The Experts below are selected from a list of 117 Experts worldwide ranked by ideXlab platform

Frans Charles Servatius Ramaekers - One of the best experts on this subject based on the ideXlab platform.

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto)

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto) chemically. It became obvious that the expression of NSP-C, as detected by immunofluorescence microscopy and Western blotting, is most prominent in cell lines with a high degree of neuronal differentiation, such as LA-N-5. Such highly differentiated cells also express other neural and Neuroendocrine markers, such as neural cell adhesion molecule (NCAM), neurofilament Proteins, synaptophysin, and chromogranin. NSP-A was observed in all cell lines to a different extent. However, no clear correlation was observed with the degree of neuronal differentiation as defined by other neuronal and Neuroendocrine markers or morphology. NSP-B could not be detected. The induction of neuronal differentiation with nerve growth factor, dbcAMP, and retinoic acid in the rat pheochromocytoma cell line PC12 and the human teratocarcinoma cell line hNT2, respectively, induced the expression of NSP-A and NSP-C in these cell lines parallel to the induction of neurofilament Protein expression. It is concluded that NSP-C expression, in particular, is strongly correlated with neuronal differentiation.

  • Neuroendocrine-Specific Protein (NSP)-reticulons as independent markers for non-small cell lung cancer with Neuroendocrine differentiation. An in vitro histochemical study
    Histochemistry and Cell Biology, 1997
    Co-Authors: Nicole H. M. Senden, Ilona Linnoila, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons have recently been discovered and were shown to exhibit a restricted, Neuroendocrine/neural-Specific expression pattern. These Protein aggregates are anchored to the membranes of the endoplasmic reticulum and occur in small cell lung cancer (SCLC), but not in typical non-SCLC. In the current study we have examined the occurrence of NSP-reticulons in non-SCLC cell lines known to express Neuroendocrine features (non-SCLC-NE). NSP-reticulon expression was observed in all three non-SCLC-NE cell lines studied, albeit with variable intensity and in varying numbers of cells. Western blot analysis confirmed the presence of NSP-reticulon expression in these non-SCLC-NE cell lines, and showed that they were predominantly of the NSP-A type. When compared to conventional Neuroendocrine markers, NSP-reticulons revealed a distinct staining profile, showing only partial overlap with the other markers. The non-SCLC-NE cell lines combined these Neuroendocrine characteristics with some features of non-SCLC. We conclude that NSP-reticulon expression is restricted to lung carcinoma cells with a Neuroendocrine phenotype and predict that these constituents may become clinically relevant markers for the detection of Neuroendocrine differentiation in solid tumours.

  • A comparison of NSP-reticulons with conventional Neuroendocrine markers in immunophenotyping of lung cancers
    The Journal of Pathology, 1997
    Co-Authors: Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Adriaan P. De Bruïne, S. S. Wagenaar, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons are endoplasmic reticulum-associated Protein complexes, which have been identified as markers for Neuroendocrine differentiation. In this study, the expression of two members of the family of NSP-reticulons, NSP-A and NSP-C, have been investigated in different types of lung cancer and compared with the expression patterns of five conventional Neuroendocrine markers, the neural cell adhesion molecule (NCAM), synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. NSP-A and NSP-C antibodies were reactive with most carcinoid tumour and small cell lung carcinoma (SCLC) cases, while atypical carcinoid tumours showed a variable expression. In the total group of Neuroendocrine tumours, a high concordance of expression was found between NSP-A and NSP-C, while their expression correlated well with NCAM and synaptophysin positivity. Chromogranin A, Leu-7, and neurofilament Proteins were shown to be expressed to a limited extent in these Neuroendocrine tumours. In a selected group of non-SCLCs known to exhibit Neuroendocrine features, NSP-A expression was detected at much higher frequency than NSP-C. In virtually all NSP-A positive cases, this expression was associated with one or more of the other neuroendocine markers. NSP-A expression showed a stronger correlation with conventional Neuroendocrine markers than NCAM. In detecting Neuroendocrine differentiation in non-SCLC, NSP-A is more sensitive than synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. It is concluded that NSP-reticulons are valuable markers in the diagnosis of Neuroendocrine differentiation in non-SCLC and should be used in conjunction with NCAM. © 1997 John Wiley & Sons, Ltd.

  • Neuroendocrine Specific Protein c nsp c subcellular localization and differential expression in relation to nsp a
    European Journal of Cell Biology, 1996
    Co-Authors: Nicole H. M. Senden, Anton J. M. Roebroek, Jos L V Broers, H J K Van De Velde, E D J Timmer, J E Boers, Frans Charles Servatius Ramaekers
    Abstract:

    : A mouse monoclonal antibody RNL-4, as well as rabbit polyclonal antiserum POL-8 were raised against a synthetic peptide, encompassing the first twenty unique amino-terminal amino acid residues of NSP-C. The Specificity of both immunoreagents was established in an ELISA assay using the synthetic peptide and by their immunoreactivity to NSP-C fusion Proteins. Immunofluorescence analysis of COS-1 cells, transfected with NSP-C cDNA, showed staining of the endoplasmic reticulum with RNL-4 and POL-8. No cross-reactivity of these reagents with NSP-A or NSP-B was seen. Immunohistochemical studies in normal human tissues showed expression of NSP-C in tissues of neural and Neuroendocrine origin, i.e. neurons of the central and peripheral nervous system, the neurohypophysis, adrenal medulla, adenohypophysis, pars intermedia, and in sporadic neuroedocrine cells of the lung. Expression of NSP-C was found in several small cell lung cancer (SCLC) cell lines, in non-SCLC cell lines with Neuroendocrine features, but not in typical non-SCLC cell lines. Also, in a neuroblastoma cell line NSP-C expression was observed. Immunoblotting and immunoprecipitation studies with RNL-4 and POL-8 identified the 23 kDa NSP-C polypeptide in these cell lines. Immunofluorescence microscopy showed that also in these cell lines NSP-C is located at the endoplasmic reticulum, as shown before for NSP-A and NSP-B. In some of the cell lines coexpression of NSP-A and NSP-C was observed, while in others only one of the two could be detected. The differential expression of NSP-A and NSP-C in these cell lines is confirmed by immunoblotting and was also evident at the mRNA level. When NSP-A and NSP-C were coexpressed, the number of NSP-C-positive cells was always less than the number of NSP-A-positive cells. A partial colocalization of NSPs was observed in the endoplasmic reticulum. Cell fractionation studies revealed that both Proteins are retained in the membranous fraction of the cell, from which they can be solubilized by Triton X-100. Immunoprecipitation analyses under native conditions indicate that NSP-C does not need to associate with NSP-A to form high molecular weight NSP-reticulons.

Jos L V Broers - One of the best experts on this subject based on the ideXlab platform.

  • Neuroendocrine Specific Protein c a marker of neuronal differentiation is reduced in brain of patients with down syndrome and alzheimer s disease
    Biochemical and Biophysical Research Communications, 2000
    Co-Authors: Jos L V Broers, Nigel J Cairns, Gert Lubec
    Abstract:

    Abstract Neuroendocrine-Specific Protein C (NSP-C) is found in neural and Neuroendocrine cells and associated with the endoplasmic reticulum. Its expression was found to correlate with the degree of neuronal differentiation. As the neuropathological findings in Down syndrome (DS) includes deficits of differentiation, and we detected a downregulated sequence with 100% homology with NSP-C homolog mRNA in temporal cortex of patients with DS as well as Alzheimer's disease (AD) using differential display-polymerase chain reaction (DD-PCR), we decided to examine the Protein levels of NSP-C in temporal, frontal cortex and cerebellum of DS and AD. To normalize NSP-C versus neuronal density, we also determined neuron-Specific enolase (NSE) levels and calculated the ratios. NSP-C was significantly reduced in DS (temporal and frontal cortex) and AD (frontal cortex) compared to controls. The significant decrease of NSP-C in DS was even more pronounced when related to NSE levels. Impaired differentiation in DS brain may well be due to absolutely and relatively decreased NSP-C levels in temporal and frontal cortex. As NSP-C was also reduced in AD frontal cortex, NSP-C deficits in these disorders may be reflecting neurodegenerative changes rather than a primary and Specific finding of DS or AD pathogenesis.

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto)

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto) chemically. It became obvious that the expression of NSP-C, as detected by immunofluorescence microscopy and Western blotting, is most prominent in cell lines with a high degree of neuronal differentiation, such as LA-N-5. Such highly differentiated cells also express other neural and Neuroendocrine markers, such as neural cell adhesion molecule (NCAM), neurofilament Proteins, synaptophysin, and chromogranin. NSP-A was observed in all cell lines to a different extent. However, no clear correlation was observed with the degree of neuronal differentiation as defined by other neuronal and Neuroendocrine markers or morphology. NSP-B could not be detected. The induction of neuronal differentiation with nerve growth factor, dbcAMP, and retinoic acid in the rat pheochromocytoma cell line PC12 and the human teratocarcinoma cell line hNT2, respectively, induced the expression of NSP-A and NSP-C in these cell lines parallel to the induction of neurofilament Protein expression. It is concluded that NSP-C expression, in particular, is strongly correlated with neuronal differentiation.

  • Neuroendocrine-Specific Protein (NSP)-reticulons as independent markers for non-small cell lung cancer with Neuroendocrine differentiation. An in vitro histochemical study
    Histochemistry and Cell Biology, 1997
    Co-Authors: Nicole H. M. Senden, Ilona Linnoila, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons have recently been discovered and were shown to exhibit a restricted, Neuroendocrine/neural-Specific expression pattern. These Protein aggregates are anchored to the membranes of the endoplasmic reticulum and occur in small cell lung cancer (SCLC), but not in typical non-SCLC. In the current study we have examined the occurrence of NSP-reticulons in non-SCLC cell lines known to express Neuroendocrine features (non-SCLC-NE). NSP-reticulon expression was observed in all three non-SCLC-NE cell lines studied, albeit with variable intensity and in varying numbers of cells. Western blot analysis confirmed the presence of NSP-reticulon expression in these non-SCLC-NE cell lines, and showed that they were predominantly of the NSP-A type. When compared to conventional Neuroendocrine markers, NSP-reticulons revealed a distinct staining profile, showing only partial overlap with the other markers. The non-SCLC-NE cell lines combined these Neuroendocrine characteristics with some features of non-SCLC. We conclude that NSP-reticulon expression is restricted to lung carcinoma cells with a Neuroendocrine phenotype and predict that these constituents may become clinically relevant markers for the detection of Neuroendocrine differentiation in solid tumours.

  • A comparison of NSP-reticulons with conventional Neuroendocrine markers in immunophenotyping of lung cancers
    The Journal of Pathology, 1997
    Co-Authors: Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Adriaan P. De Bruïne, S. S. Wagenaar, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons are endoplasmic reticulum-associated Protein complexes, which have been identified as markers for Neuroendocrine differentiation. In this study, the expression of two members of the family of NSP-reticulons, NSP-A and NSP-C, have been investigated in different types of lung cancer and compared with the expression patterns of five conventional Neuroendocrine markers, the neural cell adhesion molecule (NCAM), synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. NSP-A and NSP-C antibodies were reactive with most carcinoid tumour and small cell lung carcinoma (SCLC) cases, while atypical carcinoid tumours showed a variable expression. In the total group of Neuroendocrine tumours, a high concordance of expression was found between NSP-A and NSP-C, while their expression correlated well with NCAM and synaptophysin positivity. Chromogranin A, Leu-7, and neurofilament Proteins were shown to be expressed to a limited extent in these Neuroendocrine tumours. In a selected group of non-SCLCs known to exhibit Neuroendocrine features, NSP-A expression was detected at much higher frequency than NSP-C. In virtually all NSP-A positive cases, this expression was associated with one or more of the other neuroendocine markers. NSP-A expression showed a stronger correlation with conventional Neuroendocrine markers than NCAM. In detecting Neuroendocrine differentiation in non-SCLC, NSP-A is more sensitive than synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. It is concluded that NSP-reticulons are valuable markers in the diagnosis of Neuroendocrine differentiation in non-SCLC and should be used in conjunction with NCAM. © 1997 John Wiley & Sons, Ltd.

Nicole H. M. Senden - One of the best experts on this subject based on the ideXlab platform.

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto)

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto) chemically. It became obvious that the expression of NSP-C, as detected by immunofluorescence microscopy and Western blotting, is most prominent in cell lines with a high degree of neuronal differentiation, such as LA-N-5. Such highly differentiated cells also express other neural and Neuroendocrine markers, such as neural cell adhesion molecule (NCAM), neurofilament Proteins, synaptophysin, and chromogranin. NSP-A was observed in all cell lines to a different extent. However, no clear correlation was observed with the degree of neuronal differentiation as defined by other neuronal and Neuroendocrine markers or morphology. NSP-B could not be detected. The induction of neuronal differentiation with nerve growth factor, dbcAMP, and retinoic acid in the rat pheochromocytoma cell line PC12 and the human teratocarcinoma cell line hNT2, respectively, induced the expression of NSP-A and NSP-C in these cell lines parallel to the induction of neurofilament Protein expression. It is concluded that NSP-C expression, in particular, is strongly correlated with neuronal differentiation.

  • Neuroendocrine-Specific Protein (NSP)-reticulons as independent markers for non-small cell lung cancer with Neuroendocrine differentiation. An in vitro histochemical study
    Histochemistry and Cell Biology, 1997
    Co-Authors: Nicole H. M. Senden, Ilona Linnoila, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons have recently been discovered and were shown to exhibit a restricted, Neuroendocrine/neural-Specific expression pattern. These Protein aggregates are anchored to the membranes of the endoplasmic reticulum and occur in small cell lung cancer (SCLC), but not in typical non-SCLC. In the current study we have examined the occurrence of NSP-reticulons in non-SCLC cell lines known to express Neuroendocrine features (non-SCLC-NE). NSP-reticulon expression was observed in all three non-SCLC-NE cell lines studied, albeit with variable intensity and in varying numbers of cells. Western blot analysis confirmed the presence of NSP-reticulon expression in these non-SCLC-NE cell lines, and showed that they were predominantly of the NSP-A type. When compared to conventional Neuroendocrine markers, NSP-reticulons revealed a distinct staining profile, showing only partial overlap with the other markers. The non-SCLC-NE cell lines combined these Neuroendocrine characteristics with some features of non-SCLC. We conclude that NSP-reticulon expression is restricted to lung carcinoma cells with a Neuroendocrine phenotype and predict that these constituents may become clinically relevant markers for the detection of Neuroendocrine differentiation in solid tumours.

  • A comparison of NSP-reticulons with conventional Neuroendocrine markers in immunophenotyping of lung cancers
    The Journal of Pathology, 1997
    Co-Authors: Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Adriaan P. De Bruïne, S. S. Wagenaar, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons are endoplasmic reticulum-associated Protein complexes, which have been identified as markers for Neuroendocrine differentiation. In this study, the expression of two members of the family of NSP-reticulons, NSP-A and NSP-C, have been investigated in different types of lung cancer and compared with the expression patterns of five conventional Neuroendocrine markers, the neural cell adhesion molecule (NCAM), synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. NSP-A and NSP-C antibodies were reactive with most carcinoid tumour and small cell lung carcinoma (SCLC) cases, while atypical carcinoid tumours showed a variable expression. In the total group of Neuroendocrine tumours, a high concordance of expression was found between NSP-A and NSP-C, while their expression correlated well with NCAM and synaptophysin positivity. Chromogranin A, Leu-7, and neurofilament Proteins were shown to be expressed to a limited extent in these Neuroendocrine tumours. In a selected group of non-SCLCs known to exhibit Neuroendocrine features, NSP-A expression was detected at much higher frequency than NSP-C. In virtually all NSP-A positive cases, this expression was associated with one or more of the other neuroendocine markers. NSP-A expression showed a stronger correlation with conventional Neuroendocrine markers than NCAM. In detecting Neuroendocrine differentiation in non-SCLC, NSP-A is more sensitive than synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. It is concluded that NSP-reticulons are valuable markers in the diagnosis of Neuroendocrine differentiation in non-SCLC and should be used in conjunction with NCAM. © 1997 John Wiley & Sons, Ltd.

  • Neuroendocrine Specific Protein c nsp c subcellular localization and differential expression in relation to nsp a
    European Journal of Cell Biology, 1996
    Co-Authors: Nicole H. M. Senden, Anton J. M. Roebroek, Jos L V Broers, H J K Van De Velde, E D J Timmer, J E Boers, Frans Charles Servatius Ramaekers
    Abstract:

    : A mouse monoclonal antibody RNL-4, as well as rabbit polyclonal antiserum POL-8 were raised against a synthetic peptide, encompassing the first twenty unique amino-terminal amino acid residues of NSP-C. The Specificity of both immunoreagents was established in an ELISA assay using the synthetic peptide and by their immunoreactivity to NSP-C fusion Proteins. Immunofluorescence analysis of COS-1 cells, transfected with NSP-C cDNA, showed staining of the endoplasmic reticulum with RNL-4 and POL-8. No cross-reactivity of these reagents with NSP-A or NSP-B was seen. Immunohistochemical studies in normal human tissues showed expression of NSP-C in tissues of neural and Neuroendocrine origin, i.e. neurons of the central and peripheral nervous system, the neurohypophysis, adrenal medulla, adenohypophysis, pars intermedia, and in sporadic neuroedocrine cells of the lung. Expression of NSP-C was found in several small cell lung cancer (SCLC) cell lines, in non-SCLC cell lines with Neuroendocrine features, but not in typical non-SCLC cell lines. Also, in a neuroblastoma cell line NSP-C expression was observed. Immunoblotting and immunoprecipitation studies with RNL-4 and POL-8 identified the 23 kDa NSP-C polypeptide in these cell lines. Immunofluorescence microscopy showed that also in these cell lines NSP-C is located at the endoplasmic reticulum, as shown before for NSP-A and NSP-B. In some of the cell lines coexpression of NSP-A and NSP-C was observed, while in others only one of the two could be detected. The differential expression of NSP-A and NSP-C in these cell lines is confirmed by immunoblotting and was also evident at the mRNA level. When NSP-A and NSP-C were coexpressed, the number of NSP-C-positive cells was always less than the number of NSP-A-positive cells. A partial colocalization of NSPs was observed in the endoplasmic reticulum. Cell fractionation studies revealed that both Proteins are retained in the membranous fraction of the cell, from which they can be solubilized by Triton X-100. Immunoprecipitation analyses under native conditions indicate that NSP-C does not need to associate with NSP-A to form high molecular weight NSP-reticulons.

Anton J. M. Roebroek - One of the best experts on this subject based on the ideXlab platform.

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto)

  • Neuronal differentiation is accompanied by NSP-C expression
    Cell and Tissue Research, 1998
    Co-Authors: Jurgen Hens, Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Frans Charles Servatius Ramaekers, Ronny Nuydens, Hugo Geerts, Jos L V Broers
    Abstract:

    Neuroendocrine-Specific Protein (NSP) reticulons are expressed in neural and Neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto) chemically. It became obvious that the expression of NSP-C, as detected by immunofluorescence microscopy and Western blotting, is most prominent in cell lines with a high degree of neuronal differentiation, such as LA-N-5. Such highly differentiated cells also express other neural and Neuroendocrine markers, such as neural cell adhesion molecule (NCAM), neurofilament Proteins, synaptophysin, and chromogranin. NSP-A was observed in all cell lines to a different extent. However, no clear correlation was observed with the degree of neuronal differentiation as defined by other neuronal and Neuroendocrine markers or morphology. NSP-B could not be detected. The induction of neuronal differentiation with nerve growth factor, dbcAMP, and retinoic acid in the rat pheochromocytoma cell line PC12 and the human teratocarcinoma cell line hNT2, respectively, induced the expression of NSP-A and NSP-C in these cell lines parallel to the induction of neurofilament Protein expression. It is concluded that NSP-C expression, in particular, is strongly correlated with neuronal differentiation.

  • Neuroendocrine-Specific Protein (NSP)-reticulons as independent markers for non-small cell lung cancer with Neuroendocrine differentiation. An in vitro histochemical study
    Histochemistry and Cell Biology, 1997
    Co-Authors: Nicole H. M. Senden, Ilona Linnoila, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons have recently been discovered and were shown to exhibit a restricted, Neuroendocrine/neural-Specific expression pattern. These Protein aggregates are anchored to the membranes of the endoplasmic reticulum and occur in small cell lung cancer (SCLC), but not in typical non-SCLC. In the current study we have examined the occurrence of NSP-reticulons in non-SCLC cell lines known to express Neuroendocrine features (non-SCLC-NE). NSP-reticulon expression was observed in all three non-SCLC-NE cell lines studied, albeit with variable intensity and in varying numbers of cells. Western blot analysis confirmed the presence of NSP-reticulon expression in these non-SCLC-NE cell lines, and showed that they were predominantly of the NSP-A type. When compared to conventional Neuroendocrine markers, NSP-reticulons revealed a distinct staining profile, showing only partial overlap with the other markers. The non-SCLC-NE cell lines combined these Neuroendocrine characteristics with some features of non-SCLC. We conclude that NSP-reticulon expression is restricted to lung carcinoma cells with a Neuroendocrine phenotype and predict that these constituents may become clinically relevant markers for the detection of Neuroendocrine differentiation in solid tumours.

  • A comparison of NSP-reticulons with conventional Neuroendocrine markers in immunophenotyping of lung cancers
    The Journal of Pathology, 1997
    Co-Authors: Nicole H. M. Senden, Helgi J. K. Van De Velde, Anton J. M. Roebroek, Erika D J Timmer, Jos L V Broers, Adriaan P. De Bruïne, S. S. Wagenaar, Frans Charles Servatius Ramaekers
    Abstract:

    Neuroendocrine-Specific Protein (NSP)-reticulons are endoplasmic reticulum-associated Protein complexes, which have been identified as markers for Neuroendocrine differentiation. In this study, the expression of two members of the family of NSP-reticulons, NSP-A and NSP-C, have been investigated in different types of lung cancer and compared with the expression patterns of five conventional Neuroendocrine markers, the neural cell adhesion molecule (NCAM), synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. NSP-A and NSP-C antibodies were reactive with most carcinoid tumour and small cell lung carcinoma (SCLC) cases, while atypical carcinoid tumours showed a variable expression. In the total group of Neuroendocrine tumours, a high concordance of expression was found between NSP-A and NSP-C, while their expression correlated well with NCAM and synaptophysin positivity. Chromogranin A, Leu-7, and neurofilament Proteins were shown to be expressed to a limited extent in these Neuroendocrine tumours. In a selected group of non-SCLCs known to exhibit Neuroendocrine features, NSP-A expression was detected at much higher frequency than NSP-C. In virtually all NSP-A positive cases, this expression was associated with one or more of the other neuroendocine markers. NSP-A expression showed a stronger correlation with conventional Neuroendocrine markers than NCAM. In detecting Neuroendocrine differentiation in non-SCLC, NSP-A is more sensitive than synaptophysin, chromogranin A, Leu-7, and neurofilament Proteins. It is concluded that NSP-reticulons are valuable markers in the diagnosis of Neuroendocrine differentiation in non-SCLC and should be used in conjunction with NCAM. © 1997 John Wiley & Sons, Ltd.

  • Neuroendocrine Specific Protein c nsp c subcellular localization and differential expression in relation to nsp a
    European Journal of Cell Biology, 1996
    Co-Authors: Nicole H. M. Senden, Anton J. M. Roebroek, Jos L V Broers, H J K Van De Velde, E D J Timmer, J E Boers, Frans Charles Servatius Ramaekers
    Abstract:

    : A mouse monoclonal antibody RNL-4, as well as rabbit polyclonal antiserum POL-8 were raised against a synthetic peptide, encompassing the first twenty unique amino-terminal amino acid residues of NSP-C. The Specificity of both immunoreagents was established in an ELISA assay using the synthetic peptide and by their immunoreactivity to NSP-C fusion Proteins. Immunofluorescence analysis of COS-1 cells, transfected with NSP-C cDNA, showed staining of the endoplasmic reticulum with RNL-4 and POL-8. No cross-reactivity of these reagents with NSP-A or NSP-B was seen. Immunohistochemical studies in normal human tissues showed expression of NSP-C in tissues of neural and Neuroendocrine origin, i.e. neurons of the central and peripheral nervous system, the neurohypophysis, adrenal medulla, adenohypophysis, pars intermedia, and in sporadic neuroedocrine cells of the lung. Expression of NSP-C was found in several small cell lung cancer (SCLC) cell lines, in non-SCLC cell lines with Neuroendocrine features, but not in typical non-SCLC cell lines. Also, in a neuroblastoma cell line NSP-C expression was observed. Immunoblotting and immunoprecipitation studies with RNL-4 and POL-8 identified the 23 kDa NSP-C polypeptide in these cell lines. Immunofluorescence microscopy showed that also in these cell lines NSP-C is located at the endoplasmic reticulum, as shown before for NSP-A and NSP-B. In some of the cell lines coexpression of NSP-A and NSP-C was observed, while in others only one of the two could be detected. The differential expression of NSP-A and NSP-C in these cell lines is confirmed by immunoblotting and was also evident at the mRNA level. When NSP-A and NSP-C were coexpressed, the number of NSP-C-positive cells was always less than the number of NSP-A-positive cells. A partial colocalization of NSPs was observed in the endoplasmic reticulum. Cell fractionation studies revealed that both Proteins are retained in the membranous fraction of the cell, from which they can be solubilized by Triton X-100. Immunoprecipitation analyses under native conditions indicate that NSP-C does not need to associate with NSP-A to form high molecular weight NSP-reticulons.

Jiangang Yuan - One of the best experts on this subject based on the ideXlab platform.

  • molecular characterization of human reticulon 3 as a potential marker during the differentiation of human neuroblastoma sh sy5y cells
    Progress in Natural Science, 2004
    Co-Authors: Hongbo Yang, Xiaozhong Peng, Boqin Qiang, Xiaowei Huang, Jiangang Yuan
    Abstract:

    Abstract Neuroendocrine-Specific Protein (NSP) -reticulons are endoplasmic reticulum-associated Protein complexes, which are localized in the endoplasmic reticulum (ER) and identified as markers for Neuroendocrine differentiation. In the present study, human reticulon 3 gene (hRTN3) was cloned and its expression pattern in a variety of tissues was investigated. Truncated hRTN3s corresponding to the C-terminal (hRTN3-C) domain were expressed and purified. hRTN3 mRNA was down-regulated during the differentiation of human neuroblastoma cell line SH-SY5Y induced by all-trans-retinoic acid (RA), which suggests that, like other members of the reticulon family, hRTN3 is a potential marker for Neuroendocrine differentiation.

  • overexpression of human reticulon 3 hrtn3 in astrocytoma
    Clinical Neuropathology, 2004
    Co-Authors: Xiaowei Huang, Xiaozhong Peng, Boqin Qiang, Hongbo Yang, Jiangang Yuan
    Abstract:

    : Neuroendocrine-Specific Protein (NSP) reticulons (RTN) are endoplasmic reticulum-associated Protein complexes, which are localized in the endoplasmic reticulum (ER) and identified as markers for Neuroendocrine differentiation. At least 4 different RTN genes have been identified in mammals, but in most cases, the functions of the encoded Proteins except mammalian RTN4-A and RTN4-B are still elusive. In the present study, the expression of human reticulon 3 (hRTN3) in a variety of tissues was investigated. Northern blotting hybridization revealed higher expression of hRTN3 in normal brain and some endocrine-related organs such as thyroid relative to other non-endocrine organs. Twelve surgical specimens meeting the histological criteria for astrocytoma grade II, III and IV were investigated by in situ hybridization and immunohistochemical analysis, which demonstrated the overexpression of hRTN3 in astrocytoma tumor cells, while in non-cancerous brain tissues hRTN3 was mainly expressed in neurons and almost no signals in glia cells could be detected. The differential expression of hRTN3 between astrocytoma and non-cancerous tissue may provide insight into the progress of astrocytoma.