Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
|
pubmed:dateCreated |
1993-12-7
|
pubmed:abstractText |
Bacterial beta-galactosidase is widely used as a marker for gene expression and in cell tracing experiments. In a survey of three transgenic mouse lines expressing beta-galactosidase in the central nervous system (CNS) under the control of different promoters, we find substantial variation in the intracellular distribution of the lacZ protein. In line M beta P5, transgene beta-galactosidase expression is driven by a promoter/enhancer fragment from the oligodendrocyte-specific myelin basic protein gene; however, electron microscopy of histochemically stained preparations reveals transgene expression not only in oligodendrocytes but also in some neurons. Immunofluorescence and immunoperoxidase staining show the beta-galactosidase protein distributed throughout the perikaryal cytoplasm of oligodendrocytes and in processes reaching to myelin sheaths. By contrast, immunoreactive protein appears restricted in neurons to one or a few small perikaryal immunoreactive granules. The granules are visible in the electron microscope as amorphous inclusion bodies of moderate electron density and lack a limiting membrane. Histochemical staining patterns with X-gal and Bluo-gal echoed the protein distribution: diffuse distribution of enzyme protein yielded cells filled with substrate, while punctate enzyme distribution yielded restricted or punctate histochemical staining. Examination of two other lines using different promoter/enhancers to drive expression in the CNS showed both diffuse and punctate beta-galactosidase immunolocalization and histochemical staining. The amount of protein synthesized or other properties, yet unidentified, intrinsic to the target cells may determine the intracellular distribution of beta-galactosidase. In retroviral marking studies, clone members have been identified as those cells filled with X-gal reaction product. This approach may underestimate both clone size and the minimum number of divisions separating the members of each clone.
|
pubmed:grant | |
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Sep
|
pubmed:issn |
0360-4012
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
1
|
pubmed:volume |
36
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
88-98
|
pubmed:dateRevised |
2007-11-14
|
pubmed:meshHeading |
pubmed-meshheading:8230324-Animals,
pubmed-meshheading:8230324-Bacterial Proteins,
pubmed-meshheading:8230324-Cell Line,
pubmed-meshheading:8230324-Central Nervous System,
pubmed-meshheading:8230324-Gene Expression Regulation,
pubmed-meshheading:8230324-Genes, Synthetic,
pubmed-meshheading:8230324-Mice,
pubmed-meshheading:8230324-Mice, Inbred C57BL,
pubmed-meshheading:8230324-Mice, Inbred DBA,
pubmed-meshheading:8230324-Mice, Transgenic,
pubmed-meshheading:8230324-Nerve Tissue Proteins,
pubmed-meshheading:8230324-Neuroglia,
pubmed-meshheading:8230324-Neurons,
pubmed-meshheading:8230324-Recombinant Fusion Proteins,
pubmed-meshheading:8230324-Stem Cells,
pubmed-meshheading:8230324-Subcellular Fractions,
pubmed-meshheading:8230324-beta-Galactosidase
|
pubmed:year |
1993
|
pubmed:articleTitle |
Intracellular distribution of transgenic bacterial beta-galactosidase in central nervous system neurons and neuroglia.
|
pubmed:affiliation |
Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029-6574.
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
|