Amelia Younossi-Hartenstein

EARLY NEUROGENESIS OF THE DROSOPHILA BRAIN

We have studied the formation of the neuroblasts of the Drosophila brain which segregate from the procephalic neurectoderm. The expression domains of the segment polarity gene engrailed (en) allow one to subdivide the procephalic neuroectoderm into tritocerebral, deuterocerebral, and protocerebral neuromeres. Based upon the expression pattern of the proneural gene lethal of scute (lsc), as well as the pattern of brain neuroblast segregation, the protocerebral and deuterocerebral neuromeres can be further subdivided into a central, anterior, and posterior domain. A total of 75–80 neuroblasts segregate in a stereotyped pattern from the procephalic neurectoderm of each side during stages 9–11. With respect to their position and the expression of the markers asense (ase) and seven‐up (sup), 23 small groups of one to five neuroblasts each were identified. The first eight groups (Pc1–4, Dc1–3, Dp1), collectively called SI/II neuroblasts in analogy to the subpopulation of ventral neuroblasts which appear at the same stage), arise from the central domain of the protocerebral and deuterocerebral neurectoderm, respectively. Later groups form anteriorly and posteriorly from the earlier ones, leading to a centrifugal growth of the procephalic neuroblast population. SIII neuroblasts (Pa1–4, Pp1–2, Dp2) arise during stage 10, SIV neuroblasts (Pa5–6, Pp3–4, Da1, T1–2) during early stage 11, and SV neuroblasts (Pp5, Pdm) during late stage 11 and early stage 12. The dorsomedial domain of the procephalic neurectoderm represents a special case. Unlike other procephalic neuroblasts which delaminate from the surface ectoderm as individual cells, cells of the dorsomedial protocerebral domain are internalized during stage 12 as large, coherent clusters by a movement which can be best characterized as a combination of mass‐delamination and invagination.

The behaviour of Drosophila adult hindgut stem cells is controlled by Wnt and Hh signalling

The intestinal tract maintains proper function by replacing aged cells with freshly produced cells that arise from a population of self-renewing intestinal stem cells (ISCs). In the mammalian intestine, ISC self renewal, amplification and differentiation take place along the crypt–villus axis, and are controlled by the Wnt and hedgehog (Hh) signalling pathways. However, little is known about the mechanisms that specify ISCs within the developing intestinal epithelium, or about the signalling centres that help maintain them in their self-renewing stem cell state. Here we show that in adult Drosophila melanogaster, ISCs of the posterior intestine (hindgut) are confined to an anterior narrow segment, which we name the hindgut proliferation zone (HPZ). Within the HPZ, self renewal of ISCs, as well as subsequent proliferation and differentiation of ISC descendants, are controlled by locally emanating Wingless (Wg, a Drosophila Wnt homologue) and Hh signals. The anteriorly restricted expression of Wg in the HPZ acts as a niche signal that maintains cells in a slow-cycling, self-renewing mode. As cells divide and move posteriorly away from the Wg source, they enter a phase of rapid proliferation. During this phase, Hh signal is required for exiting the cell cycle and the onset of differentiation. The HPZ, with its characteristic proliferation dynamics and signalling properties, is set up during the embryonic phase and becomes active in the larva, where it generates all adult hindgut cells including ISCs. The mechanism and genetic control of cell renewal in the Drosophila HPZ exhibits a large degree of similarity with what is seen in the mammalian intestine. Our analysis of the Drosophila HPZ provides an insight into the specification and control of stem cells, highlighting the way in which the spatial pattern of signals that promote self renewal, growth and differentiation is set up within a genetically tractable model system.

Embryonic origin and differentiation of the Drosophila heart

We have followed the normal development of the different cell types associated with the Drosophila dorsal vessel, i.e. cardioblasts, pericardial cells, alary muscles, lymph gland and ring gland, by using several tissue-specific markers and transmission electron microscopy. Precursors of pericardial cells and cardioblasts split as two longitudinal rows of cells from the lateral mesoderm of segments T2-A7 (“cardiogenic region”) during stage 12. The lymph gland and dorsal part of the ring gland (corpus allatum) originate from clusters of lateral mesodermal cells located in T3 and T1/dorsal ridge, respectively. Cardioblast precursors are strictly segmentally organized; each of T2-A6 gives rise to six cardioblasts. While moving dorsally during the stages leading up to dorsal closure, cardioblast precursors become flattened, polarized cells aligned in a regular longitudinal row. At dorsal closure, the leading edges of the cardioblast precursors meet their contralateral counterparts. The lumen of the dorsal vessel is formed when the trailing edges of the cardioblast precursors of either side bend around and contact each other. The amnioserosa invaginates during dorsal closure and is transiently attached to the cardioblasts; however, it does not contribute to the cells associated with the dorsal vessel and degenerates during late embryogenesis. We describe ultrastructural characteristics of cardioblast differentiation and discuss similarities between cardioblast development and capillary differentiation in vertebrates.

Early development of the Drosophila brain: IV. Larval neuropile compartments defined by glial septa.

In this study, we have analyzed the architecture of the brain neuropile of the Drosophila larva, which is formed by two main structural elements: long axon tracts and terminal axonal/dendritic arborizations carrying synapses. By using several molecular markers expressed in neurons and glial cells, we show that the early larval neuropile is subdivided by glial sheaths into numerous compartments. The three‐dimensional layout of these compartments and their relationship to the pattern of long axon tracts described in the accompanying article (Nassif et al. [2003] J. Comp. Neurol 417–434) was modeled by using a three‐dimensional illustration computer software. On the basis of their location relative to each other and to long axon tracts, larval brain compartments can be identified with compartments defined by structural and functional criteria for the adult fly brain. We find that small precursors of most of the compartments of the adult central brain can be identified in the early larva. Changes in brain compartmental organization occurring during larval growth are described. Neuropile compartments, representing easily identifiable landmark structures, will assist in future analyses of Drosophila brain development in which the exact location of neurons and their axonal trajectories is of importance. J. Comp. Neurol. 455:435–450, 2003.

Embryonic origin of the imaginal discs of the head of Drosophila melanogaster

The embryonic development of the primordia of the Drosophila head was studied by using an enhancer trap line expressed in these structures from embryonic stage 13 onward. Particular attention was given to the question of how the adult head primordia relate to the larval head segments. The clypeo-labral bud to the stage 13 embryo is located at a lateral position in the labrum adjacent to the labral sensory complex (“epiphysis”). Both clypeo-labral bud and sensory complex are located anterior to the engrailed-expression domain of the labrum. Throughout late embryogenesis and the larval period, the clypeo-labral bud forms integral part of the epithelium lining the roof of the atrium. The labial disc originates from the lateral labial segment adjacent to the labial sensory complex (“hypophysis”). It partially overlaps with the labial en-domain. After head involution, the labial disc forms a small pocket in the ventro-lateral wall of the atrium. The eye-antenna disc develops from a relatively large territory occupying the dorso-posterior part of the procephalic lobe, as well as parts of the dorsal gnathal segments. Cells in this territory are greatly reduced in number by cell death during stages 12–14. After head involution, the presumptive eye-antenna disc occupies a position in the lateral-posterior part of the dorsal pouch. Evagination of this tissue occurs during the first hours after hatching. In the embryo, no en-expression is present in the presumptive eye-antenna disc. en-expression starts in three separate regions in the third instar larva.

Development of the Drosophila entero-endocrine lineage and its specification by the Notch signaling pathway

In this paper we have investigated the developmental-genetic steps that shape the entero-endocrine system of Drosophila melanogaster from the embryo to the adult. The process starts in the endoderm of the early embryo where precursors of endocrine cells and enterocytes of the larval midgut, as well as progenitors of the adult midgut, are specified by a Notch signaling-dependent mechanism. In a second step that occurs during the late larval period, enterocytes and endocrine cells of a transient pupal midgut are selected from within the clusters of adult midgut progenitors. As in the embryo, activation of the Notch pathway triggers enterocyte differentiation and inhibits cells from further proliferation or choosing the endocrine fate. The third step of entero-endocrine cell development takes place at a mid-pupal stage. Before this time point, the epithelial layer destined to become the adult midgut is devoid of endocrine cells. However, precursors of the intestinal midgut stem cells (pISCs) are already present. After an initial phase of symmetric divisions which causes an increase in their own population size, pISCs start to spin off cells that become postmitotic and express the endocrine fate marker, Prospero. Activation of Notch in pISCs forces these cells into an enterocyte fate. Loss of Notch function causes an increase in the proliferatory activity of pISCs, as well as a higher ratio of Prospero-positive cells.

The embryonic development of the polyclad flatworm Imogine mcgrathi

Abstract In this paper we describe the embryonic development of the polyclad flatworm Imogine mcgrathi. Imogine is an indirect developer that hatches as a planctonic Goette’s larva after an embryonic period of approximately 7 days. Light and electron microscopic analyses of sections of staged embryos were combined with antibody stainings of wholemounted embryos to reconstruct the origin and movement of the primordia of the various organ systems, with particular emphasis on the nervous system. We introduce a system of morphologically defined stages aimed at facilitating future studies and cross-species comparisons among flatworm embryos. Imogine embryos undergo typical spiral cleavage. Micromere quartets 1–3 form an irregular double layer of mesenchymal cells that during gastrulation expands over micromere quartet 4. Micromere 4d divides into several large mesendodermal precursors whose position defines the ventral pole of the embryo. These cells, along with the animal micromeres that obtained a sub-surface position during cleavage, form a deep layer of cells that gives rise to all internal structures, including the nervous system, musculature, nephridia, and gut. Micromeres 4a–c are large yolky cells that are incorporated into the lumen of the gut, but do not themselves contribute to the gut epithelium. Shortly after gastrulation, cell differentiation sets in. Cells located at the surface adopt epithelial characteristics and form cilia that result in continuous movement of the post-gastrula stage embryo. Deep cells at the lateral margins of the embryo become organized into a protonephridial tube. A cluster of approximately 50 deep cells at the anterior pole forms the brain, in which we have identified sets of founder neurons of the brain commissure and the dorsal and ventral connectives. The early differentiating neurons, along with other cells forming stabilized microtubules (ciliated cells of the epidermis, gut and protonephridia; apical gland cells) could be analyzed in detail because of their labeling with an antibody against acetylated α-tubulin. Our findings indicate that, despite significant differences in the cleavage pattern and arrangement of blastomeres in the early embryo, morphogenesis and organ formation of a polyclad embryo follows a pattern that is very similar to the pattern observed by us and others in phylogenetically more evolved rhabdocoel flatworms.

A novel tissue in an established model system: the Drosophila pupal midgut

The Drosophila larval and adult midguts are derived from two populations of endodermal progenitors that separate from each other in the early embryo. As larval midgut cells differentiate into an epithelial layer, adult midgut progenitors (AMPs) remain as small clusters of proliferating, undifferentiated cells attached to the basal surface of the larval gut epithelium. During the first few hours of metamorphosis, AMPs merge into a continuous epithelial tube that overgrows the larval layer and differentiates into the adult midgut; at the same time, the larval midgut degenerates. As shown in this paper, there is a second, transient pupal midgut that develops from the AMPs at the beginning of metamorphosis and that intercalates between the adult and larval midgut epithelia. Cells of the transient pupal midgut form a multilayered tube that exhibits signs of differentiation, in the form of septate junctions and rudimentary apical microvilli. Some cells of the pupal midgut develop as endocrine cells. The pupal midgut remains closely attached to the degenerating larval midgut cells. Along with these cells, pupal midgut cells are sequestered into the lumen where they form the compact “yellow body.” The formation of a pupal midgut has been reported from several other species and may represent a general feature of intestinal metamorphosis in insects.

Embryonic development of the nervous system of the rhabdocoel flatwormMesostoma lingua (Abildgaard, 1789)

We have analyzed the embryonic development of the Mesostoma nervous system, using a combination of histology, transmission electron microscopy, and wholemount immunohistochemistry. Neural progenitors are formed at an early stage when the Mesostoma embryo constitutes a multilayered mesenchymal mass of cells. A neurectoderm as in vertebrates or arthropods is absent. Only after neurons in the deep layers of the embryo have started differentiating do superficial cells reorganize into an epithelium that will give rise to the epidermis. Neurons are clustered in two anterior, bilaterally symmetric brain hemispheres. An antibody against acetylated beta‐tubulin (anti‐acTub) that labels neurotubules reveals an invariant pattern of pioneer neurons in the brain of midstage embryos. Pioneer neurons are grouped in several small clusters at characteristic positions. They pioneer several commissural tracts of the brain and two pairs of ventral and dorsal connectives, respectively. J. Comp. Neurol. 416:461–474, 2000.

Lineage‐based analysis of the development of the central complex of the drosophila brain

Most neurons of the central complex belong to 10 secondary (larvally produced) lineages. In the late larva, undifferentiated axon tracts of these lineages form a primordium in which all of the compartments of the central complex can be recognized as discrete entities. Four posterior lineages (DPMm1, DPMpm1, DPMpm2, and CM4) generate the classes of small‐field neurons that interconnect the protocerebral bridge, fan‐shaped body, noduli, and ellipsoid body. Three lineages located in the anterior brain, DALv2, BAmv1, and DALcl2, form the large‐field neurons of the ellipsoid body and fan‐shaped body, respectively. These lineages provide an input channel from the optic tubercle and connect the central complex with adjacent anterior brain compartments. Three lineages in the posterior cortex, CM3, CP2, and DPMpl2, connect the posterior brain neuropil with specific layers of the fan‐shaped body. Even though all of the compartments of the central complex are prefigured in the late larval brain by the axon tracts of the above‐mentioned lineages, the neuropil differentiates during the first 2 days of the pupal period when terminal branches and synapses of secondary neurons are formed. During this phase the initially straight horizontal layers of the central complex bend in the frontal plane, which produces the characteristic shape of the fan‐shaped and ellipsoid body. Our analysis provides a comprehensive picture of the lineages that form the central complex, and will facilitate future studies that address the structure or function of the central complex at the single cell level. J. Comp. Neurol. 519:661–689, 2011.