Co-localisation, heterophilic interactions and regulated expression of IgLON family proteins in the chick nervous system

Lodge, Anthony P.; Howard, Mark; McNamee, Christine J.; Moss, Darren M.

doi:10.1016/s0169-328x(00)00184-4

Cited by 47 publications

(53 citation statements)

References 27 publications

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“…Antisera recognizing chicken LAMP, OBCAM and CEPU-1 were described previously (Lodge et al, 2000). α1LAMP-Fc, α2OBCAM-Fc and α2βCEPU-1-Fc (GenBank accession numbers for the respective IgLON sequences are Q98919, Q98892 and Q90773) were prepared either from stably transfected J558L mouse myeloma cells as described previously or from calciumphosphate-mediated transient transfection of HEK 293 human embryonic kidney cells, using methods described elsewhere (Chen and Okayama, 1988).…”

Section: Methodsmentioning

confidence: 99%

“…Immunofluorescence microscopy CHO cells were live stained with IgLON-Fc or IgLON-specific antisera, essentially as described previously (Lodge et al, 2000). Cells grown on coverslips were incubated at room temperature for 20 minutes in blocking buffer [0.12 M sodium phosphate buffer (pH 7.4), 1% bovine serum albumin (BSA)] containing antisera (1:100) or IgLON-Fc (25 µg ml -1 ) and binding was detected by incubation with Texas-Red-conjugated anti-human IgG or goat anti-rat IgG (1:100; both from Jackson ImmunoResearch).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were extracted in 20 mM Tris-HCl pH 7.6, 2 mM EGTA, 1% NP-40 and Complete™ (Roche) at 37°C for 15 minutes and centrifuged at 50,000 g at room temperature for 15 minutes. For dot blots, 2 µl of supernatant was spotted on nitrocellulose and then processed as for a western blot (Lodge et al, 2000).…”

Section: Transient Transfection Of Singly Transfected Cho Cellsmentioning

confidence: 99%

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Diglons are heterodimeric proteins composed of IgLON subunits, and Diglon-CO inhibits neurite outgrowth from cerebellar granule cells

Reed

McNamee

Rackstraw

et al. 2004

Journal of Cell Science

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IgLONs are a family of four cell adhesion molecules belonging to the Ig superfamily that are thought to play a role in cell-cell recognition and growth-cone migration. One member of the family, opioid-binding cell-adhesion molecule (OBCAM), might act as a tumour suppressor. Previous work has shown that limbic-system-associated protein (LAMP), CEPU-1/Neurotrimin and OBCAM interact homophilically and heterophilically within the family. Here, we show that, based on their relative affinities, CEPU-1 might be both a homo- and a heterophilic cell adhesion molecule, whereas LAMP and OBCAM act only as heterophilic cell adhesion molecules. A binding assay using recombinant IgLONs fused to human Fc showed that IgLONs are organized in the plane of the membrane as heterodimers, and we propose that IgLONs function predominantly as subunits of heterodimeric proteins (Diglons). Thus, the four IgLONs can form six Diglons. Furthermore, although singly transfected cell lines have little effect on neurite outgrowth, CHO cell lines expressing both CEPU-1 and OBCAM (Diglon-CO) inhibit neurite outgrowth from cerebellar granule cells.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Transient Transfection Of Singly Transfected Cho Cellsmentioning

confidence: 99%

See 1 more Smart Citation

Diglons are heterodimeric proteins composed of IgLON subunits, and Diglon-CO inhibits neurite outgrowth from cerebellar granule cells

Reed

McNamee

Rackstraw

et al. 2004

Journal of Cell Science

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“…Members of the pan-retina cell-cell adhesion molecules group are expressed in many cell types in the retina to mediate nonspecific retinal cell adhesion. Examples of this group include the neural cell adhesion molecule (N-CAM) (Daniloff et al, 1986), L1 (Chung et al, 2004), P84 and its interacting partner Integrin Associated Protein (IAP) (Mi et al, 2000), N-cadherin (Malicki et al, 2003), Neuroligin (von Kriegstein and Schmitz, 2003), N-Syndecan (Inatani et al, 2002), and LAMP, an isoform of IgLON (Lodge et al, 2000). Members of the specific cell-cell adhesional molecules group display regional expression patterns in the retina and are likely involved in regional adhesion functions that are important for specific retinal layers.…”

Section: Introductionmentioning

confidence: 99%

Nok plays an essential role in maintaining the integrity of the outer nuclear layer in the zebrafish retina

Wei

Zou

Takechi

et al. 2006

Experimental Eye Research

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Proper visual function of the vertebrate retina requires the maintenance of the integrity of the retinal outer nuclear layer (ONL), which is often affected in many blinding human retinal diseases.While the structural integrity of the ONL has long been considered to be maintained primarily through the outer limiting membrane (OLM), we have little knowledge on the development and maintenance of the OLM itself. Here, by analyzing the adhering properties of photoreceptors in zebrafish N-cad and nok mutants, we demonstrated for the first time that the nok gene is essential for the establishment and/or maintenance of the OLM. In addition, our results imply the possibility that Nok, Crumbs, and their associated proteins may constitute a type of photoreceptor-photoreceptor junctional complex that has not been described before. Thus, our study provides novel insights into the mechanisms by which the integrity of the ONL is maintained in the vertebrate retina.

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“…on May 10, 2018. by guest www.bloodjournal.org From proteins have been shown to be involved in cell-to-cell adhesion and to interact homophilically and heterophilically within the family with various binding affinities. [23][24][25][26] Using the optimized washing conditions described previously, we compared the DropArray technology with conventional 384-well plates using a regular automated washing procedure. Here, COS7 (adherent), HEK293T (semiadherent), and HEK293S (suspension-adapted) were transiently transfected with LSAMP, HNT, or OPCML constructs.…”

mentioning

confidence: 99%

Application of a new wall-less plate technology to complex multistep cell-based investigations using suspension cells

Quiñones¹,

Moore

Nicholes³

et al. 2013

Blood

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• The DropArray technology is compatible with the retention of suspension cells in multistep procedures thus enabling novel assay methods.• This technology enabled visualization and quantification of specific killing events triggered by bispecific antibodies engaging T cells. IntroductionThe demonstration that single cells could be grown in vitro 1 combined with the development of specific growth media 2 and later the establishment of the first cell lines [3][4][5][6] definitively marks the birth of cell culture as a critical research tool. Since then, investigators have developed a myriad of in vitro cellular models to further the understanding of various normal and pathologic cellular processes as well as to screen and characterize potential therapeutic modalities. All this progress occurred in concert with the development of many technologies that have impacted the investigator's ability to establish specific culture conditions and measure or visualize different cellular signals. As a result, cell-based assays are today almost universally used in biology research laboratories.During the past decade, cell-based assays have become fundamental and irreplaceable tools in the drug discovery and development industry. This trend was driven mainly by 2 rationales. First, the completion of the human genome, 7 combined with advances in functional genomics and proteomics, 8 has led to the need to evaluate thousand of potential new targets. Second, the high attrition rate of therapeutic candidates at the preclinical and clinical stages generated the need for more biologically relevant highthroughput screening approaches, bringing cell-based assay technologies to the forefront of the drug-discovery strategy. As a result, investigators' ability to develop rapid, flexible, robust, and costeffective high-throughput cell-based assays became of paramount importance.Despite significant technological progress in enabling technologies such as molecular labeling and the advent of high content screening approaches, cell-based assays, in part because of their well-plate format, continue to have major limitations. Although wells are an efficient and simple strategy to segregate experimental conditions in formats from 6-to 1536-well plates, they have major restrictions when suspension, loosely adherent, and in some cases fully adherent cells are used, especially with high-throughput formats such as 96-, 384-, and 1536-well plates. Microwell plates not only limit the use of certain cells but also significantly reduce the spectrum of experimental procedures that can be implemented in high-throughput cell-based screenings. Because the addition and removal of reagents to the wells could definitively compromise the cells, various technologies have been developed to circumvent these limitations. The homogeneous assay, developed in recent years, provides a convenient "add, mix, and read" approach to explore a broad spectrum of biologic events from cell viability/ Submitted July 31, 2012; accepted November 24, 2012. Prepublished online as Bl...

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Co-localisation, heterophilic interactions and regulated expression of IgLON family proteins in the chick nervous system

Cited by 47 publications

References 27 publications

Diglons are heterodimeric proteins composed of IgLON subunits, and Diglon-CO inhibits neurite outgrowth from cerebellar granule cells

Diglons are heterodimeric proteins composed of IgLON subunits, and Diglon-CO inhibits neurite outgrowth from cerebellar granule cells

Nok plays an essential role in maintaining the integrity of the outer nuclear layer in the zebrafish retina

Application of a new wall-less plate technology to complex multistep cell-based investigations using suspension cells

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