Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.

The human GFAP splice variants GFAPDelta164 and GFAPDeltaexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jinte Middeldorp, Simone A van den Berge, Eleonora Aronica, Dave Speijer, Elly M Hol
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d36ebe57eec64cc4b8ff8902b462e214
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:d36ebe57eec64cc4b8ff8902b462e214
record_format	dspace
spelling	oai:doaj.org-article:d36ebe57eec64cc4b8ff8902b462e2142021-11-25T06:28:20ZSpecific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.1932-620310.1371/journal.pone.0007663https://doaj.org/article/d36ebe57eec64cc4b8ff8902b462e2142009-11-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19888461/?tool=EBIhttps://doaj.org/toc/1932-6203The human GFAP splice variants GFAPDelta164 and GFAPDeltaexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's disease brains. In this study we aimed at further investigating the neuronal GFAP+1 expression and we started by affinity purifying the GFAP+1 antibody. The purified antibody resulted in a loss of neuronal GFAP+1 signal, although other antibodies directed against the amino- and carboxy-terminus of GFAPalpha still revealed GFAP-immunopositive neurons, as described before. With an in-depth analysis of a western blot, followed by mass spectrometry we discovered that the previously detected neuronal GFAP+1 expression was due to cross-reactivity of the antibody with neurofilament-L (NF-L). This was confirmed by double-label fluorescent immunohistochemistry and western blotting with the unpurified GFAP+1 antibody and an antibody against NF-L. Our data imply that NF-L can accumulate in some tangle-like structures in Alzheimer brains. More importantly, the purified GFAP+1 antibody clearly revealed a specific subtype of astrocytes in the adult human brain. These large astrocytes are present throughout the brain, e.g., along the subventricular zone, in the hippocampus, in the striatum and in the spinal cord of controls, Alzheimer, and Parkinson patients. The presence of a specific GFAP-isoform suggests a specialized function of these astrocytes.Jinte MiddeldorpSimone A van den BergeEleonora AronicaDave SpeijerElly M HolPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 11, p e7663 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jinte Middeldorp Simone A van den Berge Eleonora Aronica Dave Speijer Elly M Hol Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
description	The human GFAP splice variants GFAPDelta164 and GFAPDeltaexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's disease brains. In this study we aimed at further investigating the neuronal GFAP+1 expression and we started by affinity purifying the GFAP+1 antibody. The purified antibody resulted in a loss of neuronal GFAP+1 signal, although other antibodies directed against the amino- and carboxy-terminus of GFAPalpha still revealed GFAP-immunopositive neurons, as described before. With an in-depth analysis of a western blot, followed by mass spectrometry we discovered that the previously detected neuronal GFAP+1 expression was due to cross-reactivity of the antibody with neurofilament-L (NF-L). This was confirmed by double-label fluorescent immunohistochemistry and western blotting with the unpurified GFAP+1 antibody and an antibody against NF-L. Our data imply that NF-L can accumulate in some tangle-like structures in Alzheimer brains. More importantly, the purified GFAP+1 antibody clearly revealed a specific subtype of astrocytes in the adult human brain. These large astrocytes are present throughout the brain, e.g., along the subventricular zone, in the hippocampus, in the striatum and in the spinal cord of controls, Alzheimer, and Parkinson patients. The presence of a specific GFAP-isoform suggests a specialized function of these astrocytes.
format	article
author	Jinte Middeldorp Simone A van den Berge Eleonora Aronica Dave Speijer Elly M Hol
author_facet	Jinte Middeldorp Simone A van den Berge Eleonora Aronica Dave Speijer Elly M Hol
author_sort	Jinte Middeldorp
title	Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
title_short	Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
title_full	Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
title_fullStr	Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
title_full_unstemmed	Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
title_sort	specific human astrocyte subtype revealed by affinity purified gfap antibody; unpurified serum cross-reacts with neurofilament-l in alzheimer.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/d36ebe57eec64cc4b8ff8902b462e214
work_keys_str_mv	AT jintemiddeldorp specifichumanastrocytesubtyperevealedbyaffinitypurifiedgfapantibodyunpurifiedserumcrossreactswithneurofilamentlinalzheimer AT simoneavandenberge specifichumanastrocytesubtyperevealedbyaffinitypurifiedgfapantibodyunpurifiedserumcrossreactswithneurofilamentlinalzheimer AT eleonoraaronica specifichumanastrocytesubtyperevealedbyaffinitypurifiedgfapantibodyunpurifiedserumcrossreactswithneurofilamentlinalzheimer AT davespeijer specifichumanastrocytesubtyperevealedbyaffinitypurifiedgfapantibodyunpurifiedserumcrossreactswithneurofilamentlinalzheimer AT ellymhol specifichumanastrocytesubtyperevealedbyaffinitypurifiedgfapantibodyunpurifiedserumcrossreactswithneurofilamentlinalzheimer
_version_	1718413711802105856

Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.

Ejemplares similares