Forrest Haun

Lesion of the habenular efferent pathway produces anxiety and locomotor hyperactivity in rats: a comparison of the effects of neonatal and adult lesions

Recent studies have implicated the habenula in modulating states of arousal and chronic responses to stress. We examined whether lesion of the habenula efferent pathway, the fasciculus retroflexus (FR), at either 3 (P3) or 70 (P70) days of age affects stress-related anxiety (elevated plus-maze test) and activity levels (open-field test) in rats tested as adults. Both P3- and P70-lesioned rats showed chronically elevated plasma levels of corticosterone. Rats receiving FR lesions as neonates (P3) exhibited greater open arm avoidance on the elevated plus-maze than controls 2 months postoperatively, suggesting a heightened state of anxiety. In contrast, P70-lesioned rats behaved similarly to controls on the plus-maze, but showed increased locomotion and increased grooming in the open field, effects not observed in P3-lesioned rats. When an additional stressful condition was imposed (5 days of social isolation plus 24 h food deprivation) before testing, both FR-lesion groups showed an attenuation of the normal behavioral responses (decreased open-arm entries/time in open arms, increased freezing). The effects of FR lesions on activity and behavioral indices of anxiety may be due to disruption of lateral habenular projections to dopaminergic neurons in the ventral tegmentum and/or projections to regions containing high concentrations of benzodiazepine receptors, the median and dorsal raphe and dorsal periaqueductal gray. Behavioral differences observed as a function of lesion age suggest differential capabilities of P3- and P70-lesioned rats to utilize compensatory mechanisms to correct FR lesion-induced deficits.

Cortical transplants reveal CNS trophic interactions in situ

Five days after transplanting fetal rat posterior cortex into newborn host rats with posterior cortex lesions, the hosts dorsal lateral geniculate nucleus (dLGN) contains 2-5 times as many neurons as the dLGN of lesion-only controls. This effect is temporary and restricted to neurons created later in dLGN neurogenesis. Similar transplants of cerebellar plate are ineffective. These findings suggest that intracranial transplants of CNS tissue can be a source of specific trophic support to particular host neurons.

Diffusible proteins prolong survival of dorsal lateral geniculate neurons following occipital cortex lesions in newborn rats

Removal of the occipital cortex in newborn rats results in the rapid and nearly complete degeneration of the dorsal lateral geniculate nucleus (dLGN) in 5 days. In previous studies we have shown that transplants of embryonic posterior cortex neurons, which are allowed to develop in culture for 5 days prior to transplantation into the site of the lesion, prolong the survival of a particular population of host dLGN neurons for an additional week. In this study we tested the possibility that the transplant cells synthesize diffusible proteins which are responsible for this neurotrophic effect. Culture medium conditioned by explants of embryonic occipital cortex and diencephalon was concentrated by vacuum dialysis or ultrafiltration through membranes with at least a 10-kDa cut-off. This concentrated medium was loaded into polyacrylamide or sodium alginate gels which were then implanted into the cavity of the lesion. Five days after implantation, the alginate-conditioned-medium implants result in a 3-fold increase in dLGN survival compared to unconditioned medium controls, while a two-fold increase in survival of the nucleus is found with the polyacrylamide-conditioned-medium implants. Proteolysis of the conditioned medium eliminates all neurotrophic activity. The results suggest that the death of dLGN neurons following the cortical lesion is due to the loss of diffusible proteinaceous neurotrophic factors--factors that may operate during normal in vivo development of the geniculocortical pathway.

Dissociation of behavioral changes in rats resulting from lesions of the habenula versus fasciculus retroflexus and their possible anatomical substrates.

Lesions in either the habenula or its primary efferent pathway, the fasciculus retroflexus (FR), impaired avoidance responding. However, lesions of only the FR provided a persistent elevation of locomotor activity. Immunocytochemical study of the interpeduncular nucleus (IPN) through injection of retrograde tracers into the IPN and the overlying ventral tegmental area indicated that habenular lesions spared both rostral habenula and forebrain projections to the caudal midbrain, but these projections were axotomized by FR lesions. Rostral sparing of the habenula resulted in normal peptidergic staining in the IPN, and normal cholinergic innervation was absent. Performance of individual rats in behavioral tests was consistent with variations in anatomical sparing. Such considerations may account for previous discrepancies in functional effects of habenular lesions.

Rescue of both rapidly and slowly degenerating neurons in the dorsal lateral geniculate nucleus of adult rats by a cortically derived neuron survival factor

We investigated the death of dorsal lateral geniculate nucleus (dLGN) neurons after lesions to the visual cortex of adult rats and the effects of supplying target-derived neurotrophic molecules to the lesion cavity. The neurotrophic factor is retrieved from cocultures of the embryonic primordia of the geniculocortical pathway and its survival promoting properties for different populations of dLGN neurons (based on their time of origin) have been documented in previous studies of neonatal rats with occipital cortex lesions. In the present study, rats were exposed to [3H]thymidine on E14 or E15/16 to label either earlier or later generated dLGN neurons. When animals were at least 45 days old we made discrete lesions to the principal projection zones in area 17 of these two dLGN populations. Counts of surviving labeled cells show a relatively rapid death of E15/16 dLGN neurons in control animals, with a maximal loss by 2 weeks postlesion. The death of E14 dLGN neurons is more protracted, with a maximal loss by 2 months postlesion. A 2-week infusion of the CM fraction rescues the majority of the neurons that would otherwise die in both populations compared to the controls which receive a similarly prepared fraction of unconditioned medium. Moreover, this CM fraction can sustain E14-generated dLGN neurons up to 6 weeks after the neurotrophic factor(s) is no longer being supplied exogenously. Thus the rescue of axotomized adult dLGN neurons appears to be permanent, at least for the early generated population. These findings are consistent with the idea that target-derived molecules have a role in the survival of mature neurons, as they are known to have for developing neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Different populations of dorsal lateral geniculate nucleus neurons have concentration-specific requirements for a cortically derived neuron survival factor

A macromolecular fraction of conditioned culture medium (CM) derived from explant cocultures of embryonic rat posterior cortex and caudal thalamus is able to support the survival of neurons in the dorsal lateral geniculate nucleus (dLGN) of newborn rats following ablation of dLGN cortical target areas. In the present study we tested whether the survival-promoting activity of this target-derived neurotrophic agent was concentration dependent and whether different subpopulations of dLGN neurons were equally responsive. With the starting concentration of the CM fraction designated X, increasing concentration results in a progressive falloff in trophic activity so that at 200X overall dLGN survival is similar to that seen in unconditioned medium (UM) controls. In contrast, diluting the fraction produces an increase in activity until maximal survival is achieved at 0.2X. Further dilutions result in a decline in trophic activity until control values are reached at 0.001X. Two populations of neurons within the dLGN, defined by their time of origin, respond in a specific manner to the different concentrations. Neurons generated during the early stages of neurogenesis (E14) have maximal survival (25.8%) at 0.05X, whereas those neurons generated later (E15/16) are maximally supported (30.7% survival) at 10X, a 200-fold difference in concentration. While it is possible that separate neurotrophic and neurotoxic molecules exist for each of these populations of dLGN neurons, the most parsimonious interpretation of the data is that a single cortically derived neurotrophic factor exists whose production is strictly controlled during development to achieve maximal effect on different populations of thalamic neurons that may be functionally distinct.

Trophic Effects of Transplants Following Damage to the Cerebral Cortex

One of the remarkable differences between developing and mature brains is their response to injury. A specific lesion to developing nervous system pathways usually results in rapid and profound neuron loss while the same lesion in adults may produce only gradual cell loss or no cell death at all.I4 This observation is particularly intriguing because of its apparent conflict with the well-known behavioral observation that functional recovery from damage to the central nervous system (CNS) is generally more likely if the damage occurs to developing rather than mature CNS systems?6 Several explanations have been offered for this difference in the response of differentaged neurons to lesions. First, the fact that younger neurons are less likely to have formed sufficient numbers of sustaining inputs or sustaining collateral axons may make them much more vulnerable when those few inputs or collaterals that have developed are damaged.’” In the CNS, collateral development is likely to be especially important to damaged neurons because these cells often have more widespread connections than neurons in the peripheral nervous system (PNS). A second suggestion relates to the “inherent metabolic capabilities” of younger neurons versus mature neurons! As the neuron matures, so does its protein synthetic machinery, which may allow the cell to maintain itself for longer periods after lesions. Metabolic maturity seems likely to play an important role in the survival of PNS neurons after axotomy because mature synthetic processes appear to be required to support regeneration. A third idea, which may in fact encompass the other two, is that lesions to afferent or target structures separate the cell from sources of specific trophic factors supplied by these connecting cells. Developing neurons may depend more critically on these factors for both survival and continued differentiation. Mature neurons may no longer require such trophic agents,’ or may have a much lower “rate of utilization” of the factors? Support for this “trophic hypothesis,” at least for developing neurons, has come mainly from studies with nerve growth factor (NGF) in the PNS. NGF is a specific trophic agent for sensory and sympathetic ganglion cells; it is essential for the survival and differentiation of these cells during development. It is unique among the putative trophic factors that have been suggested or even partially isolated because its actions are demonstrable in vivo. Exogenous NGF can rescue naturally degenerating ganglion

The role of the habenula-interpeduncular pathway in modulating levels of circulating adrenal hormones

The fasciculus retroflexus (FR) is the major pathway by which the medial and lateral habenular nuclei project to the interpeduncular nucleus (IPN) and ventral tegmentum. Recent work has suggested that the habenula-interpeduncular system may be involved in the regulation of states of arousal. Bilateral FR lesions have been shown to disrupt chronically, and habenula transplants have been shown to restore normal sleep patterns in rats [J. NeuroscL, 12 (1992) 3282-3290]. In this study, we examined whether FR lesions and habenula cell transplants would also modify chronically the circulating plasma levels of the stress-related hormones, norepinephrine (NE), epinephrine (EPI) and corticosterone. When plasma samples were obtained via retro-orbital eye-bleed during anesthesia, animals with FR lesions had significantly increased levels of plasma NE, EPI and corticosterone 2-3 months postoperatively compared to unoperated controls. Transplants of embryonic habenula cells placed near the denervated IPN in FR-lesioned animals restored levels of NE and EPI to normal, but did not attenuate elevated corticosterone levels. When plasma samples were obtained in conscious animals via indwelling arterial cannulae, FR-lesioned rats likewise exhibited increased basal levels of corticosterone but plasma levels of catecholamines were similar to those of unoperated controls. Differences in our results obtained using the two methods of blood sampling may be explained by the effects of anesthesia and stress associated with the eye-bleed method. Thus, the effect of FR lesions in increasing plasma levels of catecholamines may not reflect a difference in basal hormone levels, but a heightened sympathetic adrenomedullary response to stress. While these results indicate that the integrity of the habenular efferent pathway is important in modulating circulating levels of hormones associated with the stress response, two separate mechanisms appear to control its interactions with sympathetic-adrenal medullary and adrenocortical pathways.

Transient and Permanent Patterns of Expression of the Low-Affinity Neurotrophin Receptor in the Interpeduncular Nucleus of the Rat

There is a prominent cholinergic projection from the medial habenular to the interpeduncular nucleus (IPN) which develops postnatally. In this study we examined the developmental course of expression of the low-affinity neurotrophin receptor (LANR), a receptor that binds to all members of the neurotrophin family, in the IPN. Three systems express LANR in the IPN. The cholinergic habenular axons that project to the intermediate and central subnuclei of the IPN are immunoreactive only during the time that the axons are growing and forming characteristic crest synapses in the intermediate subnuclei. The dorsomedial (DM) subnuclei, which are neither cholinergic targets nor contain cholinergic neurons, develop LANR immunoreactivity postnatally and the expression remains high in the adult. The walls of the prominent system of arterioles and venules that penetrate the IPN and ascend through the intermediate subnuclei are strongly immunoreactivity during the time of active angiogenesis and retain detectable but diminished levels of immunoreactivity in the adult. The LANR immunoreactivity seen in the vessels is likely to be associated with the peripheral sympathetic axons that innervate the smooth muscle in the vessels. These three systems within the same nucleus, which differ in phenotype, develop neurotrophin receptor immunoreactivity contemporaneously but their levels of expression differ in the adult, suggesting that they are regulated in different ways, possibly by different members of the neurotrophin family.

Transplant-Mediated Recovery of Innervation and Function: The Habenulo-Interpeduncular System of the Rat

Transplants of fetal cells into host brains can be used to examine the development of specific afferent projections and can also provide a means of repairing brain damage and thereby restoring function after injury. Restoration of function mediated by transplanted tissue provides compelling evidence that the implanted cells have integrated functionally in the host brain. We have used transplants of fetal tissue into host brains whose interpeduncular nuclei have been denervated by bilateral lesions to the fasciculus retroflexus, firstly to examine some of the features regulating innervation of the interpeduncular nucleus in the rat and, secondly, to characterize the functional organization of this system.