The early development of major projections from caudal levels of the spinal cord to the brainstem and cerebellum in the gray short-tailed Brazilian opossum, Monodelphis domestica.

The Brazilian short-tailed opossum, Monodelphis domestica, is born 14-15 days after copulation and is available for experimentation at stages of development corresponding to those which occur in utero in placental mammals. In the present study, we took advantage of the opossum's embryology to study the development of projections from caudal levels of the spinal cord to the brainstem and cerebellum using axonal tracing methods. In all cases, a 2-3 day survival time was used for axonal transport. When injections of Fast blue (FB) were made into caudal levels of the thoracic cord at postnatal day (PD) 1 or 2, axonal labeling could not be identified at supraspinal levels. When injections were made at PD3, however, labeled axons were found in the fasciculus gracilis at caudal medullary levels, within the ventrolateral medulla and pons, within an incipient inferior cerebellar peduncle, and within the cerebellar anlage. The dorsal root origin of at least some of the axons within the fasciculus gracilis was evidenced by the transganglionic transport of cholera toxin conjugated to horseradish peroxidase from the hindlimbs. After FB injections at PD7, a few labeled axons could be traced from the fasciculus gracilis into the nucleus gracilis and from the ventrolateral pathway to the inferior olive. Generally comparable results were obtained using wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). In cases injected with FB at PD9, the pattern of brainstem labeling was adult-like. Although labeled axons were present within the cerebellum of animals injected with FB on PD3, they were limited to the marginal zone. Axonal labeling was present within an identifiable internal granular layer in cases injected with either FB or WGA-HRP at PD16, and it appeared to be limited to specific bands which foreshadowed those seen at later stages of development and in the adult animal. In some cases, labeled axons were present within the molecular layer where they were not seen in the adult animal. Our results provide a timetable for the normal development of projections from caudal levels of the spinal cord to the brainstem and cerebellum in Monodelphis and show that such development occurs postnatally rather than prenatally, as in placental mammals.

The origins of supraspinal projections to the cervical and lumbar spinal cord at different stages of development in the gray short-tailed Brazilian opossum, Monodelphis domestica.

We have used the retrograde transport of Fast blue (FB) to study the origins of supraspinal projections to the lumbar and cervical spinal cord at different stages of development in the Brazilian, short-tailed opossum, Monodelphis domestica. Monodelphis was chosen for study because its young are born in a very immature state, 14-15 days after copulation, making it possible to manipulate its nervous system in an embryonic state without intra-uterine surgery. When injections of FB were made into the lumbar cord at postnatal day (PD) 1, neurons were labeled within several areas of the reticular formation (the retroambiguus nucleus, the ventral and dorsal reticular nuclei of the medulla, the gigantocellular reticular nucleus, the lateral paragigantocellular reticular nucleus, and the pontine reticular nucleus), the presumptive coeruleus complex, and the lateral vestibular nucleus. In many cases, labeled neurons were also found within the caudal raphe and the presumptive interstitial nucleus of the medial longitudinal fasciculus. The results of immunocytochemical studies provided evidence for catecholaminergic and serotoninergic neurons in the brainstem at PD1 and for axons of both phenotypes in the spinal cord. By PD3, labeled neurons were found within the ventral gigantocellular and ventral pontine nuclei of the reticular formation, the spinal trigeminal nucleus, and the presumptive paraventricular nucleus of the hypothalamus. When injections were made at PD4, neurons were also labeled within the medial and inferior vestibular nuclei, the red nucleus, the mesencephalic nucleus of the trigeminal nerve, the presumptive nucleus of Edinger-Westphal and the lateral hypothalamus. By at least PD7, the pattern of supraspinal labeling was similar to that obtained at older ages and in the adult animal. When FB was injected into the cervical cord at PD1, neurons were labeled in all of the areas labeled by lumbar injections at the same age and in larger numbers. In addition, labeled neurons were found within the ventral gigantocellular and spinal trigeminal nuclei. When cervical injections were made at PD15, labeled neurons were found within the deep cerebellar nuclei and amygdala and by PD17 they were also present within the superior colliculus and cerebral cortex. In some cases, cortical labeling was present outside the areas labeled by comparable injections in adult animals.(ABSTRACT TRUNCATED AT 400 WORDS)