Triggering receptor expressed in myeloid (TREM) cells 2, a by cells, osteoclasts and microglia, is known to play protective role bones brain. Mutations of the (or its coupling protein, DAP12) sustain fact genetic disease affecting two organs, polycystic lipomembraneous osteodysplasia with sclerosing leukoencephalopathy (PLOSL or Nasu-Hakola disease). So far, specific agonist(s) TREM2 have not been identified (their) transduction mechanisms are largely unknown. Heat shock protein 60 (Hsp60)...

Enlargeosomes are cytoplasmic organelles discharged by regulated exocytosis, identified immunofluorescence of their membrane marker, desmoyokin/Ahnak, but never revealed at the ultrastructural level. Among numerous enlargeosome‐positive cells, richest and most extensively characterized those a PC12 clone, PC12‐27, defective classical neurosecretion. By using immunoperoxidase labeling formaldehyde‐fixed, Triton‐X‐100‐permeabilized PC12‐27 we have now enlargeosomes as small vesicles scattered...

Abstract Cultured astrocytes exhibit a flat/epitelioid phenotype much different from the star‐like of tissue astrocytes. Upon exposure to treatments that affect small GTPase Rho and/or its effector ROCK, however, flat undergo stellation, with restructuring cytoskeleton and outgrowth processes lamellipodia, assuming closer exhibited in situ . The mechanisms this change are known only part. Using ROCK blocker drug Y27632, which induces rapid (tens min), dose‐dependent reversible stellations,...

Abstract The expression of two cytosolic, high affinity Ca 2+ ‐binding proteins, calbindin‐28 and calretinin, has been investigated in the cerebellum hippocampus young old rats (from 12 days to 30 months) by combining immunofluorescence Western blotting. Three markers, calreticulin (the major binding protein within lumen endoplasmic reticulum), MAP‐2 (a microtubule concentrated neuronal dendrites) synaptophysin (an integral synaptic vesicles), were studied parallel. In cerebellar cortex a...

The mechanisms governing the fast, regulated exocytosis of enlargeosomes have been unknown, except for participation annexin-2 in a pre-fusion step. We investigated whether any SNAREs are involved. In PC12-27 cells, which enlargeosome-rich, expressed exhibited various distributions (trans-Golgi network, scattered puncta, plasma membrane); however, only VAMP4 was colocalized discrete puncta with enlargeosome marker desmoyokin. organelle, revealed by capacitance increases and surface...

Unpredictable chronic mild stress (CMS) is among the most popular protocols used to induce depressive-like behaviors such as anhedonia in rats. Differences CMS often result variable degree of vulnerability, and mechanisms behind resilience are great interest neuroscience due their involvement development psychiatric disorders, including major depressive disorder. Expression likely driven by long-term alterations corticolimbic system downregulation dopamine (DA) signaling. Although we have a...

Significance Much of the available information on NGF-induced neural cell differentiation has been first obtained by study a cultured line, pheochromocytoma PC12. Using PC12 cells, we now demonstrate two new aspects process, possibly important also in neurons: ( i ) direct binding, activation, and signaling NGF receptor TrkA adhesion protein, L1CAM, when administered as soluble construct surface-exposed adjacent cells; ii expression, exposed for few hours to treatment, an exocytic vesicle,...

Neurite outgrowth is known as a slow (days) process occurring in nerve cells and neurons during neurotrophin treatment upon transfer to culture, respectively. Using Y27632, drug that induces activation of Rac1, downstream step the signaling cascade, we have identified new form outgrowth, which rapid (<1 hour) extensive (>500 μm2 surface enlargement/single cell/first hour). However, this takes place only (PC12-27 SH-SY5Y cells, embryonic neonatal neurons) rich an exocytic...

Ideally, elucidating the role of specific brain circuits in animal behavior would require ability to measure activity at all involved synapses, possibly with unrestricted field view, thus even those boutons deeply located into brain. Here, we introduce and validate an efficient scheme reporting synaptic vesicle cycling vivo. This is based on SynaptoZip, a genetically encoded molecule deploying vesicular lumen bait moiety designed capture upon exocytosis labeled alien peptide, Synbond. The...

Enlargeosomes, a new type of widely expressed cytoplasmic vesicles, undergo tetanus toxin-insensitive exocytosis in response to cytosolic Ca 2+ concentration ([Ca ] i ) rises. Cell biology enlargeosomes is still largely unknown. By combining immunocytochemistry (marker desmoyokin-Ahnak, d/A) capacitance electrophysiology the enlargeosome-rich, neurosecretion-defective clone PC12-27, we show that 1) two responses, cell surface enlargement and d/A appearance, occur with similar kinetics same...

Abstract Sixteen clones, recently isolated from the PC12 nerve cell line, were analysed for a variety of markers and activities. Two endoplasmic reticulum (ER) luminal markers, chaperone protein BiP major Ca 2+ storage calreticulin, as well 40‐kD rough ER membrane marker plus‐end‐directed mirotubule motor protein, kinesin, found to be expressed at similar levels. These results suggest that size ER, function microtubules capacity rapidly exchanging store do not change substantially among...

Extracellular vesicles (EVs), a broad term for the lipid microparticles known as microvesicles and exosomes, are discharged by cells into their surrounding space. Microvesicles upon outward plasma membrane budding, while exosomes secreted after multivesicular body (MVB) fusion with membrane. The majority of information regarding EV biology comes from studies performed in non-polarized cells. Here we characterize release polarized We found substantial asymmetry number composition EVs produced...

A peptide corresponding to amino acid sequence 188–201 of the α‐subunit Torpedo AChR binds α‐Bgtx. The S‐S bridge between Cys 192 and 193 is essential for binding as Tyr in position 189. same human AChR, which instead has a Thr 189, α‐Bgtx with much lower efficiency. Monoclonal antibodies raised against recognize indicating that synthetic native molecule share some immunological epitopes. With computer graphics energy refinement molecular model this been elaborated.

In neurons and neurosecretory (nerve) cells, neurite outgrowth requires surface enlargement sustained by exocytosis of specific but poorly characterized vesicles. A canonical, relatively slow form is known to require the v-SNARE Ti-VAMP. Recently, we have identified a new, rapid form, triggered activation Rac1 enlargeosomes (v-SNARE: VAMP4). By parallel study various pheochromocytoma PC12 cell clones exhibiting either single or both forms outgrowth, show that expression enlargeosomes, their...

Abstract The effect of stress on animal behavior and brain activity has been attracting growing attention in the last decades. Stress dramatically affects several aspects behavior, including motivation cognitive functioning, used to model human pathologies such as post‐traumatic disorder. A key question is whether alters plastic potential synaptic circuits. In this work, we evaluated if dopamine (DA)‐dependent plasticity medial prefrontal cortex (mPFC). On male adolescent rats, characterized...

Astrocytes' organisation affects the functioning and fine morphology of brain, both in physiological pathological contexts. Although many aspects their role have been characterised, complex functions remain, to a certain extent, unclear with respect contribution brain cell communication. Here, we studied effects nanotopography microconfinement on primary hippocampal rat astrocytes. For this purpose, fabricated nanostructured zirconia surfaces as homogenous substrates micrometric patterns,...

Artificially grown neuronal cultures of brain cells have been used for decades in the attempt to reproduce and study vitro complexity circuits. It soon became evident that this alone was insufficient, because random architecture these artificial networks. Important groundwork therefore resulted development methods confine adhesion at specific locations match predefined network topologies connectivity. Despite notable progress neural circuitry engineering, there is still need micropatterned...