Glycosphingolipids are controlled by the spatial organization of their metabolism and transport specificity. Using immunoelectron microscopy, we localize to Golgi stack glycosyltransferases that produce glucosylceramide (GlcCer), lactosylceramide (LacCer), GM3. GlcCer is synthesized on cytosolic side must translocate across lumen for LacCer synthesis. However, only very little natural translocates in vitro. As reaches cell surface when vesicular trafficking inhibited, it a post-Golgi...

In axons, proper localization of proteins, vesicles, organelles, and other cargoes is accomplished by the highly regulated coordination kinesins dyneins, molecular motors that bind to translocate them along microtubule (MT) tracks. Impairment axonal transport implicated in pathogenesis multiple neurodegenerative disorders including Alzheimer's Huntington's diseases. To understand how MT-based cargo motility delineate its role neurodegeneration, it critical analyze detailed dynamics moving...

Little is known about the release and intercellular transport of Wnt proteins from mammalian cells. Lipoproteins may act as carriers for movement gradient formation lipid-linked morphogens Wingless Hedgehog in Drosophila. To investigate whether such a mechanism can occur mammals, we have studied cultured Wnt3a associated with lipoproteins culture medium not extracellular vesicles or exosomes. Although was both high-density (HDL) low-density lipoproteins, only HDL allowed mouse fibroblasts....

Dynamin 1 (Dyn1) and Dyn2 are neuronal ubiquitously expressed isoforms, respectively, of the multidomain GTPase required for clathrin-mediated endocytosis (CME). Although they 79% identical, Dyn1 not fully functionally redundant. Through direct measurements basal assembly-stimulated activities, membrane binding, self-assembly, fission on planar curved templates, we have shown that is an efficient curvature generator, whereas primarily a sensor. Using Dyn1/Dyn2 chimeras, identified...

Adrenoleukodystrophy (ALD) is an X-linked neurodegenerative disorder characterized by elevated very long chain fatty acid (VLCFA) levels, reduced activity of peroxisomal VLCFA-CoA ligase, and variable phenotypic expression. A putative gene for ALD was recently identified surprisingly encodes a protein (ALDP) that belongs to family transmembrane transporters regulated or activated ATP (the ABC proteins). We have examined genomic DNA from probands mutations in the gene. detected large...

Dynamin-2 (Dyn2) is ubiquitously expressed and catalyzes membrane fission during clathrin-mediated endocytosis in nonneuronal cells. We have previously shown that Dyn2 inefficiently generates curvature only mediates of highly curved membranes. This led to the hypothesis other endocytic accessory proteins (EAPs) generate needed sculpt a sufficiently narrow neck trigger assembly fission. Candidates for this activity are EAPs bind dynamin proline/arginine-rich domain (PRD) through their SH3...

Aging-related memory impairment and pathological disorders such as Alzheimer’s disease differ between males females, yet little is known about how aging-related changes in the transcriptome chromatin environment sexes hippocampus. To investigate this question, we compared accessibility landscape gene expression/alternative splicing pattern of young adult aged mouse hippocampus both females using ATAC-seq RNA-seq. We detected significant aging-dependent expression genes involved immune...

Analyses of RNA from different developmental stages Schistosoma mansoni showed stage-specific expression heat-shock protein 70 (hsp70), which is regulated by a program and stress. The program, common to hsp70 other genes (e.g. paramyosin), refers constitutive in miracidia sporocyst adult worm but not cercariae, the termination gene transcription during sporocyst/cercaria transformation. Stress induction, specific hsp70, transient accumulation high levels mRNA cercariae/schistosomula...

Melanosomes are lysosome‐related organelles that coexist with lysosomes in mammalian pigment cells. Melanosomal and lysosomal membrane proteins share similar sorting signals their cytoplasmic tail, raising the question how they segregated. We show control melanocytes, melanosomal enzymes tyrosinase‐related protein 1 (Tyrp1) tyrosinase follow an intracellular Golgi to melanosome pathway, whereas absence of glycosphingolipids, observed pass over cell surface. Unexpectedly, lysosome‐associated...

Sphingolipids are considered to play a key role in protein sorting and membrane trafficking. In melanocytic cells, of lysosomal melanosomal proteins requires the sphingolipid glucosylceramide (GlcCer). This information is located lumenal domain proteins. We found that two processes dependent on pH, sialylation acid lipase (LAL) activity were aberrant GM95 melanocyte which do not produce glycosphingolipids. Using fluorescence lifetime imaging microscopy (FLIM), we pH trans ‐Golgi network...

The aim of this study was to evaluate the screening policies cystic fibrosis (CF) in Jewish population. prevalence mutations that account for CF Israel have been defined past by determining frequency affected individuals. This is a population-based and is, therefore, different from previous patient-based studies. We found D1152H, W1089X, 405 + IG-->A were present some ethnic groups which no patients carrying these reported. These facts necessitate reevaluation policy regarding Israel....

Understanding the mechanisms by which molecular motors coordinate their activities to transport vesicular cargoes within neurons requires quantitative analysis of motor/cargo associations at single vesicle level. The goal this protocol is use fluorescence microscopy correlate ("map") position and directionality movement live cargo composition relative amounts associated with same cargo. "Cargo mapping" consists imaging fluorescently labeled moving in axons cultured on microfluidic devices,...

X-linked adrenoleukodystrophy (ALD) is a peroxisomal disorder characterized by impaired β-oxidation of very-long-chain fatty acids (VLCFAs). This probably due to reduced activation the VLCFAs and results in demyelination nervous system adrenocortical insufficiency. The ALD gene localized on Xq28, has 10 exons encodes protein 745 amino with significant homology membrane PMP70. Characterizing disease causing mutations importance prenatal diagnosis because 12-20% women who are obligatory...

Abstract Pulsed laser photolysis experiments are carried out in lipid bilayer vesicles which the fluorescence of excited pyrene (P), pyrene‐ethanol (P‐CH 2 OH) and pyrene‐monosulphonate (P‐SO − 3 ) quenched by N.N‐diethylaniline (DEA). The observed photochemical patterns basically different from those encountered homogeneous solutions: No detectable exciplex emission is observed, yield separated ions P + DEA small. Two alternative explanations given for complete lack an vesicles. One...

Understanding the mechanisms by which molecular motors coordinate their activities to transport vesicular cargoes within neurons requires quantitative analysis of motor/cargo associations at single vesicle level. The goal this protocol is use fluorescence microscopy correlate ("map") position and directionality movement live cargo composition relative amounts associated with same cargo. "Cargo mapping" consists imaging fluorescently labeled moving in axons cultured on microfluidic devices,...

Abstract Aging-related memory impairment and pathological disorders such as Alzheimer’s disease differ between males females, yet little is known about how aging-related changes in the transcriptome chromatin environment sexes hippocampus. To investigate this question, we compared accessibility landscape gene expression/alternative splicing pattern of young adult aged mouse hippocampus both females using ATAC-seq RNA-seq. We detected significant aging-dependent expression genes involved...