Presence of neuronal proteins in serially cultured cells from autopsy human brain.
Cells that contain the high (NF-H) and medium (NF-M) molecular weight neurofilaments and neuron-specific enolase (NSE) have been cultured from human brain (Brodmann frontal area 9 and hippocampus) obtained at routine autopsy. Three of these cultures (from a 52-year-old man with vascular dementia, a 75-year-old man with Alzheimer's disease, and an 80-year-old normal woman) have been propagated to passage 12-14 and studied in detail. Cellular morphology was pleomorphic and consistent with a relatively low level of differentiation, by both light microscopic and ultrastructural examination. Immunocytochemical reactivity in a characteristic filamentous pattern was observed with monoclonal antibodies to nonphosphorylated epitopes of NF-H and NF-M as well as with redundant antibody probes specific for NF-H and NF-M. Immunoblotting studies confirmed the presence of NF-H and NF-M. Immunocytochemistry and Immunoblotting also demonstrated the presence of NSE, but not of glial fibrillary acidic protein or other glial markers. Immunocytochemical studies also indicated the presence of proline hydroxylase, an enzyme found in fibroblasts but not neurons. These studies indicate that cells can be cultured from autopsy human brain which are not neurons but nevertheless express molecules used as markers (NF-H, NF-M, and NSE) of adult neurons in vivo and in culture.
Blass
J P
JP
Altschul Laboratory for Dementia Research, Burke Medical Research Institute, Cornell University Medical College, White Plains, NY 10605.
Markesbery
W R
WR
Ko
L W
LW
DeGiorgio
L
L
Sheu
K F
KF
Darzynkiewicz
Z
Z
D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology; D001921:Brain / Q000166:cytology* / Q000201:enzymology / Q000473:pathology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005260:Female; D005434:Flow Cytometry; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism
D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D010751:Phosphopyruvate Hydratase