Amot and Yap1 regulate neuronal dendritic tree complexity and locomotor coordination in mice.

The angiomotin (Amot)-Yes-associated protein 1 (Yap1) complex plays a major role in regulating the inhibition of cell contact, cellular polarity, and cell growth in many cell types. However, the function of Amot and the Hippo pathway transcription coactivator Yap1 in the central nervous system remains unclear. We found that Amot is a critical mediator of dendritic morphogenesis in cultured hippocampal cells and Purkinje cells in the brain. Amot function in developing neurons depends on interactions with Yap1, which is also indispensable for dendrite growth and arborization in vitro. The conditional deletion of Amot and Yap1 in neurons led to a decrease in the complexity of Purkinje cell dendritic trees, abnormal cerebellar morphology, and impairments in motor coordination. Our results indicate that the function of Amot and Yap1 in dendrite growth does not rely on interactions with TEA domain (TEAD) transcription factors or the expression of Hippo pathway-dependent genes. Instead, Amot and Yap1 regulate dendrite development by affecting the phosphorylation of S6 kinase and its target S6 ribosomal protein.

Two Independent Functions of Collier/Early B Cell Factor in the Control of Drosophila Blood Cell Homeostasis.

Blood cell production in the Drosophila hematopoietic organ, the lymph gland, is controlled by intrinsic factors and extrinsic signals. Initial analysis of Collier/Early B Cell Factor function in the lymph gland revealed the role of the Posterior Signaling Center (PSC) in mounting a dedicated cellular immune response to wasp parasitism. Further, premature blood cell differentiation when PSC specification or signaling was impaired, led to assigning the PSC a role equivalent to the vertebrate hematopoietic niche. We report here that Collier is expressed in a core population of lymph gland progenitors and cell autonomously maintains this population. The PSC contributes to lymph gland homeostasis by regulating blood cell differentiation, rather than by maintaining core progenitors. In addition to PSC signaling, switching off Collier expression in progenitors is required for efficient immune response to parasitism. Our data show that two independent sites of Collier/Early B Cell Factor expression, hematopoietic progenitors and the PSC, achieve control of hematopoiesis.

The muscle pattern of the Drosophila abdomen depends on a subdivision of the anterior compartment of each segment.

In the past, segments were defined by landmarks such as muscle attachments, notably by Snodgrass, the king of insect anatomists. Here, we show how an objective definition of a segment, based on developmental compartments, can help explain the dorsal abdomen of adult Drosophila. The anterior (A) compartment of each segment is subdivided into two domains of cells, each responding differently to Hedgehog. The anterior of these domains is non-neurogenic and clones lacking Notch develop normally; this domain can express stripe and form muscle attachments. The posterior domain is neurogenic and clones lacking Notch do not form cuticle; this domain is unable to express stripe or form muscle attachments. The posterior (P) compartment does not form muscle attachments. Our in vivo films indicate that early in the pupa the anterior domain of the A compartment expresses stripe in a narrowing zone that attracts the extending myotubes and resolves into the attachment sites for the dorsal abdominal muscles. We map the tendon cells precisely and show that all are confined to the anterior domain of A. It follows that the dorsal abdominal muscles are intersegmental, spanning from one anterior domain to the next. This view is tested and supported by clones that change cell identity or express stripe ectopically. It seems that growing myotubes originate in posterior A and extend forwards and backwards until they encounter and attach to anterior A cells. The dorsal adult muscles are polarised in the anteroposterior axis: we disprove the hypothesis that muscle orientation depends on genes that define planar cell polarity in the epidermis.

A short receptor downregulates JAK/STAT signalling to control the Drosophila cellular immune response.

The posterior signalling centre (PSC), a small group of specialised cells, controls hemocyte (blood cell) homeostasis in the Drosophila larval hematopoietic organ, the lymph gland. This role of the PSC is very reminiscent of the "niche," the micro-environment of hematopoietic stem cells in vertebrates. We have recently shown that the PSC acts in a non-cell-autonomous manner to maintain janus tyrosine kinase/signal transducers and activators of transcription (JAK/STAT) signalling in hematopoietic progenitors (prohemocytes), thereby preserving the multipotent character necessary for their differentiation into lamellocytes, a cryptic and dedicated immune cell type required to fight specific immune threats such as wasp parasitism. In this report, on the basis of a knock out generated by homologous recombination, we show that a short type I cytokine-related receptor CG14225/Latran is required for switching off JAK/STAT signalling in prohemocytes. This is a prerequisite to massive differentiation of lamellocytes upon wasp parasitisation. In vivo and cell culture assays indicate that Latran forms heteromers with Domeless, the Drosophila type I cytokine signalling receptor related to mammalian GP130, and antagonises Domeless activity in a dose-dependent manner. Our analysis further shows that a primary immune response to wasp parasitism is a strong decrease in cytokine mRNA levels in the lymph gland, followed by an increase in the latran/domeless ratio. We propose that this sequence of events culminates in the complete inhibition of residual JAK/STAT signalling by Latran. JAK/STAT activity has been associated with several human diseases including leukaemia while knock-out studies in mice point to a central role of this pathway in hematopoiesis and regulation of immune functions. The specific function of Drosophila Latran is, to our knowledge, the first in vivo example of a role for a nonsignalling receptor in controlling a dedicated immune response, and thus raises the question of whether short, nonsignalling receptors also control specific aspects of vertebrate cellular immunity.

Hematopoietic progenitors and hemocyte lineages in the Drosophila lymph gland.

The Drosophila lymph gland (LG) is a model system for studying hematopoiesis and blood cell homeostasis. Here, we investigated the patterns of division and differentiation of pro-hemocytes in normal developmental conditions and response to wasp parasitism, by combining lineage analyses and molecular markers for each of the three hemocyte types. Our results show that the embryonic LG contains primordial hematopoietic cells which actively divide to give rise to a pool of pro-hemocytes. We found no evidence for the existence of bona fide stem cells and rather suggest that Drosophila pro-hemocytes are regulated as a group of cells, rather than individual stem cells. The fate-restriction of plasmatocyte and crystal cell progenitors occurs between the end of embryogenesis and the end of the first larval instar, while Notch activity is required for the differentiation of crystal cells in third instar larvae only. Upon parasitism, lamellocyte differentiation prevents crystal cell differentiation and lowers plasmatocyte production. We also found that a new population of intermediate progenitors appears at the onset of hemocyte differentiation and accounts for the increasing number of differentiated hemocytes in the third larval instar. These findings provide a new framework to identify parameters of developmental plasticity of the Drosophila lymph gland and hemocyte homeostasis in physiological conditions and in response to immunological cues.

Ontogeny of the Drosophila larval hematopoietic organ, hemocyte homeostasis and the dedicated cellular immune response to parasitism.

Over the years, the fruit fly Drosophila melanogaster has become a major invertebrate model to study developmental and evolutionary aspects of both humoral and cellular aspects of innate immunity. Drosophila hematopoiesis which supplies three types of circulating hemocytes, occurs in two spatially and temporally distinct phases during development. The first embryonic phase is described in detail in accompanying reviews in this Int. J. Dev. Biol. Special Issue. The second phase takes place at the end of larval development in a specialised hematopoietic organ, termed the lymph gland. We review here recent studies on the ontogeny of the lymph gland, focusing on the formation and role of the Posterior Signalling Center which acts as a niche for hematopoietic progenitors. We then report recent progress in understanding the dedicated cellular immune response of Drosophila larvae against parasitization by Hymenopterae, a common threat for many Dipterae. This response involves the differentiation of lamellocytes, a cryptic cell fate, revealing the high degree of plasticity of Drosophila hematopoiesis. We end up by integrating studies in Drosophila within a more general picture of insect hematopoiesis and hemocyte homeostasis.

The hematopoietic niche: a Drosophila model, at last.

The niche provides a specialised microenvironment necessary for maintenance of stem cells in a non differentiated state. While the hematopoietic stem cell (HSC) niche in vertebrates was the first to be recognized, Drosophila niches supporting germline stem cells were characterised first. Recent evidence for the existence of a niche maintaining hematopoietic precursors in Drosophila opens the way to study in vivo the niche/hematopoietic precursors interactions. The availability of a large collection of cell markers, mutants and sophisticated genetic tools makes Drosophila an attractive model for investigating the cellular and molecular mechanisms that are involved in these interactions.

Control of blood cell homeostasis in Drosophila larvae by the posterior signalling centre.

Drosophila haemocytes (blood cells) originate from a specialized haematopoietic organ-the lymph gland. Larval haematopoietic progenitors (prohaemocytes) give rise to three types of circulating haemocytes: plasmatocytes, crystal cells and lamellocytes. Lamellocytes, which are devoted to encapsulation of large foreign bodies, only differentiate in response to specific immune threats, such as parasitization by wasps. Here we show that a small cluster of signalling cells, termed the PSC (posterior signalling centre), controls the balance between multipotent prohaemocytes and differentiating haemocytes, and is necessary for the massive differentiation of lamellocytes that follows parasitization. Communication between the PSC and haematopoietic progenitors strictly depends on the PSC-restricted expression of Collier, the Drosophila orthologue of mammalian early B-cell factor. PSC cells act, in a non-cell-autonomous manner, to maintain JAK/STAT signalling activity in prohaemocytes, preventing their premature differentiation. Serrate-mediated Notch signalling from the PSC is required to maintain normal levels of col transcription. The key role of the PSC in controlling blood cell homeostasis is reminiscent of interactions between haematopoietic progenitors and their micro-environment in vertebrates, thus further highlighting the interest of Drosophila as a model system for studying the evolution of haematopoiesis and cellular innate immunity.

Rheological properties of erythrocytes in patients suffering from erysipelas. Examination with LORCA device.

The rheological properties of erythrocytes: elongation and the aggregation, as well as basic peripheral blood parameters: RBC number, hematocrit, MCV, MCH, MCHC reticulocytes, fibrinogen level, ESR, and glucose level were studied in a group of erysipelas patients. The concentration of malonyl dialdehyde (MDA) in erythrocytes was also measured, as an indicator of oxidative stress exerted. The study involved 18 erysipelas patients and 18 healthy subjects. The rheological properties and the aggregation of erythrocytes were studied with the use of a LORCA instrument (Laser-assisted Optical Rotational Cell Analyser). Differences were found in elongation index (EI) between the blood control (C) and patients (P), with an increase of this value only at 0.58 Pa shear stress and at 1.13 Pa shear stress. Upstroke were used as the indexes of RBC deformability, and the following aggregation parameters: intensity of light scattering in the prior disaggregation (Isc dis) - C - 43.0 au (arbitrary units), P - 44.8 au, time when cells are round and not aggregated (Isc top) - C - 51.6 au, P - 49.3 au, the amplitude of aggregation, the difference between Isc max and Isc min (AMP) - C - 39.9 au, P - 28.7 au, aggregation index (AI) - C - 64.6%, P - 70.0%, time for reach one half of the maximum aggregation (t(1/2)) - C - 2.0 s, P - 1.5 s, threshold shear stress, the lowest force that breaks the aggregation formed (Y(thr)) in P exceed 170% control group were measured. A decrease in the elasticity of erythrocytes in erysipelas patients was associated with the simultaneous increase of MDA - C - 0.08 nM/mgHb, P - 0.11 nM/mgHb content in the membranes of red blood cells. Basic peripheral blood parameters studied in patients with erysipelas did not differ significantly from the control group except for WBC, fibrinogen (C - 3.8 g/l, P - 7.8 g/l), and ESR (C - 6.1 mm/h, P - 45.4 mm/h) which were significantly higher.