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.
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 HAP1(474-577) 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 HAP1(474-577), 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.
Fujinaga
Ryutaro
R
Division of Neuroanatomy, Department of Neuroscience, Yamaguchi University School of Medicine, Minami-Kogushi 1-1-1, Ube, Yamaguchi, 755-8505, Japan.
Yanai
Akie
A
Nakatsuka
Hirokazu
H
Yoshida
Kumiko
K
Takeshita
Yukio
Y
Uozumi
Kanako
K
Zhao
Changjiu
C
Hirata
Kazuko
K
Kokubu
Keiji
K
Nagano
Mamoru
M
Shinoda
Koh
K
D000818:Animals; D001324:Autoantigens / Q000097:blood / Q000276:immunology*; D015415:Biomarkers / Q000032:analysis / Q000097:blood; D002460:Cell Line; D017353:Gene Deletion; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D046529:Microscopy, Electron, Transmission; D016253:Microscopy, Immunoelectron; D009419:Nerve Tissue Proteins / Q000097:blood / Q000235:genetics / Q000276:immunology*; D009830:Olfactory Bulb / Q000378:metabolism; D010926:Placental Hormones / Q000097:blood / Q000276:immunology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D001324:Autoantigens; D015415:Biomarkers; C490757:Hap1 protein, rat; D007106:Immune Sera; D009419:Nerve Tissue Proteins; D010926:Placental Hormones; D020033:Protein Isoforms; C064203:placental antigen X-P2