A Myelin Proteolipid Protein-LacZ Fusion Protein Is Developmentally Regulated and Targeted to the Myelin Membrane in Transgenic Mice

Transgenic mice were generated with a fusion gene carrying a portion of the murine myelin proteolipid protein (PLP) gene, including the first intron, fused to the E. coli LacZ gene. Three transgenic lines were derived and all lines expressed the transgene in central nervous system white matter as measured by a histochemical assay for the detection of β-galactosidase activity. PLP-LacZ transgene expression was regulated in both a spatial and temporal manner, consistent with endogenous PLP expression. Moreover, the transgene was expressed specifically in oligodendrocytes from primary mixed glial cultures prepared from transgenic mouse brains and appeared to be developmentally regulated in vitro as well. Transgene expression occurred in embryos, presumably in pre- or nonmyelinating cells, rather extensively throughout the peripheral nervous system and within very discrete regions of the central nervous system. Surprisingly, beta-galactosidase activity was localized predominantly in the myelin in these transgenic animals, suggesting that the NH_2-terminal 13 amino acids of PLP, which were present in the PLP-LacZ gene product, were sufficient to target the protein to the myelin membrane. Thus, the first half of the PLP gene contains sequences sufficient to direct both spatial and temporal gene regulation and to encode amino acids important in targeting the protein to the myelin membrane

A Deficiency of Ceramide Biosynthesis Causes Cerebellar Purkinje Cell Neurodegeneration and Lipofuscin Accumulation

Sphingolipids, lipids with a common sphingoid base (also termed long chain base) backbone, play essential cellular structural and signaling functions. Alterations of sphingolipid levels have been implicated in many diseases, including neurodegenerative disorders. However, it remains largely unclear whether sphingolipid changes in these diseases are pathological events or homeostatic responses. Furthermore, how changes in sphingolipid homeostasis shape the progression of aging and neurodegeneration remains to be clarified. We identified two mouse strains, flincher (fln) and toppler (to), with spontaneous recessive mutations that cause cerebellar ataxia and Purkinje cell degeneration. Positional cloning demonstrated that these mutations reside in the Lass1 gene. Lass1 encodes (dihydro)ceramide synthase 1 (CerS1), which is highly expressed in neurons. Both fln and to mutations caused complete loss of CerS1 catalytic activity, which resulted in a reduction in sphingolipid biosynthesis in the brain and dramatic changes in steady-state levels of sphingolipids and sphingoid bases. In addition to Purkinje cell death, deficiency of CerS1 function also induced accumulation of lipofuscin with ubiquitylated proteins in many brain regions. Our results demonstrate clearly that ceramide biosynthesis deficiency can cause neurodegeneration and suggest a novel mechanism of lipofuscin formation, a common phenomenon that occurs during normal aging and in some neurodegenerative diseases

Experiential Education and the Work Environment Abroad: Student Work Abroad Program

Johnson & Wales University College of Business provides students with curricula that incorporate both industry experience and classroom learning. One example of this complementary approach, known as the SWAP program, is predicated on the idea that students who participate in experiential learning activities develop an increased knowledge and understanding of organizational function, professional networking, management and leadership responsibilities, organizational and professional performance expectations and initiatives, and the ability to examine and analyze classroom learning in a real, not simulated, business environment

Comparison of monoamine concentrations in the brains of adult male and female Japanese quail.

A fluorometric assay measuring brain tissue concentrations of norepinephrine, dopamine, and serotonin has been validated for Japanese quail. Accuracy, precision, specificity, and parallelism were determined. The sensitivity of the assays was 6 ng/tube, which allowed individual assay of 1 to 2 mg hypothalamic tissue. In Experiment 1, relatively large areas of brain from adult, reproductively active males and females were found to differ significantly in norepinephrine content in optic lobes and for dopamine in right telencephalon. A microdissection technique was used in Experiment 2 to sample small portions of hypothalamic tissue. Sex differences were observed for norepinephrine in the sections containing the lobus paraolfactorius and the preoptic, anterior, and medial hypothalamus. Differences in monoamine content were most apparent when smaller areas dissected by microdissection were analyzed. These results give evidence for sex differences in the monoamine content in specific areas of the brain