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Microglia in the cerebral wall of the human telencephalon at second trimester.

We have recently begun to gain a clearer understanding of the phasing and patterns of colonization of the developing human brain by microglia. In this study we investigated the distribution, morphology and phenotype of microglia specifically within the wall of the human telencephalon from 12 to 24 gestational weeks (gw), a period that corresponds to the development of thalamocortical fibres passing through the transient subplate region of the developing cerebral wall. Sections from a total of 45 human fetal brains were immunoreacted to detect CD68 and MHC class II antigens and histochemically reacted with RCA-1 and tomato lectins. These markers were differentially expressed by anatomically discrete populations of microglia in the cerebral wall: two cell populations were noted during the initial phase of colonization (12-14 gw): (i) CD68++ RCA-1+ MHC II- amoeboid cells aligned within the subplate, and (ii) RCA-1++ CD68- MHC II- progenitors in the marginal layer and lower cortical plate that progressively ramified within the subplate, without seemingly passing through an 'amoeboid' state. At this stage microglia were largely absent from the germinal layers and the intermediate zone. From 14 to 15 gw, however, MHC class II positive cells were also detected within germinal layers and in the corpus callosum, and these cells, which coexpressed CD68 antigen (a marker associated with phagocytosis), further populated the lower half of the telencephalon from 18 to 24 gw. These findings are discussed in relation to developmental events that take place during the second trimester within the wall of the telencephalon.

Antigens, CD↗

Early neuronal and glial determination from mouse E10.5 telencephalon embryonic stem cells: an in vitro study.

We report here a novel in vitro model for differentiating neuronal and glial cells from mouse embryonic day 10 telencephalon stem cells. At this developmental stage, the telencephalon consists of a single layer of neuroepithelial stem cells. We used various markers of proliferation and differentiation (Ki-67, nestin, BrdU, Tuj-1 and GFAP) to follow proliferative progenitors and to identify neuronal and glial derivatives. Neuronal derivatives were obtained from nestin+ progenitors. GFAP+ astrocytic derivatives were detected after only 72 h of culture. Both neuronal and glial derivatives were generated close to nestin-positive aggregates. In addition, we were able to manipulate neuronal determination of telencephalon stem cells by gene transient transfection as demonstrated by RP42 gene overexpression. These observations suggest that this in vitro model is of potential use for studying early steps in neuronal or glial determination from embryonic stem cells, an issue of key importance for adult brain cell therapy approaches.

Animals↗

The effects of telencephalic lesions on visually mediated prey orienting behavior in the leopard frog (Rana pipiens). II. The effects of limited lesions to the telencephalon.

Unilateral removal of the telencephalon in the leopard frog, Rana pipiens, produces a contralateral deficit in visual prey orienting behavior [Patton and Grobstein, 1997]. In mammals, such deficits are most commonly associated with damage to the isocortex, a pallial derived structure. In contrast, we here report that in leopard frogs, lesions that remove substantial areas of one telencephalic lobe, including virtually the entire pallium, have no discernible effect on visual orienting behavior. Restricted lesions to the ventrocaudal telencephalon, however, produce an effect that closely resembles that produced by the complete removal of one telencephalic lobe. The 'critical area' that is both included in all lesions that are effective in producing a severe deficit and excluded from all ineffective lesions includes a portion of the caudal striatum. The striatum is known to play a significant role in anuran vision. It thus seems likely that the deficit produced by unilateral removal of the telencephalon in the leopard frog is due specifically to the removal of the caudal striatum. Unilateral lesions to the striatum have previously been shown to produce a contralateral deficit in visual orienting behavior in cats, and a role for the striatonigral pathway in the production of the visual orienting deficit that follows visual cortex lesions has been proposed. The current findings call attention to the possible general importance of the striatum in the control of vertebrate visual orienting behaviors.

Animals↗

Identification and isolation of a full-length clone of mouse GMFB (Gmfb), a putative intracellular kinase regulator, differentially expressed in telencephalon.

We identified new transcribed sequences, using a differential display paradigm to select genes expressed in proliferating neuroblasts from mouse telencephalon at 10 days of embryonic development. In this systematic search, we isolated a 361-bp partial 3' untranslated region (3' UTR) homologous to the 3' UTR of the human gene encoding a putative intracellular kinase regulator, glia maturation factor beta (GMFB). We cloned a full-length, 4,311-bp mouse cDNA containing a 270-bp 5' UTR, a 3,615-bp 3' UTR, and an open reading frame of 426 nucleotides encoding a putative 142 amino-acid protein, identical to human GMFB, with the exception of two amino acids. This 4.3-kb transcript is present in a variety of adult tissues and is developmentally regulated as shown by Northern blot analysis. Differential expression in telencephalon was demonstrated by quantification of radioactive relative RT-PCR and confirmed by in situ hybridization. The isolation of this full-length clone of mouse Gmfb should facilitate investigation of the intracellular mechanisms involved in the development of telencephalon.

3' Untranslated Regions↗

Notch3 signaling promotes radial glial/progenitor character in the mammalian telencephalon.

The Notch signaling pathway is known to influence cell fate in the developing mammalian nervous system. Previous work in the mouse telencephalon has shown that activated Notch1 promotes radial glial and astrocytic character in vivo, and fibroblast growth factor (FGF)-responsive neural progenitor character in vitro. In light of studies suggesting that Notch3 can antagonize Notch1, we tested the effects of activated Notch3 (NICD3) in the mouse telencephalon. Infection of embryonic day 9.5 telencephalic progenitors in vivo with NICD3 promoted radial glial/progenitor character embryonically and astrocyte fate postnatally. In addition, expression of NICD3 in telencephalic progenitors in vitro increased neurosphere frequency in FGF2, but was incompatible with neurosphere growth in epidermal growth factor (EGF). Thus, in the developing telencephalon, Notch1 and Notch3 function similarly, and may activate similar signaling cascades. Consistent with this notion, expression of an activated form of the Notch effector CBF1 (CBF1-VP16), or of the pathway target Hes5 promoted radial glial/progenitor character in vivo. Interestingly, unlike NICD1 and NICD3, CBF1-VP16 and Hes5 did not inhibit neurosphere growth in EGF, suggesting that this effect may be mediated at least in part by CBF1/Hes-independent signaling.

Animals↗

Distribution of carboxylic esterases in the telencephalon and diencephalon of a microchiropteran bat (Taphozous melanopogon Temminck).

The contribution deals with the histoenzymological makeup of telencephalon and diencephalon of a microchiropteran bat which is considered to have a primitive type of brain. Three esterases, namely acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and nonspecific esterase (NSE), have been studied. In comparison with other mammalian brains the telencephalon and diencephalon of the bat demonstrate a high concentration of enzyme, especially in the nucleus caudatus and putamen, stria medullaris and the nuclei of the thalamus. AChE concentration is higher in neurons, while BChE is seen in almost all fibrous areas of the telencephalon and diencephalon. NSE activity is prominent in neurons and neuropil of neocortex, thalamus and hypothalamus. All the above data indicate that the brain of echolocating microchiropteran bats is metabolically much more active than that of other mammals. This is specifically true of the auditory system which is considerably specialized in these animals; this is also confirmed by the present histoenzymological study.

Acetylcholinesterase↗

Tlx and Pax6 co-operate genetically to establish the pallio-subpallial boundary in the embryonic mouse telencephalon.

We have examined the role of Tlx, an orphan nuclear receptor, in dorsal-ventral patterning of the mouse telencephalon. Tlx is expressed broadly in the ventricular zone, with the exception of the dorsomedial and ventromedial regions. The expression spans the pallio-subpallial boundary, which separates the dorsal (i.e. pallium) and ventral (i.e. subpallium) telencephalon. Despite being expressed on both sides of the pallio-subpallial boundary, Tlx homozygous mutants display alterations in the development of this boundary. These alterations include a dorsal shift in the expression limits of certain genes that abut at the pallio-subpallial boundary as well as the abnormal formation of the radial glial palisade that normally marks this boundary. The Tlx mutant phenotype is similar to, but less severe than, that seen in Small eye (i.e. Pax6) mutants. Interestingly, removal of one allele of Pax6 on the homozygous Tlx mutant background significantly worsens the phenotype. Thus Tlx and Pax6 cooperate genetically to regulate the establishment of the pallio-subpallial boundary. The patterning defects in the Tlx mutant telencephalon result in a loss of region-specific gene expression in the ventral-most pallial region. This correlates well with the malformation of the lateral and basolateral amygdala in Tlx mutants, both of which have been suggested to derive from ventral portions of the pallium.

Amygdala↗

Mash1 regulates neurogenesis in the ventral telencephalon.

Previous studies have shown that mice mutant for the gene Mash1 display severe neuronal losses in the olfactory epithelium and ganglia of the autonomic nervous system, demonstrating a role for Mash1 in development of neuronal lineages in the peripheral nervous system. Here, we have begun to analyse Mash1 function in the central nervous system, focusing our studies on the ventral telencephalon where it is expressed at high levels during neurogenesis. Mash1 mutant mice present a severe loss of progenitors, particularly of neuronal precursors in the subventricular zone of the medial ganglionic eminence. Discrete neuronal populations of the basal ganglia and cerebral cortex are subsequently missing. An analysis of candidate effectors of Mash1 function revealed that the Notch ligands Dll1 and Dll3, and the target of Notch signaling Hes5, fail to be expressed in Mash1 mutant ventral telencephalon. In the lateral ganglionic eminence, loss of Notch signaling activity correlates with premature expression of a number of subventricular zone markers by ventricular zone cells. Therefore, Mash1 is an important regulator of neurogenesis in the ventral telencephalon, where it is required both to specify neuronal precursors and to control the timing of their production.

Animals↗

Wnt and Bmp signalling cooperatively regulate graded Emx2 expression in the dorsal telencephalon.

Pattern formation of the dorsal telencephalon is governed by a regionalisation process that leads to the formation of distinct domains, including the future hippocampus and neocortex. Recent studies have implicated signalling proteins of the Wnt and Bmp gene families as well as several transcription factors, including Gli3 and the Emx homeobox genes, in the molecular control of this process. The regulatory relationships between these genes, however, remain largely unknown. We have used transgenic analysis to investigate the upstream mechanisms for regulation of Emx2 in the dorsal telencephalon. We have identified an enhancer from the mouse Emx2 gene that drives specific expression of a lacZ reporter gene in the dorsal telencephalon. This element contains binding sites for Tcf and Smad proteins, transcriptional mediators of the Wnt and Bmp signalling pathway, respectively. Mutations of these binding sites abolish telencephalic enhancer activity, while ectopic expression of these signalling pathways leads to ectopic activation of the enhancer. These results establish Emx2 as a direct transcriptional target of Wnt and Bmp signalling and provide insights into a genetic hierarchy involving Gli3, Emx2 and Bmp and Wnt genes in the control of dorsal telencephalic development.

Animals↗

Embryogenesis of the rat telencephalon--a morphologic and stereologic analysis.

Comparative researches of borderline between telencephalon neuroepithelium and its surrounding mesenchyme in successive early developing stages lack in literature. The aim of this investigation was to carry out systematic morphologic and stereologic analyses of rat telencephalon in early developmental stages. We analysed semithin (1 microm) serial sections of rat embryonic brain from the 12th (E12) to the 15th (E15) day of gestation. The surface densities (SV) of an external mesenchymal surface, an internal mesenchymal surface and a neuroepithelial (ventricular) surface were examined stereologically and compared. The surface density of the external mesenchymal surface was the biggest at E12 (4,05 mm-1) and the least at E15 (1,87 mm-1)-p<0,0005. The surface density of the internal mesenchymal surface was the biggest at E12 (4,02 mm-1) and the least at E15 (2,69 mm-1)-p<0,0005. The surface density of the internal neuroepithelial surface was the biggest at E12 (3,31 mm-1) and the least at E15 (2,01 mm-1)-p<<0,0005. Our stereological examines give objective numerical proof of significant morphogenetic changes in telencephalon shape described by morphologic analyses. The major advantage of stereological methods is the possibility to carry out the estimation procedures in specified, well-defined brain regions or layers.

Animals↗

Signs of normal proliferation in the telencephalon of adult male songbirds (serinus serinus), as shown by PCNA-positivity.

The immunocytochemical expression of Proliferating Cell Nuclear Antigen (PCNA) (a cycline that coadjuvates DNA polymerase delta) becomes appreciable in the cell cycle when DNA synthesis occurs; hence cells in the S phase can be revealed by means of monoclonal antibodies. Therefore, PCNA can be considered a marker of proliferation, and numerous literature reports have demonstrated the reliability of the PCNA test. Since normal neurogenic events can still occur in the brain tissue of adult homeothermic vertebrates (especially songbirds), we evaluated if the persistence of spontaneous proliferation could be revealed in adult male songbirds (Serinus serinus) using the PCNA marker, the same test we used previously to study the persistence of natural proliferation in the encephalon of adult heterothermic vertebrates. The patterns of PCNA positivity showed normal proliferation in the telencephalon of the adult male Serinus serinus. This activity was shown by cells interposed among the epithelial cells lining the lateral side of each ventricular cavity, both in correspondence to the apical tracts and declivities of the ependyma and arranged, here and there, either in groups or slightly separated. As in our previous studies on PCNA expression and persistence of spontaneous encephalic proliferation in adult poikilothermal vertebrates (in the telencephalon of Podarcis, Triturus and Rana, and in the telencephalon, mesencephalon and cerebellum of Carassius), the results of the present research largely agree with the findings of previous Authors, usually obtained with different methods. This agreement confirms the reliability of the PCNA test used for this type of investigation.

Animals↗

Microscopic structure of the brain of Barbus meridionalis Risso. I. Telencephalon.

The telencephalon of Barbus meridionalis is composed of two oval telencephalic lobes separated by a medial ventricle and joined exclusively by the anterior commissure. Histologically, the dorsal and ventral telencephalic areas are well defined, the limiting plane between them being a cell free zone. In the ventral area it is possible to distinguish the following parts: dorsal, supracommissural, postcommissural, lateral, intermedia and the nucleus entopenduncularis. In the dorsal area there are three peripheral longitudinal zones--medialis, dorsalis and lateralis--together with a central zone; caudally situated is a posterior dorsolateral region which is the main receptor site for olfactory fibers and closely associated to this is the nucleus tenia. The olfactory projections to the telencephalon course along two bundles: the medial and the lateral olfactory tracts. The telencephalon projects to diencephalic regions via three large bundles: the lateral and medial telencephalic bundles and the strio-medullary system.

Animals↗

Differential growth of the cell production systems in the lateral wall of the developing mouse telencephalon.

Three major cell production systems were identified in the lateral wall of the mouse telencephalon. These were morphologically evident as the medial and lateral elevations and the pallial crescent. Each was originally derived from a small, circumscribed, subset of ventricular cells. These formed the parent populations of large and proliferatively complex precursor pools which gave rise to the large numbers and considerable variety of neuron populations of the telencephalon. An attempt was made to identify the major neuron groups derived from each system by using the ventricular cell processes as a guide to the site of neuron origin. The proliferative changes occurring in the two elevations, the variety of their neuronal output, the early loss of a radial structure and the diversity of the final adult configurations were considered to represent a more complex series of changes than the corresponding events in the cortical tissue, which was generated from the pallial crescent where neurons accumulate within an adaptation of the original radial structure. It was considered that the number and complexity of the changes in the genome required to produce and organise such subcortical diversity was of a greater order of magnitude than those evident in the cortical areas. It was suggested that the genesis of the mammalian basal telencephalon should be considered as one of the major evolutionary achievements in the conglomerate of changes which occurred during the transition from the reptilian to the mammalian grade of organisation in the forebrain.

Animals↗

The acetylcholine system in the brain of cyclostomes with special references to the telencephalon.

The distribution of cholinergic neurons is studied in the brains of cyclostomes (Lampetra fluviatilis and Myxine glutinosa) representing an early stage of vertebrate evolution. Histochemical localization of acetylcholinesterase (AChE) is complemented by quantitative determinations of acetylcholine (ACh), cholinacetyltransferase (ChAT) and AChE. The determinations are carried out in homogenates of whole brains, the brain regions telencephalon, diencephalon, mesencephalon and medulla oblongata. The telencephalon is further subdivided and studied in more detail.The transmitter distribution found in cyclostomes is compared to data from other vertebrates. Observations on the ACh-distribution in the telencephalon are discussed with the problems of identifying telencephalic homologies.

Acetylcholine↗

Neuronal and non-neuronal aromatase in primary cultures of developing zebra finch telencephalon.

Estrogenic metabolites of circulating androgens have important effects on the organization and activation of neural circuits controlling reproductive behavior and physiology in males of many vertebrate species. Previous studies indicate that aromatase, the enzyme that converts androgens to estrogens, is expressed most abundantly in neurons in limbic brain regions. Songbirds are unique in that aromatase is expressed at unusually high levels throughout the telencephalon of both males and females. We assume that estrogens formed in the telencephalon itself masculinize neural circuits controlling song, since the brain is a major source of circulating estrogens in adult males. However, the cellular localization of telencephalic aromatase in songbirds remains unknown. We have established primary cultures from telencephalons of developing zebra finches and found aromatase activity (conversion of 3H-androstenedione or 3H-testosterone to 3H-estrone plus 3H-estradiol) at some of the highest levels reported for brain tissue of any species. Both neurons and glia were identified in these cultures based on cell morphology and labeling by specific immunohistochemical markers. However, when culture conditions were manipulated to reduce the incidence of either neurons or glia by varying the age of cultures or their plating density, treating with the neurotoxin kainic acid, physically shaking off loosely attached neurons, or preparing cultures in media that encouraged enrichment of neurons, high levels of aromatase persisted. Furthermore, Northern blot analysis of total RNA extracted from enriched neuronal or glial cultures indicated the presence of aromatase mRNA in both cell preparations. In situ hybridization with a zebra finch aromatase cDNA probe conjugated to digoxigenin showed the cultures contained darkly labeled neurons and lightly labeled non-neuronal cells, presumably astrocytes. We conclude that aromatase is expressed in both neuronal and non-neuronal cells in these cultures, suggesting that both cell types may also express the enzyme in vivo. The presence of aromatase outside of neurons suggests that glia may be targets of estrogen action or that glia may supply some estrogen to the estrogen-sensitive neural circuits in this species.

Aging↗

The 5-HT3 receptor is present in different subpopulations of GABAergic neurons in the rat telencephalon.

The type 3 serotonin receptor (5-HT3R) is a ligand-gated ion channel whose presence in the CNS has been established by radioligand binding, in situ hybridization, and immunohistochemical analysis. To analyze further the role of the 5-HT3R in the CNS, we used in situ hybridization and immunocytochemistry to determine that 5-HT3R-expressing neurons are mainly GABA-containing cells in the rat telencephalon. We determined that 5-HT3R/GABA-containing neurons do not exhibit somatostatin immunoreactivity but often contain cholecystokinin (CCK) immunoreactivity. 5-HT3R-expressing cells with CCK immunoreactivity were observed in the neocortex, olfactory cortex, hippocampus, and amygdala. The 5-HT3R/CCK interneurons represent between 35 and 66% of the total population of CCK-containing cells in the neocortex. Further characterization of the 5-HT3R/GABAergic neurons was based on their calcium-binding protein immunoreactivity and showed that these neurons lack parvalbumin (PV) and represent a subpopulation of calbindin (CB)-containing interneurons that were preferentially present in the CA1-CA3 subfield of the hippocampus. Although some 5-HT3R/GABAergic neurons with calretinin (CR) were found in the neocortex, olfactory cortex, hippocampus, and amygdala, these neurons were more often present in the agranular insular and piriform cortices. We conclude that the neuronal expression of the 5-HT3R is selective within the GABA neuron population in the rat telencephalon. These 5-HT3R-expressing interneurons might contain CCK, CB, and CR. We suggest that serotonin through the 5-HT3R may regulate GABA and CCK neurotransmission in the telencephalon.

Animals↗

Distribution of neurofilaments in the telencephalon and mesencephalon of the adult and developing Gallotia galloti lizard.

The location and chronology during development of the immunoreactivity due to the presence of neurofilaments (NF) in telencephalon and mesencephalon of the lizard Gallotia galloti has been studied. For this purpose we have used two antibodies recognizing both phosphorylated and non phosphorylated neurofilaments (NF), a polyclonal Ab (NF 005), and a commercial monoclonal antibody (NF-200). The study was completed by using the Bielschowsky technique. During ontogeny, the anti-NF 005 immunoreactivity appeared at E40 in some tracts in mesencephalon and increased in intensity in isolated nerve fibers, tracts and commissurae till adult. However, a weak staining appeared in some neurons. In telencephalon, the reactivity was detected only in adult specimens. It was clearly more abundant in mesencephalon than in telencephalon, which could indicate that a greater complexity and functional importance exist in the lizard midbrain in relation to other primitive regions as the basal nuclei and cortical areas. In contrast to young specimens, the monoclonal anti-NF 200 was detected in neuronal perikarya, dendrites and axons in adults. Thus, in lizards, both antibodies highly recognized phosphorylated and non-phosphorylated forms of proteins of NF (NF-H). In mammals, these forms of proteins are implicated in axonal maturation. The presence of these NF in reptiles, identified for the first time, proved to be phylogenetically stable. The anti-NF immunoreactivity distribution occurs both caudo-rostrally and from the ventral to the dorsal regions.

Animals↗

Retinoic acid synthesis for the developing telencephalon.

The small lipid retinoic acid is known to promote neuronal differentiation in vitro and to act as a teratogen in the embryonic brain, but very little is known about the natural role of endogenously synthesized retinoic acid in forebrain development. Retinoic acid is synthesized mainly by three retinaldehyde dehydrogenases. We show here where the retinaldehyde dehydrogenases for the developing telencephalon are expressed and how their expression patterns change over developmental time. Retinoic acid diffusing from the retinaldehyde dehydrogenase sites is likely to influence the early telencephalon before the beginning of neurogenesis, as well as differentiation and radial migration of neurons into the cerebral cortex. Because of its diffusible character, retinoic acid represents a unique tool for the coordination of growth processes over an intermediate distance range in the developing telencephalon.

Aldehyde Oxidoreductases↗