Distribution of neuropeptide K-immunoreactivity in the rat central nervous system

The distribution of neuropeptide K (NPK), a 36-residue amidated peptide originally isolated from porcine brain, is described in the rat CNS by immunohistochemical methods. Antibodies were generated in rabbits to N-terminus and C-terminus regions of the peptide and the distribution of immunoreactive cell bodies and fibers was mapped in colchicine-treated and normal rat brains. Major areas of cell body staining included the medial habenular nucleus, the ventromedial nucleus of the hypothalamus, the interpeduncular nucleus, the lateral dorsal tegmental nucleus, the nucleus raphe pallidus, and the nucleus of the solitary tract. Some of the areas of dense NPK-fiber immunoreactivity included the ventral pallidum, the caudate-putamen, certain areas of the hypothalamus, the central and medial amygdaloid nuclei, the entopeduncular nucleus, the habenular nuclei, the substantia nigra pars reticulata, the caudal part of the spinal nucleus of the trigeminal nerve, the nucleus of the solitary tract and the dorsal horn of the spinal cord. A striking similarity exists between this pattern of immunoreactive staining and that described for substance P, suggesting that the tachykinin systems do not exist independently in the brain. The possible roles for multiple tachykinins in the brain are discussed.     

大白鼠脚内核的传入性联系

众所周知,肉食动物和大白鼠的脚内核,相当于灵长类的内侧苍白球(Nagy et al.1978;Fox and Schmitz 1944);它们的细胞形态、传入及传出均相同。早期以及近年来的一些研究工作者,虽然在研究其他核团的投射时,联系到一些本核团的传入,但是尚缺乏对本核团传人的系统研究。本实验即是应用辣根过氧化物酶的逆行传递法来研究大白鼠脚内核的传入性联系。     

Substance P-like immunoreactivity in the parietal eye visual system of the lizard Uta stansburiana

Summary We examined the parietal eye visual system of the iguanid lizard Uta stansburiana for the presence of substance P-like immunoreactivity by use of both immunofluorescence and peroxidase-antiperoxidase techniques. In the parietal eye no substance P-containing somata were found; however, its plexiform layer contained small (ca. 1 m diam) immunoreactive fibers. These fibers apparently originate outside the parietal eye. Immunoreactive fibers also were found in the parietal nerve, the dorsal sac, and the leptomeninx of the pineal gland. No labeled somata were observed in any of these regions in either normal or colchicine treated animals. Previously we demonstrated that a system of centrifugal fibers to the parietal eye originates from neurons in the dorsal sac (Engbretson et al. 1981). The apparent absence of substance P-containing neurons in the dorsal sac suggests that the substance P-containing fibers in the parietal eye are not the previously observed centrifugal fibers. The source of the substance P-containing fibers in the parietal eye is unknown. The pars dorsolateralis of the left medial habenular nucleus receives a dense substance P-positive projection. No such projection was seen in the right habenula. Simultaneous visualization of the terminals of ganglion cells of the parietal eye (labeled with orthograde intraaxonally transported horseradish peroxidase) and substance P-like immunofluorescence showed that the locus of habenular immunoreactivity is distinct from the projection field of the parietal eye. Thus the substance P-positive terminals in the habenula do not originate in the parietal eye. Transection of the parietal nerve confirmed this conclusion.     

Distribution of somatostatinlike immunoreactivity in the brain of the little brown bat (Myotis lucifugus)

The distribution of somatostatinlike immunoreactive (SLI) perikarya, axons, and terminals was mapped in subcortical areas of the brain of the little brown bat, Myotis lucifugus, using light microscopic immunocytochemistry. A preponderance of immunoreactivity was localized in reticular, limbic, and hypothalamic areas including: 1) in the forebrain: the bed nucleus of the stria terminalis; lateral preoptic, dorsal, anterior, lateral and posterior hypothalamic areas; amygdaloid, periventricular, arcuate, supraoptic, suprachiasmatic, ventromedial, dorsomedial, paraventricular, lateral and medial mammillary, and lateral septal nuclei; the nucleus of the diagonal band of Broca and nucleus accumbens septi; 2) in the midbrain: the periaqueductal gray, interpeduncular, dorsal and ventral tegmental, pretectal, and Edinger-Westphal nuclei; and 3) in the hindbrain: the superior central and parabrachial nuclei, nucleus incertus, locus coeruleus, and nucleus reticularis gigantocellularis. Other areas containing SLI included the striatum (caudate nucleus and putamen), zona incerta, infundibulum, supramammillary and premammillary nuclei, medial and dorsal lateral geniculate nuclei, entopeduncular nucleus, lateral habenular nucleus, central medial thalamic nucleus, central tegmental field, linear and dorsal raphe nuclei, nucleus of Darkschewitsch, superior and inferior colliculi, nucleus ruber, substantia nigra, mesencephalic nucleus of V, inferior olivary nucleus, inferior central nucleus, nucleus prepositus, and deep cerebellar nuclei. While these results were similar in some respects to those previously reported in rodents, they also provided interesting contrasts.     

Efferent projections from the lateral geniculate nucleus to the pineal complex of the Mongolian gerbil (Meriones unguiculatus)

Summary The intergeniculate leaflet of the lateral geniculate nucleus is considered to modulate circadian activity rhythms probably mediated by a direct neuronal connection to the suprachiasmatic nucleus. The present study in the gerbil demonstrates, by anterograde tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L), the existence of an additional neuronal projection from a subportion of the lateral geniculate nucleus, involving the intergeniculate leaflet, directly to the pineal gland. PHA-L-immunoreactive nerve fibers originating from perikarya at the injection site were located under the optic tract projecting towards the midsagittal plane. Delicate PHA-L-immunoreactive nerve fibers were observed in the posterior paraventricular thalamic nucleus, precommissural nucleus, olivary pretectal nucleus, anterior and posterior pretectal nuclei, and posterior commissure. Single fibers could be followed from the caudal part of the medial habenular nucleus and the pretectal area into the rostral part of the deep pineal gland. Other fibers continued through the posterior commissure into the contralateral hemisphere to terminate in the same structures as on the ipsilateral side. From the posterior commissure, small bundles of thick fibers entered the deep pineal gland where they arborized among the endocrine cells. A few nerve fibers were observed in the habenular commissure and the pineal stalk, but no fibers were identified in the superficial pineal. This direct geniculo-pineal connection suggests that the pineal gland is directly influenced by the optic system.     

Prolonged expression of c-Fos protein in the lateral habenular nucleus of the Japanese monkey (Macaca fuscata) after eye enucleation

To elucidate the effect of traumatic stress on the lateral habenular nucleus, we investigated the time course of the expression of c-Fos protein in this nucleus of the Japanese monkey (Macaca fuscata) after enucleation of one eye using c-Fos protein immunocytochemistry. c-Fos protein-like immunoreactive neurons were significantly increased; the increase started 1 h after the enucleation and remained high for 3-9 h in the lateral habenular nucleus on both sides. These results suggest that the prolonged expression of c-Fos protein occurred in the lateral habenular nucleus after traumatic stress through multiple transsynaptic activations.     

Organization of the ferret lateral cervical nucleus and cervicothalamic tract

To elucidate the organization of the ferret spinocervicothalamic pathway (SCTP), we examined the lateral cervical nucleus (LCN) and the termination of the cervicothalamic tract (CTT) in this species. In thionin-stained sections, the ferret LCN appeared as an easily delineated column of cells in the dorsolateral funiculus from about mid-C3 to the rostral end of C1, with most cells located in the C1 and C2 segments. In transverse sections, the LCN was elongated along a dorsolateral to ventromedial axis and in the rostral half of C2 and caudal half of C1 continuous with the neck of the dorsal horn. The number of ferret LCN cells was estimated to 2,500-3,700, with an average of 3,340. Substance P-like immunoreactive fibers located preferentially in the ventromedial part of the LCN, whereas serotonin-like immunoreactive fibers were found throughout the nucleus. Anterograde transport of wheat germ agglutinin-horseradish peroxidase conjugate and biotinylated dextran amine demonstrated that the ferret CTT terminates extensively in the peripheral parts of the ventral posterior lateral nucleus. Sparser termination was evident in the ventral posterior inferior nucleus, in the medial nucleus of the posterior complex, and in the medial part of the magnocellular medial geniculate nucleus. Thus, although the LCN is significantly smaller in ferrets than in cats and raccoons, the organization of the LCN and of the cervicothalamic tract is closely similar in the three species. These findings indicate a conserved general organization of the SCTP among carnivores.     

Organization of the ferret lateral cervical nucleus and cervicothalamic tract

To elucidate the organization of the ferret spinocervicothalamic pathway (SCTP), we examined the lateral cervical nucleus (LCN) and the termination of the cervicothalamic tract (CTY) in this species. In thionin-stained sections, the ferret LCN appeared as an easily delineated column of cells in the dorsolateral funiculus from about mid-C3 to the rostral end of C1, with most cells located in the C1 and C2 segments. In transverse sections, the LCN was elongated along a dorsolateral to ventromedial axis and in the rostral half of C2 and caudal half of C1 continuous with the neck of the dorsal horn. The number of ferret LCN cells was estimated to 2,500-3,700, with an average of 3,340. Substance P-like immunoreactive fibers located preferentially in the ventromedial part of the LCN, whereas serotonin-like immunoreactive fibers were found throughout the nucleus. Anterograde transport of wheat germ agglutinin-horseradish peroxidase conjugate and biotinylated dextran amine demonstrated that the ferret CTT terminates extensively in the peripheral parts of the ventral posterior lateral nucleus. Sparser termination was evident in the ventral posterior inferior nucleus, in the medial nucleus of the posterior complex, and in the medial part of the magnocellular medial geniculate nucleus. Thus, although the LCN is significantly smaller in ferrets than in cats and raccoons, the organization of the LCN and of the cervicothalamic tract is closely similar in the three species. These findings indicate a conserved general organization of the SCTP among carnivores.     

Distribution of oxytocin and vasopressin neurons in the diencephalon of the Japanese horseshoe bat, Rhinolophus ferrumequinum. An immunohistochemical study.

The distribution of oxytocin (OXT) and vasopressin (VP) neurons in the diencephalon of the hibernating Japanese horseshoe bat, Rhinolophus ferrumequinum, was immunohistochemically investigated by the avidin-biotin complex method. Magnocellular OXT and VP neurons were localized mainly in the paraventricular nucleus and the supraoptic nucleus. In addition to these main nuclei, both kinds of magnocellular neurons were also found in the periventricular nucleus, perifornical area and lateral hypothalamic area. Extensively distributed parvocellular neurons containing only VP were observed in the rostral and middle portions of the suprachiasmatic nucleus. The size of OXT and VP magnocellular neurons was almost equal in the paraventricular and ventromedial supraoptic nuclei, whereas VP neurons were significantly larger than OXT neurons in the dorsolateral supraoptic nucleus. The OXT and VP cells in the ventral supraoptic nucleus showed a distinctive elliptical shape. Both OXT and VP fibers were distributed in the lateral habenular nucleus, stria medullaris thalami, lateral preoptic area, stria terminalis, and medial and supracapsular part of the bed nucleus of the stria terminalis. Moreover, OXT fibers were found in the substantia nigra, and VP fibers were noted in the nucleus reunions and the paraventricular nucleus of the thalamus.     

Morphology of neurons and axon terminals associated with descending and ascending pathways of the lateral forebrain bundle in Rana esculenta

Summary The cells of origin of afferent and efferent pathways of the lateral forebrain bundle were studied with the aid of the cobalt-filling technique. Ascending afferents originated from the lateral thalamic nucleus, central thalamic nucleus, posterior tuberculum and the cerebellar nucleus. They terminated in the anterior entopeduncular nucleus, amygdala and the striatum. Telencephalic projection neurons, which are related to the lateral forebrain bundle, were located mainly in the ventral striatum and the anterior entopeduncular nucleus, but were not so numerous in the dorsal striatum. Irrespective of their location, most of the neurons projecting axons into the lateral forebrain bundle had piriform or pyramidal perikarya. Long apical dendrites usually arborized in a narrow space, whereas widely arborizing secondary dendrites originated from short dendritic trunks. The other neurons that contributed to the lateral forebrain bundle were fusiform or multipolar cells. Striatal efferents terminated in the pretectal area and in the anterodorsal, anteroventral and posteroventral tegmental nuclei.     

Substance P-like immunoreactivity in the trigeminal sensory nuclei of an infrared-sensitive snake,Agkistrodon blomhoffi

Summary With the peroxidase-antiperoxidase immunohistochemical method we ascertained the presence of substance P-like immunoreactivity (SPLI) in fibers and cell bodies of the trigeminal sensory system of the pit viper, Agkistrodon blomhoffi. There are a few SPLI fibers each in the principal sensory nucleus and the main neuropil of the lateral descending nucleus (i.e., the infrared sensory nucleus); a moderate number in the descending nucleus; and a large number in the caudal subnucleus, the medial edges of the interpolar subnucleus, and the marginal neuropil of the lateral descending nucleus. About 30% of the cell bodies in the ophthalmic and maxillo-mandibular ganglia show SPLI, and of the two craniocervical ganglia, the proximal ganglion has many more cells with SPLI than the distal ganglion. The SPLI distribution in the common trigeminal sensory system is similar to that of mammals, and suggests that the function of this system is also similar. In the infrared sensory system, the differing distribution in the main and marginal neuropils suggests separate functions for these two structures in the system.     

电刺激大鼠外侧缰核对延髓甩尾相关细胞的活动和甩尾反射的影响

在浅麻醉大鼠上,在延髓腹内侧结构内观察到三种具有不同放电类型的细胞,即乃尾前放电骤停的撤反应细胞,甩尾前放电骤增的给反应细胞和甩尾无关的中性细胞。电刺激外侧缰核可抑制撤反应细胞的自发放电,加强给反应细胞自发放电,从而易化两类细胞的甩尾相关反应,同时易化伤害刺激引起的甩尾反射。实验结果说明,外侧缰核对节段性防御反射有易化作用,这种易化作用可能是通过延髓内撤反应和反应细胞的协同活动而实现的。     

Central connections of the olfactory bulb in the goldfish,Carassius auratus

Summary The central connections of the goldfish olfactory bulb were studied with the use of horseradish peroxidase methods. The olfactory bulb projects bilaterally to ventral and dorsolateral areas of the telencephalon; further targets include the nucleus praeopticus periventricularis and a caudal olfactory nucleus near the nucleus posterior tuberis in the diencephalon, bilaterally. The contralateral bulb and the anterior commissure also receive an input from the olfactory bulb. Contralateral projections cross in rostral and caudal portions of the anterior commissure and in the habenular commissure. Retrogradely labeled neurons are found in the contralateral bulb and in three nuclei in the telencephalon bilaterally; the neurons projecting to the olfactory bulb are far more numerous on the ipsilateral side than in the contralateral hemisphere. Afferents to the olfactory bulb are found to run almost entirely through the lateral part of the medial olfactory tract, while the bulb efferents are mediated by the medial part of the medial olfactory tract and the lateral olfactory tract. Selective tracing of olfactory sub-tracts reveals different pathways and targets of the three major tract components. Reciprocal connections between olfactory bulb and posterior terminal field suggest a laminated structure in the dorsolateral telencephalon.     

Preganglionic neurons of the sphenopalatine ganglia reside in the dorsal facial area of the medulla in cats

Stimulation of the sphenopalatine ganglion (SPG), a parasympathetic ganglion of the facial nerve, or the dorsal facial area (DFA), an area in the lateral tegmental field just dorsal to the facial nucleus, induces an increase in blood flow of the common carotid artery (CCA). This study attempted to clarify the anatomical and functional relationships between the SPG and the DFA, and to demonstrate putative serotonergic (5-HT) and substance P (SP) innervations to the neurons of the DFA in regulation of the CCA blood flow in cats. Horseradish peroxidase (HRP), a retrograde tracer, was injected in the SPG. All HRP-labeled neurons were distributed in the reticular areas dorsal and lateral to the superior olivary nucleus and the facial nucleus, extending from the caudal half of the superior olivary nucleus to the rostral 3/4 of the facial nucleus on the HRP-injected side. They were grouped into five clusters, namely lateral circumference of the superior olivary nucleus, dorsal circumference of the superior olivary nucleus, lateral circumference of the facial nucleus, dorsal circumference of the facial nucleus, and the DFA. The percentage of HRP-neurons in each cluster was 0.5 +/- 0.1% (mean +/- S.E., n=6), 15.2 +/- 1.9%, 23.7 +/- 0.9%, 52.5 +/- 1.7%, and 8.3 +/- 0.7%, respectively. Glutamate stimulation of the DFA (at 5.0 to 7.0 mm rostral to the obex, 2.8 to 4.0 mm lateral to the midline, and 2.5 to 3.5 mm ventral to the dorsal surface of the medulla), but not other areas, resulted in the increased CCA blood flow. The 5HT- and SP-immunoreactive nerve terminals abutted on the ChAT-immunoreactive cell body (preganglionic neurons) in the DFA. In conclusion, parasympathetic preganglionic neurons in the DFA project fibers to the SPG, are innervated by 5HT- and SP-like nerve terminals, and are responsible for regulation of the CCA blood flow. They may be also important in regulation of the cerebral blood flow.     

CSF-contacting and other somatostatin-immunoreactive neurons in the brains of Anguilla anguilla,Phoxinus phoxinus,and Salmo gairdneri (Teleostei)

Summary A system of somatostatin-immunoreactive neurons was demonstrated in the brains of the eel, Anguilla anguilla, the European minnow, Phoxinus phoxinus, and the rainbow trout, Salmo gairdneri, by means of the light-microscopic indirect immunoperoxidase technique.In the anterior periventricular nucleus, somatostatin-immunoreactive cerebrospinal fluid (CSF)-contacting neurons display intensely stained intraventricular dendritic protrusions, perikarya, and axonal processes. The latter taper into a somatostatin-immunoreactive fiber plexus extending to the infundibulum, the proximal neurohypophysis, and the lateral and mammillary recesses.In addition, somatostatin-immunoreactive neurons were demonstrated in the magnocellular preoptic, entopeduncular and dorsolateral thalamic nuclei, further in the pretectal area and the ventrolateral tegmentum. Somatostatin-immunoreactive fiber bundles project via the stria medullaris toward the habenular nucleus; they also course in the dorsomedial-ventrolateral direction at the level of the pretectal-tegmental area, and within the ventral and dorsal tegmentum.The presence of somatostatin in a variety of different neurons of the teleost brain is discussed in connection with their tentative inhibitory function. The CSF-contacting neurons of the anterior periventricular nucleus are supposed to function as sensors that pass information from the CSF to the somatostatin system of the hypothalamus and/or other components of the neuroendocrine apparatus.Dedicated to Professor Helmut Leonhardt, Kiel, on the occasion of his 65^th birthdayThis investigation was supported by grants from the Deutsche Forschungsgemeinschaft to A.O. (Ok 1/25-3) and H.W.K. (Ko 758/2-2)On leave of absence from the second Department of Anatomy, Semmelweis OTE, Budapest, Hungary     

Patterns of olfactory projections in the desert iguana,Dipsosaurus dorsalis

The neural organization of the olfactory system in the desert iguana, Dipsosaurus dorsalis, has been investigated by using the Fink-Heimer technique to trace the efferents of the main and accessory olfactory bulbs, and Golgi preparations to determine the spatial relations between olfactory afferents and neurons in the primary olfactory centers. The accessory olfactory bulb projects to the ipsilateral nucleus sphericus via the accessory olfactory tract. The main olfactory bulb projects to the ipsilateral telen-cephalon via four tracts. The medial olfactory tract projects to the rostral continuation of medial cortex and to the septum. The intermediate olfactory tract projects to the olfactory tubercle and retrobulbar formation. The lateral olfactory tract projects to the rostral part of lateral cortex. The intermediate and lateral olfactory tracts also merge caudally to form the stria medullaris, which crosses the midline in the habenular commissure and distributes fibers to the contralateral hemisphere via two tracts. The lateral corticohabenular tract terminates in the contralateral lateral cortex. The anterior olfactohabenular tract terminates in the contralateral olfactory tubercle, retrobulbar formation and septum. The relation of olfactory afferents to neurons in the medial cortex, lateral cortex, nucleus sphericus, and septum corresponds to a pattern of organization that is typical of many olfactorecipient structures. Such structures are trilaminar, with neurons whose somata are situated in the intermediate layer (layer 2) sending spine-laden dendrites into an outer, molecular layer (layer 1). Olfactory afferents intersect the distal segments of these dendrites. By contrast, other olfactorecipient structures in Dipsoaurus deviate from the familiar pattern. Olfactory afferents intersect somata lying in layer 2 of the retrobulbar formation. Olfactory afferents include some fibers which course perpendicularly to the surface of the olfactory tubercle and extend deep to layer 2.     

Development of the habenulointerpeduncular tract of rats

Development of the habenulointerpeduncular tract has been carried out on fixed brains obtained from 21 day rat embryos and from neonatal animals on the 0 and 9 days of postnatal development by DiI tracing method (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) along neuronal membranes. The marker was inserted into the nuclei of the habenula, the interpeduncular nucleus, and into the area of raphe nuclei. Neurons and fibers that contained DiI were identified on vibratome sections by fluorescent and confocal microscopy. We have found that reciprocal connections between the lateral habenular nucleus and raphe nuclei are formed in the prenatal period by stage E21. Raphe nuclei innervating neurons were located in dorso- and ventrocaudal parts of the lateral habenular nucleus. Projections of the medial habenular nucleus onto interpeduncular nucleus were found only in the postnatal period from P2. Neurons that provide a source of these projections form characteristic groups inside the medial habenular nucleus. Therefore, the present study for the first time describes heterogenic formation of different projection systems that are included in the habenulointerpeduncular tract of rats at perinatal ontogenesis.     

Projection of cervical dorsal root fibers to the medulla oblongata in the brush-tailed possum, Trichosurus vulpecula

This study describes the projection of cervical spinal afferent nerve fibers to the medulla in the brush-tailed possum, a marsupial mammal. After single dorsal roots (between C2 and T1) were cut in a series of animals, the Fink-Heimer method was used to demonstrate the projection fields of fibers entering the CNS via specific dorsal roots. In the high cervical spinal cord, afferent fibers from each dorsal root form a discrete layer in the dorsal funiculus. The flattened laminae from upper cervical levels are lateral and those from lower cervical levels are medial within the dorsal columns. All afferent fibers at this level are separated from gray matter by the corticospinal fibers in the dorsal funiculus. All cervical roots project throughout most of the length of the well-developed main cuneate nucleus in a loosely segmentotopic fashion. Fibers from rostral roots enter more lateral parts of the nucleus, and fibers from lower levels pass to more medial areas; but terminal projection fields are typically large and overlap extensively. At more rostral medullary levels, fibers from all cervical dorsal roots also reach the external cuneate nucleus. The spatial arrangement here is more complex and more extensively overlapped than in the cuneate nucleus. Rostral cervical root fibers reach ventral and ventrolateral areas of the external cuneate nucleus and continue to its rostral pole; more caudal root fibers project to more dorsal and medial regions within the nucleus. These results demonstrate that projection patterns of spinal afferents in this marsupial are similar to those seen in the few placental species for which detailed data concerning this system are available.     

Primary projections of the trigeminal nerve in two species of sturgeon: Acipenser oxyrhynchus and Scaphirhynchus platorynchus

Horseradish peroxidase histochemical studies of afferent and efferent projections of the trigeminal nerve in two species of chondrostean fishes revealed medial, descending and ascending projections. Entering fibers of the trigeminal sensory root project medially to terminate in the medial trigeminal nucleus, located along the medial wall of the rostral medulla. Other entering sensory fibers turn caudally within the medulla, forming the trigeminal spinal tract, and terminate within the descending trigeminal nucleus. The descending trigeminal nucleus consists of dorsal (DTNd) and ventral (DTNv) components. Fibers of the trigeminal spinal tract descend through the lateral alar medulla and into the dorsolateral cervical spinal cord. Fibers exit the spinal tract throughout its length, projecting to the ventral descending trigeminal nucleus (DTNv) in the medulla and to the funicular nucleus at the obex. Retrograde transport of HRP through sensory root fibers also revealed an ascending bundle of fibers that constitutes the neurites of the mesencephalic trigeminal nucleus, cell bodies of which are located in the rostral optic tectum. Retrograde transport of HRP through motor root fibers labeled ipsilateral cells of the trigeminal motor nucleus, located in the rostral branchiomeric motor column.     

The origin of central pinealopetal nerve fibers in the Mongolian gerbil as demonstrated by the retrograde transport of horseradish peroxidase

The location of perikarya and nerve fibers projecting via the habenular and posterior commissures from the brain into the pineal organ of the Mongolian gerbil was investigated by the use of the retrograde horseradish peroxidase (HRP)-tracing method. After microiontophoretic or hydraulic injection of the tracer into the superficial pineal gland via a glass micropipette, and after survival periods of 6 to 48 h, the animals were transcardially perfused and the brains processed for the histochemical demonstration of the enzyme. In the pineal stalk 15 to 20 nerve fibers, including 4 to 7 myelinated elements, were traced back to the brain. HRP-labeled perikarya were located in the medial and lateral habenular nuclei as well as in the nucleus of the posterior commissure. Few fibers projected rostrally to perikarya in the paraventricular nucleus of the hypothalamus. A striking and persistent finding was the labeling of fibers that, in the habenular area, bent laterad and continued ventral to the optic tract. These fibers originated from perikarya located in the dorsal nucleus of the lateral geniculate body. These results strongly suggest a central innervation of the pineal organ in the Mongolian gerbil originating from hypothalamic and limbic areas of the brain as well as from the optic system.