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Further characterization of autoantibodies to GABAergic neurons in the central nervous system produced by a subset of children with autism.

Wills S, Rossi CC, Bennett J, Martinez Cerdeño V, Ashwood P, Amaral DG, Van de Water J - Mol Autism (2011)

Bottom Line: Autoantibody-positive cells rarely expressed calretinin.Some cell populations stained in the primate (such as the Golgi neurons in the cerebellum) were not as robustly immunoreactive in the mouse brain.Further, these findings confirm the autoantibody-targeted cells to be a subpopulation of GABAergic interneurons.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 451 Health Sciences Drive, Suite 6510 GBSF, Davis, CA 95616, USA. javandewater@ucdavis.edu.

ABSTRACT

Background: Autism is a neurodevelopmental disorder characterized by impairments in social interaction and deficits in verbal and nonverbal communication, together with the presence of repetitive behaviors or a limited repertoire of activities and interests. The causes of autism are currently unclear. In a previous study, we determined that 21% of children with autism have plasma autoantibodies that are immunoreactive with a population of neurons in the cerebellum that appear to be Golgi cells, which are GABAergic interneurons.

Methods: We have extended this analysis by examining plasma immunoreactivity in the remainder of the brain. To determine cell specificity, double-labeling studies that included one of the calcium-binding proteins that are commonly colocalized in GABAergic neurons (calbindin, parvalbumin or calretinin) were also carried out to determine which GABAergic neurons are immunoreactive. Coronal sections through the rostrocaudal extent of the macaque monkey brain were reacted with plasma from each of seven individuals with autism who had previously demonstrated positive Golgi cell staining, as well as six negative controls. In addition, brain sections from adult male mice were similarly examined.

Results: In each case, specific staining was observed for neurons that had the morphological appearance of interneurons. By double-labeling sections with plasma and with antibodies directed against γ-aminobutyric acid (GABA), we determined that all autoantibody-positive neurons were GABAergic. However, not all GABAergic neurons were autoantibody-positive. Calbindin was colabeled in several of the autoantibody-labeled cells, while parvalbumin colabeling was less frequently observed. Autoantibody-positive cells rarely expressed calretinin. Sections from the mouse brain processed similarly to the primate sections also demonstrated immunoreactivity to interneurons distributed throughout the neocortex and many subcortical regions. Some cell populations stained in the primate (such as the Golgi neurons in the cerebellum) were not as robustly immunoreactive in the mouse brain.

Conclusions: These results suggest that the earlier report of autoantibody immunoreactivity to specific cells in the cerebellum extend to other regions of the brain. Further, these findings confirm the autoantibody-targeted cells to be a subpopulation of GABAergic interneurons. The potential impact of these autoantibodies on GABAergic disruption with respect to the etiology of autism is discussed herein.

No MeSH data available.


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Photomicrographs of the CA3 region of the hippocampus and the dentate gyrus of the macaque monkey. (A) Nissl-stained section showing the major layers of the CA3 region of the hippocampus. Deep to superficial layers include so, stratum oriens; pcl, pyramidal cell layer; sl, stratum lucidum, sr, stratum radiatum, sl-m, stratum lacunosum-moleculare. (B) Section of the CA3 region of the hippocampus stained immunohistochemically with plasma from a representative AU subject. Numerous labeled neurons are observed in the stratum radiatum (black arrows) and the adjacent stratum lacunosum-moleculare (open arrows). Only background level staining is observed in the pyramidal cell layer, but there are a variety of labeled neurons in stratum oriens (white arrows). (C) Nissl-stained section showing the layers of the dentate gyrus. Layers include pl, polymorphic layer; gcl, granule cell layer; and ml, molecular layer. (D) Section of the dentate gyrus stained immunohistochemically with plasma from a child with autism. Numerous labeled neurons (black arrows) are shown in the polymorphic cell layer, whereas fewer positive cells (open arrows) are shown in the molecular layer. Calibration bars, 100 μm. The bar in B applies to A, and the bar in D applies to C.
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Figure 6: Photomicrographs of the CA3 region of the hippocampus and the dentate gyrus of the macaque monkey. (A) Nissl-stained section showing the major layers of the CA3 region of the hippocampus. Deep to superficial layers include so, stratum oriens; pcl, pyramidal cell layer; sl, stratum lucidum, sr, stratum radiatum, sl-m, stratum lacunosum-moleculare. (B) Section of the CA3 region of the hippocampus stained immunohistochemically with plasma from a representative AU subject. Numerous labeled neurons are observed in the stratum radiatum (black arrows) and the adjacent stratum lacunosum-moleculare (open arrows). Only background level staining is observed in the pyramidal cell layer, but there are a variety of labeled neurons in stratum oriens (white arrows). (C) Nissl-stained section showing the layers of the dentate gyrus. Layers include pl, polymorphic layer; gcl, granule cell layer; and ml, molecular layer. (D) Section of the dentate gyrus stained immunohistochemically with plasma from a child with autism. Numerous labeled neurons (black arrows) are shown in the polymorphic cell layer, whereas fewer positive cells (open arrows) are shown in the molecular layer. Calibration bars, 100 μm. The bar in B applies to A, and the bar in D applies to C.

Mentions: As illustrated in Figure 6, there are populations of plasma-labeled cells in the hippocampus and dentate gyrus. We have previously evaluated the distribution of GABAergic neurons in the macaque monkey hippocampal formation [36,37]. The population of autoantibody-positive cells in the hippocampal formation was morphologically heterogeneous. In the CA3 region of the hippocampus (Figures 6A and 6B), labeled neurons were located mainly in the stratum oriens, stratum radiatum and stratum lacunosum-moleculare. Relatively few labeled neurons were observed in the pyramidal cell layer or the stratum lucidum. The dendrites of labeled neurons were well-stained and often had a horizontal orientation, particularly in stratum oriens and stratum lacunosum-moleculare. In the dentate gyrus (Figures 6C and 6D), the majority of labeled cells were located in the polymorphic layer. Again, they had variable morphologies. Some appeared to be typical dentate basket cells, but others had dendrites that were mainly oriented parallel to the granule cell layer. There were a few small, round, labeled cells in the molecular layer of the dentate gyrus.


Further characterization of autoantibodies to GABAergic neurons in the central nervous system produced by a subset of children with autism.

Wills S, Rossi CC, Bennett J, Martinez Cerdeño V, Ashwood P, Amaral DG, Van de Water J - Mol Autism (2011)

Photomicrographs of the CA3 region of the hippocampus and the dentate gyrus of the macaque monkey. (A) Nissl-stained section showing the major layers of the CA3 region of the hippocampus. Deep to superficial layers include so, stratum oriens; pcl, pyramidal cell layer; sl, stratum lucidum, sr, stratum radiatum, sl-m, stratum lacunosum-moleculare. (B) Section of the CA3 region of the hippocampus stained immunohistochemically with plasma from a representative AU subject. Numerous labeled neurons are observed in the stratum radiatum (black arrows) and the adjacent stratum lacunosum-moleculare (open arrows). Only background level staining is observed in the pyramidal cell layer, but there are a variety of labeled neurons in stratum oriens (white arrows). (C) Nissl-stained section showing the layers of the dentate gyrus. Layers include pl, polymorphic layer; gcl, granule cell layer; and ml, molecular layer. (D) Section of the dentate gyrus stained immunohistochemically with plasma from a child with autism. Numerous labeled neurons (black arrows) are shown in the polymorphic cell layer, whereas fewer positive cells (open arrows) are shown in the molecular layer. Calibration bars, 100 μm. The bar in B applies to A, and the bar in D applies to C.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Photomicrographs of the CA3 region of the hippocampus and the dentate gyrus of the macaque monkey. (A) Nissl-stained section showing the major layers of the CA3 region of the hippocampus. Deep to superficial layers include so, stratum oriens; pcl, pyramidal cell layer; sl, stratum lucidum, sr, stratum radiatum, sl-m, stratum lacunosum-moleculare. (B) Section of the CA3 region of the hippocampus stained immunohistochemically with plasma from a representative AU subject. Numerous labeled neurons are observed in the stratum radiatum (black arrows) and the adjacent stratum lacunosum-moleculare (open arrows). Only background level staining is observed in the pyramidal cell layer, but there are a variety of labeled neurons in stratum oriens (white arrows). (C) Nissl-stained section showing the layers of the dentate gyrus. Layers include pl, polymorphic layer; gcl, granule cell layer; and ml, molecular layer. (D) Section of the dentate gyrus stained immunohistochemically with plasma from a child with autism. Numerous labeled neurons (black arrows) are shown in the polymorphic cell layer, whereas fewer positive cells (open arrows) are shown in the molecular layer. Calibration bars, 100 μm. The bar in B applies to A, and the bar in D applies to C.
Mentions: As illustrated in Figure 6, there are populations of plasma-labeled cells in the hippocampus and dentate gyrus. We have previously evaluated the distribution of GABAergic neurons in the macaque monkey hippocampal formation [36,37]. The population of autoantibody-positive cells in the hippocampal formation was morphologically heterogeneous. In the CA3 region of the hippocampus (Figures 6A and 6B), labeled neurons were located mainly in the stratum oriens, stratum radiatum and stratum lacunosum-moleculare. Relatively few labeled neurons were observed in the pyramidal cell layer or the stratum lucidum. The dendrites of labeled neurons were well-stained and often had a horizontal orientation, particularly in stratum oriens and stratum lacunosum-moleculare. In the dentate gyrus (Figures 6C and 6D), the majority of labeled cells were located in the polymorphic layer. Again, they had variable morphologies. Some appeared to be typical dentate basket cells, but others had dendrites that were mainly oriented parallel to the granule cell layer. There were a few small, round, labeled cells in the molecular layer of the dentate gyrus.

Bottom Line: Autoantibody-positive cells rarely expressed calretinin.Some cell populations stained in the primate (such as the Golgi neurons in the cerebellum) were not as robustly immunoreactive in the mouse brain.Further, these findings confirm the autoantibody-targeted cells to be a subpopulation of GABAergic interneurons.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 451 Health Sciences Drive, Suite 6510 GBSF, Davis, CA 95616, USA. javandewater@ucdavis.edu.

ABSTRACT

Background: Autism is a neurodevelopmental disorder characterized by impairments in social interaction and deficits in verbal and nonverbal communication, together with the presence of repetitive behaviors or a limited repertoire of activities and interests. The causes of autism are currently unclear. In a previous study, we determined that 21% of children with autism have plasma autoantibodies that are immunoreactive with a population of neurons in the cerebellum that appear to be Golgi cells, which are GABAergic interneurons.

Methods: We have extended this analysis by examining plasma immunoreactivity in the remainder of the brain. To determine cell specificity, double-labeling studies that included one of the calcium-binding proteins that are commonly colocalized in GABAergic neurons (calbindin, parvalbumin or calretinin) were also carried out to determine which GABAergic neurons are immunoreactive. Coronal sections through the rostrocaudal extent of the macaque monkey brain were reacted with plasma from each of seven individuals with autism who had previously demonstrated positive Golgi cell staining, as well as six negative controls. In addition, brain sections from adult male mice were similarly examined.

Results: In each case, specific staining was observed for neurons that had the morphological appearance of interneurons. By double-labeling sections with plasma and with antibodies directed against γ-aminobutyric acid (GABA), we determined that all autoantibody-positive neurons were GABAergic. However, not all GABAergic neurons were autoantibody-positive. Calbindin was colabeled in several of the autoantibody-labeled cells, while parvalbumin colabeling was less frequently observed. Autoantibody-positive cells rarely expressed calretinin. Sections from the mouse brain processed similarly to the primate sections also demonstrated immunoreactivity to interneurons distributed throughout the neocortex and many subcortical regions. Some cell populations stained in the primate (such as the Golgi neurons in the cerebellum) were not as robustly immunoreactive in the mouse brain.

Conclusions: These results suggest that the earlier report of autoantibody immunoreactivity to specific cells in the cerebellum extend to other regions of the brain. Further, these findings confirm the autoantibody-targeted cells to be a subpopulation of GABAergic interneurons. The potential impact of these autoantibodies on GABAergic disruption with respect to the etiology of autism is discussed herein.

No MeSH data available.


Related in: MedlinePlus