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Mario F Wullimann

Publications and source records attributed to Mario F Wullimann.

15 recordsLinked to original sources

A phylotypic stage in vertebrate brain development: GABA cell patterns in zebrafish compared with mouse.

A recent comparison of early forebrain gene expression in mouse and zebrafish revealed highly comparable expression patterns of developmentally relevant genes, for example, of proneural (Neurogenin1, NeuroD, Mash1/Zash1a) genes involved in neurogenesis at a particular time window (mouse: embryonic day 12.5/13.5; zebrafish: 3 days). Here we extend this analysis to the description of gamma-aminobutyric acid (GABA) cell patterns in the early postembryonic zebrafish brain (i.e., during early secondary neurogenesis). We find again an astonishing degree of correspondences of GABA cell patterns between zebrafish and mouse during this previously established critical time window, for example, regarding absence of GABA cells in certain forebrain regions (pallium, dorsal thalamus, eminentia thalami) or with respect to the spatiotemporal occurrence of GABA cells (e.g., late cerebellar GABA cells). Furthermore, there is perfect correlation with previously established proneural gene expression patterns (i.e., absence of Mash1/Zash1a gene expression in GABA-cell-free forebrain regions) between mouse and zebrafish. The available information in additional vertebrate species, especially in Xenopus, is also highly consistent with our analysis here and suggests that a "phylotypic stage" of neurogenesis during vertebrate brain development may be present.

Animals↗

Secondary neurogenesis in the brain of the African clawed frog, Xenopus laevis, as revealed by PCNA, Delta-1, Neurogenin-related-1, and NeuroD expression.

After primary neurogenesis in the Xenopus laevis embryo, a massive new surge of neurogenesis and related neurogenic and proneural gene expression occurs in the spinal cord at the beginning of the larval period (starting at Stage 46), which corresponds to well-documented secondary neurogenesis in larval zebrafish central nervous system development. Here, we document related neural proliferation and gene expression patterns in the brain of Xenopus, in various embryonic and larval stages, showing the distribution of proliferative cells (immunostaining of cells containing the proliferating cell nuclear antigen; the auxiliary protein of DNA polymerase delta; PCNA), and the activity of some critical genes expressed during neurogenesis (i.e., Delta-1, Neurogenin-related-1, NeuroD). This study reveals that the early larval stage in Xenopus (Stage 48) displays patterns of proliferation (PCNA), as well as of neurogenic (Delta-1) and proneural (Ngnr-1; NeuroD) gene expression that are qualitatively almost identical to those seen in the 3-day postembryonic zebrafish or the 12.5/13.5-day embryonic mouse. Furthermore, a comparable bauplan of early proliferation zones (including their neuromeric organization) as described in the postembryonic zebrafish apparently exists in tetrapods (Xenopus). Altogether, the data presented suggest a common brain bauplan on the level of early proliferation patterns and neurogenic/proneural gene activity in anamniotes, if not vertebrates.

Animals↗

The adult central nervous cholinergic system of a neurogenetic model animal, the zebrafish Danio rerio.

The central nervous cholinergic system of the zebrafish (Danio rerio), a model animal for neurogenetics, is documented here using immunohistochemical methods for visualizing choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme. Neuronal cell bodies containing ChAT are present in the telencephalon (lateral nucleus of ventral telencephalic area), preoptic region (anterior/posterior parvocellular and magnocellular preoptic nuclei), diencephalon (habenula, dorsal thalamus, posterior tuberculum), mesencephalon (Edinger-Westphal (EW) nucleus, oculomotor nerve nucleus, rostral tegmental nucleus, tectal type XIV neurons), isthmic region (nucleus lateralis valvulae, secondary gustatory-viscerosensory nucleus, nucleus isthmi (NI), perilemniscal nucleus, superior reticular nucleus (SRN)) and rhombencephalon (trochlear, trigeminal, abducens, facial, glossopharyngeal-vagal motor nerve nuclei, rostral and caudal populations of octavolateralis efferent neurons). In addition, some ChAT positive neurons are present in the rhombencephalic reticular formation, the central gray, and in cells accompanying the descending trigeminal tract. Obvious ChAT positive terminal fields are present in the supracommissural nucleus of area ventralis telencephali and the medial zone of area dorsalis telencephali, parvocellular superficial pretectal nucleus, torus semicircularis, medial octavolateralis nucleus, facial, glossopharyngeal, and vagal lobes, and in the inferior lobe (around the periventricular nucleus of the lateral recess and in the diffuse nucleus). The identification of all central nervous cholinergic systems provided here in this model system is pivotal for future detailed studies of their development and maintenance, e.g., with regard to the zebrafish ventral telencephalic and isthmic superior reticular neuronal populations, likely representing the homologues of at least part of the cholinergic basal forebrain and pedunculopontine/laterodorsal tegmental ascending activating systems of mammals, respectively.

Animals↗

Connections of the ventral telencephalon (subpallium) in the zebrafish (Danio rerio).

Connections of the medial precommissural subpallial ventral telencephalon, i.e., dorsal (Vd, interpreted as part of striatum) and ventral (Vv, interpreted as part of septum) nuclei of area ventralis telencephali, were studied in the zebrafish (Danio rerio) using two tracer substances (DiI or biocytin). The following major afferent nuclei to Vd/Vv were identified: medial and posterior pallial zones of dorsal telencephalic area, and the subpallial supracommissural and postcommissural nuclei of the ventral telencephalic area, the olfactory bulb, dorsal entopeduncular, anterior and posterior parvocellular preoptic and suprachiasmatic nuclei, anterior, dorsal and central posterior dorsal thalamic, as well as rostrolateral nuclei, periventricular nucleus of the posterior tuberculum, posterior tuberal nucleus, various tuberal hypothalamic nuclei, dorsal tegmental nucleus, superior reticular nucleus, locus coeruleus, and superior raphe nucleus. Efferent projections of the ventral telencephalon terminate in the supracommissural nucleus of area ventralis telencephali, the posterior zone of area dorsalis telencephali, habenula, periventricular pretectum, paracommissural nucleus, posterior dorsal thalamus, preoptic region, midline posterior tuberculum (especially the area dorsal to the posterior tuberal nucleus), tuberal (midline) hypothalamus and interpeduncular nucleus. Strong reciprocal interconnections likely exist between septum and preoptic region/midline hypothalamus and between striatum and dorsal thalamus (dopaminergic) posterior tuberculum. Regarding ascending activating/modulatory systems, the pallium shares with the subpallium inputs from the (noradrenergic) locus coeruleus, and the (serotoninergic) superior raphe, while the subpallium additionally receives such inputs from the (dopaminergic) posterior tuberculum, the (putative cholinergic) superior reticular nucleus, and the (putative histaminergic) caudal hypothamalic zone.

Amino Acids↗

Identification and morphogenesis of the eminentia thalami in the zebrafish.

This study documents early zebrafish brain expression patterns (2-5 days postfertilization) of proliferating neural (PCNA) as well as early-determined (Pax6, Zash-1a, Zash-1b, neurogenin1, neuroD) and differentiating (Hu-proteins) neuronal cells. These patterns are used to outline the spatiotemporal local dynamics of secondary neurogenesis as well as neuronal migration and differentiation in the region of the eminentia thalami. The analysis presented not only allows identification for the first time of the eminentia thalami in the zebrafish model system (because it forms a neurogenin1/neuroD-guided locus of neurogenesis in contrast to adjacent preoptic region and ventral thalamus) but furthermore shows that the entopeduncular complex is a derivative of the embryonic zebrafish eminentia thalami, which has never been reported for a teleost before. An analysis of the relevant literature shows that the mammalian entopeduncular nucleus/avian paleostriatum primitivum/reptilian globus pallidus clearly are part of the basal ganglia (i.e., the pallidum). In amniote embryos, an anterior entopeduncular area is recognized at the base of the medial ganglionic eminence (i.e., the future pallidum; part of alar plate of prosomere 5), separate from the more posterior eminentia thalami (alar prosomere 4). There is a comparable periventricular eminentia thalami in (young and adult) amphibians and teleosts. However, the migrated anterior entopeduncular nucleus of anuran amphibians likely is homologous to part of the pallidum of other vertebrates and has no developmental relationship to the eminentia thalami. In contrast, the migrated teleostean entopeduncular complex does not correspond to a pallidal division but is indeed the adult derivative of the early-recognized eminentia thalami as shown in this study.

Animals↗

Anatomy of neurogenesis in the early zebrafish brain.

The detailed architecture of postembryonic (i.e. secondary, as opposed to primary) neurogenesis in the zebrafish brain at 2 days postfertilization was investigated by studying expression domains of various proneural basic helix-loop-helix genes (i.e. neuroD=nrd, neurogenin1=ngn1, Zash-1b) and neurogenic genes (i.e. Notch-1a, deltaA) on the level of in situ-hybridized sectioned material and compared with brain sections of the same age immunostained for PCNA (a proliferation marker) or for Hu-proteins (marker for early neuronal differentiation). Whereas both Notch-1a and deltaA domains are present in all proliferative zones of the brain, only the more diffuse and scattered deltaA expression appears to extend additionally into the adjacent postmitotic gray matter. Zash-1b is the first achaete-scute orthologue shown here to be expressed exclusively in all proliferative central nervous zones (except for the eye). The ngn1 and nrd genes are typically both expressed in overlapping fashion in many-but not all-brain regions, with ngn1 being more restricted towards the ventricular proliferative zones and nrd extending more laterally. This fact as well as comparisons with PCNA- and Hu-immunostains indicate that a great proportion of ngn1-positive cells are mitotic, but some appear to extend into the postmitotic gray matter where the nrd-domains lie. A comparison of the relative extents of PCNA- (proliferative), nrd- (freshly determined) and Hu-positive (differentiating) cell populations allows to determine the relative maturation state of a given brain part. Similar to findings in late embryonic amniote brains, expression of nrd is absent (from 2 to 5 days) in the zebrafish subpallium, ventral preoptic region, ventral thalamus and hypothalamus. These four regions are also free of ngn1 expression (with the exception of an unusual peripheral ngn1 domain in the preoptic region), indicating that a neurogenetic network not involving nrd and ngn1 is at work there. Our characterization of locally distinct patterns and dynamics of secondary neurogenesis in the entire early (2 dpf) postembryonic zebrafish brain delivers the blueprint for more specialized functional studies.

Aging↗

Development of the catecholaminergic system in the early zebrafish brain: an immunohistochemical study.

Tyrosine hydroxylase-containing cells (TH cells) were investigated immunohistochemically in early and late postembryonic zebrafish brain sections (at 2 and 5 days postfertilization [dpf]) yielding an improved neuroanatomical resolution of spatiotemporal developmental dynamics of the catecholaminergic system. Additionally, double-immunolabel preparations for visualizing TH cells and cells containing the proliferating cell nuclear antigen (PCNA cells) were carried out allowing for a prosomeric interpretation of early forebrain TH cell clusters. Many TH cell populations recently described in the adult zebrafish brain could be identified in the present study by location and cell type already in the 5 dpf (e.g. eight of 12 adult diencephalic TH cell populations) and 2 dpf (e.g. five of 12 adult TH cell populations) zebrafish brain. Early and adult diencephalic TH cells are restricted to the pretectum (P1) and ventral thalamus (P3) in the alar plate, and to various TH groups in the basal plate posterior tuberculum (P3), as well as to various populations in the hypothalamus (secondary prosencephalon). The alar plate ventral thalamic and most anterodorsal posterior tubercular TH cell populations range among the earliest detectable ones. There was no indication of migration of TH cells from the midbrain-hindbrain boundary or anterior neural ridge into the diencephalon.

Animals↗

Hypothalamic inferior lobe and lateral torus connections in a percomorph teleost, the red cichlid (Hemichromis lifalili).

The neuroanatomic connections of the inferior lobe and the lateral torus of the percomorph Hemichromis lifalili were investigated by 1,1', dioctadecyl-3,3,3',3'-tetramethylindo-carbocyanine perchlorate (DiI) tracing. The inferior lobe and the lateral torus both receive afferents from the secondary gustatory nucleus. Additional afferents reach the inferior lobe from the nucleus glomerulosus, nucleus suprachiasmaticus, dorsal and central posterior thalamic nucleus, nucleus lateralis valvulae, magnocellular part of the magnocellular nucleus of the preoptic region, caudal nucleus of the preglomerular region, posterior tuberal nucleus, area dorsalis of the telencephalon, and a tegmental nucleus (T2). Efferents from the inferior lobe and the lateral torus terminate in the dorsal hypothalamic neuropil and corpus mamillare. Furthermore, the inferior lobe projects to the medial nucleus of the lateral tuberal hypothalamus and perhaps makes axo-axonal synapses in the tractus tectobulbaris rectus. The inferior lobe and the torus lateralis have reciprocal connections with the preglomerular tertiary gustatory nucleus and posterior thalamic nucleus and are also mutually interconnected. The inferior lobe is also reciprocally connected with the medial nucleus of the preglomerular region, reticular formation and sparsely with the anterior dorsal thalamic and the ventromedial thalamic nuclei. Thus, whereas the lateral torus is exclusively connected with the gustatory system, the inferior lobe is of a multisensory nature. In comparison with the goldfish (Carassius auratus), the connectivity pattern of the inferior lobe of Hemichromis lifalili reflects its specialization with respect to the visual system, as it receives qualitative (i.e., dorsal posterior, anterior, and ventromedial thalamic nuclei) as well as quantitative (i.e., nucleus glomerulosus) additional visual input.

Animals↗

BrdU-, neuroD (nrd)- and Hu-studies reveal unusual non-ventricular neurogenesis in the postembryonic zebrafish forebrain.

In the postembryonic zebrafish forebrain, subpial locations of neurogenesis do exist in the early cerebellar external granular layer, and--unusually among vertebrates--in the primordial pretectal (M1) and preglomerular (M2) Anlagen as shown here with 5-bromo-2'-deoxyuridine (BrdU)/Hu-immunocytochemistry and in situ hybridization of neuroD. An intermediate BrdU incubation time of 12-16 h reveals in addition to proliferative ventricularly located cells those in M1 and M2. This BrdU saturation-labeling shows--in conjunction with a Hu-assay demonstrating earliest neuronal differentiation--that proliferating cells in M1 and M2 represent neuronal progenitors. This is demonstrated by single BrdU-labeled and double BrdU-/Hu-labeled cells in these aggregates. Further, expression of NeuroD--a marker for freshly determined neuronal cells--confirms this unusual subpial postembryonic forebrain neurogenesis.

Animals↗

Expression of Zash-1a in the postembryonic zebrafish brain allows comparison to mouse Mash1 domains.

Four areas in the late embryonic murine forebrain, i.e. the subpallium (striatum), the preoptic region, the ventral thalamus, and the hypothalamus, have been described to express the basic helix-loop-helix (bHLH) gene mammalian achaete-scute homolog Mash1 (Ascl1, Mouse Genome Informatics) in a complementary fashion to another bHLH gene, neurogenin1 (ngn1) (Neurod3, Mouse Genome Informatics), which is expressed in directly adjacent forebrain regions. We report here that the four regions previously identified as subpallium, preoptic region, ventral thalamus and hypothalamus (i.e. ventral inferior lobe) in the postembryonic zebrafish brain show Zash-1a expression at 3 days postfertilization (dpf), whereas none of those areas express the bHLH gene neuroD (nrd) between 2 and 5 dpf. This indicates that two well established alternative genetic pathways involved in neurogenesis in the amniote (mammalian) brain are present in homologous phenotypic locations in the anamniote (zebrafish) brain as well and that these pathways possibly act similarly in the generation of different neuronal phenotypes (e.g. subpallial GABAergic interneurons versus pallial glutamatergic projection neurons, or dopaminergic neurons versus other neurotransmitter phenotypes). Furthermore, previous initial identification of early postembryonic brain subdivisions in the zebrafish is strongly corroborated by these expression patterns.

Animals↗

The teleostean forebrain: a comparative and developmental view based on early proliferation, Pax6 activity and catecholaminergic organization.

An improved comparative interpretation of the teleostean forebrain suggests that the dorsal tier (Vd,Vc) and ventral tier (Vv,Vl) nuclei of the ventral telencephalic area (subpallium) represent the striatum and septum, respectively. Among other arguments, a dopaminergic innervation originating in the diencephalic posterior tubercle reaches Vd and dense efferents of Vv project to the midline hypothalamus in the adult zebrafish subpallium. The adult area dorsalis telencephali represents the teleostean pallium. Regulatory genes typically expressed in the early amniote subpallium (e.g., Dlx-1) are also restricted to the presumptive zebrafish ventral telencephalic area. Further, early Pax6 protein distribution in the zebrafish telencephalon corresponds to the migrating stream noted at the pallial-subpallial boundary in amniotes, but a ventricular, radial glia-based expression in the pallium is absent. The peripherally migrated, adult diencephalic preglomerular complex of the basal plate posterior tubercle (early: M2) provides sensory inputs to the pallium. Early Pax6 protein distribution indicates that at least part of M2 may directly originate from alar plate ventral thalamic Pax6-expressing cells. Dopaminergic cells of the basal plate posterior zebrafish forebrain (P1-P3) are restricted to the ventral thalamic prosomere (P3), including those forming the adult ascending dopaminergic system. Moreover, the latter likely depend developmentally on the dorsally adjacent alar plate Pax6-expressing cells.

Animals↗

Expression domains of neuroD (nrd) in the early postembryonic zebrafish brain.

NeuroD acts as one of the earliest transcription factors promoting differentiation in newborn neurons immediately after determination by neurogenin1 (ngn1). Here, we show the expression of neuroD in the 2 days postfertilization (dpf) zebrafish brain by in situ hybridization on sections. Generally, neuroD expressing cells lie one to several cell rows away from the ventricular surface, i.e., they seem to flank proliferative cells. However, most cells of the gray matter lie even more peripheral and are unstained. Highly specific local patterns of neuroD expressing cells are observed along the entire rostrocaudal zebrafish brain axis at that age with characteristic neuroD expression free zones (e.g., preoptic region and hypothalamus). Interestingly, these regions where neuroD expression is absent display a higher proliferative activity compared to neuroD-positive areas. To date it is an open question, whether this negative correlation of neuroD-expression and proliferation has a causal background.

Animals↗

Connections of the ventral telencephalon and tyrosine hydroxylase distribution in the zebrafish brain (Danio rerio) lead to identification of an ascending dopaminergic system in a teleost.

We studied the connections and catecholaminergic organization of the subpallium in the zebrafish, in particular to demonstrate the origin of the ascending dopaminergic system of teleosts, by using the tracers DiI or biocytin in combination with tyrosine hydroxylase (TH) immunohistochemistry. Retrogradely labeled cells were found in the olfactory bulb, the area dorsalis telencephali, the preoptic region, the dorsal and ventral thalamus, the posterior tubercle, the preglomerular region, and the medulla oblongata. Moreover, the zebrafish subpallium has strong reciprocal connections with the tuberal hypothalamus. Double-labeled cells (for TH and tracer) were identified in two locations of the rostral posterior tubercle: small round neurons in its periventricular nucleus and large pear-shaped cells adjacent to it. These double-labeled cells of the posterior tubercle presumably represent the teleostean dopaminergic system ascending to the striatum.

Afferent Pathways↗