Rostral Diencephalon Research Articles

The Stuff that Dreams Are Made on: The Case of Peduncular Hallucinosis

This study describes five patients with the syndrome of peduncular hallucinosis (PH): hallucinations that occur after lesions in the rostral pons, midbrain, or diencephalon — caused in these cases by acute ischemic or hemorrhagic stroke. The hallucinations were primarily visual, or both visual and auditory, and consisted of vivid scenes that involved people, animals, or objects experienced in the present. Each patient had several distinct hallucinated events. They were more likely to occur in the evening or at night but also occurred during the day. Each …

Zinc-finger genesFezandFez-likefunction in the establishment of diencephalon subdivisions

Fez and Fez-like (Fezl) are zinc-finger genes that encode transcriptional repressors expressed in overlapping domains of the forebrain. By generating Fez;Fezl-deficient mice we found that a redundant function of Fez and Fezl is required for the formation of diencephalon subdivisions. The caudal forebrain can be divided into three transverse subdivisions: prethalamus (also called ventral thalamus), thalamus (dorsal thalamus) and pretectum. Fez;Fezl-deficient mice showed a complete loss of prethalamus and a strong reduction of the thalamus at late gestation periods. Genetic marker analyses revealed that during early diencephalon patterning in Fez;Fezl-deficient mice, the rostral diencephalon (prospective prethalamus) did not form and the caudal diencephalon (prospective thalamus and pretectum) expanded rostrally. Fez;Fezl-deficient mice also displayed defects in the formation of the zona limitans intrathalamica (ZLI), which is located on the boundary between the prethalamus and thalamus. Fez and Fezl are expressed in the region rostral to the rostral limit of Irx1 expression, which marks the prospective position of the ZLI. Transgene-mediated misexpression of Fezl or Fez caudal to the ZLI repressed the caudal diencephalon fate and affected the formation of the Shh-expressing ZLI. These data indicate that Fez and Fezl repress the caudal diencephalon fate in the rostral diencephalon, and ZLI formation probably depends on Fez/Fezl-mediated formation of diencephalon subdivisions.

Morphogenesis and regionalization of the medaka embryonic brain.

We examined the morphogenesis and regionalization of the embryonic brain of an acanthopterygian teleost, medaka (Oryzias latipes), by in situ hybridization using 14 gene probes. We compared our results with previous studies in other vertebrates, particularly zebrafish, an ostariophysan teleost. During the early development of the medaka neural rod, three initial brain vesicles arose: the anterior brain vesicle, which later developed into the telencephalon and rostral diencephalon; the intermediate brain vesicle, which later developed into the caudal diencephalon, mesencephalon, and metencephalon; and the posterior brain vesicle, which later developed into the myelencephalon. In the late neural rod, the rostral brain bent ventrally and the axis of the brain had a marked curvature at the diencephalon. In the final stage of the neural rod, ventricles began to develop, transforming the neural rod into the neural tube. In situ hybridization revealed that the brain can be divided into three longitudinal zones (dorsal, intermediate, and ventral) and many transverse subdivisions, on the basis of molecular expression patterns. The telencephalon was subdivided into two transverse domains. Our results support the basic concept of neuromeric models, including the prosomeric model, which suggests the existence of a conserved organization of all vertebrate neural tubes. Our results also show that brain development in medaka differs from that reported in other vertebrates, including zebrafish, in gene-expression patterns in the telencephalon, in brain vesicle formation, and in developmental speed. Developmental and genetic programs for brain development may be somewhat different even among teleosts.

Developmental expression of a novel Ftz-F1 homologue, ff1b (NR5A4 ), in the zebrafish Danio rerio

The Ftz-F1 genes encode orphan receptors of the nuclear receptor superfamily. The mammalian Ftz-F1 homologue, SF-1, has been found to be essential for the proper development of the adrenal-gonadal axis and it also plays a critical role in mammalian sex-determination. We report here the isolation and characterisation of a novel zebrafish Ftz-F1 gene, ff1b. Whole-mount in situ hybridization revealed onset of expression in the developing rostral diencephalon at 22 h post-fertilization (h.p.f.). Later, at 30 h.p.f., transcripts could be detected in the anterior regions of the pancreatic anlagen. Expression in both locations peaks at 36 h.p.f. and disappears at around 48 h.p.f.

Responses of Single Neurons in the Toad’s Caudal Ventral Striatum to Moving Visual Stimuli and Test of Their Efferent Projection by Extracellular Antidromic Stimulation/Recording Techniques

Previous work in anuran amphibians has shown that activity in the caudal ventral striatum correlates with visuomotor activity: orienting responses toward prey fail to occur after striatal lesions. Thus it has been suggested that the striatum influences visually guided behavior. Therefore, the present study investigates visual response properties from neurons recorded in the striatum. Extracellular recordings of 104 single neurons of the cane toad’s (Bufo marinus) caudal ventral striatum (STR) reveal five different response properties: resting discharge activity uninfluenced by the visual test stimuli (group STR1, 24.0%); resting discharge activity increased by any moving visual object (STR2, 31.7%); preference to moving compact objects (STR3, 15.4%); preference to certain configurational moving objects (STR4a and b, 13.5%), and resting activity reduced by visual stimuli (STR5, 15.4%). The receptive fields of these neurons encompassed the contralateral (46%) or the entire field of vision (54%). Of the neurons recorded in the striatum, 34% responded to electrical stimuli applied in the rostral diencephalon to the ipsilateral lateral forebrain bundle (LFB) which connects the striatum with the optic tectum (e.g. either directly or via pretectum or tegmentum). Various electrically driven STR neurons (40%) have axons that project caudally through the LFB, which was suggested by their antidromic activation in response to electrical stimuli applied to the LFB in the rostral diencephalon. In the present study, the main striatal output is mediated by ‘motion detectors’ (STR2) and ‘compact object perceivers’ (STR3). It is suggested that the caudal ventral striatum is involved in visual attentional processes that allow the translation of perception into action.

Cardiovascular regulatory actions of the hypocretins in brain

The hypocretins, also known as the orexins, are alternate translation products of a single gene. The recognition of their production in neurons of the rostral diencephalon, and their axonal localization in brain sites known to be important in the control of appetite, led to the demonstration of their orexogenic actions. However, these peptides are not as potent as other appetite stimulating neuropeptides and they have been localized in areas of brain more related to cardiovascular function. We verified the orexogenic actions of hypocretin-1 (Hcrt-1) and hypocretin-2 (Hcrt-2) in an ad libitum feeding model and identified the threshold dose to be 1 nmol when given into the lateral cerebroventricle (i.c.v.). Even at threshold doses for feeding, both Hcrt-1 and Hcrt-2 given i.c.v. into conscious, unrestrained rats stimulated significant elevations in mean arterial blood pressure, that appeared dose related. These elevations were relatively long lasting, mirroring the time course of a pressor dose of angiotensin II (0.1 nmol i.c.v.); however, the magnitude of blood pressure elevation to hypocretin did not equal that of A II. These data suggest an additional, non-appetitive action of the hypocretins and indicate that the peptide and receptor mapping studies may have predicted important roles for the peptides in the central nervous system control of cardiovascular function.

Differential patterning of ventral midline cells by axial mesoderm is regulated by BMP7 and chordin.

Ventral midline cells in the neural tube have distinct properties at different rostrocaudal levels, apparently in response to differential signalling by axial mesoderm. Floor plate cells are induced by sonic hedgehog (SHH) secreted from the notochord whereas ventral midline cells of the rostral diencephalon (RDVM cells) appear to be induced by the dual actions of SHH and bone morphogenetic protein 7 (BMP7) from prechordal mesoderm. We have examined the cellular and molecular events that govern the program of differentiation of RDVM cells under the influence of the axial mesoderm. By fate mapping, we show that prospective RDVM cells migrate rostrally within the neural plate, passing over rostral notochord before establishing register with prechordal mesoderm at stage 7. Despite the co-expression of SHH and BMP7 by rostral notochord, prospective RDVM cells appear to be specified initially as caudal ventral midline neurectodermal cells and to acquire RDVM properties only at stage 7. We provide evidence that the signalling properties of axial mesoderm over this period are regulated by the BMP antagonist, chordin. Chordin is expressed throughout the axial mesoderm as it extends, but is downregulated in prechordal mesoderm coincident with the onset of RDVM cell differentiation. Addition of chordin to conjugate explant cultures of prechordal mesoderm and neural tissue prevents the rostralization of ventral midline cells by prechordal mesoderm. Chordin may thus act to refine the patterning of the ventral midline along the rostrocaudal axis.

Localization of nitric oxide-synthesizing neurons sending projections to the dorsal raphe nucleus of the rat

The origin of the nerve fibers immunoreactive for neuronal nitric oxide synthase (nNOS) in the rat dorsal raphe nucleus (DRN) was determined by combining the use of cholera toxin subunit b (CTb) as a retrograde tracer and nNOS immunohistochemistry with a monoclonal anti-nNOS antibody. Double labeled CTb-nNOS cell bodies were distributed from the rostral diencephalon to the caudal medulla oblongata, in about 20 areas of the brain. Several of the areas displaying double labeled cells are known for their involvement in the control of the sleep–wake cycle and/or transmission of nociception.

Sites of origin and patterns of migration of vasotocin/mesotocin neurons in developing brain of the chick

Vasotocin/mesotocin (VT/MT) producing neurons are known to migrate extensively during development of the hypothalamus. Birthdating studies as well as immunohistochemical studies suggested the possibility that VT/MT producing neurons originate from specific sites of the neural tube. Furthermore, a relationship between the site of origin and the eventual fate of VT/MT cells has been suggested. This study proposes to identify the sites of origin of VT/MT cells and to establish whether magnocellular and parvocellular VT/MT, and neuromodulatory and neurosecretory VT/MT arise from common or different areas of the developing neural tube. To do so, the embryological distribution of VT/MT producing neurons of the chick was studied with immunohistochemistry. Analysis of the youngest brains in which VT/MT cells could be detected (embryonic day 7.25, E7.25) suggested the presence of two separate sites of origin. The first site was located in the hypothalamic anlage, next to the third ventricle, and the second in the mesencephalon, next to the fourth ventricle. Three-dimensional reconstructions of the location of VT/MT cells throughout development substantiated the hypothesis that diencephalic VT/MT cells originate from the first site while mesencephalic ones originate from the second site. Mesencephalic VT/MT producing cells were confined to the nucleus of Edinger-Westphal and were only detectable during a brief period in development (E7.25-E10). Diencephalic VT/MT producing neurons were noted to form two main paths from their sites of origin to the rostral diencephalon. Quantitative analysis confirmed this caudal to rostral displacement. Magnocellular and parvocellular VT/MT+ cells were intermingled at the diencephalic site of origin as well as in the migratory paths. Neuromodulatory and neurosecretory VT/MT cells of the diencephalon appeared to be derived from a common diencephalic site of origin. These studies support the hypothesis that while specific groups of progenitors may be important in allowing their offspring to produce VT/MT, they do not appear to influence the morphological attributes (magnocellular vs. parvocellular), nuclear locations, or functional characteristics of these cells.

Catecholamines and Indoleamines in the Central Nervous System of a Urodele Amphibian: A Microdissection Study with Emphasis on the Distribution of Epinephrine (Part 2 of 2)

Individual brain nuclei and regions of the central nervous system of adult male roughskin newts (Taricha granulosa) were microdissected and the concentrations of norepinephrine, epinephrine, 3,4-dihydroxyphenylacetic acid, dopamine, 5-hydroxyindoleacetic acid, and serotonin were determined using high performance liquid chromatography (HPLC) with electrochemical detection. The pattern of distribution of these catecholamines and indoleamines revealed many similarities between this urodele and other vertebrates. The highest concentrations of biogenic amines were observed in brainstem, hypothalamic, and basal forebrain structures; the lowest concentrations were observed in the internal granule layer of the olfactory bulb and pallial structures of the telencephalon. High concentrations of catecholamines and indoleamines were found in hypothalamic periventricular regions that are known to include cerebrospinal fluid-contacting, monoamine-containing neuronal cell bodies. The rostral diencephalon, which included the preoptic recess organ, had high concentrations of the primary catecholamines, norepinephrine and dopamine, and extremely high concentrations of the secondary catecholamine epinephrine. The dorsomedial infundibular hypothalamic region, which included the paraventricular organ, had high concentrations of dopamine and serotonin. The lateral infundibular hypothalamic region, which included the nucleus infundibularis dorsalis, had high concentrations of each of the biogenic amines. The results revealed unique patterns of distribution for each of the catecholamines and indoleamines studied, and provided evidence that regions of the hypothalamus that include cerebrospinal fluid-contacting, monoamine-containing neuronal cell bodies are focal regions for the metabolism of multiple biogenic amines.

Catecholamines and indoleamines in the central nervous system of a urodele amphibian: a microdissection study with emphasis on the distribution of epinephrine.

Individual brain nuclei and regions of the central nervous system of adult male roughskin newts (Taricha granulosa) were microdissected, and the concentrations of norepinephrine, epinephrine, 3,4-dihydroxyphenylacetic acid, dopamine, 5-hydroxyindoleacetic acid, and serotonin were determined using high performance liquid chromatography (HPLC) with electrochemical detection. The pattern of distribution of these catecholamines and indoleamines revealed many similarities between this urodele and other vertebrates. The highest concentrations of biogenic amines were observed in brainstem, hypothalamic, and basal forebrain structures; the lowest concentrations were observed in the internal granule layer of the olfactory bulb and pallial structures of the telencephalon. High concentrations of catecholamines and indoleamines were found in hypothalamic periventricular regions that are known to include cerebrospinal fluidcontacting, monoamine-containing neuronal cell bodies. The rostral diencephalon, which included the preoptic recess organ, had high concentrations of the primary catecholamines, norepinephrine and dopamine, and extremely high concentrations of the secondary catecholamine epinephrine. The dorsomedial infundibular hypothalamic region, which included the paraventricular organ, had high concentrations of dopamine and serotonin. The lateral infundibular hypothalamic region, which included the nucleus infundibularis dorsalis, had high concentrations of each of the biogenic amines. The results revealed unique patterns of distribution for each of the catecholamines and indoleamines studied, and provided evidence that regions of the hypothalamus that include cerebrospinal fluid-contacting, monoamine-containing neuronal cell bodies are focal regions for the metabolism of multiple biogenic amines.

Unusual brain nucleus in the diencephalon of a perciform (nototheniid) fish from Antarctica.

This cytoarchitectonic analysis on the rostral diencephalon of antarctic perciform fishes confirms the previously established perciform diencephalic body plan. However, it resulted in the detection of an additional, large and conspicuous brain nucleus (unusual nucleus, UN) in the antarctic nototheniid Pleuragramma antarcticum. This nucleus has not been seen before in any other perciform. The UN is probably homologous to a visually related nucleus (named nucleus rostrolateralis and lateral entopeduncular nuclear area, respectively), that has so far only been found in two most primitive (osteoglosso-form) and one most highly derived (atherinoform) species of teleost. Our present results indicate that this brain region, which belongs to the ancient diencephalic organization of ray-finned fishes, is expressed in the brains of fishes due to their particular requirements regarding life styles rather than due to their phylogenetical relationships.

Notes on the organization of the rostral diencephalon of the Atherinomorph Swordtail-Fish Xiphophorus helleri

By using conventional histological methods, the nuclear organization of the rostral diencephalon of the swordtail-fish, Xiphophorus helleri (Poecilidae, Atherinomorpha, Teleostei) was analyzed and compared with those from other teleost species. The subdivisions of the entopeduncular region, especially in their relative position and cytoarchitectonic structure, reveal clear differences between the various teleost orders. A new nucleus, the lateral entopeduncular nuclear area (leNA), found within this region, is described.

Cellular distribution of immunoreactive somatostatin in the basal forebrain of horses and ponies

Summary In other experimental models, somatostatin (SRIF) helps coordinate central nervous system (CNS) activity associated with the response to stress. It also plays an important role in the effects of exercise, stress and bacterial infections on growth hormone (GH) release and it may interact with specific SRIF receptors found on lymphocytes. Pituitary GH is a potent immunomodulatory hor- mone which is important to normal function of the thymus and a wide range of immune defense mechanisms. This experiment mapped the distribution of immunoreactive (ir) SRIF in the basal forebrain of horses and ponies. Results are compared to those for other species in order to determine if the anatomical substrate for stress-dependant SRIF effects may be similar to that described for other animal models. Immersion fixed brain tissues were obtained from animals submitted for euthanasia and/or necropsy. Alternate sections were stained for ir-SRIF using characterized polyclonal SRIF antibody and the peroxidase-anti-peroxidase technique, as previously described. Distribution of SRIF cells and fibers was mapped on projection drawings following light microscopic examination. Tissues in this analysis included cell groups at the border between the telencephalon and the rostral diencephalon, the hypothalamus, caudate nucleus (n) subthalamus and thalamus. At rostral levels, ir-SRIF was widely dispersed throughout medial tissues. Caudally, pronounced immunoreactivity was found in ventromedial tissues. Reactive perikarya were localized in the caudate n, olfactory tuberculum, n of the lamina terminalis, rostral preoptic n, suprachiasmatic n, periventricular n, supraoptic n, paraventricular n, arcuate n and ventromedial n. Numerous SRIF fibers extended into the lateral septum, all preoptic n, paraventricular n, rostral hypothalamic n, dorsomedial n, ventromedial n and mammillary n. Reactive fibers were also found in midline thalamic n, perifomical areas, the zona incerta and the habenular n. This distribution is similar to that described in most vertebrates. It reflects the role of SRIF in multiple neurocrine interactions as well as its neuroendocrine activity. Results from this experiment suggest that equine SRIF is likely to participate in neural and endocrine pathways similar to that for other mammalian species. Similar localization patterns also suggest that the coordination of SRIF cell functions within the equine CNS may be comparable to that in other species. This suggests that studies in other species are likely to facilitate the characterization of control mechanisms for SRIF secretion in the equine species.

The diencephalon and optic tectum of the longnose gar, Lepisosteus osseus (L.): cytoarchitectonics and distribution of acetylcholinesterase.

The cytoarchitecture of nuclei in the diencephalon and the distribution of acetylcholinesterase (AChE) in the diencephalon and optic tectum were analyzed in the longnose gar, Lepisosteus osseus, a non-teleost actinopterygian fish. Nuclei were identified in the preoptic area, thalamus, posterior tubercle, hypothalamus, synencephalon, and pretectum which are homologous to like-named nuclei in teleosts and other non-teleost actinopterygians. Of particular note, a nucleus in the rostral diencephalon, nucleus rostrolateralis, which has previously been identified only in the osteoglossomorph Pantodon, is present in the long-nose gar. The posterior pretectal nucleus, previously identified in teleosts and in the bowfin Amia, is also present in gars. The small size of the posterior pretectal nucleus in gars supports the hypothesis that this nucleus was small plesiomorphically. The distribution of AChE in the diencephalon and optic tectum corresponds in most respects to that found in teleosts. The superficial pretectal nuclei, including the posterior pretectal nucleus, are strongly positive for AChE. In contrast, most of the nuclei within the preglomerular complex are negative for AChE. Acetylcholinesterase is present in some of the fibers in the optic tracts and in most retinorecipient nuclei, as well as in some other nuclei and tracts.

Reciprocal connections between the ‘nucleus rotundus’ and the dorsal lateral telencephalon in the weakly electric fish Gnathonemus petersii

Injections of horseradish peroxidase into the dorsal lateral telencephalon of the weakly electric mormyrid fish Gnathonemus petersii resulted in retrogradely filled cells and anterogradely labelled terminals in the ‘nucleus rotundus’ of the ipsilateral rostral diencephalon. This connection courses via the lateral part of the lateral forebrain bundle. The present results suggest a particularly close relationship between the Dla cell group of the lateral telencephalon and the ‘nucleus rotundus’.

Retinal Projections in the Freshwater Butterfly Fish, <i>Pantodon buchholzi</i> (Osteoglossoidei) (Part 2 of 2)

The freshwater butterfly fish, Pantodon buchholzi, is a member of the most primitive radiation of teleosts. The retinofugal projections were studied in this fish with autoradiographic and horseradish peroxidase (HRP) methods, and the cytoarchitecture of the retinorecipient regions in the diencephalon and pretectum was analyzed with Bodian-, cresylecht-violet- and acetylcholinesterase-reacted sections. The rostral diencephalon of Pantodon contains a large retinorecipient nucleus, not previously identified in any other fish, i.e. nucleus rostrolateralis. Other nuclei that are described correspond to those previously recognized in other species. The majority of retinorecipient nuclei are positive for acetylcholinesterase, particularly those in the pretectum, as has been found in other species of teleosts. Most of the retinofugal fibers decussate in the optic chiasm. Some fibers project via the axial optic tract to preoptic nuclei and a region in the rostral hypothalamus. Fibers leave the medial optic tract to terminate in nucleus rostrolateralis and in dorsal and ventral thalamic nuclei, accessory optic and tubercular nuclei, periventricular and central pretectal nuclei, and sparsely in the deep tectal fascicle and terminal field. Dorsal optic tract fibers project to the dorsal accessory optic nucleus, superficial and central pretectal nuclei, and superficial and deep tectal layers. Ventral optic tract fibers project to the superficial pretectum, accessory optic nuclei, posterior tuberculum, nucleus corticalis in the central pretectum, and superficial tectal layer. Fibers that remain in the ipsilateral optic tract project to most of the targets reached by contralaterally projecting fibers. A few fibers in the contralateral medial optic tract redecussate via the posterior commissure to reach the ipsilateral periventricular pretectum. No labeled retinopetal cells caudal to the olfactory bulb were identified in any of the HRP cases.

The distribution of microglia and cell death in the fetal rat forebrain

The appearance and distribution of microglia in the fetal and early postnatal rat forebrain have been examined with the aid of a peroxidase-conjugated lectin derived from Griffonia simplicifolia. This distribution has in turn been correlated with that of pyknotic figures in the same Nissl-counterstained sections. Round and ameboid microglia may be recognised in the fetal forebrain as early as E11, at a stage when the telencephalic vesicles are beginning to develop. By E13, concentrations of round microglia are found at the dorsal and rostral limits of the diencephalic vesicle (dorsal lamina terminalis) and in the adjacent medial walls of the telencephalic vesicles. These cells are often seen to have pyknotic material within their cytoplasm. Microglia remain concentrated in this region until E17. From E15, blood vessels and round and ameboid microglia concentrate in the region of the future hippocampus and appear to be drawn into the hippocampal fissure as the cortical plate folds to form Ammon's horn. At E15, ameboid microglia are also concentrated in the developing fornix, which first becomes apparent at this age. Microglia remain concentrated in the septomesocortical junction area, and may contribute to the concentrations of microglia previously reported in the region of the developing corpus callosum and cavum septi pellucidi. Microglia probably concentrate in the dorsal lamina terminalis and medial telencephalon at E13 in response to the cell death noted in this region, but other concentrations of microglia in the forebrain are not accompanied by similar aggregations of cell death. These findings indicate that the junction of the telencephalon and rostral diencephalon attracts concentrations of microglia from E13 throughout fetal and early postnatal life, coincident with the infolding of the hippocampus (E13-E19) and several days before the development of the corpus callosum (from E19 onwards).

Retinal Projections in the Freshwater Butterfly Fish, <i>Pantodon buchholzi</i> (Osteoglossoidei) (Part 1 of 2)

The freshwater butterfly fish, Pantodon buchholzi, is a member of the most primitive radiation of teleosts. The retinofugal projections were studied in this fish with autoradiographic and horseradish peroxidase (HRP) methods, and the cytoarchitecture of the retinorecipient regions in the diencephalon and pretectum was analyzed with Bodian-, cresylecht-violet- and acetylcholinesterase-reacted sections. The rostral diencephalon of Pantodon contains a large retinorecipient nucleus, not previously identified in any other fish, i.e. nucleus rostrolateralis. Other nuclei that are described correspond to those previously recognized in other species. The majority of retinorecipient nuclei are positive for acetylcholinesterase, particularly those in the pretectum, as has been found in other species of teleosts. Most of the retinofugal fibers decussate in the optic chiasm. Some fibers project via the axial optic tract to preoptic nuclei and a region in the rostral hypothalamus. Fibers leave the medial optic tract to terminate in nucleus rostrolateralis and in dorsal and ventral thalamic nuclei, accessory optic and tubercular nuclei, periventricular and central pretectal nuclei, and sparsely in the deep tectal fascicle and terminal field. Dorsal optic tract fibers project to the dorsal accessory optic nucleus, superficial and central pretectal nuclei, and superficial and deep tectal layers. Ventral optic tract fibers project to the superficial pretectum, accessory optic nuclei, posterior tuberculum, nucleus corticalis in the central pretectum, and superficial tectal layer. Fibers that remain in the ipsilateral optic tract project to most of the targets reached by contralaterally projecting fibers. A few fibers in the contralateral medial optic tract redecussate via the posterior commissure to reach the ipsilateral periventricular pretectum. No labeled retinopetal cells caudal to the olfactory bulb were identified in any of the HRP cases.

Ontogeny of immunoreactive gonadotropin-releasing hormone neuronal systems in amphibians

The ontogeny of gonadotropin-releasing hormone (GnRH) systems was investigated in 3 anuran amphibians (genus Rana) by means of immunocytochemical (ICC) techniques and antibodies generated against 3 different forms of GnRH. Antisera that recognize primarily chicken II and mammalian GnRHs revealed two anatomically and developmentally distinct GnRH systems. One system, referred to here as the forebrain-spinal cord system, contained GnRH immunoreactive (ir) fibers extending from the rostral diencephalon through the ventromedial brainstem to the spinal cord. Intensity of labeling was robust in the youngest, premetamorphic tadpoles, but decreased with age. GnRH immunolabeling in the hypothalamic-pituitary tract was not detected until late prometamorphosis and increased with age. Development of GnRHir in the hypothalamic-pituitary tract coincided with first appearance of GnRHir in the terminal nerve in R. catesbeiana, but not in R. cascadae or R. aurora, suggesting species differences. Comparisons of results obtained with antisera to different forms of GnRH support the interpretation that the forebrain-spinal cord system, hitherto undescribed in amphibians, develops first and synthesizes a non-mammalian, chicken II-like GnRH, and that the hypothalamic-pituitary system develops later and synthesizes primarily mammalian GnRH. We speculate that the forebrain-spinal cord system may represent a GnRH innervation of frog sympathetic ganglia, and that the two GnRH systems are chemically and embryonically distinct.