Two families of cell-adhesion molecules, predominantly presynaptic neurexins and postsynaptic neuroligins, are important for the formation functioning synapses in brain, mutations several genes encoding these transmembrane proteins have been found autism patients. However, very little is known about how targeted to which mechanisms regulate this process. Using various epitope-tagged primary hippocampal neurons wild-type knock-out mice vitro transgenic animals vivo , we show that trafficked...

Neurexophilin 3 (Nxph3) is a specific ligand of synaptic α-neurexins that are essential for efficient neurotransmitter release. Previous biochemical work demonstrated Nxph3 interacts with an extracellular domain in tight complex; however, no information available on the localization or functional role brain. Here, we generated lacZ reporter gene knock-in mice to investigate distribution at single-cell level and knockout examine its importance. expression was restricted mostly...

α-Neurexins constitute a family of neuronal cell surface molecules that are essential for efficient neurotransmission, because mice lacking two or all three α-neurexin genes show severe reduction synaptic release. Although analyses knock-outs and transgenic rescue animals suggested an involvement voltage-dependent Ca 2+ channels, it remained unclear whether α-neurexins have general role in -dependent exocytosis how they may affect channels. Here we by membrane capacitance measurements from...

Cognitive decline is a debilitating aspect of aging and neurodegenerative diseases such as Alzheimer9s disease are closely associated with mitochondrial dysfunction, increased reactive oxygen species, neuroinflammation, astrogliosis. This study investigated the effects decreased antioxidant response specifically in astrocytes on cognitive performance neuronal function C57BL/6J mice using tamoxifen-inducible astrocyte-specific knockout manganese superoxide dismutase (aSOD2-KO), matrix that...

Astrocytic dysfunction is central to age-related neurodegenerative diseases. However, the mechanisms leading astrocytic are not well understood. We identify that among diverse cellular constituents of brain, murine and human astrocytes enriched in expression CBS. Depleting CBS causes mitochondrial dysfunction, increases production reactive oxygen species (ROS) decreases bioenergetics can be partially rescued by exogenous H

Protein tyrosine phosphatase MEG2 (PTP-MEG2) is a unique nonreceptor associated with transport vesicles, where it facilitates membrane trafficking by dephosphorylation of the N-ethylmaleimide-sensitive fusion factor. In this study, we identify neurotrophin receptor TrkA as novel cargo whose to cell surface requires PTP-MEG2 activity. addition, also substrate PTP-MEG2, which dephosphorylates both Tyr-490 and Tyr-674/Tyr-675 TrkA. As result, overexpression down-regulates NGF/TrkA signaling...

Postsynaptic AMPA-type glutamate receptors (AMPARs) are among the major determinants of synaptic strength and can be trafficked into out synapses. Neuronal activity regulates AMPAR trafficking during plasticity to induce long-term changes in strength, including potentiation (LTP) depression (LTD). Rab family GTPases regulate most membrane eukaryotic cells; particularly, Rab11 its effectors implicated mediating postsynaptic insertion LTP. To explore function Rab11Fip5, a neuronal effector...

Oxytocin plays critical roles in the brain as a neuromodulator, regulating social and other affective behavior. However, regulatory mechanisms controlling oxytocin receptor (OXTR) signaling neurons remain unexplored. In this study, we have identified robust rapid-onset desensitization of OXTR response multiple regions mouse brain. Both cell autonomous spiking presynaptic activation undergo similar agonist-induced desensitization. G-protein-coupled kinases (GRK) GRK2, GRK3, GRK6 are recruited...

Spinocerebellar ataxia (SCA) is a neurodegenerative disorder characterized by and cerebellar atrophy. A number of different mutations gives rise to types SCA with characteristic ages onset, symptomatology, rates progression. type 34 (SCA34) caused in ELOVL4 (ELOngation Very Long-chain fatty acids 4), acid elongase essential for biosynthesis Long Chain Saturated Polyunsaturated Fatty Acids (VLC-SFA VLC-PUFA, resp., ≥28 carbons), which have important functions the brain, skin, retina,...

Elongation of very long fatty acids-4 (ELOVL4) mediates biosynthesis chain-fatty acids (VLC-FA; ≥28 carbons). Various mutations in this enzyme result spinocerebellar ataxia-34 (SCA34). We generated a rat model human SCA34 by knock-in naturally occurring c.736T>G, p.W246G mutation the Elovl4 gene. Our previous analysis homozygous W246G mutant ELOVL4 rats (MUT) revealed early-onset gait disturbance and impaired synaptic transmission plasticity at parallel fiber-Purkinje cell (PF-PC)...

Elongation of the Very-Long-Chain Fatty Acids-4 (ELOVL4) enzyme that is expressed in neuronal tissues, sperm, and testes mediates biosynthesis very-long-chain polyunsaturated fatty acids (VLC-PUFAs) from dietary long chain PUFAs (LC-PUFAs). The VLC-PUFAs are critical for reproductive function. Therefore, mutations ELOVL4 affect VLC-PUFA contribute to retinal degenerative diseases including Autosomal Dominant Stargardt-like Macular Dystrophy (STGD3). Recent studies have also shown not only a...

Oxytocin receptor (OXTR) is expressed in a distinct population of neurons the lateral septum (LS), among other brain regions, and responsible for regulating various social non-social behaviors, including reward processing, feeding, memory, anxiety, fear. The LS serves as key link between cortical subcortical yet synaptic inputs that drive OXTR-expressing have not been characterized. Here, we established retrograde anterograde viral tracing mouse to map input connections intermediate part...

Presynaptic Ca2+-influx through voltage-gated Ca2+ channels (VGCC) is a key signal for synaptic vesicle release. Synaptic neurexins can partially determine the strength of transmission by regulating VGCCs. However, it unknown whether modulate via all VGCC subtypes similarly. Here, we performed live cell imaging boutons from primary hippocampal neurons with Ca2+-indicator. We used expression inactive and active Cre recombinase to compare control conditional knockout lacking either or selected...

AMPA receptors (AMPAR) are organized into supramolecular complexes in association with other membrane proteins that provide exquisite regulation of their biophysical properties and subcellular trafficking. Proline-rich transmembrane protein 1 (PRRT1), also named as SynDIG4, is a component native AMPAR multiple brain regions. Deletion PRRT1 leads to altered surface levels phosphorylation status AMPARs, well impaired forms synaptic plasticity. Here, we have investigated the mechanisms...

Presynaptic Ca2+-influx through voltage-gated Ca2+-channels (VGCC) is a key signal for synaptic vesicle release. Synaptic neurexins can partially determine the strength of transmission by regulating VGCCs. However, it unknown whether modulate via all VGCC subtypes similarly. Here, we performed live cell imaging boutons from primary hippocampal neurons with Ca2+-indicator. We used expression inactive and active Cre recombinase to compare control conditional knockout lacking either or selected...

Abstract Spinocerebellar ataxia 34 (SCA34) is an autosomal dominant disease that arises from point mutations in the fatty acid elongase, Elongation of Very Long Chain Fatty Acids 4 (ELOVL4), which essential for synthesis Chain-Saturated (VLC-SFA) and Chain-Polyunsaturated (VLC-PUFA) (28–34 carbons long). SCA34 considered a neurodegenerative disease. However, novel rat model (SCA34-KI rat) with knock-in W246G ELOVL4 mutation causes human shows early motor impairment aberrant synaptic...

Very long chain-saturated and -polyunsaturated fatty acids (VLC-SFA VLC-PUFA, respectively) are a functionally important class of containing 28 carbons or more in their acyl chain. They synthesized by the elongation very acids-4 (ELOVL4) enzyme, expressed mainly brain, retina, skin, testes, meibomian gland, where these found (Agbaga et al., 2008). Further, organs exhibit tissue-specific VLC-PUFA VLC-SFA biosynthesis incorporation into complex lipids for specific functions. In Meibomian...

Presynaptic Ca2+ influx through voltage-gated channels (VGCCs) is a key signal for synaptic vesicle release. Synaptic neurexins can partially determine the strength of transmission by regulating VGCCs. However, it unknown whether modulate via all VGCC subtypes similarly. Here, we performed live cell imaging boutons from primary hippocampal neurons with indicator. We used expression inactive and active Cre recombinase to compare control conditional knockout lacking either or selected neurexin...

Bioluminescence resonance energy transfer (BRET) is widely employed for real-time monitoring of G protein-coupled receptor activity, interactions, and trafficking in heterologous cell lines, yet its use neuronal systems remains limited. Here, we present a protocol to apply BRET assays primary cultures from mouse embryos. We describe steps key concepts generating plasmid constructs lentivirus preparations, plating lentiviral transduction cultured neurons 96-well plates, data collection...