Habenula and thalamus cell transplants mediate different specific patterns of innervation in the interpeduncular nucleus.

Innervation of specific peptidergic and cholinergic compartments of the interpeduncular nucleus (IPN) was investigated using embryonic cell suspension transplants immunoreactive for substance P (SP) and ChAT. In both neonatal and adult host rats, the IPN was first denervated of its normal SP and cholinergic input from the medial habenula by bilateral lesions of the fasciculi retroflexi (FR). In adult hosts, transplants of embryonic habenular cells placed near the denervated IPN mediated a return of the normal pattern of SP staining restricted to habenula-target subnuclei, plus an increase in staining intensity of SP cells intrinsic to the IPN. There was no recovery of ChAT staining. A similar pattern of SP staining resulted following habenular transplants into neonatal hosts, but in addition there was a partial recovery of normal ChAT staining in cholinergic subnuclei and anomalous ChAT staining in normally peptidergic subnuclei. Control transplants of embryonic thalamus cells placed into adult hosts produced a surprising pattern of ChAT staining in the IPN identical to that seen with habenula transplants placed into neonatal hosts; the adult IPN was thus able to support reinnervation mediated by an aberrant cholinergic source while being refractory to its normal habenular cholinergic afferents. This pattern of results implies regulation by the IPN of habenular SP and cholinergic innervation, and some interaction between the maturing normal cholinergic afferents and their targets that is missing when these afferent sources are abnormal.

Habenula and thalamus cell transplants restore normal sleep behaviors disrupted by denervation of the interpeduncular nucleus.

The preceding companion study (Eckenrode et al., 1992) showed that cell suspension transplants of fetal habenula cells placed near the interpeduncular nucleus (IPN) following lesions of the fasciculus retroflexus (FR) restore the normal pattern of substance P (SP) staining in habenular target subnuclei of the IPN in both perinatal and adult hosts, and restore ChAT staining in the IPN of perinatal hosts. Similarly placed transplants of fetal thalamus cells only restore ChAT staining in the IPN of adult hosts. In this study, we examined the functional significance of these restored staining patterns. We used a behavioral measure of the integrity of REM-stage and non-REM-stage sleep, the "flower pot" test, and assayed (1) normal adult rats, (2) FR-lesioned control animals (neonatal or adult operates), (3) animals receiving FR lesions and transplants of fetal habenula cells (perinatal or adult hosts), and (4) animals receiving FR lesions and transplants of fetal thalamus cells (adult hosts). FR lesions decrease markedly the muscle atonia component of REM sleep and reduce duration of sleep episodes. Transplants that restore SP staining in the IPN (habenular transplants into either perinatal or adult lesion hosts) restore normal frequency of REM atonia; transplants that restore ChAT staining (habenular transplants into perinatal hosts or thalamic transplants into adult hosts) restore normal duration of sleep episodes. The number of SP-immunoreactive cells in the transplants predicts recovery of REM atonia, and the number of ChAT cells in habenular (but not thalamic) transplants predicts restoration of sleep duration.(ABSTRACT TRUNCATED AT 250 WORDS)

Opiate binding sites in the interpeduncular nucleus of the rat: normal distribution and the effects of fasciculus retroflexus lesions.

Opiate receptors have been localized autoradiographically to many regions of the rat central nervous system. The interpeduncular nucleus has an especially high concentration of these receptors. We used [3H]naloxone and [125I] [D-Ala2,MePhe4,Glyol5]enkephalin as ligands to map the distribution of opiate receptors among the subnuclei of the interpeduncular nucleus. The rostral subnucleus contains label that is densest dorsally. More caudally, high densities of opiate binding sites are found in the lateral, rostral, and central subnuclei. The dorsal subnucleus contains a moderate density of binding sites and the intermediate subnuclei contain almost none. Opiate receptors have also been localized to the medial habenulae and the fasciculi retroflexi, which provide a major afferent input to the interpeduncular nucleus. Lesions of the fasciculi retroflexi decreased the density of opiate binding sites in the rostral and lateral subnuclei of the interpeduncular nucleus. There were no changes seen in the dorsal, intermediate or central subnuclei. These results suggest that a minority of opiate receptors in the interpeduncular nucleus are located presynaptically on fasciculi retroflexi axons. Immunocytochemical studies have demonstrated that the rat interpeduncular nucleus contains substance P, serotonin and enkephalin, and each has a distinct subnuclear distribution. Although the opiate binding sites have a wider distribution than substance P, serotonin, or enkephalin individually, the pattern of opiate binding most closely parallels substance P distribution. The combined distribution of substance P, serotonin, and enkephalin is equivalent to that of the opiate binding sites.

Acetylcholine in the interpeduncular nucleus of the rat: normal distribution and effects of deafferentation.

We studied the cholinergic projection to the interpeduncular nucleus (IPN) by examining localization of choline acetyltransferase (ChAT) in the habenula, fasciculus retroflexus (FR) and among the subnuclei of the IPN of the rat, using and antibody raised against ChAT. ChAT-containing neurons were present in the ventral portion of the medial habenula, ChAT-stained axons were present in the FR and ChAT-stained axons and terminals were present in the rostral, central and intermediate subnuclei of the IPN. No ChAT staining was seen in the lateral or dorsal subnuclei. The pattern of ChAT localization was thus complementary to the pattern of the habenular substance P projection to the IPN. Lesions of the FR eliminated all ChAT from the IPN while lesions of the stria medullaris produced a modest decrease. Unilateral FR lesions indicated that the FR projection to the central and rostral subnuclei is largely bilateral and symmetrical and that to the intermediate subnuclei is largely ipsilateral. We found no evidence of lesion-induced plasticity, i.e. replacement of ChAT immunoreactivity, by surviving FR axons in these adult brains.

Normal development and effects of early deafferentation on choline acetyltransferase, substance P and serotonin-like immunoreactivity in the interpeduncular nucleus.

The normal postnatal development and response to neonatal fasciculus retroflexus (FR) lesions of serotonin, substance P (SP), and choline acetyltransferase (ChAT) distribution are described for the rat interpeduncular nucleus (IPN). Serotonin-, SP- and ChAT-containing axons differed in development, distribution, and response to deafferentation. Serotonergic axons and cell bodies were present at birth. SP was present in the FR and in the lateral subnuclei by 3 days of age but did not appear in the rostral or dorsal subnuclei until 7-14 days. Intrinsic SP perikarya were not seen until 17 days of age. The development of ChAT was late, appearing only during the second week of life and not reaching adult patterns and density until after 21 days of age. The pattern of development of cytochrome oxidase and Bodian silver staining are also described. Both cytochrome oxidase and Bodian staining paralleled the patterns of localization and development of ChAT staining. Bilateral neonatal FR lesions resulted in a permanent loss of ChAT and cytochrome oxidase staining throughout the IPN and of SP in the lateral and rostral subnuclei. No changes were seen in the serotonergic system. Following unilateral lesions, the pattern of SP loss and replacement paralleled that seen after adult lesions. The pattern of replacement of ChAT differed from that after adult lesions in that there was partial replacement in the ipsilateral intermediate subnucleus following neonatal lesions. This result suggests that late developing cholinergic axons can innervate the contralateral intermediate nucleus to a much greater extent following infant lesions than following adult lesions.