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A Novel Marker for Purkinje Cells, KIAA0864 Protein. An Analysis Based on a Monoclonal Antibody HFB-16 in Developing Human Cerebellum

Yasuhiro Nakamura, Munehiko Yamamoto, Eriko Oda, Yonehiro Kanemura, Eri Kodama, Atsuyo Yamamoto, Hideyuki Yamamoto, Kenji Miyado, Hirotaka James Okano, Ryouji Fukagawa, Koichi Higaki, Mami Yamasaki and Hideyuki Okano

Department of Pathology, St. Mary's Hospital, Kurume, Japan (YN,RF,KH); Department of Chemistry, Kurume University School of Medicine, Kurume, Japan (MY,EO); Tissue Engineering Research Center, National Institute of Advanced Industrial Science and Technology, Amagasaki, Hyogo, Japan (YK,EK,AY,HY); Institute for Clinical Research (YK,MY), and Department of Neurosurgery (MY), Osaka National Hospital, Osaka, Japan; Department of Reproductive Biology and Pathology, National Center for Child Health and Development, Tokyo, Japan (KM); Department of Physiology, Keio University School of Medicine, Tokyo, Japan (HJO,HO); and Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan (HJO,HO)

Correspondence to: Yasuhiro Nakamura, MD, Department of Pathology, St. Mary's Hospital, 422, Tsubukuhon-machi, Kurume-shi, Japan 830-8543. E-mail: naka{at}st-mary-med.or.jp

In the search for immunohistochemical markers of the developing human brain, a monoclonal antibody, HFB-16, was raised against homogenates from the cerebrum of a 15-gestational-week-old (GW) human fetus and screened on paraffin-embedded human embryonic brain specimens. This antibody was particularly useful as a marker for Purkinje cells in the developing human cerebellum. Positive immunoreactivities with HFB-16 first appeared in the Purkinje cell layer at 17 GW. From 20 to 24 GW, positive immunoreactivities were found above the lamina dissecans. After 25 GW, dendrites of Purkinje cells were found with the HFB-16 antibody, and the nerve fibers of the Purkinje cells became positive after 35 GW. Neurons in the dentate nucleus and external and internal granular layers reacted negatively to this antibody. After 1 year, when the external granular layer faded out, the dendrites of the Purkinje cells reached the pial surface of the cerebellum, and nerve fibers began to develop in the white matter. This antibody was also useful for characterization of components in heterotopic neurons found in various anomaly syndromes such as trisomy 13. Expressional cloning indicated the antigen against HFB-16 to be human KIAA0864 protein, which is supposed to be an alternative splicing product of p116Rip, whose function has not yet been elucidated. The antigenicity of the KIAA0864 protein was confirmed using human cDNA of the KIAA0864 protein, a protein expression vector, and an HFB-16 antibody. (J Histochem Cytochem 53:423–430, 2005)

Key Words: Purkinje cell • monoclonal antibody • cerebellar development • KIAA0864 protein • immunohistochemistry • expressional cloning

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