Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis
Abstract In Alzheimer’s disease (AD), microglia are affected by disease processes, but may also drive pathogenesis. AD pathology-associated microglial populations have been identified with single-cell RNA-Seq, but have not been validated in human brain tissue with anatomical context. Here, we quanti...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-10-01
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| Series: | Acta Neuropathologica Communications |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40478-020-01047-9 |
| _version_ | 1818976718887583744 |
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| author | Molly E. V. Swanson Emma L. Scotter Leon C. D. Smyth Helen C. Murray Brigid Ryan Clinton Turner Richard L. M. Faull Mike Dragunow Maurice A. Curtis |
| author_facet | Molly E. V. Swanson Emma L. Scotter Leon C. D. Smyth Helen C. Murray Brigid Ryan Clinton Turner Richard L. M. Faull Mike Dragunow Maurice A. Curtis |
| author_sort | Molly E. V. Swanson |
| collection | DOAJ |
| description | Abstract In Alzheimer’s disease (AD), microglia are affected by disease processes, but may also drive pathogenesis. AD pathology-associated microglial populations have been identified with single-cell RNA-Seq, but have not been validated in human brain tissue with anatomical context. Here, we quantified myeloid cell markers to identify changes in AD pathology-associated microglial populations. We performed fluorescent immunohistochemistry on normal (n = 8) and AD (n = 8) middle temporal gyri, co-labelling the pan-myeloid cell marker, Iba1, with one of 11 markers of interest (MOIs): CD45, HLA-DR, CD14, CD74, CD33, CD206, CD32, CD163, P2RY12, TMEM119, L-Ferritin. Novel image analyses quantified the single-cell abundance of Iba1 and each MOI. Each cell was gated into one Iba1-MOI population (Iba1low MOIhigh, Iba1high MOIhigh, or Iba1high MOIlow) and the abundance of each population was compared between AD and control. Triple-labelling of L-Ferritin and Iba1 with a subset of MOIs was performed to investigate L-Ferritin-MOI co-expression on Iba1low cells. Iba1low MOIhigh myeloid cell populations delineated by MOIs CD45, HLA-DR, CD14, CD74, CD33, CD32, and L-Ferritin were increased in AD. Further investigation of the Iba1low MOIhigh populations revealed that their abundances correlated with tau, but not amyloid beta, load in AD. The Iba1low microglial population highly expressed L-Ferritin, reflecting microglial dysfunction. The L-Ferritinhigh CD74high HLA-DRhigh phenotype of the Iba1low population mirrors that of a human AD pathology-associated microglial subpopulation previously identified using single-cell RNA-Seq. Our high-throughput immunohistochemical data with anatomical context support the microglial dysfunction hypothesis of AD. |
| first_indexed | 2024-12-20T16:16:18Z |
| format | Article |
| id | doaj.art-fd4bdb9a974b40289fbe32b437cd3db9 |
| institution | Directory Open Access Journal |
| issn | 2051-5960 |
| language | English |
| last_indexed | 2024-12-20T16:16:18Z |
| publishDate | 2020-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Acta Neuropathologica Communications |
| spelling | doaj.art-fd4bdb9a974b40289fbe32b437cd3db92022-12-21T19:33:48ZengBMCActa Neuropathologica Communications2051-59602020-10-018111610.1186/s40478-020-01047-9Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysisMolly E. V. Swanson0Emma L. Scotter1Leon C. D. Smyth2Helen C. Murray3Brigid Ryan4Clinton Turner5Richard L. M. Faull6Mike Dragunow7Maurice A. Curtis8Department of Anatomy and Medical Imaging, Faculty of Medical and Health Science, University of AucklandCentre for Brain Research, Faculty of Medical and Health Science, University of AucklandCentre for Brain Research, Faculty of Medical and Health Science, University of AucklandDepartment of Anatomy and Medical Imaging, Faculty of Medical and Health Science, University of AucklandDepartment of Anatomy and Medical Imaging, Faculty of Medical and Health Science, University of AucklandDepartment of Anatomical Pathology, LabPlus, Auckland City HospitalDepartment of Anatomy and Medical Imaging, Faculty of Medical and Health Science, University of AucklandCentre for Brain Research, Faculty of Medical and Health Science, University of AucklandDepartment of Anatomy and Medical Imaging, Faculty of Medical and Health Science, University of AucklandAbstract In Alzheimer’s disease (AD), microglia are affected by disease processes, but may also drive pathogenesis. AD pathology-associated microglial populations have been identified with single-cell RNA-Seq, but have not been validated in human brain tissue with anatomical context. Here, we quantified myeloid cell markers to identify changes in AD pathology-associated microglial populations. We performed fluorescent immunohistochemistry on normal (n = 8) and AD (n = 8) middle temporal gyri, co-labelling the pan-myeloid cell marker, Iba1, with one of 11 markers of interest (MOIs): CD45, HLA-DR, CD14, CD74, CD33, CD206, CD32, CD163, P2RY12, TMEM119, L-Ferritin. Novel image analyses quantified the single-cell abundance of Iba1 and each MOI. Each cell was gated into one Iba1-MOI population (Iba1low MOIhigh, Iba1high MOIhigh, or Iba1high MOIlow) and the abundance of each population was compared between AD and control. Triple-labelling of L-Ferritin and Iba1 with a subset of MOIs was performed to investigate L-Ferritin-MOI co-expression on Iba1low cells. Iba1low MOIhigh myeloid cell populations delineated by MOIs CD45, HLA-DR, CD14, CD74, CD33, CD32, and L-Ferritin were increased in AD. Further investigation of the Iba1low MOIhigh populations revealed that their abundances correlated with tau, but not amyloid beta, load in AD. The Iba1low microglial population highly expressed L-Ferritin, reflecting microglial dysfunction. The L-Ferritinhigh CD74high HLA-DRhigh phenotype of the Iba1low population mirrors that of a human AD pathology-associated microglial subpopulation previously identified using single-cell RNA-Seq. Our high-throughput immunohistochemical data with anatomical context support the microglial dysfunction hypothesis of AD.http://link.springer.com/article/10.1186/s40478-020-01047-9MicrogliaAlzheimer’s diseaseTauAmyloid betaDysfunctionImmunohistochemistry |
| spellingShingle | Molly E. V. Swanson Emma L. Scotter Leon C. D. Smyth Helen C. Murray Brigid Ryan Clinton Turner Richard L. M. Faull Mike Dragunow Maurice A. Curtis Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis Acta Neuropathologica Communications Microglia Alzheimer’s disease Tau Amyloid beta Dysfunction Immunohistochemistry |
| title | Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis |
| title_full | Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis |
| title_fullStr | Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis |
| title_full_unstemmed | Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis |
| title_short | Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis |
| title_sort | identification of a dysfunctional microglial population in human alzheimer s disease cortex using novel single cell histology image analysis |
| topic | Microglia Alzheimer’s disease Tau Amyloid beta Dysfunction Immunohistochemistry |
| url | http://link.springer.com/article/10.1186/s40478-020-01047-9 |
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