Regulatory signals and signal molecules in early neurogenesis of Drosophila melanogaster

Summary The ectodermal germ layer of Drosophila melanogaster gives rise to two major cell lineages, the neural and the epidermal. Progenitor cells for each of these lineages arise from groups of cells, whose elements must decide between taking on either fate. Commitment of the progenitor cells to one of the developmental fates implies two factors. One is intrinsic to the ectodermal cells and determines a propensity to take on neural fate; this factor is probably represented by the products of the so-called proneural genes, which are differentially distributed throughout the ectoderm. The other factor in the cells' decision to adopt one of the two alternative fates is intercellular communication, which is mediated by the products of the so-called neurogenic genes. Two types of interactions, one inhibiting and the other stimulating neural development, have been inferred. We discuss here the assumed role of various neurogenic genes, in particular Notch and Delta, in these processes.Offprint requests to: J.A. Campos-Ortega     

Endocytosis-independent mechanisms of Delta ligand proteolysis

Delta proteins function as cell surface ligands for Notch receptors in a highly conserved signal transduction mechanism. Delta activates Notch by "trans-endocytosis", whereby endocytosis of Delta that is in complex with Notch on a neighboring cell induces activating cleavages in Notch. Alternatively, proteolysis of Delta renders the ligand inactive by dissociating the extracellular and cytosolic domains. How proteolysis and trans-endocytosis cooperate in Delta function is not well understood. We now show that Drosophila Delta proteolysis occurs independent of and prior to endocytosis in neuroblasts and ganglion mother cells in vivo and cells in culture. Delta cleavage occurs at two novel sites that we identify in the juxtamembrane (JM) and transmembrane (TM) domains. In addition to the previously identified Kuzbanian ADAM protease, which acts on the JM domain, proteolysis in the TM domain is facilitated by a thiol-sensitive aspartyl protease that is distinct from Presenilin. Furthermore, cleavage in the TM domain is upregulated in the presence of Notch. Overall, Drosophila Delta proteolysis differs from the conventional regulated intramembrane proteolysis (RIP) mechanism by two criteria: (1) TM-domain processing of Delta is not sensitive to Presenilin, and (2) TM and JM domain cleavages occur independently of each other. Altogether, these data support a model whereby proteolysis can modulate Delta ligand activity independently of endocytosis.     

The Plasticity of the DRG Neurons Belonging to Different Subpopulations After Dorsal Rhizotomy

SUMMARY 1. The plasticity of sensory neurons following the injury to their axons is very important for prognosis of recovery of afferent fibers with different modality. It is evident that the response of dorsal root ganglion (DRG) neurons after peripheral axotomy is different depending on the deficiency in neurotrophic factors from peripheral region. The loss of cells appears earlier and is more severe in B-cells (small, dark cells with unmyelinated axons) than in A-cells (large, light cells with myelinated axons).2. We studied using immunohistochemical methods the response of DRG neurons to dorsal rhizotomy and combined injury of central and peripheral neuronal processes. A quantitative analysis of DRG neurons tagged by the selective markers isolectin B4 (IB4) and the heavy molecular component of the neurofilament triplet (NF200) antibody, selective for subpopulations of small and large/medium DRG neurons, respectively, was performed after dorsal rhizotomy, peripheral axotomy, and their combination.3. The number of NF200⁺-neurons is reduced substantially after both dorsal rhizotomy and peripheral axotomy, while the decrease of IB4⁺-neurons is observed only in combined injury, i.e., dorsal rhizotomy accompanied with sciatic nerve injury.4. Our results show that distinct subpopulations of DRG neurons respond differently to the injury of their central processes. The number of NF200⁺-neurons decreases to greater degree following dorsal rhizotomy in comparison to IB4⁺-neurons.     

Nervous control of egg progression into the common oviduct and genital chamber of the stick-insect Carausius morosus

In Carausius morosus, egg-laying activity is nocturnal. When an oöcyte leaves the ovariole (ovulation) during the day, it remains in a chamber which is located between the genital valves until the evening. The admission of another egg into the common oviduct is possible only when the egg chamber is empty. Nerve cord transection between the 7th and 8th abdominal ganglia interrupts the reflex which controls the progression of the egg through the common oviduct.Sensory hairs on the walls of the egg chamber, which is innervated by the 8th abdominal ganglion, are described. The information perceived by these sensory hairs enters the 8th ganglion, then travels to the 7th which innervates the oviducts. Present experimental results together with the observed existence of numerous anastomoses between the segmental nerves from different ganglia, suggest that though the normal reflex travels via the nerve cord, a lateral pathway is also possible.The effector is the common oviduct. The muscular sheath of this organ shows a special feature which is related to the control of egg passage: the muscle fibres no longer form a complete layer as they do in the lateral oviducts but rather form a half-sleeve of circular fibres around the dorsal face. The contraction of this half-cylindrical muscle prevents the progression of eggs.     

Role of Notch pathway in terminal follicle cell differentiation during Drosophila oogenesis

 During Drosophila oogenesis the body axes are determined by signaling between the oocyte and the somatic follicle cells that surround the egg chamber. A key event in the establishment of oocyte anterior-posterior polarity is the differential patterning of the follicle cell epithelium along the anterior-posterior axis. Both the Notch and epithelial growth factor (EGF) receptor pathways are required for this patterning. To understand how these pathways act in the process we have analyzed markers for anterior and posterior follicle cells accompanying constitutive activation of the EGF receptor, loss of Notch function, and ectopic expression of Delta. We find that a constitutively active EGF receptor can induce posterior fate in anterior but not in lateral follicle cells, showing that the EGF receptor pathway can act only on predetermined terminal cells. Furthermore, Notch function is required at both termini for appropriate expression of anterior and posterior markers, while loss of both the EGF receptor and Notch pathways mimic the Notch loss-of-function phenotype. Ectopic expression of the Notch ligand, Delta, disturbs EGF receptor dependent posterior follicle cell differentiation and anterior-posterior polarity of the oocyte. Our data are consistent with a model in which the Notch pathway is required for early follicle cell differentiation at both termini, but is then repressed at the posterior for proper determination of the posterior follicle cells by the EGF receptor pathway. Received: 5 November 1998 / Accepted: 14 December 1998     

兴义维蚋幼虫扭动摄食方式的神经-肌肉结构基础

蚋类幼虫为河流清洁者,可作为水污染的指标.本文用抗辣根过氧化物酶抗体(anti-HRP)和鬼笔环肽(phalloidin),通过免疫荧光显示兴义维蚋Simulium xingyiense幼虫腹神经索及外周神经与肌肉连接结构.结果表明幼虫腹神经索由11对神经节及连接的神经索组成.每对体神经节由两团神经胞体聚合而成,细胞核分布呈无头无尾的Φ形.在1～5腹神经节两侧,纵神经连接横神经呈现\"梯形结构\";虫体腹部肌肉分横肌、纵肌和斜肌3类肌群.神经节中部发出两对侧神经入腹肌,再继续分支,其神经末梢呈链珠状突触结构连接肌肉,增加神经与肌肉的接触面积.这种神经-肌肉的连接方式可能与兴义维蚋幼虫扭动摄食行为有关.本研究初步阐述了兴义维蚋幼虫腹神经索及腹部神经与肌肉连接的细微结构基础.     

Neurexin-1 is required for synapse formation and larvae associative learning in Drosophila

Neurexins are highly polymorphic cell-surface adhesive molecules in neurons. In cultured mammalian cell system, they were found to be involved in synaptogenesis. Here, we report for the first time that Drosophila neurexin is required for synapse formation and associative learning in larvae. Drosophila genome encodes a single functional neurexin (CG7050; Neurexin-1 or Nrx-1), which is a homolog of vertebrate alpha-neurexin. Neurexin-1 is expressed in central nervous system and highly enriched in synaptic regions of the ventral ganglion and brain. Neurexin-1 null mutants are viable and fertile, but have shortened lifespan. The synapse number is decreased in central nervous system in Neurexin-1 null mutants. In addition, Neurexin-1 null mutants exhibit associative learning defect in larvae.     

节肢动物肠道及腹神经索的降解实验*

节肢动物的内部器官组织, 如消化系统和神经系统等精细结构在寒武纪化石中的保存特征对探索早期生命和地球环境的演化具有重要意义。近年来, 其原始形态与保存机理被广泛讨论, 说明此类型软躯体组织的保存行为还需多方面的实验和理论探索。实验埋藏学作为埋藏学的分支, 为解释生物化石化过程中解剖结构的变化及相应特异埋藏化石库的形成提供了重要理论支撑和实验技术手段。本文以节肢动物南美白对虾的肠道和腹神经索为材料, 在不同温度及组合条件下进行埋藏学实验, 以观察此类生物软组织的降解过程。实验结果显示, 肠道和腹神经索在不同温度下的降解趋势相似, 但降解速率不同: 肠道和腹神经索在25℃条件下的降解速率和降解程度明显大于其在10℃条件。同一温度条件下, 肠道与腹神经索的降解程度存在早期差异; 10℃条件下, 反应产生的微生物膜形态及规模有所不同, 肠道-腹神经索组合产生的微生物膜规模远大于肠道、腹神经索的单独处理组, 且难以辨认。由此可见, 温度是影响肠道和腹神经索降解速度和程度的重要因素。此外, 微生物在生物内部器官组织降解过程中起到了重要的生化作用。生物器官组织的降解过程, 以及不同条件下微生物膜的形成对化石的形态特征尤其是内部精细结构的保存解释具有重要影响。     

The relationship between the ventral nerve cord,body size and phylogeny in ground beetles (Coleoptera: Carabidae)

The ventral nerve cord in the family Carabidae (considered in the widest sense! exhibits variations in the degree of fusion of thoracic and abdominal ganglia. There are usually three discrete thoracic ganglia and between one and seven discrete abdominal ganglia, the number differing between tribes. Of the 44 tribes and 177 species examined, 38 tribes contained species showing no differences in the degree of ventral nerve cord consolidation. However, the remaining six tribes showed variations in the degree of ventral nerve cord consolidation between genera (Lebiini, Cychrini, Nebriini, Scaritini, Licinini and Brachinini), whilst one genus showed variations between species (Leistus, Nebriini). No variation in ventral nerve cord consolidation was observed in conspecifics. The degree of ventral nerve cord consolidation is inversely proportional to overall body length. With respect to phylogeny, the degree of consolidation of the nerve cord docs not consistently support the traditional Carabinae-Harpalinae subfamily division. However, the Harpalinae always have four or less discrete abdominal ganglia (with the sole exception of the Broscinij, whilst the Carabinae exhibit almost the whole range of variations. Thus the Harpalinae (or the major pari of it) may be a monophyletic group, but this is not true of the Carabinae. Trends in the degree of ventral nerve cord consolidation for the various tribes were noted, and phylogenetic implications were evaluated wherever possible.     

The relationship between the ventral nerve cord, body size and phylogeny in ground beetles (Goleoptera: Garabidae)

The ventral nerve cord in the family Carabidae (considered in the widest sense! exhibits variations in the degree of fusion of thoracic and abdominal ganglia. There are usually three discrete thoracic ganglia and between one and seven discrete abdominal ganglia, the number differing between tribes. Of the 44 tribes and 177 species examined, 38 tribes contained species showing no differences in the degree of ventral nerve cord consolidation. However, the remaining six tribes showed variations in the degree of ventral nerve cord consolidation between genera (Lebiini, Cychrini, Nebriini, Scaritini, Licinini and Brachinini), whilst one genus showed variations between species (Leistus , Nebriini). No variation in ventral nerve cord consolidation was observed in conspecifics. The degree of ventral nerve cord consolidation is inversely proportional to overall body length. With respect to phylogeny, the degree of consolidation of the nerve cord docs not consistently support the traditional Carabinae-Harpalinae subfamily division. However, the Harpalinae always have four or less discrete abdominal ganglia (with the sole exception of the Broscinij, whilst the Carabinae exhibit almost the whole range of variations. Thus the Harpalinae (or the major pari of it) may be a monophyletic group, but this is not true of the Carabinae. Trends in the degree of ventral nerve cord consolidation for the various tribes were noted, and phylogenetic implications were evaluated wherever possible.     

Discovery and implications of transcellular mitophagy

The mitochondrial quality control system regulating mitochondria biogenesis, dynamics, and degradation has been extensively studied because of its roles in normal cell homeostasis and dysfunction due to aging or disease. Mitochondria degradation is generally thought to occur by autophagy and has therefore been viewed as a cell-autonomous process. In a recent study, we demonstrated that a large fraction of retinal ganglion cell mitochondria undergo lysosomal degradation within the astrocytes of the optic nerve head. It will be important to determine whether other neurons with long axons also use transcellular mitophagy, or transmitophagy, as a primary mitochondrial quality control mechanism either under normal physiological conditions or in disease. The elucidation of the underlying molecular mechanisms is necessary to determine whether defects in transmitophagy are involved in pathogenesis and whether it should become a therapeutic target.     

Isolation of the abdomen releases oviposition behaviour in females of the cricket, Acheta domesticus

Upon isolation, abdomens of adult female house crickets (Acheta domesticus) produced abdominal and ovipositor movements characteristic of normal oviposition. Oviposition behaviour was thus released even in reproductively mature or immature virgins where under normal conditions it was never observed. Decapitation was not sufficient to release oviposition behaviour but transection of the ventral nerve cord between the thorax and the abdomen of immobilized females evoked the response. These observations indicate that the motor programmes for certain components of the oviposition sequence reside in the abdominal ganglia. Moreover, the prerequisite circuitry for ovipositional posturing of the abdomen and ovipositor appears to be functional prior to sexual maturity and insemination, primed by mating, and subject to inhibition by the thoracic ganglia.     

Dll1 and Dll4 function sequentially in the retina and pV2 domain of the spinal cord to regulate neurogenesis and create cell diversity

Signalling mediated by Notch receptors is known to have multiple functions during vertebrate neural development, regulating processes like progenitor differentiation and cell type diversification. Various Notch ligands are expressed in the developing nervous system and their activities might contribute to this multiplicity of functions. Here, we show that two Delta-like genes, Dll1 and Dll4, are sequentially expressed in differentiating neurons of the embryonic mouse retina and spinal cord's pV2 domain, with Dll1 starting to be expressed before Dll4. Analysis of Dll1 mutants reveals this gene is necessary and sufficient to maintain a pool of progenitors in the embryonic neuroepithelium. Accordingly, in the spinal cord domains where Dll1 is the only expressed Notch ligand, its inactivation leads to an increased rate of neurogenesis and premature differentiation of neural progenitors. In contrast, in the pV2 domain and retina where Dll1 is co-expressed with Dll4, progenitors are not exhausted and cell diversity is maintained. Together, our results support a model where Dll1 and Dll4 are part of a unique genetic circuitry that regulates subsequent steps of neurogenesis in the retina and pV2 domain: while Dll1 serves to prevent the untimely differentiation of neural progenitors, Dll4 might function to generate diversity within the population of differentiating neurons.