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Anti-human placental antigen complex X-P2 (hPAX-P2) anti-serum recognizes C-terminus of huntingtin-associated protein 1A common to 1B as a determinant marker for the stigmoid body

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Abstract

The anti-serum against an unknown human placental antigen complex X-P2 (hPAX-P2) immunohistochemically recognizes three putative molecules (hPAX-P2S, hPAX-P2N, and hPAX-P2R), each of which is associated with the stigmoid bodies (STBs), necklace olfactory glomeruli (NOGs), or reticulo-filamentous structures (RFs) in the rat brain. The STBs also contain huntingtin-associated protein 1 (HAP1), and the HAP1-cDNA transfection induces STB-like inclusions in cultured cells. In order to clarify the relationship between hPAX-P2S and HAP1 isoforms (A/B), we performed Western blotting, immuno-histo/cytochemistry for light- and electron-microscopy and pre-adsorption tests with HAP1 deletion fragments. The results showed that the anti-hPAX-P2 anti-serum recognizes HAP1474–577 of HAP1A/B in Western blotting and strongly immunostains HAP1A-induced STB-like inclusions but far weakly detects HAP1B-induced diffuse structures in HAP1-transfected HEK 293 cells. In the rat brain, immunoreactivity of the anti-hPAX-P2 anti-serum for the STBs was eliminated by pre-adsorption with HAP1474–577, whereas no pre-adsorption with any different HAP1 fragments can suppress immunoreactivity for the NOGs and RFs, which were not immunoreactive to anti-HAP1 anti-serum. These findings indicate that hPAX-P2S, which is distinct from hPAX-P2N and hPAX-P2R, is identical with STB-constituted HAP1 and that the HAP1-induced/immunoreactive inclusions correspond to the hPAX-P2-immunoreactive STBs previously identified in the brain.

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Acknowledgments

This study was supported by Grant-in-Aids for Young Scientists (B) (No. 17700333) and for Scientific Research on Priority Areas (No.17052016) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) and a Grant-in-Aid for Scientific Research (C) (No.17500231) from the Japan Society for the Promotion of Science (JSPS). We cordially thank Prof. Yoshio Osawa (Hauptman-Wookward Medical Research Institute Inc., Buffalo, NY, USA) for gifting us anti-hPAX-P2 anti-serum. We are grateful to Mr. Chikahisa Matsuo, Mr. Jun Oba, and Mrs. Yumiko Matsuzaki for their technical assistance and administrative helps with this work, and to Dr. Ken-ichi Nakahama for his excellent technical help in the early stages of the present study. We would also like to acknowledge the technical expertise of the Institute for Biomedical Research and Education, Yamaguchi University Science Research Center and the DNA Core facility of the Center for Gene Research, Yamaguchi University, supported by a Grant-in-Aid from the Ministry of Education, Science, Sports, and Culture of Japan.

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Authors and Affiliations

  1. Division of Neuroanatomy, Department of Neuroscience, Yamaguchi University School of Medicine, Minami-Kogushi 1-1-1, Ube, Yamaguchi, 755-8505, Japan

    Ryutaro Fujinaga, Akie Yanai, Hirokazu Nakatsuka, Kumiko Yoshida, Yukio Takeshita, Kanako Uozumi, Changjiu Zhao, Kazuko Hirata, Keiji Kokubu & Koh Shinoda

  2. Department of Otolaryngology, Yamaguchi University School of Medicine, Minami-Kogushi 1-1-1, Ube, Yamaguchi, 755-8505, Japan

    Kazuko Hirata

  3. Department of Anatomy and Neurobiology, Kinki University School of Medicine, Osaka-sayama, Osaka, 589-8511, Japan

    Mamoru Nagano

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  1. Ryutaro Fujinaga
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  2. Akie Yanai
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  3. Hirokazu Nakatsuka
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  4. Kumiko Yoshida
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  5. Yukio Takeshita
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  6. Kanako Uozumi
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  7. Changjiu Zhao
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  10. Mamoru Nagano
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  11. Koh Shinoda
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Correspondence to Koh Shinoda.

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Fujinaga, R., Yanai, A., Nakatsuka, H. et al. Anti-human placental antigen complex X-P2 (hPAX-P2) anti-serum recognizes C-terminus of huntingtin-associated protein 1A common to 1B as a determinant marker for the stigmoid body. Histochem Cell Biol 128, 335–348 (2007). https://doi.org/10.1007/s00418-007-0315-5

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