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Branched connections to the septum and to the entorhinal cortex from the hippocampus, amygdala, and diencephalon in the rat.

Neuronal cell populations giving origin to bifurcating projections to the septum and the entorhinal cortex were studied in the rat by means of double retrograde labeling using the fluorescent tracers Fast Blue and Diamidino Yellow. Double labeled pyramidal neurons were consistently detected in the temporal level of the CA1 area and subiculum of the hippocampal formation, where they represented at least 50% of the cells retrogradely labeled from the entorhinal injections. Double labeled neurons were also detected in the amygdala, where they prevailed in the basal complex. Scattered double labeled neurons were observed in a number of hypothalamic nuclei, with a slight predominance in the preoptic region. Finally, a few double labeled cells were detected in the midline thalamus, and especially in the thalamic paraventricular nucleus. In all these structures, double labeled neurons were located ispilaterally to the injection sites. The present data indicate that the septum and entorhinal cortex are tightly interconnected by axonal bifurcations deriving from a variety of telencephalic and diencephalic sources.

Amygdala↗

CyNodal, the Japanese newt nodal-related gene, is expressed in the left side of the lateral plate mesoderm and diencephalon.

The nodal and nodal-related genes play fundamental roles during deuterostome left-right axis formation. Several of these genes show left-sided expression in the lateral plate mesoderm and brain region. We have isolated the nodal-related gene, CyNodal, from Cynops pyrrhogaster. CyNodal mRNA is detected at the marginal zone and left side of several tissues. The left-sideness of CyNodal mRNA expression is highly conserved throughout vertebrate evolution. However, CyNodal mRNA expression shows little variation from the Xenopus nodal-related gene 1, in that CyNodal gene expression in the left lateral plate mesoderm shifts from posterior to anterior at least twice.

Amino Acid Sequence↗

Expression of Lrrn1 marks the prospective site of the zona limitans thalami in the early embryonic chicken diencephalon.

An unknown chicken gene selected from a published substractive hybridization screen (GenBank Accession No. ; [Christiansen, J.H., Coles, E.G., Robinson, V., Pasini, A., Wilkinson, D.G., 2001. Screening from a subtracted embryonic chick hindbrain cDNA library: identification of genes expressed during hindbrain, midbrain and cranial neural crest development. Mech. Dev. 102, 119-133.]) was deemed of interest because of its dynamic pattern of expression across the forebrain and midbrain regions. A 528bp fragment cloned from early chick embryo cDNA and used for in situ hybridization corresponded to part of the 3' untranslated region of the chicken gene Leucine-rich repeat neuronal protein 1 (Lrrn1). The expression of this gene, mapped in the embryonic chick brain between stages HH10 and HH26, apparently preconfigures the zona limitans thalami site before overt formation of this boundary structure. Apart of colateral expression in the forebrain, midbrain and hindbrain basal plate, the most significant expression of Lrrn1 was found early on across the entire alar plate of midbrain and forebrain (HH10). This unitary domain soon divides at HH14 into a rostral part, across alar secondary prosencephalon and prospective alar prosomere 3 (prethalamus; caudal limit at the prospective zona limitans), and a caudal part in alar prosomere 1 (pretectum) and midbrain. The rostral forebrain domain later downregulates gradually most extratelencephalic signal of Lrrn1, but the rostral shell of zona limitans retains expression longer. Expression in the caudal alar domain also changes by downregulation within its pretectal subdomain. Caudally, the midbrain domain ends at the isthmo-mesencephalic junction throughout the studied period. Embryonic Lrrn1 signal also appears in the somites and in the otic vesicle.

Amino Acid Sequence↗

The distribution of FOS-immunoreactive neurons in the brainstem, midbrain and diencephalon of fetal sheep in response to acute hypoxia in mid and late gestation.

FOS immunohistochemistry was used to map the distribution of neuronal pathways activated by hypoxia in fetal sheep. Conscious pregnant sheep were exposed to hypoxia (7-9% O2, 1-2% CO2, balance N2) for 2 h at either 100-105 days (n=5) or 130-133 days gestation (n=5); term is approximately 147 days. The hypoxia caused cessation of breathing movements at both fetal ages, and increased FOS staining in the medulla (area postrema, dorsal motor nucleus of vagus, nucleus solitary tract, ventrolateral medulla); pons (locus coeruleus and subcoeruleus, lateral and medial parabrachial nuclei); midbrain (habenula, periaqueductal grey, substantia nigra, areas ventrolateral to Red Nucleus); and hypothalamus (anterior and lateral hypothalamic areas, paraventricular and supraoptic nuclei). The results were essentially the same at both gestational ages, except that hypoxia increased FOS-staining in the habenula only in the older fetuses. The presence of FOS protein in pontomedullary cardiorespiratory nuclei at 100-105 days gestation indicates that the peripheral chemoreceptors respond to hypoxia at this early age, and in the subcoeruleus and medial parabrachial regions of the pons is consistent with lesion studies suggesting these areas mediate the inhibition of fetal breathing in response to hypoxia. FOS staining in the ventrolateral periaqueductal grey and habenula was unexpected, and suggests that pathways normally involved in response to noxious stimuli, or which are part of the hypothalamic 'defense' response are activated by hypoxia in the fetus. Some FOS-labelling could arise secondarily as a consequence of the cardiovascular and endocrine responses to hypoxia.

Animals↗

Extracellular signal-regulated kinases 1 and 2 phosphorylated neurons in the tele- and diencephalon of rat after visceral pain stimulation: an immunocytochemical study.

We aimed at verifying whether extracellular signal-regulated kinases (erks) 1 and 2 are activated, i.e. phosphorylated, in forebrain neurons after visceral pain stimulation (VPS). Ether and urethane anaesthetized rats received an intraperitoneal injection of acetic acid or were left untreated (ECT, UCT). After 2 h the animals were perfused. Paraffin embedded brain sections immunoreacted with an antibody selective for the phosphorylated erks. The light microscope analysis revealed only a few labelled neurons in ECT, while in UCT, positive cells were detected. In VPS rats (VPSR) positive cells were mainly distributed in regions, such as the hypothalamic anterior and thalamic paraventricular midline nuclei, amygdala, hippocampal and parahippocampal, insular and perirhinal cortex, involved in nociception and/or visceral activities. Our data suggest an association of erks activation with the emotional component of nociception; moreover, they show that erks activation is not suppressed by anaesthesia.

Acetic Acid↗

Reciprocal connections between the preglomerular nucleus and the central posterior/prepacemaker nucleus in the diencephalon of weakly electric fish, Apteronotus leptorhynchus.

The central posterior/prepacemaker nucleus of gymnotiform fish is a bilateral cell group located in the dorsal thalamus. This complex consists of approximately 10,000 neurons which can be divided into several subpopulations. One subpopulation comprised of a few hundreds of neurons projects to the pacemaker nucleus in the medulla oblongata, thus constituting the prepacemaker nucleus portion of this complex. By employing in vitro tract-tracing techniques, we have, in the present investigation, examined the pattern of connectivity formed by the central posterior/prepacemaker nucleus with a diencephalic cell group, the preglomerular nucleus. As demonstrated by anterograde and retrograde tracing, a subpopulation of several hundreds of neurons located in the central posterior/prepacemaker nucleus project to the ipsi- and contralateral preglomerular nucleus. Double-labelling experiments revealed that at least a fraction of these neurons also innervate the pacemaker nucleus. In the preglomerular nucleus, a large number of neurons give rise to projections that terminate in the ipsilateral central posterior/prepacemaker nucleus. The reciprocal connection between the central posterior/prepacemaker nucleus and the preglomerular nucleus may be used to relay sensory information directly conveyed to one of the two nuclei indirectly to the other nucleus. The existence of at least some central posterior/prepacemaker nucleus neurons projecting to both the preglomerular nucleus and the pacemaker nucleus may provide the morphological basis for the transmission of an efference copy of electromotor information produced by neurons in the central posterior/prepacemaker nucleus to the preglomerular nucleus.

Animals↗

Developmental expression of nitric oxide synthase in the rat diencephalon with special reference to the thalamic paratenial nucleus.

Using immunohistochemistry and in situ hybridization the distribution of nitric oxide synthase (NOS) was investigated in the rat brain during pre- and postnatal development. At E15 weak NOS-like immunoreactivity (NOS-LI) could be seen in the differentiation field of the anterior hypothalamus. At E17 strong NOS-LI was observed in the developing neurons of the hypothalamic paraventricular nucleus, supraoptic nucleus, anterodorsal nucleus and lateral hypothalamic areas. In the thalamic paratenial nucleus a strong NOS-LI was observed in these neurons at E17, E18 and P1 with a weaker intensity at P3, P7, P9 and P15, whereas at P30 and in adult rats no NOS-positive neurons could be detected. NOS expression at E17 and P3 was verified by in situ hybridization. These results suggest that NO may have a developmental role at least in one of the regions studied, the thalamic paratenial nucleus.

Aging↗

Ontogenic profile of the expression of the mu opioid receptor gene in the rat telencephalon and diencephalon: an in situ hybridization study.

The developmental profile of mu (mu) opioid receptor gene expression has been characterized in the embryonic, postnatal and adult rat brain by in situ hybridization histochemistry. By ED12, mu opioid receptor mRNA was detectable in the deep neuroepithelium of the cortical plate. In the developing rat central nervous system (ED13-PD40), transcripts were seen over numerous telencephalic and diencephalic structures, such as the olfactory bulb, caudate-putamen, nucleus accumbens, amygdaloid complex, hippocampal formation, hypothalamus and thalamus. In the vast majority of brain regions examined, the developmental profile of the mu opioid receptor gene expression is similar to that of its translated protein as established using receptor autoradiography. Once a hybridization signal is detected in the prenatal period, it gradually increased to reach maximal levels during the second and third postnatal weeks. By the end of the third postnatal week, mu opioid receptor mRNA levels decreased to reach amounts seen in adulthood. Our study demonstrates that mu opioid receptor gene expression is seen very early on in the embryonic rat brain with transient increases observed during the critical period of neurogenesis, neuronal migration and synaptogenesis, suggesting a role of this opioid receptor subtype in brain developmental processes.

Animals↗

Distribution of melanin-concentrating hormone (MCH)-like immunoreactivity in neurons of the diencephalon of sheep.

An immunohistochemical study with an antiserum raised against salmon melanin concentrating-hormone has demonstrated the presence of numerous melanin concentrating-hormone-immunoreactive neurons in the lateral hypothalamic areas of the sheep. The pattern of distribution of these perikarya is similar to that of rodents and primates. In sheep, however, melanin concentrating-hormone-immunoreactive neurons appeared to form two gatherings: the first is situated ventromedially to the internal capsule and the second in the dorsolateral hypothalamus. In these areas, numerous immunostained perikarya are observed. Compared to the rats, labelled neurons extended more caudally in the ventral tegmental area and more rostrally above the optic chiasma. Compared to primates, these neurons are less numerous in the periventricular area. In our study, dense networks of melanin concentrating-hormone-immunoreactive varicose fibers were observed in the supramamillary nucleus, the lateral hypothalamus, the nucleus medialis thalami and nucleus reuniens and in the bed nucleus of the stria terminalis.

Animals↗

Expression of a novel aristaless related homeobox gene 'Arx' in the vertebrate telencephalon, diencephalon and floor plate.

We have isolated a novel homeobox gene that is expressed in the vertebrate central nervous system and which shows striking similarity to the Drosophila al gene in the homeodomain (85% identity) and in a 17 amino acid-sequence near the carboxyl-terminus. This gene was designated Arx (aristaless related homeobox gene) in consideration of its structural similarity to the al gene. Arx was highly conserved between mouse and zebrafish. Neuromeric expression in the forebrain and longitudinal expression in the floor plate were observed in mouse and zebrafish. The expression of Arx in the ganglionic eminence and ventral thalamus overlapped regionally with that of Dlx1, but the cell layer where Arx is expressed differed from that of the Dlx1. This gene was also found to be expressed in the dorsal telencephalon (presumptive cerebral cortex) of mouse embryos. The structure and expression pattern of Arx with respect to any possible relationship to al and Dlx1, as well as the function of Arx in the floor plate are discussed.

Amino Acid Sequence↗

Mesotocin gene expression in the diencephalon of domestic fowl: cloning and sequencing of the MT cDNA and distribution of MT gene expressing neurons in the chicken hypothalamus.

This study focuses on the structure and expression of the mesotocin (MT) gene in the chicken hypothalamus. Using an anchored and nested RT-PCR strategy, combined with circular RACE-PCR, the full length sequence of the chicken MT cDNA was obtained. The cDNA and derived amino acid sequences conformed to the structure of the oxytocin-like gene family. However, unlike most mammalian species, there does not appear to be frequent gene conversion between the MT and AVT cDNA sequences. A single specific hybridization signal of 1.2 kb was detected by Southern analysis of chicken genomic DNA, indicating only a single gene copy in the chicken genome. Northern analysis of hypothalamic RNA revealed a single band at approximately 0.6 kb. Using the same probe for in situ hybridization histochemistry, MT-mRNA was demonstrated to be predominantly localized in the parvocellular, magnocellular and periventricular subgroups of the paraventricular nucleus and, when compared to the distribution of neurons containing arginine-vasotocin (AVT)-mRNA in the same region, with far fewer neurons expressing the MT gene in the lateral subgroups. Only few and scattered neurons expressing the MT gene were found in the ventral and external subgroups of the supraoptic nucleus in which many neurons contain AVT transcripts, as demonstrated in consecutive sections. In all nuclei investigated, the intensity of AVT and MT hybridization signals per cell was approximately equal. No specific labelling for MT-mRNA was found in the bed nucleus of the stria terminalis, nor the nucleus accumbens. Using immunocytochemical detection of AVT and in situ hybridization for neurons expressing MT-mRNA, some neurons were found to contain both AVT and MT gene products in the paraventricular nucleus but not in the supraoptic nucleus.

Amino Acid Sequence↗

Distribution of tyrosine hydroxylase-immunoreactive neurones in the diencephalon and mesencephalon of the coypu (Myocastor coypus).

The aim of the present study was to examine the catecholaminergic neurones located within the midbrain of the coypu, a South American hystricomorph rodent. The neuronal distribution of the catecholaminergic systems and morphological parameters of the immunostained cell bodies and fibres were investigated, using an immunohistochemical method. The brains of five coypu were fixed, immersed in gelatine-glycerol and cut in 40-micron slices using a freezing microtome. Samples were processed with ultrasound-based antigen retrieval and stained with labelled antityrosine hydroxylase monoclonal antibody. An image analyser was used to measure the neuronal bodies. The catecholaminergic neurones of the tuberoinfundibular system were mainly observed in the arcuate and periventricular nuclei with their axons projecting towards to the median eminence; they represented 28% of the global population of tyrosine hydroxylase-immunoreactive cells observed. Significant morphological differences were observed in comparison with the other two studied systems. Fifty per cent of total catecholaminergic neurones were detected in the nigrostriatal system distributed in the reticular and compact substance nigra. Most neuronal bodies had a fusiform aspect. The immunoreactive neurones of the mesolimbic system represented 22% of the total population. They were distributed around the interpeduncular nucleus. Two types of morphologically different catecholaminergic systems of the brain were established: hypothalamic neurones located in the periventricular and arcuate nuclei and mesencephalic neurones located in the substance nigra and interpeduncular nuclei. These systems showed morphological and probably physiological-pharmacological differences.

Animals↗