Observations on the development of brainstem-spinal systems in the North American oppossum.

The North American oppossum is born 12 to 13 days after conception and and is available for 90 days or more in an external pouch where it can be observed and experimentally manipulated. It is of particular interest that the hindlimbs of the newborn opossum are very immature and remain immobile for a week or more after birth. Degeneration techniques reveal that immature brainstem axons are present within the marginal zone of the lumbosacral cord before hindlimb movements begin (our state I) and material processed for formaldehyde induced fluorescence shows that some of them transport monoamines. Several lines of evidence suggest that part of the fluorescent axons arise within the nucleus locus coeruleus. At this early stage the electron microscope reveals that all brainstem-spinal axons are small (0.1--0.4 micrometer in diameter) and unmyelinated. By the time random hindlimb movements can be observed (stage II), brainstem axons, including those transporting monoamines, can be demonstrated to have grown into limited areas of the intermediate zone of the lumbosacral cord and to arise from most of the areas contributing to them in the adult animal (horseradish peroxidase technique). Such axons are still immature and it is not yet clear that they have formed synaptic terminals. Brainstem axons continue to grow into the intermediate zone of the lumbosacral cord for some time and come to occupy all of their adult territories before thoracic transection produces obvious change in hindlimb motility (beginning of stage III). It is still another 20 days or so before thoracic transection produces spinal shock comparable to that in the adult animal. The relatively mature use of the hindlimbs and the full expression of spinal shock correlate with changes in the technique and survival time needed to demonstrate degenerating brainstem axons in experimental material.

Synaptogenesis in the ventromedial hypothalamus of the prenatal mouse.

Mouse embryos from day 13 to day 19 of gestation (E13 through E19) were removed by Caesarean section and their brains were prepared for electron microscopy. Coronal sections were examined in three planes through the ventromedial hypothalamus: anterior, the level at which the optic tracts pass posterolaterally to become partially enclosed in the major brain mass; middle, the level at which the floor of the third ventricle begins to widen and flatten; posterior, the level at which the most anterior infundibulum appears. Three types of junctional complexes were examined. Close junctions are identified as straight, parallel areas of two apposed membranes which appear more electron-dense than immediately adjacent regions. The membranes are separated by a clearly visible cleft. Unlike synapses, no clear synaptic vesicles are found in either of the adjacent profiles unless randomly distributed and accompanied by ribosomes or glycogen. Close junctions are seen most frequently on day E15, then decrease in number through E19. Their participation in synaptogenesis is discussed from temporal, morphological, distributional and quantitative perspectives and is provisionally rejected. Immature synapses show only the minimal membrane specialization found in close junctions, but vesicles are present, ribosomes absent in at least one of the adjacent cytoplasms. Their appearance peaks on E17-E18, paralleling and slightly preceding that of the mature synapses. Their evolution from close junctions has only weak temporal support. Mature synapses display the cytoplasmic densities which immature synapses lack. They attain their greatest prenatal numbers on E18, then decrease in number on E19. The conclusion is advanced that synaptogenesis does--or at least can--occur without the prior appearance of avesicular regions of increased membrane density.