Andrea Buchstaller's research while affiliated with Tufts University and other places

Publications (19)

Article
Full-text available
Recent advances in genomics and proteomics have generated a change in emphasis from hypothesis-based to discovery-based investigations. Genomic and proteomic studies based on differential expression microarrays or comparative proteomics often provide many potential candidates for functionally important roles in normal and diseased cells. High throu...
Article
Full-text available
An interaction of growth cone axonin-1 with the floor-plate NgCAM-related cell adhesion molecule (NrCAM) was shown to play a crucial role in commissural axon guidance across the midline of the spinal cord. We now provide evidence that axonin-1 mediates a guidance signal without promoting axon elongation. In an in vitro assay, commissural axons grew...
Article
Directed growth cone movement is crucial for the correct wiring of the nervous system. This movement is governed by the concerted actions of cell surface receptors, signaling proteins, cytoskeleton-associated molecules, and molecular motors. In order to investigate the molecular basis of growth cone motility, we applied a new technique to functiona...
Article
Neural cell adhesion molecules composed of immunoglobulin and fibronectin type III-like domains have been implicated in cell adhesion, neurite outgrowth, and fasciculation. Axonin-1 and Ng cell adhesion molecule (NgCAM), two molecules with predominantly axonal expression exhibit homophilic interactions across the extracellular space (axonin- 1/axon...
Article
Full-text available
Neural cell adhesion molecules composed of immunoglobulin and fibronectin type III-like domains have been implicated in cell adhesion, neurite outgrowth, and fasciculation. Axonin-1 and Ng cell adhesion molecule (NgCAM), two molecules with predominantly axonal expression exhibit homophilic interactions across the extracellular space (axonin- 1/axon...
Article
Recent investigations on the molecular interactions between the neuronal cell adhesion molecules axonin-1 and NgCAM have brought intriguing novel results suggesting that tetrameric complexes of the cell adhesion molecules axonin-1 and NgCAM could represent functional units of cellular recognition, capable of eliciting distinctive intracellular sign...
Article
The axonal surface glycoproteins neuronglia cell adhesion molecule (NgCAM) and axonin-1 promote cell-cell adhesion, neurite outgrowth and fasciculation, and are involved in growth cone guidance. A direct binding between NgCAM and axonin-1 has been demonstrated using isolated molecules conjugated to the surface of fluorescent microspheres. By expres...
Article
The axonal surface glycoproteins neuronglia cell adhesion molecule (NgCAM) and axonin-1 promote cell-cell adhesion, neurite outgrowth and fasciculation, and are involved in growth cone guidance. A direct binding between NgCAM and axonin-1 has been demonstrated using isolated molecules conjugated to the surface of fluorescent microspheres. By expres...
Article
Growth cones at the tips of growing axons move along predetermined pathways to establish synaptic connections between neurons and their distant targets. To establish their orientation, growth cones continuously sample for, and respond to, guidance information provided by cell surfaces and the extracellular matrix. To identify specific guidance cues...
Article
Cell adhesion molecules expressed on the axonal membrane have been thought to be involved in the guidance of axons to their target area. In the chick, axonin-1 and NgCAM have been shown to promote, through reciprocal interactions, neurite outgrowth in vitro. We have recently shown that chick retinal ganglion cells (RGC) express both proteins as ear...
Article
Full-text available
Neural cell adhesion molecules of the immunoglobulin superfamily mediate cellular interactions via homophilic binding to identical molecules and heterophilic binding to other family members or structurally unrelated cell-surface glycoproteins. Here we report on an interaction between axonin-1 and Nr-CAM/Bravo. In search for novel ligands of axonin-...
Article
Polyclonal antibodies were raised to synthetic peptides having amino acid sequences corresponding with the N- or C-terminal part of the gamma-aminobutyric acidA (GABAA) receptor alpha 5-subunit. These anti-peptide alpha 5(2-10) or anti-peptide alpha 5(427-433) antibodies reacted specifically with GABAA receptors purified from the brains of 5-10-day...
Article
: Polyclonal antibodies were raised to synthetic peptides having amino acid sequences corresponding with the N- or C-terminal part of the γ-aminobutyric acidA (GABAA) receptor α5-subunit. These anti-peptide α5(2–10) or anti-peptide α5(427–433) antibodies reacted specifically with GABAA receptors purified from the brains of 5–10-day-old rats in an e...
Article
beta 2- and beta 3-subunits of GABAA receptors purified from the brains of 5-10-day-old rats were investigated in the intact or completely N-deglycosylated state using the beta-subunit-specific monoclonal antibody bd-17 and polyclonal antibodies directed against synthetic amino acid sequences specific for the GABAA receptor beta 2- or beta 3-subuni...
Article
GABAA receptors purified from the brains of 5- to 10-day-old rats were completely N- and O-deglycosylated using N-glycanase and/or neuraminidase plus O-glycanase. Intact or completely deglycosylated receptors were subjected to SDS-polyacrylamide gel electrophoresis and Western blot analysis. Polyclonal antibodies directed against synthetic aminoaci...
Article
GABAA receptors purified from the brains of 5- to 10-day-old rats were completely N- and O-deglycosylated using N-glycanase and/or neuraminidase plus O-glycanase. Intact or completely deglycosylated receptors were subjected to SDS-polyacrylamide gel electrophoresis and Western blot analysis. Polyclonal antibodies directed against synthetic aminoaci...

Citations

... important roles in axon growth and guidance (Buchstaller et al., 1996;Westphal et al., 2010;Enriquez-Barreto et al., 2012). As expected, these ABC3 substrates were expressed in NM axons on E9 (Figures 9A,B-B ,C-C ). ...
... While CALI inactivates target proteins inside a 2 mm laser spot, micro-CALI focuses a laser beam through the optics of an inverted microscope such that inactivation occurs within À10 ìm diameters (reviewed in ref. 37). Micro-CALI allows inactivation of proteins in one cell at a time, and even fractions of a cell, to allow for direct phenotypic observation of functional inactivation of a given protein. ...
... 86 They were raised in rabbits, either against synthetic peptides corresponding to specific sequences of the respective GABA A receptor subunits, coupled to keyhole-limpet hemocyanin (subunits a1-a3, a5, a6 and g3) or against fusion proteins derived from DNA constructs of the maltose-binding protein gene with specific sequences of the S. Pirker et al. 816 Abbreviations used in the figures 7 facial receptor subunit genes and transcribed in Escherichia coli (subunits a4, b1-b3, g1, g2 and d). 54,86 All antibodies were affinity purified and have been extensively characterized by various methods, including immunoprecipitation, western blotting and immunocytochemistry. 10,25,28,37,40,54,56,58,60,62,81,92,93,99,103 Controls were prepared using the primary antisera preadsorbed with 10 mg/ml of the respective synthetic peptide or fusion protein (24 h, 4ЊC). When preadsorbed antibodies were included in the incubation, no staining (subunits a2, a3, a6, g1) or a marked reduction of staining intensity (subunits a1, a4, a5, b1, b2, b3, g2, g3, d) was obtained with few exceptions: slight residual staining of somata was observed in the brainstem with the preadsorbed a5-subunit antibody; faint staining was also observed with the g2-subunit antibody in the cortex, with the g3subunit antibody in Purkinje cells, and with the d-subunit antibody in the cerebellum (granule cells, cerebellar nuclei) and the brainstem after preadsorbing with the respective peptide. ...
... Domains Ig3-4 of SAX-7S are necessary for its function in neuronal maintenance L1 family members play diverse roles via homophilic interactions through their extracellular domains which mediate homophilic cell adhesion (Brummendorf and Rathjen 1996;Brummendorf et al. 1998;Hortsch 2000;Haspel and Grumet 2003), and mutating different extracellular Ig-like domains of vertebrate L1 perturbs its homophilic and/or heterophilic binding in in vitro assays (Holm et al. 1995;Zhao and Siu 1995;Montgomery et al. 1996;Felding-Habermann et al. 1997;Blaess et al. 1998;Kunz et al. 1998;De Angelis et al. 1999;Oleszewski et al. 1999;Haspel et al. 2000;Castellani et al. 2002;De Angelis et al. 2002) and neurite outgrowth (Appel et al. 1993). In C. elegans, neuronal expression of an SAX-7S recombinant version lacking Ig5-6 domains rescued AIY/AVK neuronal soma position defects of sax-7(nj48) mutants, whereas a recombinant version lacking Ig3-4 domains does not (Pocock et al. 2008). ...
... 327,328 Upon irradiation, the excited chromophore generates 1 O 2 in the immediate vicinity of the target protein, resulting in oxidative damage and functional impairment 329 while other biomolecules remain largely unaffectedthis may even enable selective destruction of one subunit of multiprotein complexes 330 or inactivation of the POI only in a defined area when focused laser light is used. 331 This exquisite spatiotemporal control relies on the exceedingly short lifetime and diffusion distance of 1 O 2 . ...
... Endo H-resistant and -sensitive 3 was detected in control cells and those expressing Plic-1 ( Fig. 2A). For comparison, samples were also treated with N-glycosidase-F, which cleaves all N-linked glycans, resulting in the detection of a single band of 52 kDa ( Fig. 2A) (17,28). We measured the levels of Endo H-sensitive 3 subunits in the absence or presence of Plic-1. ...
... The adjoining parietal cortex was used for comparison. GABA A Rs exist as pentameric subunit complexes which can be allosterically modulated by numerous pharmacological agents (Rabow et al., 1995 ;Sieghart, 1992a, b, c, d;Sieghart, 1995;Sieghart et al., 1992;Wafford et al., 1993;Yu et al., 2006). Molecular cloning has revealed 6-α, 4-β, 3-γ, 1-δ, 1-ε, 1-π, 1-θ, and 3-ρ GABA A receptor subunits Sieghart, 2008,2009;Rabow et al., 1995;Rudolph et al., 2001;Sieghart, 1995;Wafford and Ebert, 2006). ...
... Several potential GABA A R subunit N-glycosylation sites have been identified by the presence of an Nglycosylation consensus sequence in an extracellular protein domain (Chen et al, 2012;Farriol-Mathis et al, 2004). Nglycosylation of GABA A R subunits has been confirmed in vivo using rodent models (Buchstaller et al, 1991a;Buchstaller et al, 1991b) and cell culture lines (Buller et al, 1994;Connolly et al, 1996;Lo et al, 2010;Tanaka et al, 2008); however, which subunits are N-glycosylated in human brain has not been characterized. This study aims to elucidate if alterations of N-glycosylation in schizophrenia are specific to the glutamatergic system or are more generalized, by characterizing the N-glycosylation pattern of GABA A R subunits in normal human cortex and identifying potential alterations in schizophrenia. ...
... Polymerization of this receptor is well documented. Smears are expected because of the glycosylation of the GABA A R b-3 subunit [58] and this modification is known to even shift electrophoretic mobility resulting into different apparent molecular weights. Figure 2A shows that no band can be observed for fractions 1-4 and the entire lanes containing these fraction were transferred into the 2-DE run (shown for fraction 1; Fig. 2B); Western blotting revealed the presence of the subunit b-3 in a broad range of apparent molecular weight (Fig. 2C), which it corresponded to 1-DE BN-PAGE Western blot result (shown in Fig. 1). ...
... In the postnatal cerebellum, CNTN2 is transiently expressed on pre-migratory granule cells (Yamamoto et al., 1986). CNTN2 is involved in many physiological neuronal processes, such as the interaction with glial cells (Suter et al., 1995), structural integrity of retinal ganglion cell axons (Chatzopoulou et al., 2008), axonal guidance (Baeriswyl and Stoeckli, 2008;Furley et al., 1990;Wolman et al., 2008) and synapse formation (Leshchyns'ka and Sytnyk, 2016). Consistent with the alteration of synaptic function and axonal loss in pre-symptomatic AD cases (Mattson, 2004), CNTN2 has been widely associated with the disease pathogenesis and is known to interact with AD associated proteins. ...