dc.contributor.author |
Highet, Blake |
|
dc.contributor.author |
Vikas Anekal, Praju |
|
dc.contributor.author |
Ryan, Brigid |
|
dc.contributor.author |
Murray, Helen |
|
dc.contributor.author |
Coppieters, Natacha |
|
dc.contributor.author |
Victor Dieriks, Birger |
|
dc.contributor.author |
Singh-Bains, Malvindar K |
|
dc.contributor.author |
Mehrabi, Nasim F |
|
dc.contributor.author |
Faull, Richard LM |
|
dc.contributor.author |
Dragunow, Michael |
|
dc.contributor.author |
Curtis, Maurice A |
|
dc.coverage.spatial |
England |
|
dc.date.accessioned |
2021-12-13T01:38:25Z |
|
dc.date.available |
2021-12-13T01:38:25Z |
|
dc.date.issued |
2021-5 |
|
dc.identifier.citation |
Journal of neurochemistry 157(4):1270-1283 May 2021 |
|
dc.identifier.issn |
0022-3042 |
|
dc.identifier.uri |
https://hdl.handle.net/2292/57752 |
|
dc.description.abstract |
In situ hybridization (ISH) is a powerful tool that can be used to localize mRNA expression in tissue samples. Combining ISH with immunohistochemistry (IHC) to determine cell type provides cellular context of mRNA expression, which cannot be achieved with gene microarray or polymerase chain reaction. To study mRNA and protein expression on the same section we investigated the use of RNAscope® ISH in combination with fluorescent IHC on paraffin-embedded human brain tissue. We first developed a high-throughput, automated image analysis workflow for quantifying RNA puncta across the total cell population and within neurons identified by NeuN<sup>+</sup> immunoreactivity. We then applied this automated analysis to tissue microarray (TMA) sections of middle temporal gyrus tissue (MTG) from neurologically normal and Alzheimer's Disease (AD) cases to determine the suitability of three commonly used housekeeping genes: ubiquitin C (UBC), peptidyl-prolyl cis-trans isomerase B (PPIB) and DNA-directed RNA polymerase II subunit RPB1 (POLR2A). Overall, we saw a significant decrease in total and neuronal UBC expression in AD cases compared to normal cases. Total expression results were validated with RT-qPCR using fresh frozen tissue from 5 normal and 5 AD cases. We conclude that this technique combined with our novel automated analysis pipeline provides a suitable platform to study changes in gene expression in diseased human brain tissue with cellular and anatomical context. Furthermore, our results suggest that UBC is not a suitable housekeeping gene in the study of post-mortem AD brain tissue. |
|
dc.format.medium |
Print-Electronic |
|
dc.language |
eng |
|
dc.publisher |
Wiley |
|
dc.relation.ispartofseries |
Journal of neurochemistry |
|
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
|
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
|
dc.rights.uri |
https://authorservices.wiley.com/author-resources/Journal-Authors/licensing/self-archiving.html |
|
dc.subject |
Humans |
|
dc.subject |
Alzheimer Disease |
|
dc.subject |
Cyclophilins |
|
dc.subject |
DNA-Directed RNA Polymerases |
|
dc.subject |
Ubiquitin C |
|
dc.subject |
RNA, Messenger |
|
dc.subject |
Immunohistochemistry |
|
dc.subject |
In Situ Hybridization, Fluorescence |
|
dc.subject |
Gene Expression Profiling |
|
dc.subject |
Genes, Essential |
|
dc.subject |
Aged |
|
dc.subject |
Aged, 80 and over |
|
dc.subject |
Middle Aged |
|
dc.subject |
Female |
|
dc.subject |
Male |
|
dc.subject |
High-Throughput Screening Assays |
|
dc.subject |
Workflow |
|
dc.subject |
Transcriptome |
|
dc.subject |
Alzheimer's Disease |
|
dc.subject |
automated analysis workflow |
|
dc.subject |
housekeeping genes |
|
dc.subject |
immunohistochemistry |
|
dc.subject |
in situ hybridization |
|
dc.subject |
ubiquitin C (UBC) |
|
dc.subject |
Aged |
|
dc.subject |
Aged, 80 and over |
|
dc.subject |
Alzheimer Disease |
|
dc.subject |
Cyclophilins |
|
dc.subject |
DNA-Directed RNA Polymerases |
|
dc.subject |
Female |
|
dc.subject |
Gene Expression Profiling |
|
dc.subject |
Genes, Essential |
|
dc.subject |
High-Throughput Screening Assays |
|
dc.subject |
Humans |
|
dc.subject |
Immunohistochemistry |
|
dc.subject |
In Situ Hybridization, Fluorescence |
|
dc.subject |
Male |
|
dc.subject |
Middle Aged |
|
dc.subject |
RNA, Messenger |
|
dc.subject |
Transcriptome |
|
dc.subject |
Ubiquitin C |
|
dc.subject |
Workflow |
|
dc.subject |
Science & Technology |
|
dc.subject |
Life Sciences & Biomedicine |
|
dc.subject |
Biochemistry & Molecular Biology |
|
dc.subject |
Neurosciences |
|
dc.subject |
Neurosciences & Neurology |
|
dc.subject |
Alzheimer's Disease |
|
dc.subject |
automated analysis workflow |
|
dc.subject |
housekeeping genes |
|
dc.subject |
immunohistochemistry |
|
dc.subject |
in situ hybridization |
|
dc.subject |
ubiquitin C (UBC) |
|
dc.subject |
0601 Biochemistry and Cell Biology |
|
dc.subject |
1109 Neurosciences |
|
dc.title |
fISHing with immunohistochemistry for housekeeping gene changes in Alzheimer's disease using an automated quantitative analysis workflow. |
|
dc.type |
Journal Article |
|
dc.identifier.doi |
10.1111/jnc.15283 |
|
pubs.issue |
4 |
|
pubs.begin-page |
1270 |
|
pubs.volume |
157 |
|
dc.date.updated |
2021-11-18T00:00:58Z |
|
dc.rights.holder |
Copyright: International Society for Neurochemistry |
en |
pubs.author-url |
https://www.ncbi.nlm.nih.gov/pubmed/33368239 |
|
pubs.end-page |
1283 |
|
pubs.publication-status |
Published |
|
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
Research Support, Non-U.S. Gov't |
|
pubs.subtype |
Journal Article |
|
pubs.elements-id |
833868 |
|
dc.identifier.eissn |
1471-4159 |
|
pubs.online-publication-date |
2021-2-2 |
|