Caudal Diencephalon Research Articles

Mesodiencephalic projections to the vestibular complex in the cat.

The distribution of cells in the rostral medial mesencephalon and caudal diencephalon which project to the vestibular complex was mapped in the cat by using retrograde axonal transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Subsequent experiments using anterograde transport of WGA-HRP clarified the position of the terminations of the mesodiencephalic-derived afferents in the vestibular complex. After large injections which involved the entire vestibular complex, retrogradely labeled cells were seen in both the ipsilateral and contralateral interstitial nucleus of Cajal (INC) and were more numerous in its rostral pole. Labeled cells also occurred in the perifascicular region, both immediately adjacent to the fasciculus retroflexus and rostroventral to it. Fusiform midline cells of the Edinger-Westphal nucleus were also labeled, as well as a number of cells in the adjacent somatic portion of the oculomotor complex (OMC). Another group of labeled cells was observed within the contralateral medial terminal nucleus of the accessory optic tract (MTN) and in the posterior hypothalamic nucleus. Injections limited to subregions of the vestibular complex resulted in similar but slightly varying distributions and numbers of retrogradely labeled cells. After injections covering the caudal half of the medial vestibular nucleus (MVN) and descending vestibular nucleus (DVN), labeled cells in the INC and tegmentum dorsal to it were especially prominent, but none was seen in the MTN or OMC. Injections placed in the rostral MVN, lateral vestibular nucleus, y group, and superior vestibular nucleus resulted in a distribution of labeled cells similar to that seen following global vestibular injections, but these cells were fewer in number. After an injection confined to the y group, a small number of retrogradely labeled cells were seen in the rostral pole of the INC and immediately ventral to the fasciculus retroflexus. Projections from the rostral medial mesencephalon and caudal diencephalon to the MVN, DVN, and y group were confirmed by using anterograde transport of WGA-HRP. Direct projections from the INC-perifascicular regions and somatic neurons of the OMC to the caudal vestibular complex could play a role in eye-head coordination. Those projections from the rostral INC and MTN to the rostral vestibular complex may play a role in vertical eye movements and responses to visual stimuli which move in the vertical plane.

Comparative topography of projections from the mesodiencephalic junction to the inferior olive, vestibular nuclei, and upper cervical cord in the cat.

Distributions of neurons located in the central rostral mesencephalon and caudal diencephalon that project to the upper cervical spinal cord, vestibular nuclei, or inferior olive were studied in the cat by using retrograde axonal transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Afferent sources to all of these targets were observed in the interstitial nucleus of Cajal (INC), the region surrounding the fasciculus retroflexus (PF), and the nucleus of the fields of Forel (NFF). Three-dimensional reconstruction revealed differences in densities of cells projecting from these common areas. Spinal projecting cells were present in slightly greater numbers in the caudal two-thirds of the INC, whereas those projecting to the vestibular complex were more numerous in the rostral two-thirds of this nucleus. A relatively smaller number of olivary projecting cells were dispersed throughout the INC. Olivary afferent sources outnumber those with spinally directed or vestibularly directed axons in the PF region. In the fields of Forel, cells projecting to the vestibular nuclei or inferior olive were concentrated medially, whereas cells projecting to the spinal cord appeared both medially and laterally. Each type of afferent source was also seen in the nucleus of the posterior commissure and the posterior ventral lateral hypothalamic area. Unique sources of afferents to the inferior olive were observed in the parvicellular red nucleus (ipsilateral to the injections) and the anterior and posterior pretectal nuclei. A large number of labeled neurons was seen in the nucleus of Darkschewitsch after injections of tracer into the inferior olive, but this projection did not appear to be unique, as small numbers of labeled cells were also seen after injections into the cervical spinal cord. The Edinger-Westphal nucleus and the adjacent somatic oculomotor nucleus contained cells which projected separately to the spinal cord or the vestibular complex, and the superior colliculus contained cells which projected separately to the contralateral spinal cord or the contralateral inferior olive. In this study, it was also noted that neurons in the medial terminal nucleus of the accessory optic tract projected to the ipsilateral inferior olive or to the contralateral vestibular complex. These differences in locations and densities of cells projecting to the cervical spinal cord, vestibular complex, and inferior olive may underlie functional specializations in these areas in relation to vertical eye and head movement control and to neural systems controlling postural adjustments accompanying limb movements.

Projections of the olfactory bulb and nervus terminalis in the silver lamprey.

The connections of the olfactory bulb were traced using horseradish peroxidase. A homologue of the medial olfactory tract in gnathostomes projects to the ipsilateral septal nucleus, preoptic area and, possibly, the rostral striatum. A homologue of the lateral olfactory tract projects to the ipsilateral lateral pallium, dorsal pallium and, possibly, the medial pallium, as well as to the posterior diencephalon. A component of the lateral olfactory tract decussates in the habenular and posterior commissures and distributes to the contralateral hemisphere and caudal diencephalon. A dorsal component of secondary olfactory fibers terminates, ipsilaterally, in a dorsomedially situated neuropil that has previously been interpreted as a single glomerulus of the olfactory bulb or as an accessory olfactory bulb, as well as in the contralateral olfactory bulb after decussation in the dorsal commissure. Afferents to the olfactory bulb arise from the ipsilateral dorsal pallium, lateral pallium, a cell-poor region adjacent to the preoptic area, and the midbrain tegmentum. The extent of the secondary olfactory projections in silver lampreys could be interpreted to support the phylogenetic hypothesis that all regions of the telencephalon received secondary olfactory projections in the earliest vertebrates, but this interpretation is not unequivocal, due to questions concerning the pallial homologues in lampreys and gnathostomes. Application of horseradish peroxidase to the olfactory epithelium revealed projections to the striatum, preoptic area, hypothalamus and posterior tuberculum that are comparable to projections of the nervus terminalis in other vertebrates.

Localization of immunoreactive tyrosine hydroxylase in the goldfish brain

This report describes the distribution of tyrosine hydroxylase immunoreactive (TH-ir) structures in the brain of the goldfish (Carassius auratus). The localization of TH-ir cell groups revealed by immunocytochemical techniques is largely in accordance with catecholamine distribution previously reported in teleosts by using monoamine fluorescence; however, in the telencephalon and diencephalon, several new cell groups are elucidated. In the telencephalon, TH-ir cell bodies are observed in the olfactory bulb, area ventralis telencephali, and the central zone of the area dorsalis telencephali. TH-ir fibers and terminals are moderately dense throughout the telencephalon except for a sparse innervation of the area dorsalis, pars medialis. Immunostained cells are present in the suprachiasmatic nucleus and magnocellular and parvicellular components of the preoptic nucleus. Immunoreactive fibers from preoptic cells can be traced caudally in two main tracts to the infundibulum. Dense immunoreactivity around cells in the pituitary provides anatomical support for catecholamine involvement in the neuroendocrine axis probably via preopticohypophysial connections. At middiencephalic levels, immunoreactive cells are present in the ventral thalamus, nucleus pretectalis periventricularis, pars ventralis, and paraventricular organ pars anterioris. In the caudal diencephalon, TH-ir cells are seen within the posterior tuberal nuclei and dorsal to posterior recess. No immunostained cells are observed in the midbrain. In the hindbrain, tyrosine hydroxylase containing cells comprise three groups similar to that described using Falck-Hillarp histofluorescence (Parent et al., '78), i.e., isthmal, central medullary, and medullospinal groups. Tyrosine hydroxylase immunoreactivity is interpreted as evidence for the presence of catecholamines and not only provides an anatomical basis for the functional significance of catecholamines in teleosts, but may be useful in elucidating homologous structures in tetrapod vertebrates, although certain sites of immunoreactivity may prove to be unique to teleosts.

Cholinergic neurons of the laterodorsal tegmental nucleus: Efferent and afferent connections

The ascending projections of cholinergic neurons in the laterodorsal tegmental nucleus (TLD) were investigated in the rat by using Phaseolus vulgaris leucoagglutinin (PHA-L) and wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) anterograde tracing techniques. Two ascending pathways were identified after iontophoretic injections of PHA-L into the TLD. A long projection system courses through the dorsomedial tegmentum, caudal diencephalon, medial forebrain bundle, and diagonal band. Different branches of this system innervate the midbrain (superior colliculus, interstitial magnocellular nucleus of the posterior commissure, and anterior pretectal nucleus), the diencephalon (lateral habenular nucleus, parafascicular, anteroventral, anterodorsal, mediodorsal, and intralaminar thalamic nuclei), and the telencephalon (lateral septum and medial prefrontal cortex). The second system is shorter and more diffuse and innervates the median raphe, interpeduncular, and lateral mammillary nuclei. Retrograde tracing with WGA-HRP, combined with choline acetyltransferase immunohistochemistry, revealed that most of the TLD projections to the tectum, pretectum, thalamus, lateral septum, and medial prefrontal cortex are cholinergic. Afferents to the TLD were studied by anterograde and retrograde tracing techniques. Injection of tracers into the TLD retrogradely labelled neurons bilaterally in the midbrain reticular formation, the periaqueductal gray, the medial preoptic nucleus, the anterior hypothalamic nucleus, and the perifornical and lateral hypothalamic areas. Retrogradely labelled cells were also located bilaterally in the premammillary nucleus, paraventricular hypothalamic nucleus, zona incerta, and lateral habenular nucleus. In the telencephalon, the nucleus of the diagonal band and the medial prefrontal cortex contained retrogradely labelled neurons ipsilateral to the TLD injection site. The projections of the medial prefrontal cortex, the bed nucleus of the stria terminalis, and the lateral habenular nucleus to the TLD were confirmed in anterograde tracing studies. These findings indicate that the TLD gives rise to several ascending cholinergic projections that innervate diverse regions of the forebrain. Afferents to the TLD arise in hypothalamic and limbic forebrain regions, some of which appear to have reciprocal connections with the TLD. The latter include the lateral habenular nucleus and medial prefrontal cortex.

Cholecystokinin innervation of the ventral striatum: A morphological and radioimmunological study

Immunocytochemistry, radioimmunological assay after surgical cuts, anterograde degeneration and retrograde tracing of fluorescent dyes were used in order to elucidate the cholecystokinin-containing afferents to the ventral striatum (nucleus accumbens, olfactory tubercle and ventral part of the caudate-putamen). In agreement with the report by Hökfelt et al., 37 midbrain cholecystokinin-containing cells supply the posteromedial parts of the nucleus accumbens and olfactory tubercle, as well as the subcommissural part of caudate-putamen. Brainstem cholecystokinin afferents also reach more rostral parts of the ventral striatum including the rostrolateral olfactory tubercle. The ascending cholecystokinin axons enter the medial forebrain bundle at the meso-diencephalic border and maintain a rough medial to lateral topography at the caudal diencephalon. A second major cholecystokinin pathway, with possible origin in the piriform and medial prefrontal cortices and/or the amygdala, projects to the subcommissural caudate-putamen, the olfactory tubercle, the lateral part of the nucleus accumbens and the dorsal part of the bed nucleus of stria terminalis. Finally, the rostral part of the dorsal caudate-putamen receives a substantial cholecystokinin innervation from the basolateral amygdala and possibly from the neocortex. According to radioimmunological data, the descending telencephalic cholecystokinin system accounts for about 60% of all cholecystokinin in the rostral forebrain. The combined use of morphological and biochemical methods provided evidence for a partially overlapping distribution and possible interaction between an ascending brainstem and descending telencephalic cholecystokinin fiber systems within the striatum and related rostral forebrain areas.

A reevaluation of midbrain and diencephalic projections to the inferior olive in rat with particular reference to the rubro-olivary pathway.

Projections from the midbrain and caudal diencephalon to the inferior olivary nucleus (ION) in the rat were investigated by using anterograde and retrograde tracing techniques. Particular attention was directed to studying the projection from the red nucleus (RN) to the ION. Tritiated leucine was stereotaxically injected into the RN in six animals. The injection sites and the portion of the medulla containing the ION were processed for autoradiography. In three cases, the injections were largely confined to the RN. In these instances, no terminal labeling was noted in the ION. In the other three animals, the injections spread beyond the RN to include, in one or another of the rats, such areas as the mesencephalic reticular formation (MRF) between the RN and the periaqueductal gray (PAG), the region around the fasciculus retroflexus (FR), the prerubral area, the MRF dorsolateral to the RN, or the region dorsal to the medial geniculate body. In these animals, terminal labeling was observed ipsilaterally in the ION, chiefly in the caudal one-half of the medial accessory olive and in the dorsal lamella and lateral bend of the principal olive. Retrograde transport of lectin-conjugated horseradish peroxidase (WGA-HRP) or the fluorescent dye fast blue (FB), injected into the ION, was used to localize regions in the midbrain and caudal diencephalon containing neurons which project to the inferior olive. Following WGA-HRP injections into the ION, retrogradely labeled neurons were found in the ipsilateral ventral PAG, the MRF between the PAG and the RN, the MRF dorsolateral to the RN, and the area dorsal to the medial geniculate body. Large numbers of heavily labeled neurons were found in the area surrounding the FR. No labeled cells were noted in the RN or the nucleus of Darkschewitsch (ND). Subsequent to FB injections into the ION, fluorescent neurons were observed in the same regions which contained HRP-labeled neurons in the experiments using the enzyme as a tracer. Additionally, very small numbers of FB-labeled neurons were found in the RN and the ND. These results indicate that, in the rat, descending input from the midbrain and caudal diencephalon to the ION arises chiefly from a medially located column of neurons extending from the level of the rostral RN to the region of the FR. Little, if any, input to the ION is derived from the ND or RN in this species, unlike the cat, in which a substantial projection to the ION arises from the ND and the RN.

Stereotyped and variable growth of redirected Mauthner axons

To assay the axon tract organizing capabilities of different regions of the vertebrate CNS, Mauthner axons were redirected by grafting supernumerary hindbrains in Xenopus embryos. The 63 redirected Mauthner axons thus produced included donor axons projecting into the host CNS and host axons that grew through the graft or that were redirected in the host CNS. Two major phenomena were observed. Caudal to the optic chiasm, the Mauthner axons followed a single ipsilateral stereotyped route—the basal substrate pathway—extending in the ventral and ventrolateral marginal zone from the diencephalon to the caudal spinal cord. In contrast, rostral to the optic chiasm, these same Mauthner axons followed variable ipsilateral and contralateral routes. Even pairs of Mauthner axons entering the optic chiasm side-by-side eventually followed different routes in normal forebrains. The contrasting behaviors of the Mauthner axons growing in the rostral diencephalon and telencephalon and of the same Mauthner axons growing elsewhere suggest that there are differences in the effective guidance cues between these two regions of the developing brain. This is consistent with other types of neuroanatomical and neuroembryological evidence indicating a fundamental division between the rostral and the caudal diencephalon.

The interrelations between cell groups in the caudal diencephalon of the rat projecting to the striatum and to the medulla oblongata.

In order to investigate the topographical relationships between the caudal diencephalic cells of origin of ascending and descending projections in the rat, one fluorescent retrograde tracer was injected into the striatum and another into the medulla oblongata. The medullary injections were mainly centered in the inferior olive. Cells labeled from the striatal injections densely filled the thalamic parafascicular nucleus. Cells labeled from the medullary injections were seen ventrally to the fasciculus retroflexus in the subparafascicular nucleus. The two populations were mixed in a small area at the ventromedial border of the fasciculus retroflexus. No double labeled cells were observed. The present results indicate that caudal diencephalic cells which ascend to the striatum are different from those descending to the medulla oblongata and that they partially overlap.

Nucleus rotundus in a snake, Thamnophis sirtalis: an analysis of a nonretinotopic projection.

Nucleus rotundus, a tectorecipient thalamic nucleus in reptiles and birds, is described for the first time in a snake. The morphology of rotundal neurons and tectorotundal axons was studied at the light microscopic level by using anterograde and retrograde filling with the horseradish peroxidase (HRP). Injections of HRP in the dorsal ventricular ridge retrogradely fill neurons in rotundus. Rotundus is situated centrally in the caudal diencephalon medial to the cell plate of the retinorecipient geniculate complex and ventrolateral to the lentiform thalamic nucleus. The dendrites of rotundal neurons are long and radiate, but are confined within the cytoarchitectonically defined borders of the nucleus. Injections of HRP into the optic tectum anterogradely fill axons that project to rotundus bilaterally via the tectothalamic tract. Small injections show that axons arising from a single tectal locus distribute to all sectors of rotundus. Thus, this projection may not be retinotopically organized. However, single axons reconstructed through serial sections form spatially restricted, sheetlike terminal fields that pass caudorostrally through the entire extent of rotundus. Several hypotheses on the functional significance of such organized but nonretinotopic visual projections are discussed.

Development of the serotonergic system in the rat embryo: An immunocytochemical study

The development of central serotonergic neurons has been examined immunocytochemically utilizing an antiserum to serotonin (5-HT). Cells of the B4–B9 complex are first detected early on embryonic day 13 (E13; 7 mm crown rump length, CRL) and increase rapidly in number through E15 when they appear as bilateral columns situated from just caudal to the mesencephalic flexure to the pontine flexure. Aggregation of cells into subgroups is apparent soon after 5-HT neurons leave the ventricular zone, allowing the identification of certain subdivisions of the B4–B9 complex long before they assume their adult locations. The initial detection of 5-HT immunoreactive cells in the medulla occurs 1–2 days after the appearance of cells in the B4–B9 complex, although it has been reported that the time of origin of medullary raphe neurons (B1–B3) occurs before that of raphe neurons in the midbrain and pons (B4–B9). The first medullary 5-HT neurons, comprising the B3 subdivision occur ventro-laterally on E14 (10–11 mm CRL) at least 1–2 days before midline 5-HT neurons are visualized in the B1 and B2 groups. Thus, in contrast to cells in the B4–B9 complex, medullary 5-HT neurons complete much of their migration before they can be detected immunocytochemically, indicating that the time of onset of transmitter synthesis and storage may differ during differentiation of cells sharing a common neurotransmitter phenotype. The formation of ascending 5-HT fiber projections occurs rapidly from cells of the B4–B9 complex. Within 24 hours after the initial detection of 5-HT fiber immunoreactivity in such cells at E13, their axons are seen entering the caudal diencephalon (E14). These fibers have traversed the diencephalon and floor of the telencephalon by E15–E16 and reach the frontal neocortical pole by E17. The main ascending bundle of 5-HT axons courses through the diencephalon in the vicinity of the medial forebrain bundle, although some fibers also diverge and travel along certain pre-existing non-5HT pathways. However, examples are also found of acute directional changes in 5-HT fiber growth which do not appear to be associated with pre-formed non-5HT pathways. The pattern of ascending fiber outgrowth suggests a priority routing system which provides certain regions with 5-HT axons in a preferential sequence irrespective of the distance of these areas from 5-HT cell groups or from major bundles of ascending 5-HT fibers.

The afferent connections of the inferior olivary complex in rats: a study using the retrograde transport of horseradish peroxidase.

AbstractRetrograde transport of horseradish peroxidase (HRP) was used to define the origin of afferents to the inferior olivary complex (IOC) in rats. Using both ventral and dorsal surgical approaches to the brainstem, HRP was injected into the IOC through a micropipette affixed to the tip of a 1‐μl Hamilton syringe. After a 2‐day postoperative survival, animals were sacrificed by transcardiac perfusion with a 1% paraformaldehyde‐1.25% gluteraldehyde solution, and brains were processed according to the DeOlmos protocol (1977), using o‐dianisidine as the chromogen.Labeled cells were found at many levels of the nervous system extending from lumbar spinal cord to cerebral cortex. This wide‐ranging input from numerous regions clearly underscores the complexity of the IOC and its apparent involvement in several functions.Within the spinal cord, labeled neurons were identified from cervical to lumbar but not at sacral levels. These neurons were found contralaterally in the neck region of the dorsal horn and in the medial portions of the intermediate gray.In the caudal brainstem, reactive cells in the dorsal column nuclei, the spinal trigeminal nucleus, and the subnucleus y of the vestibular complex were observed primarily contralateral to the injection sites. Labeling within the gigantocellular, magnocellular, ventral, and lateral reticular nuclei and the nucleus prepositus hypoglossi was primarily ipsilateral. Reactive neurons in the medial and inferior vestibular nuclei were predominantly ipsilateral or contralateral to HRP injections into the caudal or rostral IOC, respectively.The dentate and interposed nuclei of the cerebellum contained small, lightly labeled neurons primarily contralateral to the injection site, while the fastigial nuclei contained a few relatively large, heavily labeled cells bilateral to caudal olivary injections.Ipsilaterally labeled mesencephalic regions included the periaqueductal gray, interstitial nucleus of Cajal, rostromedial red nucleus, ventral tegmental area, medial terminal nucleus of the accessory optic tract, nucleus of the optic tract, and the lateral deep mesencephalic nucleus. The caudal part of the pretectum and small cells of the stratum profundum of the superior colliculus were labeled predominantly contralateral to the injection.In the caudal diencephalon labeled neurons were most numerous within the nucleus of Darkschewitsch and the subparafascicular nucleus, primarily ipsilateral to olivary injections. Scattered reactive neurons were also found within the ipsilateral zone incerta. With the exception of the zona incerta, all labeled mesencephalic and diencephalic nuclei had some bilateral representation of labeled cells.No labeled neurons were identified within the basal ganglia, while numerous reactive cells were found bilaterally within layer V of the frontal and parietal cerebral cortex.

Monoamine distribution in primate brain V. Monoaminergic nuclei: Anatomy, pathways and local organization

Monoamine distribution in primate brain V. Monoaminergic nuclei: Anatomy, pathways and local organization

Monoamine distribution in primate brain. V. Monoaminergic nuclei: Anatomy, pathways, and local organization

Monoamine distribution in primate brain. V. Monoaminergic nuclei: Anatomy, pathways, and local organization

Diencephalic projections from the pontine reticular formation: Autoradiographic studies in the cat

Ascending projections to the diencephalon from the pontine reticular formation were studied in the cat by autoradiographic techniques. Projections from both rostral and caudal pontine regions ascend to the caudal diencephalon and divide into two components; a dorsal leaf terminates primarily in the thalamic intralaminar complex and a ventral leaf terminates in the subthalamic region. The relative densities of the two terminal regions vary with the injection site. Fibers originating in the caudal pons (nucleus reticularis pontis caudalis) terminate relatively heavily in the intralaminar nuclei of the dorsal thalamus, particularly the centre median, central lateral, central dorsal and paracentral nuclei, and also the dorsal medial nucleus. Relatively sparse termination occurs in the subthalamic region. In contrast, fibers from the rostral pons (nucleus reticularis pontis oralis) terminate relatively heavily in the subthalamic region, including the zona incerta, the fields of Forel, the ventral part of the thalamic reticular complex, and the lateral hypothalamus. Relatively sparse termination occurs in the dorsal thalamus, but includes the centre median, parafascicular, central lateral, paracentral and dorsal medial nuclei. These data are discussed with regard to reticular control of forebrain activity and the role of the classic dorsal and ventral components of ascending reticular projections.

Axonal patterns and sites of termination of cat superior colliculus neurons projecting in the tecto-bulbo-spinal tract.

Horseradish peroxidase was injected in the somata or axons of neurons located in the intermediate and deep layers of the superior colliculus. A group of 34 neurons with physiologically identified projection in the predorsal bundle (tectobulbo-spinal neurons, TBSNs) and two commissural tecto-tectal neurons were characterized with regard to soma-dendritic profiles, axon trajectories, collateral branching, and terminations. TBSNs belong to the class of large, multipolar, wide field neurons. They send axons through the deep white layer without generating local collaterals. Prior to decussation, all TBSNs bifurcate into an ascending branch which reaches the caudal diencephalon, and a main axon descending to the medulla or spinal cord. Regularly spaced collaterals supply a variety of structures at all rostro-caudal levels. In the midbrain, preterminal and terminal ramifications are present in the medial and lateral reticular tegmentum, in the central grey (including its supraoculo-motor zone), in the nuclei of Cajal and Dark-schewitsch and in the medial aspects of the prerubral area and the fields of Forel. Rhombencephalic targets of TBSNs include the medial pontine and bulbar reticular formation, the abducens nucleus, the nucleus reticularis tegmenti pontis and the nucleus prepositus hypoglossi. An increased density of terminal ramifications was found in several brain stem regions related to the control of eye and head movements. The widespread connections of each individual TBSN suggest that neurons of this type may provide a spatio-temporal pattern of facilitation which promotes rapid orientation of eyes, head and body towards the contralateral hemifield but does not specify the details of movement to be executed.

Effect of age and enrichment on certain brain dimensions in Brattleboro rats deficient in vasopressin

As a sequel to our first report in the series of studies on the brains of Brattleboro rats, measurements in caudal diencephalon, subcortical telencephalon, hippocampus, and pyriform cortex were made in three groups each of male heterozygous and homozygous Brattleboro rats. One group raised in standard conditions was killed at 60 days of age, another raised in standard conditions was killed at 90 days of age, and a third group raised in standard conditions for 60 days was killed at 90 days of age after 30 days of enrichment. Comparing 60- and 90-day-old standard condition animals, the heterozygous rats showed a significant increase in height of the caudal diencephalon. Comparing 90-day-old enriched animals with the 90-day-old standard group, significant increases occurred in 8 of 12 comparisons. In comparing the two types of Brattleboro rats, heterozygous rats had significantly greater brain dimensions than homozygous rats in width of the diencephalon in the 60-day-old standard group, and in witth of the telencephalon in both the 90-day-old standard and enriched groups. Differences in brain measures between heterozygous and homozygous Brattleboro rats tended to increase with age. Enrichment appeared to prevent this age-related increase in the difference between brain dimensions in the two types of Brattleboro rat. The same pattern was reflected in body weight differences, and it may be due to a greater anabolic response to enrichment in homozygous rats than in heterozygous rats. We suggest that brain abnormalities associated with congenital absence of vasopressin increase with age, and may be ameliorated by repetitive arousal as occurs in enrichment.

Identification of early neurons in the brainstem and spinal cord. II. An autoradiographic study in the mouse.

AbstractEarly neurogenesis was studied in two series of mouse embryos with tritiated thymidine (3H‐Tdr) autoradiography to determine (1) the time of origin and (2) the identity of the first postmitotic neurons. Time of origin was studied in embryos cumulatively labeled on the 9th–11th day of gestation (E8, E9, or E10, since the day of finding the vaginal plug is EO) and terminated after one cell cycle (8 hours). Autoradiographs were examined for unlabeled neurons, those which had undergone final DNA synthesis and which were usually also postmitotic. A 16‐somite embryo, cumulatively labeled from the 6‐somite stage, demonstrated 234 such cells in both alar and basal plates from the caudal diencephalon to the cervical spinal cord. Some of these young neurons, therefore, arose prior to neural tube closure, which begins at the 7‐somite stage. Data from other embryos in this series suggests that the first neurons arise at the 1–2 somite stage.Early arising neurons were identified in embryos pulse labeled with 3H‐Tdr during E8–11 and killed after postnatal day (P) 30, when neuronal types could be determined. Autoradiographs were examined for heavily labeled cells, those in final DNA synthesis at the time of treatment. Heavily labeled cells in the youngest mouse in this series (injected at E8½; ca. 8‐somite stage) were primarily small‐to medium‐sized neurons of the brainstem reticular formation, but also some small neurons of the superior olivary nucleus and large primary sensory neurons of the mesencephalic nucleus of the trigeminal nerve. The earliest neurons in the spinal cord appeared early on E9, were medium sized, and were located in the ventral horn and the intermediate zone.Comparisons between the results of this study and those of McConnell and Sechrist ('80) in the chick suggest that the youngest stages of neurogenesis are similar. The first chick nerve cells arise just before formation of the first somites; a slightly later time is indicated in the mouse. Reticular neurons are the earliestarising cells in the chick and make up the majority of the population in each of the youngest chick and mouse stages obtained. Motor and sensory neurons do not begin to originate in large numbers until later, during E3 in the chick and E9 in the mouse.

Organization of ascending serotonin systems in the adult rat brain. A radioautographic study after intraventricular administration of [ 3h]5-hydroxytryptamine

Light microscope radioautography was used to visualize ascending serotonin (5-HT) systems in adult rat brain, following prolonged latero-ventricular instillation of [ 3H]5-HT. In paraventricular and paracisternal regions, the cell bodies, axonal projections and terminal fields of 5-HT neurons were distinctly labelled, allowing comprehensive analysis of their organizational features. Two major systems of ascending 5-HT fibers could be defined: a transtegmental system originating mainly from nucleus raphe dorsalis (B-7 and B-6) and less prominently from nucleus raphe medianus (B-8), and a periventricular system predominantly issuing from the rostral pole of nucleus raphe dorsalis (B-7). The transtegmental system converges ventral-ward across the decussation of the superior cerebellar peduncles and sweeps rostral-ward, through the ventral tegmental decussation, to enter the ventral tegmental area. The periventricular system closely follows the dorsal longitudinal fasciculus of Schu¨tz. It branches off dorsally toward the superior and inferior colliculi and the subcommissural organ, but the bulk of its fibres arch abruptly ventralward, beneath the posterior commissure, to reach the dorsal hypothalamus. Along their way, these axons most likely contribute to the dense 5-HT innervation of the periventricular gray matter of the midbrain and caudal diencephalon. Both ascending 5-HT systems merge in the medial forebrain bundle area of caudal hypothalamus, wherefrom their fibres borrow several limbic pathways to reach distant territories of innervation: fasciculus retroflexus to medial habenula; striae medullaris and terminalis toward the habenula and the amygdala, respectively; fornix to hippocampus; diagonal band of Broca to septum; induseum griseum and cingulate bundle to hippocampus and neocortex. Other fibers spread more laterally, across the internal capsule, to innervate the globus pallidus and neostriatum. Finally, at the base of anterior forebrain, some 5-HT axons traverse the anterior olfactory nucleus and end in the glomerular layer of olfactory bulb. These and additional results concerning the heterogeneous distribution of 5-HT axonal varicosities in various parts of forebrain (e.g. thalamus, hypothalamus, septal area and amygdaloid complex) complement earlier data gathered by means of other neuroanatomical techniques and make it possible to present a useful model of the general organization of ascending 5-HT systems in the mammalian brain.

Substrate pathways demonstrated by transplanted Mauthner axons.

A substrate pathway is a set of aligned guidance cues. (Such cues may be either cells or molecules.) CNS substrate pathways can be demonstrated by transplanting axons to different starting locations. The stereotyped routes of transplanted axons will then demonstrate the locations of effective substrate pathways. To map CNS substrate pathways, Mauthner axons were transplanted to various unnatural locations along the CNS of Xenopus embryos. The routes of 24 experimental Mauthner axons were traced. Twenty-one of these axons grew along parts of a stereotyped route extending in the ventral marginal zone from the caudal diencephalon through the spinal cord. This ventral substrate pathway ran the length of the basal plate; thus, we call it a basal substrate pathway. One experimental Mauthner axon grew along the alar substrate pathway previously demonstrated by transplanted optic axons. The demonstrations of the alar and the basal substrate pathways suggest that during development a few long substrate pathways organize the overall layout of the long tracts of the CNS. We propose that the pattern of the earliest CNS substrate pathways is established in the neural plate and is topologically preserved as the neural plate rolls into a neural tube. This pattern may be manifest as the three-dimensional organization of the early marginal zones formed by the peripheral processes (the endfeet) of certain developing ependyma and radial glia. Subsequently, the detailed anatomy of the axon tracts and the specific terminations of individual axons are probably determined by other local chemical cues.