The Last Minutes of Oxygen Shell Burning in a Massive Star

We present the first 3D simulation of the last minutes of oxygen shell burning in an 18 solar mass supernova progenitor up to the onset of core collapse. A moving inner boundary is used to accurately model the contraction of the silicon and iron core according to a 1D stellar evolution model with a self-consistent treatment of core deleptonization and nuclear quasi-equilibrium. The simulation covers the full solid angle to allow the emergence of large-scale convective modes. Due to core contraction and the concomitant acceleration of nuclear burning, the convective Mach number increases to ~0.1 at collapse, and an l=2 mode emerges shortly before the end of the simulation. Aside from a growth of the oxygen shell from 0.51 to 0.56 solar masses due to entrainment from the carbon shell, the convective flow is reasonably well described by mixing length theory, and the dominant scales are compatible with estimates from linear stability analysis. We deduce that artificial changes in the physics, such as accelerated core contraction, can have precarious consequences for the state of convection at collapse. We argue that scaling laws for the convective velocities and eddy sizes furnish good estimates for the state of shell convection at collapse and develop a simple analytic theory for the impact of convective seed perturbations on shock revival in the ensuing supernova. We predict a reduction of the critical luminosity for explosion by 12--24% due to seed asphericities for our 3D progenitor model relative to the case without large seed perturbations.

Estilos Educativos Parentais (EMBU-A), Sintomatologia Depressiva/Ansiosa, Stress e Autoestima, numa Amostra de Adolescentes

Introdução. Os estilos educativos parentais enquanto clima educativo geral apresentam um forte impacto em diferentes resultados desenvolvimentais (e.g., sintomatologia depressiva/ansiosa e auto estima) da criança/jovem. Podem ser estudados de um ponto de vista dimensional, considerando, por exemplo, as dimensões Controlo/Sobreproteção e Suporte, ou categorial, seguindo, por exemplo, a abordagem de Baumrind, e Maccoby e Martin. Neste estudo pretendemos: verificar se existem diferenças por género e idade, nas dimensões Suporte Emocional, Sobreproteção e Rejeição (EMBU-A) (cada progenitor separadamente), na sintomatologia depressiva/ansiosa e na auto estima; avaliar se existem diferenças nas dimensões do EMBU-A, entre progenitores, na amostra total e por género; explorar associações entre as variáveis centrais do estudo (estilos educativos, sintomatologias e auto estima) e variáveis sociodemográficas, também nas três amostras; explorar diferenças entre quatro grupos criados (Pai-Filho, Pai-Filha; Mãe-Filho e Mãe-Filha) nas dimensões do EMBU-A; combinando a Sobreproteção e o Suporte Emocional (EMBU-A), e definindo os estilos educativos parentais autoritário, autoritativo, permissivo e negligente, calcular a prevalência dos mesmos na nossa amostra (por Pai e por Mãe) e explorar as suas associações com as sintomatologias e com a auto estima (separadamente, por progenitor). Metodologia. A nossa amostra é constituída por 284 adolescentes (idade média = 14,5; DP = 1,68; raparigas, n = 171, 60,2%). Todos preencheram um protocolo composto por um questionário sociodemográfico, pela Rosenberg Self- Esteem Scale (RSES), pelo DASS-21 e pelo Parental Rearing Style Questionnaire for use with Adolescents (EMBU-A/A). Resultados. De salientar o facto de os adolescentes percecionarem a mãe como apresentando valores mais elevados em todas as dimensões do EMBU-A, por comparação com o pai. Da mesma forma, ainda que com diferentes padrões de associação conforme o género, é de salientar a associação entre níveis maiores de Suporte emocional e níveis mais baixos de sintomatologia depressiva/ansiosa e stress, e a associação entre as dimensões Sobreproteção e Rejeição e níveis superiores desses resultados. Uma mais baixa escolaridade do pai associa-se a menor Suporte emocional e uma maior escolaridade da mãe a maior Sobreproteção. Uma díade constituída por progenitor e adolescente do género masculino apresenta resultados inferiores em todas as dimensões do EMBU-A, por comparação com díades formadas por mãe-filho/filha. O estilo Autoritativo é o mais prevalente na nossa amostra (em ambos os progenitores) e o estilo Autoritário é aquele que se associa a pontuações mais elevadas de psicopatologia e a uma menor auto estima. Discussão. De uma forma, genérica os resultados seguem de perto a literatura e revelam a associação entre valores mais elevados na dimensão Suporte emocional e níveis mais baixos de psicopatologia e stress, e a associação entre as dimensões Sobreproteção e Rejeição e níveis superiores desses resultados. Igualmente, o estilo educativo Autoritário está claramente associado a piores resultados nessas mesmas variáveis. São discutidas algumas implicações, no que toca a programas psicoeducativos/educação parental. / Introduction. As a general educational environment, parental rearing styles have a strong impact in different outcomes of development of the child or teenager (e.g. symptoms of depression/ anxiety and self-esteem). These can be studied from a dimensional point of view considering for example the dimensions Control/Overprotection and Support or from a categorical point of view following for example the approach of Baumrind and Maccoby and Martin. This study aims at checking whether there are differences of gender and age in the dimensions of Emotional Support, Overprotection and Rejection (EMBU-A) (each parent separately) in depressive symptoms/anxiety and self esteem; assessing whether there are differences in the dimensions of the EMBU-A, between parents in the total sample and by gender; exploring associations between the study’s main variables (rearing styles, symptomatology and self esteem) and sociodemographic variables, also in the three samples; exploring differences between the four groups created (Father-Son, Father-Daughter, Mother-Son and Mother-Daughter) in the dimensions of the EMBU-A, combining Overprotection and Emotional Support(EMBU-A) and defining the authoritarian, authoritative, indulgent, and neglectful parental rearing styles. The aim is also to estimate the prevalence of these in our sample (by Father and Mother) and explore their associations to the symptomatology and self esteem (separately, per parent). Methodology. Our sample is composed of 284 teenagers (average age = 14,5; DP = 1,68; girls, n = 171, 60,2%). All participants filled in a protocol of questionnaires consisting of a set of socio demographic questions by Rosenberg Self- Esteem Scale (RSES), by DASS-21 and by Parental Rearing Style Questionnaire for use with Adolescents (EMBU-A/A). Outcomes. It is important to emphasize the fact that teenagers perceive their mother as having higher values in all the EMBU-A dimensions compared to their father. In the same way, though with different patterns of association according to gender, it is important to emphasize the association of higher levels of Emotional Support and lower levels of depressive/anxious symptomathology and stress and the association of Overprotection and Rejection and higher levels of those outcomes. A father’s lower level of education is associated to a lower Emotional Support while a mother’s higher level of education is associated to a greater Overprotection. A dyad composed of male parent and male teenager presents lower outcomes in all EMBU-A dimensions if compared to dyads composed of mother-son/daughter. The authoritative style is the most prevalent in our sample (in both parents) and the authoritarian style is the one associated to higher scores of psychopathology and lower levels of self esteem. Debate. In general, the outcomes closely follow the literature review and reveal the association of higher values in the Emotional Support dimension and lower levels of psychopathology and stress and also the association of the dimensions of Overprotection and Rejection and higher levels of those outcomes. Equally, the authoritarian rearing style is clearly associated to the worst outcomes in those same variables. Some implications are discussed as far as psychoeducational programmes and parental rearing are concerned.

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Proliferative Defects and Formation of a Double Cortex in Mice Lacking Mltt4 and Cdh2 in the Dorsal Telencephalon

Radial glial cells (RGCs) in the ventricular neuroepithelium of the dorsal telencephalon are the progenitor cells for neocortical projection neurons and astrocytes. Here we showthatthe adherens junction proteins afadin and CDH2 are criticalforthe control of cell proliferation in the dorsal telencephalon and for the formation of its normal laminar structure. Inactivation of afadin or CDH2 in the dorsal telenceph-alon leads to a phenotype resembling subcortical band heterotopia, also known as “double cortex,” a brain malformation in which heterotopic gray matter is interposed between zones of white matter. Adherens junctions between RGCs are disrupted in the mutants, progenitor cells are widely dispersed throughout the developing neocortex, and their proliferation is dramatically increased. Major subtypes of neocortical projection neurons are generated, but their integration into cell layers is disrupted. Our findings suggest that defects in adherens junctions components in mice massively affects progenitor cell proliferation and leads to a double cortex-like phenotype.

Synthetic Polymer and Microbubble Systems for Gene Delivery to the Brain

Thesis (Ph.D.)--University of Washington, 2016-08

Role for cell-to-cell communication in stem cell specification toward pancreatic progenitors: relevance to the design of novel therapies for diabetes.

Thesis (Ph.D.)--University of Washington, 2016-08

Regenerative Cues of Myeloid Cells on Pancreatic Epithelium

Thesis (Ph.D.)--University of Washington, 2016-07

Proficiency and mechanisms of perturbation of mature T-cell homeostasis by the TCL1 family of oncogenes

In the first part of this thesis, the oncogenic potential of TCL1A family genes was comparatively evaluated by using gamma-retroviral vectors to introduce human TCL1A, MTCP1, and TML1 into hematopoietic stem cells/hematopoietic progenitor cells (HSC/HPC) of wild type mice that were transplanted into wild type recipients. TCL1A and MTCP1 recipient mice predominantly developed B-cell malignancies after a median survival of 388 days and 394 days, respectively. The presented data indicates that TCL1A and MTCP1 are oncogenes with comparable oncogenic potential and shows for the first time that MTCP1 is not only a T-cell oncogene, but is able to transform B cells as well. The third family member TML1 induced the development of immature T-cell malignancies in only a few mice. This study provides first evidence for its oncogenic function. Additionally, the transforming potential of compartment-targeted TCL1A variants was evaluated by retroviral expression of a membrane localizing myristoylated (myr-TCL1A) and a nuclear localizing (nls-TCL1A) variant. Recipients of HSC/HPC transduced with myr-TCL1A and nls-TCL1A predominantly developed B-cell malignancies after a median survival of 360 days and 349 days, respectively. There was a significantly shorter latency period for nls-TCL1A compared to the previously described generic TCL1A. Gene expression analysis revealed higher similarities between expression profiles of tumors induced by TCL1A and nls-TCL1A. Together these data implicate that TCL1A’s predominant oncogenic function might rely on its nuclear presence. The second part of this thesis aims to understand if and how TCR stimulation affects the transforming potential of TCL1A. Mature OT-1 T cells carrying monoclonal TCR’s that specifically recognize ovalbumin (OVA) were retrovirally transduced with TCL1A and repeatedly stimulated in vivo with OVA-peptides. TCR stimulated recipient mice of TCL1A transduced T cells showed a significantly accelerated leukemic outgrowth and a reduced median survival of 305 days, when compared to unstimulated recipients (417 days). These data strongly implicate a pro-leukemogenic cooperation of TCL1A and TCR signals that might be actionable in upcoming interventional designs.

Étude des gènes de fusion MLL dans les leucémies aigues humaines

Les leucémies aigues sont la conséquence d’une prolifération clonale et maligne des cellules hématopoïétiques. Elles surviennent suite à un évènement oncogénique qui se produit dans une cellule souche hématopoïétique (CSH) ou progénitrice. Cela lui confère une certaine instabilité qui engendre l’accumulation d’autres évènements génétiques et/ou épigénétiques responsables du développement clinique de la maladie. Les leucémies MLL représentent environ 10% des leucémies aigues et aujourd’hui, plus de 70 gènes de fusion ont été caractérisés. Les sangs de cordon sont une source importante de CSH et progénitrices. La purification de ces cellules et leur transformation en cellules leucémiques à l’aide de gènes de fusion MLL nous permettent de générer des leucémies aigues humaines dans des souris immunodéficientes NSG et ainsi étudier le potentiel leucémique de différents gènes de fusion MLL. Dans un premier temps, 4 gènes de fusion MLL ont été étudiés : MLL-AF9, MLL-AF4, MLL-ENL et MLL-ELL. In vitro, nous sommes capables de transformer des CSH en cellules leucémiques capables de proliférer rapidement. Les résultats in vivo nous montrent qu’il est possible de générer des leucémies avec les oncogènes MLL-AF9 et MLL-ENL. Pour les fusions MLL-ELL et MLL-AF4, bien que quelques leucémies ont pu être obtenues, plusieurs problèmes techniques nous empêchent aujourd’hui de disposer d’un modèle adéquat permettant l’étude complète de ces oncogènes. Dans un second temps, les leucémies aigues MLL-AF9 ont été étudiées dans un modèle contrôlé où les cellules souches proviennent d’un donneur unique. Grâce à ce modèle, nous avons pu démontrer que l’oncogène MLL-AF9 est suffisant pour induire le développement de la maladie. En effet aucune nouvelle mutation n’a pu être identifiée au cours du développement de la leucémie. Parmi les leucémies myéloïdes aigues (LMA) MLL-AF9 issues de ce modèle, certains gènes non mutés, dont RET, ont été identifiés comme étant de potentiels biomarqueurs de ce sous-groupe de leucémie.

Sources of antibiotic resistance in the environment

The discovery of antibiotics was a major breakthrough in medicine. However, short after their introduction in clinical practice resistant bacteria were detected. Nowadays, antibiotic resistance constitutes a serious public health problem. In hospital settings, with high resistance levels, reducing drastically the therapeutic options. Carbapenems are last-resort antibiotics used in Portugal, only in hospitals, to treat serious infections. Bacterial resistance towards this class of antibiotics has increased during last years. In Gram-negative bacteria the production of carbapenemases is a common resistance mechanism. OXA-48 is a carbapenemase of Ambler class D and represents a major concern for human health. It is frequently detected in clinical isolates of Enterobacteriaceae. There are few studies suggesting that genes encoding for OXA-48 variants originated from genes present in the chromosome of members of genus Shewanella, and have disseminated to Enterobacteriaceae members, associated with mobile genetic elements. The aim of this study was to characterize strains from different sources of Shewanella to confirm its role as OXA-48 progenitor. For this, the phylogenetic affiliation of 33 strains of Shewanella was performed by 16SrDNA and gyrB sequencing. The most common species were S. hafniensis and S. xiamenensis, but also S. aestuarii, S. baltica, S. indica, S. haliotis, S. putrefaciens, S. algidipiscicola, S. irciniae, S. algae and S. fodinae were identified. blaOXA-48-like genes were detected in 21 isolates: S. hafniensis (8/8), S. xiamenensis (5/5), S. baltica (4/4), S. algae (1/1), S. fodinae (1/1), S. putrefaciens (1/2) and S. algidipiscicola (1/2). Sequence analysis revealed that genes encoded enzymes identical to OXA-48, OXA-181 and OXA-204 but also new variants differing from OXA-48 from 2 to 81 aminoacids. Genetic context analysis revealed the C15 gene upstream and lysR gene downstream, identical to what has been identified so far flanking blaOXA-48-like genes in Shewanella spp. The assessment of antibiotic susceptibility was performed for all isolates using the disk diffusion method. In general, it was observed a great sensitivity for all antibiotics except to amoxicillin and aztreonam. Multidrug resistance was detected in only 1 isolate. Other resistance genes and the presence of integrons were not identified. Plasmids were detected in 30.3% isolates (10/ 33). These results reinforce the role of Shewanella spp. as origin of blaOXA-48-like genes.

Parámetros fenotípicos y genéticos de caracteres de la mazorca asociados al rendimiento del cultivo de maíz (Zea mays L.) cv NB-6, en postrera, Managua 2014

Con el propósito de evaluar la influencia del número de hileras en la mazorca de progenitores maternos (10, 12, 14, 16, 18, 20) sobre variables asociadas al rendimiento, estimar grados de asociación fenotípica entre rasgos y su variabilidad genética, se estableció un experimento de campo en bloques completos al azar (BCA) con cuatro réplicas, en la Finca Santa Rosa propiedad de la UNA, en época de postrera del 2014. Las variables analizadas fueron: Longitud de la mazorca (LM), diámetro de la mazorca (DM) , número de granos por hilera (NGH), peso de 100 granos (P100G), numero de hileras por mazorca ( NHM),peso de la mazorca (PM),peso en grano por mazorca (PGM), mediante los procedimientos de SAS, con estadísticos descriptivos, correlaciones de Pearson y análisis de varianza con regresión de los caracteres maternos sobre los de progenie. Los tratamientos que presentaron promedios más altos para LMp, DMp, NHMp, PMp, PGMp fueron los de 14 y 18 hileras. El tratamiento con 16 hileras mostró la más alta variación fenotípica (CV). Las correlaciones fenotípicas de progenies más destacadas fueron: PMp con PGMp y DMp; PMp con NGHp; DMp con PGMp; PMp con NGHp; NGHp con PGMp; y LMp con PMp, todas ellas positivas y altamente significativas. Las correlaciones fenotípicas entre variables de la progenie con progenitor materno más notorias fueron LM, con NHM y P100G. Del análisis de varianza, el factor NHMm fue altamente significativo para LMp y NHMp y en menor grado para DMp y P100Gp. Se estimaron niveles de variabilidad genética importante en rasgos como LMp, NHMp y NGHp, con valores de heredabilidad entre 0.36 y 0.46, los cuales son muy promisorios para programas de mejoramiento genético participativo.

The Influence of Age, Cardiorespiratory Fitness, Exercise and Sedentary Behaviours on Circulating Angiogenic Cells and Cell Surface Receptor Expression.

Cardiovascular disease (CVD) is the biggest killer of people in western civilisation. Age is a significant risk factor for the development for CVD, and treatments and therapies to address this increased risk are crucial to quality of life and longevity. Exercise is one such intervention which has been shown to reduce CVD risk. Age is also associated with endothelial dysfunction, reduced angiogenic capabilities, and reduced ability to repair the vessel wall. Circulating angiogenic cells (CACs) are a subset of circulating cells which assist in the repair and growth of the vasculature and in the maintenance of endothelial function. Reductions in these cells are observed in those with vascular disease compared to age-matched healthy controls. Exercise may reduce CVD risk by improvements in number and/or function of these CACs. Data was collected from human volunteers of various ages, cardiorespiratory fitness (CRF) levels and latent viral infection history status to investigate the effects of chronological age, CRF, viral serology and other lifestyle factors, such as sedentary behaviours and exercise on CACs. The levels of CACs in these volunteers were measured using four colour flow cytometry using various monoclonal antibodies specific to cell surface markers that are used to identify specific subsets of these CACs. In addition, the response to acute exercise of a specific subset of these CACs, termed ‘angiogenic T-cells’ (TANG) were investigated, in a group of well-trained males aged 20-40 years, using a strenuous submaximal exercise bout. Advancing age was associated with a decline in various subsets of CACs, including bone marrow-derived CD34+ progenitors, putative endothelial progenitor cells (EPCs) and also TANG cells. Individuals with a higher CRF were more likely to have higher circulating numbers of TANG cells, particularly in the CD4+ subset. CRF did not appear to modulate CD34+ progenitors or EPC subsets. Increasing sitting time was associated with reduction in TANG cells, but after correcting for the effects of fitness, sitting time no longer negatively affected the circulating number of these cells. Acute exercise was a powerful stimulus for increasing the number of TANG cells (140% increase), potentially through an SDF-1:CXCR4-dependent mechanism, but more studies are required to investigate this. Latent CMV infection was associated with higher number of TANG cells (CD8+), but only in 18-40 year old individuals, and not in an older age group (41-65 year old). The significance of this has yet to be understood. In conclusion, advancing age may contribute to increased CVD risk partly due to the observed reductions in angiogenic cells circulating in the peripheral compartment. Maintaining a high CRF may attenuate this CVD reduction by modulating TANG cell number, but potentially not CD34+ progenitor or EPC subsets. Acute exercise may offer a short window for vascular adaptation through the mobilisation of TANG cells into the circulation.

TRIB2 in normal and malignant hematopoiesis - a transcription factor network

Hematopoiesis is the tightly controlled and complex process in which the entire blood system is formed and maintained by a rare pool of hematopoietic stem cells (HSCs), and its dysregulation results in the formation of leukaemia. TRIB2, a member of the Tribbles family of serine/threonine pseudokinases, has been implicated in a variety of cancers and is a potent murine oncogene that induces acute myeloid leukaemia (AML) in vivo via modulation of the essential myeloid transcription factor CCAAT-enhancer binding protein α (C/EBPα). C/EBPα, which is crucial for myeloid cell differentiation, is commonly dysregulated in a variety of cancers, including AML. Two isoforms of C/EBPα exist - the full-length p42 isoform, and the truncated oncogenic p30 isoform. TRIB2 has been shown to selectively degrade the p42 isoform of C/EBPα and induce p30 expression in AML. In this study, overexpression of the p30 isoform in a bone marrow transplant (BMT) leads to perturbation of myelopoiesis, and in the presence of physiological levels of p42, this oncogene exhibited weak transformative ability. It was also shown by BMT that despite their degradative relationship, expression of C/EBPα was essential for TRIB2 mediated leukaemia. A conditional mouse model was used to demonstrate that oncogenic p30 cooperates with TRIB2 to reduce disease latency, only in the presence of p42. At the molecular level, a ubiquitination assay was used to show that TRIB2 degrades p42 by K48-mediated proteasomal ubiquitination and was unable to ubiquitinate p30. Mutation of a critical lysine residue in the C-terminus of C/EBPα abrogated TRIB2 mediated C/EBPα ubiquitination suggesting that this site, which is frequently mutated in AML, is the site at which TRIB2 mediates its degradative effects. The TRIB2-C/EBPα axis was effectively targeted by proteasome inhibition. AML is a very difficult disease to target therapeutically due to the extensive array of chromosomal translocations and genetic aberrations that contribute to the disease. The cell from which a specific leukaemia arises, or leukaemia initiating cell (LIC), can affect the phenotype and chemotherapeutic response of the resultant disease. The LIC has been elucidated for some common oncogenes but it is unknown for TRIB2. The data presented in this thesis investigate the ability of the oncogene TRIB2 to transform hematopoietic stem and progenitor cells in vitro and in vivo. TRIB2 overexpression conferred in vitro serially replating ability to all stem and progenitor cells studied. Upon transplantation, only TRIB2 overexpressing HSCs and granulocyte/macrophage progenitors (GMPs) resulted in the generation of leukaemia in vivo. TRIB2 induced a mature myeloid leukaemia from the GMP, and a mixed lineage leukaemia from the HSC. As such the role of TRIB2 in steady state hematopoiesis was also explored using a Trib2-/- mouse and it was determined that loss of Trib2 had no effect on lineage distribution in the hematopoietic compartment under steady-state conditions. The process of hematopoiesis is controlled by a host of lineage restricted transcription factors. Recently members of the Nuclear Factor 1 family of transcription factors (NFIA, NFIB, NFIC and NFIX) have been implicated in hematopoiesis. Little is known about the role of NFIX in lineage determination. Here we describe a novel role for NFIX in lineage fate determination. In human and murine datasets the expression of Nfix was shown to decrease as cells differentiated along the lymphoid pathway. NFIX overexpression resulted in enhanced myelopoiesis in vivo and in vitro and a block in B cell development at the pre-pro-B cell stage. Loss of NFIX resulted in disruption of myeloid and lymphoid differentiation in vivo. These effects on stem and progenitor cell fate correlated with changes in the expression levels of key transcription factors involved in hematopoietic differentiation including a 15-fold increase in Cebpa expression in Nfix overexpressing cells. The data presented support a role for NFIX as an important transcription factor influencing hematopoietic lineage specification. The identification of NFIX as a novel transcription factor influencing lineage determination will lead to further study of its role in hematopoiesis, and contribute to a better understanding of the process of differentiation. Elucidating the relationship between TRIB2 and C/EBPα not only impacts on our understanding of the pathophysiology of AML but is also relevant in other cancer types including lung and liver cancer. Thus in summary, the data presented in this thesis provide important insights into key areas which will facilitate the development of future therapeutic approaches in cancer treatment.

Dissecting signaling pathways that control glia migration in the developing Drosophila retina

The function of a complex nervous system relies on an intricate interaction between neurons and glial cells. However, as glial cells are generally born distant from the place where they settle, molecular cues are important to direct their migration. Glial cell migration is important in both normal development and disease, thus current research in the laboratory has been focused on dissecting regulatory events underlying that crucial process. With this purpose, the Drosophila eye imaginal disc has been used as a model. In response to neuronal photoreceptor differentiation, glial cells migrate from the CNS into the eye disc where they act to correctly wrap axons. To ensure proper development, attractive and repulsive signals must coordinate glial cell migration. Importantly, one of these signals is Bnl, a Fibroblast Growth Factor (FGF) ligand expressed by retinal progenitor cells that was suggested to act as a non-autonomous negative regulator of excessive glial cell migration (overmigration) by binding and activating the Btl receptor expressed by glial cells. Through the experimental results described in chapter 3 we gained a detailed insight into the function of bnl in eye disc growth, photoreceptor development, and glia migration. Interestingly, we did not find a direct correlation between the defects on the ongoing photoreceptors and the glia overmigration phenotype; however, bnl knockdown caused apoptosis of eye progenitor cells what was strongly correlated with glia migration defects. Glia overmigration due to Bnl down-regulation in eye progenitor cells was rescued by inhibiting the pro-apoptotic genes or caspases activity, as well as, by depleting JNK or Dp53 function in retinal progenitor cells. Thus, we suggest a cross-talk between those developmental signals in the control of glia migration at a distance. Importantly, these results suggest that Bnl does not control glial migration in the eye disc exclusively through its ability to bind and activate its receptor Btl in glial cells. We also discuss possible biological roles for the glia overmigration in the bnl knockdown background. Previous results in the lab showed an interaction between dMyc, a master regulator of tissue growth, and Dpp, a Transforming Growth Factor-β important for retinal patterning and for accurate glia migration into the eye disc. Thus, we became interested in understanding putative relationships between Bnl and dMyc. In chapter 4, we show that they positively cooperate in order to ensure proper development of the eye disc. This work highlights the importance of the FGF signaling in eye disc development and reveals a signaling network where a range of extra- and intra-cellular signals cooperate to non-autonomously control glial cell migration. Therefore, such inter-relations could be important in other Drosophila cellular contexts, as well as in vertebrate tissue development.

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.