Ca(2+)-triggered exocytosis is a hallmark of neurosecretory granules, but the cellular pathway leading to assembly these regulated exocytotic carriers poorly understood. Here we used pituitary AtT-20 cell line study biogenesis involved in peptide hormone secretion. We show that immature secretory granules (ISGs) freshly budded from trans-Golgi network (TGN) exhibit characteristics unregulated carriers. During subsequent maturation period they undergo an important switch become have...

The diaphanous gene is the founding member of a family Diaphanous-related formin proteins (DRFs). We identified in screen for genes that are necessary normal growth and stabilization Drosophila neuromuscular junction (NMJ). Here, we demonstrate mutations perturb synaptic at NMJ. Diaphanous protein present both presynaptically postsynaptically. However, genetic rescue experiments combination with additional interaction support conclusion dia NMJ growth. then document defects actin microtubule...

The limited resolving power of conventional diffraction-limited microscopy hinders analysis small, densely packed structural elements in cells. Expansion (ExM) provides an elegant solution to this problem, allowing for increased resolution with standard microscopes via physical expansion the specimen a swellable polymer hydrogel. Here, we apply, validate, and optimize ExM protocols that enable study Drosophila embryos, larval brains, adult body walls. We achieve lateral ∼70 nm tissues using...

Motor coordination is broadly divided into gross and fine motor control, both of which depend on proprioceptive organs. However, the channels that function specifically in control are unknown. Here we show mutations trpγ disrupt while leaving proficiency intact. The mutants unable to coordinate precise leg movements during walking, ineffective traversing large gaps due an inability making subtle postural adaptations requisite for this task. TRPγ expressed organs, required neurons glia gap...

Homeostatic plasticity functions within the nervous system to maintain normal neural functions, such as neurotransmission, predefined optimal ranges. The defined output of these neuronal processes is referred set point, which value that homeostatic defends against fluctuations. Currently, it unknown how stable points are system. In present study we used CM9 neuromuscular junctions (NMJs) in adult Drosophila investigate stability point synaptic homeostasis across lifespan fly. At fly NMJ,...

The biogenesis of peptide hormone secretory granules involves a series sorting, modification, and trafficking steps that initiate in the trans-Golgi andtrans-Golgi network (TGN). To investigate their temporal order interrelationships, we have developed pulse–chase protocol follows synthesis packaging sulfated hormone, pro-opiomelanocortin (POMC). In AtT-20 cells, sulfate is incorporated into POMC predominantly on N-linked endoglycosidase H-resistant oligosaccharides. Subcellular...

Ubiquitously reduced signaling via Methuselah (MTH), a G-protein-coupled receptor (GPCR) required for neurosecretion, has previously been reported to extend life and enhance stress resistance in flies. Whether these effects are due MTH signalling specific tissues remains unknown. We determined that expression of mth targeted the insulin-producing cells (IPCs) fly brain was sufficient oxidative resistance. Paradoxically, we discovered overexpression same similar phenotypic MTH's interaction...

Alcohol use disorders affect millions of individuals. However, the genes and signaling pathways involved in behavioral ethanol responses addiction are poorly understood. Here we identify a conserved biochemical pathway that underlies sedating effects Drosophila . Mutations Arf6 small GTPase cause hypersensitivity to ethanol-induced sedation. We show functions adult nervous system control behavior. also find Arfaptin protein directly binds activated forms Rac1 GTPases, mutants display...

Altered insulin signaling has been linked to widespread nervous system dysfunction including cognitive dysfunction, neuropathy and susceptibility neurodegenerative disease. However, knowledge of the cellular mechanisms underlying effects on neuronal function is incomplete. Here, we show that cell autonomous within Drosophila CM9 motor neuron regulates release neurotransmitter via alteration synaptic vesicle fusion machinery. This effect utilizes FOXO-dependent regulation thor gene, which...

Chemotherapy induced peripheral neuropathy (CIPN), a side effect of many anti-cancer drugs including the vinca alkaloids, is characterized by severe pain syndrome that compromises treatment in patients. Currently there are no effective treatments for this except reduction drug dose. Existing data supports model associated with CIPN result augmenting function sensory nociceptors but cellular mechanisms underlying effects on neuron not well described. Studies from animal models have suggested...

Summary The neuromuscular junction ( NMJ ) is responsible for transforming nervous system signals into motor behavior and locomotion. In the fruit fly Drosophila melanogaster , an age‐dependent decline in function occurs, analogous to experienced mice, humans, other mammals. molecular cellular underpinnings of this are still poorly understood. By specifically profiling transcriptome neurons across age using custom microarrays, we found that expression matrix metalloproteinase 1 dMMP gene...

Mutations in DCTN1, a component of the dynactin complex, are linked to neurodegenerative diseases characterized by broad collection neuropathologies. Because pleiotropic nature complex function within neuron, defining causes neuropathology DCTN1 mutants has been difficult. We combined genetic screen with cellular assays identify genes that critical for nervous system. This approach identified Drosophila homologue Arfaptin, multifunctional protein implicated membrane trafficking. find...

Because of their functional polarity and elongated morphologies, microtubule-based transport proteins organelles is critical for normal neuronal function. The proteasome required throughout the neuron highly regulated degradation a broad set protein targets whose functions underlie key physiological responses, including synaptic plasticity axonal degeneration. Molecularly, relationship between proteasomes unclear. dynein motor complex motility numerous in neurons. Here, we demonstrate that...

The accumulation of oxidative damage is strongly linked to age-dependent declines in cell function, but the contribution morbidity still debated. Many organisms seem tolerate damage, and extension health span life by augmenting antioxidant activity has been inconsistent. Here we use Drosophila model system investigate relationship among stress, span, span. oxidation-dependent dissociation Calstabin protein from ryanodine receptor shown result reduced muscle function mammals. S107 molecule...

Summary Synaptic dysfunction is considered the primary substrate for functional declines observed within nervous system during age‐related neurodegenerative disease. Dietary restriction (DR), which extends lifespan in numerous species, has been shown to have beneficial effects on many disease models. Existing data sets suggest that of DR include amelioration synaptic but evidence diet synapse lacking. Dynactin mutant flies significant increases mortality rates and exhibit progressive loss...

The generation of neuronal morphology requires transport vesicles originating from the Golgi apparatus (GA) to deliver specialized components axon and dendrites. Drosophila Arfaptin is a membrane-binding protein localized GA that required for growth presynaptic nerve terminal. Here we provide biochemical, cellular genetic evidence small GTPase Arl1 guanine-nucleotide exchange factor (GEF) Gartenzwerg are function at during synapse growth. Our data define new signaling pathway composed...