodia of growth cones arising from embryonic neurons exhibited higher initial extension rates (range: 0.07 to 0.20 micron/sec; mean = 0.13 micron/sec) than those of postnatal neurons (range: 0.01 to 0.13 micron/sec; mean = 0.09 micron/sec). These data are discussed in relation to a model proposed by Tilney and Inoue [1982] for the extension, by distal addition of G-actin to growing filaments, of another type of elongating process filled with microfilaments, the acrosomal process of Thyone sperm.

Long term (2- to 3-week) cultures of superior cervical ganglia (SCG) were established from rats and rat embryos ranging in age from 15 days of gestation (E15) to 279 days postnatal (P279). Cultures were grown on a collagen substratum and fed a serum-containing medium with added nerve growth factor. Radial outgrowth of neurites was measured as a function of time for up to 2 to 3 weeks. Computer-aided analysis generated estimates of onset time, initial rate, and subsequent changes in the rate of growth of these neurites. The explants from perinatal rats showed the fastest growth onset time (5 to 13 hr), fastest initial rate of growth (370 to 660 microns/d), and a decline in growth rate during the first 2 weeks in culture. The outgrowth from these perinatal explants was composed of many small fascicles. Neurites from the prenatal explants (E15 to E20) began to grow within 22 hr in vitro. Their rate of growth was lower initially (150 to 300 microns/d) but increased to equal the perinatal explant initial rate before again falling to an intermediate level (200 to 300 microns/d). The outgrowth from prenatal explants contained fewer larger fascicles. Postnatal explants had low initial rates of growth (70 to 176 microns/d) but exhibited an increasing growth rate in vitro, again approaching an intermediate rate of 200 to 250 microns/d after 2 to 3 weeks. Neurite outgrowth from the postnatal explants was delayed by an amount roughly correlated with the age of the animal advancing postnatal age but reached an asymptote of about 50 to 150 microns/d at about P30. The outgrowth was initially sparse but became denser with time in culture. Thus, in a culture system in which medium composition and growth substratum are held constant, marked differences can be observed in pattern, latency, initial rate, and subsequent changes in rate of neurite extension among SCG explants from different ages of rats and rat embryos.