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Sulfated proteoglycans synthesized by Neuro 2a neuroblastoma cells: Comparison between cells with and without ganglioside-induced neurites

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Abstract

Mouse neuroblastoma Neuro 2a cells are known to extend neurite-like processes in response to gangliosides added to the culture medium. We compared the structural features of proteoglycans (PG) synthesized by conventional Neuro 2a cells with those of neurite-bearing cells. Two different proteoglycans labeled with [35S]sulfate, namely, chondroitin sulfate proteoglycan (CS-PG) and heparan sulfate proteoglycan (HS-PG), were found both in the cell layer and in the culture medium of the conventional cells. CS-PG isolated from the cell layer had a Kav value of 0.38 on Sepharose CL-6B, and had CS side chains with Mr of 27,000. HS-PG in the cell layer was slightly larger (Kav of 0.33) in terms of hydrodynamic size than CS-PG, and the apparent Mr of the heparan sulfate side chains was 10,000. The structural parameters of CS-PG and HS-PG isolated from the medium were almost identical to those of the PGs in the cell layer. In addition to these PGs, single-chain HS, with an average Mr of 2,500, was observed only in the cell layer and this component was the major sulfated component in the cell layers of both control and ganglioside treated cells. The neurite-bearing cells also synthesized both CS-PG and HS-PG which were very similar in hydrodynamic size to those synthesized by the conventional cells, but the size of HS side chains was greater. Radioactivity, as35S, of each sulfated component from the gangliosideteated culture seemed to be slightly less than that of the corresponding component from the control culture. These findings indicate that the marked morphological change in Neuro 2a cells, induced by gangliosides is not accompanied by major changes in the synthesis of PGs.

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Authors and Affiliations

  1. Department of Embryology, Institute for Developmental Research, Aichi Prefecture Colony, 480-03, Kasugai, Aichi

    Kimi Watanabe & Tsuyoshi Totsuka

  2. Department of Perinatology, Institute for Developmental Research, Aichi Prefecture Colony, 480-03, Kasugai, Aichi

    Atsuhiko Oohira & Keiichi Yoshida

  3. Tokyo Research Institute, Seikagaku Kogyo Co., Ltd., Tateno, Higashiyamato, Tokyo 189, Japan

    Ritsuko Katoh-Semba

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  1. Kimi Watanabe
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  3. Ritsuko Katoh-Semba
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  4. Tsuyoshi Totsuka
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  5. Keiichi Yoshida
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Watanabe, K., Oohira, A., Katoh-Semba, R. et al. Sulfated proteoglycans synthesized by Neuro 2a neuroblastoma cells: Comparison between cells with and without ganglioside-induced neurites. Neurochem Res 14, 707–716 (1989). https://doi.org/10.1007/BF00964947

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