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PubMed1989

Evidence for substance P, serotonin and oxytocin input to medullary catecholamine neurons with diencephalic projections.

Using the retrograde transport of horseradish peroxidase (HRP) in combination with two-color immunoperoxidase staining, boutons stained with antisera to substance P (SP), serotonin (5HT) and oxytocin (OX) have been observed in contiguity with neurons in the rostral and caudal medulla that showed immunoreactivity for phenylethanolamine N-methyl transferase (PNMT) and tyrosine hydroxylase (TH), respectively, and which were backfilled with HRP injected into the diencephalon. The juxtaposition of these immunostained structures indicates that SP, 5HT and OX released from fibers in the medulla may affect the activity of adrenergic and noradrenergic medullary neurons that project to the diencephalon. Moreover, the presence of 5HT- and OX-immunoreactive processes in contiguity with medullary CA cells that send fibers to the diencephalon indicates that the raphe nuclei and the paraventricular nucleus of the hypothalamus can directly influence ascending pathways that are known to innervate the hypothalamus and appear to effect changes in vasopressin release.

Animals↗
PubMed1985

The effects of adrenalectomy on the ontogenesis of brain: noradrenaline and dopamine content.

The effect of adrenalectomy on catecholamine content in the diencephalon and the rest of the brain of male and female rats during the post-natal period was studied. Seven days after adrenalectomy, there is no change in noradrenaline or dopamine content. However, the dopamine levels of both the diencephalon and the rest of the brain decrease with age between days 45 and 60, while noradrenaline content in the diencephalon and the rest of the brain remained unchanged. Thus adrenalectomy significantly affected the developmental pattern of brain dopamine.

Adrenalectomy↗
PubMed2004

Levetiracetam protects against kainic acid-induced toxicity.

We investigated the Levetiracetam (LVT) ability to protect the brain against kainic acid (KA) induced neurotoxicity. Brain injury was induced by intraperitoneal administration of KA (10 mg/kg). Sham brain injury rats were used as controls. Animals were randomized to receive either LVT (50 mg/kg) or its vehicle (1 ml/kg) 30 min. before KA administration. Animals were sacrificed 6 hours after KA injection to measure brain malonildialdehyde (MDA), glutathione levels (GSH) and the mRNA for interleukin-1beta (IL-1beta) in the cortex and in the diencephalon. Behavioral changes were also monitored. Intraperitoneal administration of LVT decreased significantly MDA in the cortex (KA + vehicle = 0.25 +/- 0.03 nmol/mg protein; KA + LVT = 0.13 +/- 0.01 nmol/mg protein; P < 0.005), and in the diencephalons (KA + vehicle = 1,01 +/- 0.2 nmol/mg protein; KA + LVT = 0,33 +/- 0,08 nmol/mg protein; P < 0.005), prevented the brain loss of GSH in both cortex (KA + vehicle = 5 +/- 1 micromol/g protein; KA + LVT = 15 +/- 2 micromol/g protein; P < 0.005) and diencephalons (KA + vehicle = 9 +/- 0.8 micromol/g protein; KA + LVT = 13 +/- 0.3 micromol/g protein; P < 0.05), reduced brain IL-1beta mRNA and markedly controlled seizures. Histological analysis showed a reduction of cell damage in LVT treated samples. The present data indicate that LVT displays neuro-protective effects against KA induced brain toxicity and suggest that these effects are mediated, at least in part, by inhibition of lipid peroxidation.

Animals↗
PubMed2004

Altered BMP signaling disrupts chick diencephalic development.

The diencephalon is the caudal part of the forebrain and is organized into easily identifiable clusters of neurons called nuclei. Neurons in different nuclei project to discrete brain regions. Thus precise organization of the nuclei during forebrain development is necessary to build accurate neural circuits. How diencephalic development is regulated is poorly understood. BMP signaling participates in central nervous system patterning and development at many levels along the neural axis. Based on their expression we hypothesized BMPs play a role in diencephalic development. To test this hypothesis, we electroporated constitutively active and dominant negative forms of type I BMP receptors (Bmpr1a and Bmpr1b) into the embryonic chick forebrain. Ectopic induction of BMP signaling through constitutively active forms of the type I BMP receptors perturbs the normal gene expression patterns in the diencephalon and increases apoptotic cell death. These defects lead to disorganization of the diencephalic nuclei, suggesting BMP signaling is sufficient to modify diencephalic development. Loss-of-function studies, using dominant negative forms of Bmpr1a and Bmpr1b, indicate type I BMP receptors are necessary for normal eye and craniofacial development. However, they do not appear to be required for normal diencephalic development. In summary, our data indicate that while not necessary, BMP signaling via Bmpr1a and Bmpr1b, is sufficient to modify nuclear organization in the chick diencephalon.

Animals↗
PubMed2006

Sonic hedgehog from the basal plate and the zona limitans intrathalamica exhibits differential activity on diencephalic molecular regionalization and nuclear structure.

The diencephalon is the most complex area of the vertebrate brain, being particularly complex in amniotes. It has been suggested that diencephalic regionalization partially depends on local signaling mediated by sonic hedgehog (Shh). However, since the Shh gene is expressed in both the diencephalic basal plate and the zona limitans intrathalamica (ZLI), it is still unclear which of these tissues exerts morphogenetic influence on thalamic regionalization. In the present study using chick and quail embryos, we have found that although Shh from the ZLI and the basal plate induces ectopic expression of diencephalic genes in the posterior prosencephalic alar plate, only Shh originating from the ZLI can induce ectopic gene expression in the anterior alar plate, indicating that the ZLI exerts specific activity in the anterior epithelium. By introducing microbarriers between the diencephalic alar neuroepithelium and either the ZLI or the basal plate, we generated local loss of Shh expression in the ZLI, leading to alterations in molecular regionalization and subsequently, in the nuclear organization of the alar diencephalic derivatives on both sides of the ZLI. We thus demonstrate in vivo that basal signals are required to induce Shh expression in the ZLI and that Shh from the ZLI plays a pivotal role in regionalizing the alar diencephalon. The structural phenotype of Shh abolition in the ZLI consisted of a progressive pattern of alterations in diencephalic organization which were associated with the observed gradient effects in the molecular regionalization of the diencephalon. We conclude that the ZLI is a secondary organizer which exerts its morphogenetic activity through Shh.

Animals↗
PubMed2006

Clonal origin of the mammalian forebrain from widespread oriented mixing of early regionalized neuroepithelium precursors.

The forebrain is formed by remodeling and growth of the anterior neural plate. This morphogenesis occurs in response to inductive signals during gastrulation and neurulation but is poorly understood at the cellular level. Here, we have used the LaacZ method of single cells labeling to visualize, at E12.5, clones originated at early stages of mouse forebrain development. The largest clones show that single progenitors can give rise to neuroepithelial cells dispersed across the forebrain. A significant fraction of the clones, and even relatively small ones, populated both the diencephalon and the telencephalon, indicating that the clonal separation between diencephalic and telencephalic progenitors is transient and/or partial. However, two groups of large clones, populating either the diencephalon or the telencephalon, dispersed within their respective domains, suggesting an early regionalization between some diencephalic and telencephalic progenitors. Widespread oriented mixing within these territories and then clonal expansion into smaller domains probably follow this initial regionalization. These data are consistent with a model of progressive specification of forebrain domains. We propose that the ordered expansion of early regionalized progenitor pools for the diencephalon and telencephalon could establish a potential link between early inductive signals and forebrain morphogenesis.

Animals↗
PubMed1999

Expression of three Rx homeobox genes in embryonic and adult zebrafish.

The paired-class homeobox gene, Rx, is important in eye development. In this study we analyze expression patterns of three zebrafish Rx genes (Zrx1, 2, 3) in embryos and adults. All three genes show dynamic spatiotemporal patterns of expression. Zrx3 is expressed earliest, in the anteriormost region of the neural plate, in regions that give rise to ventral diencephalon and retinae. As development proceeds, Zrx3 expression is reduced in the lateral optic primordia, and is absent in the optic cup, but is retained at the ventral midline of the diencephalon, and is expressed in hypothalamus in the adult. As the neural retina begins to differentiate, Zrx3 is re-expressed in a subset of cells in the inner nuclear layer, presumably bipolar cells, and this expression is retained in the adult. In contrast, Zrx1/2 have a slightly later onset of expression, are initially coincident with Zrx3, but then become complementary, remaining on in the optic primordia but disappearing from the ventral midline of the diencephalon. Zrx1/2 are down-regulated as the retina differentiates, except in the outer nuclear layer where they continue to be expressed at high levels in cone, but not rod, photoreceptors. This is the first transcription factor described that distinguishes between cone and rod photoreceptors.

Animals↗
PubMed2004

SOX3 is required during the formation of the hypothalamo-pituitary axis.

The pituitary develops from the interaction of the infundibulum, a region of the ventral diencephalon, and Rathke's pouch, a derivative of oral ectoderm. Postnatally, its secretory functions are controlled by hypothalamic neurons, which also derive from the ventral diencephalon. In humans, mutations affecting the X-linked transcription factor SOX3 are associated with hypopituitarism and mental retardation, but nothing is known of their etiology. We find that deletion of Sox3 in mice leads to defects of pituitary function and of specific central nervous system (CNS) midline structures. Cells in the ventral diencephalon, where Sox3 is usually highly expressed, have altered properties in mutant embryos, leading to abnormal development of Rathke's pouch, which does not express the gene. Pituitary and hypothalamic defects persist postnatally, and SOX3 may also function in a subset of hypothalamic neurons. This study shows how sensitive the pituitary is to subtle developmental defects and how one gene can act at several levels in the hypothalamic-pituitary axis.

Animals↗
PubMed1998

Crossregulation between En-2 and Wnt-1 in chick tectal development.

En-1, En-2 and Wnt-1 are proposed to be essential signals for the development of the optic tectum in chick embryos. Drosophila engrailed and wingless, homologs of En (En-1 and En-2) and Wnt-1, respectively, have been shown to crossregulate each other. In the present paper, it is reported that crossregulation between En-2 and Wnt-1 is preserved in the development of the chick optic tectum. When En-2 is overexpressed by the replication competent retroviral vector, Wnt-1 is expressed ectopically at the dorsal midline of the diencephalon. When Wnt-1 is introduced extrinsically either by ectopic transplantation of mesencephalon, or by implantation of Wnt-1 producing cells, En-2 is induced ectopically at the dorsal midline of the tel-diencephalic border. Thus, ectopic expression of En-2 and Wnt-1 leads to crossregulation of each other in the chick brain. As diencephalon transdifferentiates into the optic tectum by an appropriate signal, the crossregulation of En-2 and Wnt-1 in the diencephalon may mimic the relationship required for early development in the tectum.

Animals↗
PubMed1985

gamma-Aminobutyric acid metabolism and behavioral effects after intraventricular injection of spermine in chicks.

Effects of intraventricularly injected spermine on behavior and electrocortical activity and gamma-aminobutyric acid (GABA) metabolism after a single dose of 1.13 mumol/animal were studied. Decrease in locomotor activity, sedation or sleep, and electrocortical synchronization that lasted approximately 2 h were observed. In addition spermine caused a significant increase in GABA content in diencephalon and brainstem, 30 min after administration. Concomitantly a significant increase of glutamate decarboxylase (GAD) activity was observed in cerebral hemispheres, diencephalon, and brainstem. Reduction in gamma-aminobutyrate: alpha-oxoglutarate amino-transferase (GABA-T) levels occurred in the diencephalon along with a significant increase of GABA-T in the brainstem. The present results demonstrate that spermine has the capacity to affect GABA metabolism and are in favor of the suggestion that endogenous polyamines may modulate GABAergic mechanisms.

4-Aminobutyrate Transaminase↗
PubMed1977

Diencephalic efflux of calcium ions in the monkey during exercise, thermal stress and feeding.

1. The diencephalon of the unanaesthetized macaque monkey was radio-labelled with calcium by a microinjection of 6-8 muC (45)Ca(2+) into the third cerebral ventricle through a permanently implanted cannula. Successive 5 min push-pull perfusions of the mid-line hypothalamic region with an artificial C.S.F. were carried out at a rate of 28 mul./min every 20 min. A washout curve of declining (45)Ca(2+) radioactivity was thus generated.2. When the monkey exercised strenuously on a special ;rowing machine' to obtain highly palatable banana pellets, its body temperature rose sharply. As the monkey exercised, during a sequence of push-pull perfusions, the concurrent efflux of (45)Ca(2+) ions increased markedly in the corresponding samples of diencephalic perfusate. This enhanced activity of calcium ions continued throughout a 30 min work period and persisted as long as the monkey's temperature was elevated in the interval immediately following exercise.3. Exposure of the monkey's trunk, between neck and thigh to cold air of 5 degrees C likewise augmented the amount of (45)Ca(2+) ions in the diencephalic push-pull perfusates; however, a similar exposure to air warmed to 35 degrees C failed to alter the pattern of (45)Ca(2+) efflux from the animal's diencephalon. If a fasted monkey consumed only the banana pellets but was not exercised, the level of (45)Ca(2+) in the perfusate also increased transiently, just at the onset of feeding.4. We conclude that a local change in calcium transport, binding or other cellular activity of the cation within the diencephalon could play an important role in the central mechanism underlying the set-point rise in a primate's temperature which accompanies vigorous exercise. Further, the results support the idea that this cation functions in the diencephalic control of metabolic heat production as well as in the overall processes of energy metabolism, particularly in relation to feeding.

Animals↗
PubMed1987

Distribution of dopamine in the forebrain and midbrain of the red-eared turtle, Pseudemys scripta elegans, reinvestigated using antibodies against dopamine.

The distribution of dopamine (DA) immunoreactivity in the forebrain and midbrain of the red-eared turtle, Pseudemys scripta elegans, was studied using recently developed antibodies against DA. DA-containing cells were found around the glomeruli of the olfactory bulb but not in the telencephalon proper. In the diencephalon DA cells were observed in the preoptic region, several parts of the periventricular hypothalamic nucleus, the periventricular organ, the ependymal wall of the infundibular recess, the lateral hypothalamic area and the pretectal posterodorsal nucleus. In the midbrain DA cells were found in the ventral tegmental area, the substantia nigra and the presumed reptilian homologue of the mammalian A8 cell group. Dopaminergic fibers and terminals were observed throughout the whole brain, particularly in the telencephalon and diencephalon. The olfactory tubercle, the striatum and the nucleus accumbens appear to have the most dense innervation, but the anterior olfactory nucleus and the septal area also show numerous DA fibers and terminals. Cortical areas are in general not densely innervated by DA fibers. Compared to the results obtained for a lizard, Gekko gecko, with the same antibodies, the results of the present study are very similar as regards the distribution of DA neurons, fibers and terminals. In having better developed DA cell groups in the midbrain and a stronger innervation of the striatum, Pseudemys resembles mammals more than does Gekko. In contrast, the many cerebrospinal fluid-contacting DA neurons in the hypothalamus of Pseudemys are a primitive feature of the diencephalon. The previous immunohistochemical study of Gekko, a lizard, and the present account of Pseudemys, a turtle, indicate that at least two different lines of evolution exist within the reptiles with regard to the DA innervation of the dorsal ventricular ridge. One, including turtles and, probably, crocodilians with a weak DA innervation; and another, represented by lizards, with a strong DA immunoreactivity.

Animals↗
PubMed1994

Ectopic induction and reorganization of Wnt-1 expression in quail/chick chimeras.

When grafted ectopically into the diencephalon of a chick host embryo, a portion of met-mesencephalon straddling the met-mesencephalic constriction has the capacity to induce En-2 expression in the surrounding host tissue. Subsequently, tectal and cerebellar structures, composed of both host and grafted cells, are reconstructed in this ectopic location at the expense of the host diencephalon. Previous experiments indicated that the induction of En-2 was correlated with Wnt-1 expression within the graft. The aim of the present study was: (i) to determine whether Wnt-1 expression was spatially regulated within the graft, (ii) to investigate whether host Wnt-1-expressing cells were also involved in the ectopic met-mesencephalic development and, if so, (iii) to localize these Wnt-1-positive domains in relation to the patterning of the ectopically developing met-mesencephalic territory. We studied the expression profile of Wnt-1, in relation with that of other positional markers, in quail/chick chimeras where various portions of met-mesencephalon had been grafted into the diencephalon. We found that Wnt-1 expression was reorganized within the graft, and that it was also induced in the host in contact with the graft. Moreover, these ectopic expressions of Wnt-1, in both the grafted and the surrounding host tissues, were organized in concert to form a continuous positive line at the host/graft junction, the location of which depended on the precise origin of the graft. Finally, we found that this line was frequently located at the limit between territories expressing different positional markers. We propose that Wnt-1 expression is turned on at the junction between domains of different phenotypes, and may be used as a border to stabilize these adjacent differently committed territories.

Animals↗
PubMed2000

Members of the bHLH-PAS family regulate Shh transcription in forebrain regions of the mouse CNS.

The secreted protein sonic hedgehog (Shh) is required to establish patterns of cellular growth and differentiation within ventral regions of the developing CNS. The expression of Shh in the two tissue sources responsible for this activity, the axial mesoderm and the ventral midline of the neural tube, is regulated along the anteroposterior neuraxis. Separate cis-acting regulatory sequences have been identified which direct Shh expression to distinct regions of the neural tube, supporting the view that multiple genes are involved in activating Shh transcription along the length of the CNS. We show here that the activity of one Shh enhancer, which directs reporter expression to portions of the ventral midbrain and diencephalon, overlaps both temporally and spatially with the expression of Sim2. Sim2 encodes a basic helix-loop-helix (bHLH-PAS) PAS domain containing transcriptional regulator whose Drosophila homolog, single-minded, is a master regulator of ventral midline development. Both vertebrate and invertebrate Sim family members were found sufficient for the activation of the Shh reporter as well as endogenous Shh mRNA. Although Shh expression is maintained in Sim2(-)(/)(-) embryos, it was determined to be absent from the rostral midbrain and caudal diencephalon of embryos carrying a dominant-negative transgene that disrupts the function of bHLH-PAS proteins. Together, these results suggest that bHLH-PAS family members are required for the regulation of Shh transcription within aspects of the ventral midbrain and diencephalon.

Animals↗
PubMed2001

Role of Pax3/7 in the tectum regionalization.

Pax3/7 is expressed in the alar plate of the mesencephalon. The optic tectum differentiates from the alar plate of the mesencephalon, and expression of Pax3/7 is well correlated to the tectum development. To explore the function of Pax3 and Pax7 in the tectum development, we misexpressed Pax3 and Pax7 in the diencephalon and ventral mesencephalon. Morphological and molecular marker gene analysis indicated that Pax3 and Pax7 misexpression caused fate change of the alar plate of the presumptive diencephalon to that of the mesencephalon, that is, a tectum and a torus semicircularis were formed ectopically. Ectopic tectum in the diencephalon appeared to be generated through sequential induction of Fgf8, En2 and Pax3/7. In ventral mesencephalon, which expresses En but does not differentiate to the tectum in normal development, Pax3 and Pax7 misexpression induced ectopic tectum. In normal development, Pax3 and Pax7 expression in the mesencephalon commences after Otx2, En and Pax2/5 expression. In addition, expression domain of Pax3 and Pax7 is well consistent with presumptive tectum region in a dorsoventral axis. Taken together with normal expression pattern of Pax3 and Pax7, results of misexpression experiments suggest that Pax3 and Pax7 define the tectum region subsequent to the function of Otx2 and En.

Animals↗
PubMed2002

A sonic hedgehog-dependent signaling relay regulates growth of diencephalic and mesencephalic primordia in the early mouse embryo.

Sonic hedgehog (Shh) is a key signal in the specification of ventral cell identities along the length of the developing vertebrate neural tube. In the presumptive hindbrain and spinal cord, dorsal development is largely Shh independent. By contrast, we show that Shh is required for cyclin D1 expression and the subsequent growth of both ventral and dorsal regions of the diencephalon and midbrain in early somite-stage mouse embryos. We propose that a Shh-dependent signaling relay regulates proliferation and survival of dorsal cell populations in the diencephalon and midbrain. We present evidence that Fgf15 shows Shh-dependent expression in the diencephalon and may participate in this interaction, at least in part, by regulating the ability of dorsal neural precursors to respond to dorsally secreted Wnt mitogens.

Animals↗
PubMed2003

Developmental changes in gonadotropin-inhibitory hormone in the Japanese quail (Coturnix japonica) hypothalamo-hypophysial system.

We previously isolated a novel dodecapeptide containing a C-terminal -Arg-Phe-NH(2) sequence, SIKPSAYLPLRF-NH(2) (RFamide peptide), from the Japanese quail (Coturnix japonica) brain. This novel quail peptide was shown to be located in neurons of the paraventricular nucleus (PVN) and their terminals in the median eminence (ME), and to decrease gonadotropin release from cultured anterior pituitary in adult birds. We therefore designated this peptide gonadotropin-inhibitory hormone (GnIH). Furthermore, a cDNA encoding the GnIH precursor polypeptide has been characterized. To understand the physiological roles of this peptide, in the present study we analyzed developmental changes in the expressions of GnIH precursor mRNA and the mature peptide GnIH during embryonic and posthatch ages in the quail diencephalon including the PVN and ME. GnIH precursor mRNA was expressed in the diencephalon on embryonic day 10 (E10) and showed a significant increase on E17, just before hatch. GnIH was also detected in the diencephalon on E10 and increased significantly around hatch. Subsequently, the diencephalic GnIH content decreased temporarily, and again increased progressively until adulthood. GnIH-like immunoreactive (GnIH-ir) neurons were localized in the PVN on E10, but GnIH-ir fibers did not extend to the ME. However, GnIH-ir neurons increased in the PVN on E17, just before hatch, and GnIH-ir fibers extended to the external layer of the ME, as in adulthood. These results suggest that GnIH begins its function around hatch and acts as a hypothalamic factor to regulate gonadotropin release in the bird.

Animals↗
PubMed1984

The developmental distribution of monoamines in the brain of male Japanese quail (Coturnix coturnix japonica).

Changes in the levels of the brain catecholamines, norepinephrine (NE) and dopamine (DA), were studied in the male Japanese quail during maturation. In addition the neurotransmitter, serotonin (5HT), its precursor, tryptophan ( TRYP ), and its metabolite, 5-hydroxyindoleacetic acid (5HIAA), were measured. Birds were sampled at weekly intervals between the ages of 1 and 63 days of age. Brains of the quail were dissected into cerebral hemispheres, diencephalon, brain stem, cerebellum, and optic lobes. Spectrophotofluorometric analyses of the monoamines in the brain parts were performed following the validation of this technique for use with the avian brain. Norepinephrine increased significantly (P less than .05) with age in all brain regions except the cerebellum, whereas dopamine increased significantly (P less than .05) only in the cerebral hemispheres. Serotonin levels increased significantly (P less than .05) in all brain regions except the cerebellum. The most dramatic change over maturation was observed in the cerebral hemispheres, and relatively smaller increases occurred in the diencephalon. Tryptophan decreased significantly (P less than .05) in concentration in all brain regions throughout the 9 weeks of development. However, 5HIAA did not change significantly in brain regions. Serum testosterone, as measured by testosterone radioimmunoassay, increased in conjunction with sexual maturation. In general, the monoamine levels rose with age. Additionally, the monoamine distribution appeared to increase in caudal to rostral direction. Changes in the monoamines, particularly in the diencephalon, occurred at approximately the time that increased peripheral concentrations of testosterone were also observed. This observation supports the contention that monoamines may be involved in regulating gonadotropin release during maturation.

Aging↗