Altered segmental identity and abnormal migration of motor neurons in mice lacking Hoxb-1.
AuthorsStuder M, Lumsden A, Ariza-McNaughton L, Bradley A, Krumlauf R
JournalNature
PubMed ID8967950
'Segmentation of the vertebrate hindbrain into rhombomeres is important for the anterior-posterior arrangement of cranial motor nuclei and efferent nerves. Underlying this reiterated organization, Hox genes display segmentally restricted domains of expression, such as expression of Hoxb-1 (refs 5, 6) in rhombomere 4 (r4). Here we report that absence of ... More
An anti-transferrin receptor-avidin fusion protein exhibits both strong proapoptotic activity and the ability to deliver various molecules into cancer cells.
AuthorsNg PP, Dela Cruz JS, Sorour DN, Stinebaugh JM, Shin SU, Shin DS, Morrison SL, Penichet ML
JournalProc Natl Acad Sci U S A
PubMed ID12149472
'We have developed an antibody fusion protein (anti-rat TfR IgG3-Av) with the ability to deliver different molecules into cancer cells. It consists of avidin genetically fused to the C(H)3 region of a human IgG3 specific for the rat transferrin receptor. It forms strong, noncovalent interactions with biotinylated molecules such as ... More
Expression of a lacZ transgene reveals floor plate cell morphology and macromolecular transfer to commissural axons.
AuthorsCampbell RM, Peterson AC
JournalDevelopment
PubMed ID8306884
'The floor plate is situated at the ventral midline of the neural tube and is an important intermediate target for commissural axons. During elongation, these axons converge bilaterally on the ventral midline neural tube and after crossing the floor plate make an abrupt rostral turn. Ample evidence indicates that the ... More
Sensitive and rapid detection of beta-galactosidase expression in intact cells by microinjection of fluorescent substrate.
AuthorsBrustugun OT, Mellgren G, Gjertsen BT, Bjerkvig R, Døskeland SO
JournalExp Cell Res
PubMed ID7641787
'Bacterial beta-galactosidase, coded by lacZ, is a widely used reporter for studies of transcriptional activity of eukaryotic promoters at the single cell level. Unfortunately, current detection methods, like X-gal cytochemistry, are slow, have suboptimal sensitivity, and are incompatible with cell survival. By a novel approach based on microinjection into cells ... More
Characterization of K+ currents and the cAMP-dependent modulation in cultured Drosophila mushroom body neurons identified by lacZ expression.
AuthorsWright NJ, Zhong Y
JournalJ Neurosci
PubMed ID7869080
'Electrophysiological analysis of cultured neurons provides a potential approach toward understanding the physiological defects that may contribute to abnormal behavior exhibited by mutants of the fruit fly Drosophila. However, its application has been restricted by an inability to identify a particular functional or anatomical subpopulation of neurons from the CNS. ... More
Vaccinia virus transfection of hippocampal slice neurons.
AuthorsPettit DL, Koothan T, Liao D, Malinow R
JournalNeuron
PubMed ID7718231
Here we describe a technique that uses a recombinant vaccinia virus to transfect neurons in rat hippocampal slices. This technique allows the use of molecular biological manipulations on neuronal tissue while maintaining intact synaptic function. This method should be useful in testing specific hypotheses regarding the role of synaptic proteins. ... More
Detecting enzymatic activity in cells using fluorogenic substrates.
AuthorsHaugland RP
JournalBiotech Histochem
PubMed ID8580208
Fluorogenic substrates can detect enzymatic activity associated with cells. It is difficult, however, to detect activity within a single cell or in an organelle since hydrolytic substrates yield products that rapidly leak from the cell. Several new solutions are presented including trapping the fluorescent product in membranes, in cell organelles, ... More
Potentiated transmission and prevention of further LTP by increased CaMKII activity in postsynaptic hippocampal slice neurons.
AuthorsPettit DL, Perlman S, Malinow R
JournalScience
PubMed ID7997883
Calcium-calmodulin-dependent protein kinase II (CaMKII) is a necessary component of the cellular machinery underlying learning and memory. Here, a constitutively active form of this enzyme, CaMKII(1-290), was introduced into neurons of hippocampal slices with a recombinant vaccinia virus to test the hypothesis that increased postsynaptic activity of this enzyme is ... More
Nucleic acid detection using non-radioactive labelling methods.
AuthorsMansfield ES, Worley JM, McKenzie SE, Surrey S, Rappaport E, Fortina P
JournalMol Cell Probes
PubMed ID7477006
Nucleic acid probe-based assays are now widely used in genetic research, human identification, forensics and in a broad spectrum of clinical assays in the fields of microbiology, haematology/oncology and virology. Labelled probes are used in a variety of assay formats including dot-blots, Southern blots (DNA target), Northern blots (RNA target), ... More
Verapamil inhibition of enzymatic product efflux leads to improved detection of beta-galactosidase activity in lacZ-transfected cells.
AuthorsPoot M, Arttamangkul S
JournalCytometry
PubMed ID9136753
The beta-galactosidase activity encoded by the lacZ gene of Escherichia coli is widely used to monitor successful expression of transfected genes. Fluorogenic substrates allow detection of enzyme activity in viable cells, which, subsequently, can be selected for further study on the basis of fluorescence emission. We analyzed three fluorogenic substrates ... More
Neural induction promotes large-scale chromatin reorganisation of the Mash1 locus.
AuthorsWilliams RR, Azuara V, Perry P, Sauer S, Dvorkina M, Jørgensen H, Roix J, McQueen P, Misteli T, Merkenschlager M, Fisher AG
JournalJ Cell Sci
PubMed ID16371653
Determining how genes are epigenetically regulated to ensure their correct spatial and temporal expression during development is key to our understanding of cell lineage commitment. Here we examined epigenetic changes at an important proneural regulator gene Mash1 (Ascl1), as embryonic stem (ES) cells commit to the neural lineage. In ES ... More
Human gene targeting by adeno-associated virus vectors is enhanced by DNA double-strand breaks.
AuthorsMiller DG, Petek LM, Russell DW
JournalMol Cell Biol
PubMed ID12724413
The use of adeno-associated virus (AAV) to package gene-targeting vectors as single-stranded linear molecules has led to significant improvements in mammalian gene-targeting frequencies. However, the molecular basis for the high targeting frequencies obtained is poorly understood, and there could be important mechanistic differences between AAV-mediated gene targeting and conventional gene ... More
Assembly of MHC class I molecules with biosynthesized endoplasmic reticulum-targeted peptides is inefficient in insect cells and can be enhanced by protease inhibitors.
AuthorsDeng Y, Gibbs J, Bacík I, Porgador A, Copeman J, Lehner P, Ortmann B, Cresswell P, Bennink JR, Yewdell JW
JournalJ Immunol
PubMed ID9712031
To study the requirements for assembly of MHC class I molecules with antigenic peptides in the endoplasmic reticulum (ER), we studied Ag processing in insect cells. Insects lack a class I recognition system, and their cells therefore provide a "blank slate" for identifying the proteins that have evolved to facilitate ... More
Neuropsin (OPN5)-mediated photoentrainment of local circadian oscillators in mammalian retina and cornea.