Evidence map›Paper›PMID 38981474›Full record

ArticleCell reports methods2024

Antibody-assisted selective isolation of Purkinje cell nuclei from mouse cerebellar tissue.

Luke C Bartelt, Mouad Fakhri, Grazyna Adamek, Magdalena Trybus, Anna Samelak-Czajka, Paulina Jackowiak, Agnieszka Fiszer, Craig B Lowe, Albert R La Spada, Pawel M Switonski

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Cell reports methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Luke C BarteltUniversity Program in Genetics & Genomics, Duke University Medical Center, Durham, NC 27710, USA; Departments of Pathology & Laboratory Medicine, Neurology, Biological Chemistry, and Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
Mouad FakhriDepartment of Neuronal Cell Biology, Institute of Bioorganic Chemistry, Polish Academy of Sciences, 61-704 Poznan, Poland.
Grazyna AdamekDepartment of Neuronal Cell Biology, Institute of Bioorganic Chemistry, Polish Academy of Sciences, 61-704 Poznan, Poland.
Magdalena TrybusLaboratory of Single Cell Analyses, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Anna Samelak-CzajkaLaboratory of Single Cell Analyses, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Paulina JackowiakLaboratory of Single Cell Analyses, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Agnieszka FiszerDepartment of Medical Biotechnology, Institute of Bioorganic Chemistry, Polish Academy of Sciences, 61-704 Poznan, Poland.
Craig B LoweDepartment of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
Albert R La SpadaDepartments of Pathology & Laboratory Medicine, Neurology, Biological Chemistry, and Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Department of Neurology, Duke University School of Medicine, Durham, NC 27710, USA; UCI Center for Neurotherapeutics, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: alaspada@uci.edu.
Pawel M SwitonskiDepartment of Neuronal Cell Biology, Institute of Bioorganic Chemistry, Polish Academy of Sciences, 61-704 Poznan, Poland. Electronic address: pswiton@ibch.poznan.pl.

Funding

SCA7 RETINAL DEGENERATION: MOLECULAR BASIS &THERAPYR01EY014061 · NEI · UNIVERSITY OF WASHINGTON · PI ALBERT R LA SPADA · 2002 to 2026
$8.4M
La Spada Outstanding Investigator AwardR35NS122140 · NINDS · UNIVERSITY OF CALIFORNIA-IRVINE · PI ALBERT R LA SPADA · 2021 to 2026
$7.2M
From a Long List to Causal Variants: High-Throughput Gene Regulatory Assays in Developing TissuesR35HG011332 · NHGRI · DUKE UNIVERSITY · PI LOWE, CRAIG BARRETT · 2020 to 2024
$2.3M
NEI NIH HHS R01 EY014061NHGRI NIH HHS R35 HG011332NINDS NIH HHS R35 NS122140
6 · The paper itself

Abstract

We developed a method that utilizes fluorescent labeling of nuclear envelopes alongside cytometry sorting for the selective isolation of Purkinje cell (PC) nuclei. Beginning with SUN1 reporter mice, we GFP-tagged envelopes to confirm that PC nuclei could be accurately separated from other cell types. We then developed an antibody-based protocol to make PC nuclear isolation more robust and adaptable to cerebellar tissues of any genotypic background. Immunofluorescent labeling of the nuclear membrane protein RanBP2 enabled the isolation of PC nuclei from C57BL/6 cerebellum. By analyzing the expression of PC markers, nuclear size, and nucleoli number, we confirmed that our method delivers a pure fraction of PC nuclei. To demonstrate its applicability, we isolated PC nuclei from spinocerebellar ataxia type 7 (SCA7) mice and identified transcriptional changes in known and new disease-associated genes. Access to pure PC nuclei offers insights into PC biology and pathology, including the nature of selective neuronal vulnerability.

Indexed as

Mice, Inbred C57BLPurkinje CellsAnimalsAntibodiesCell NucleusCerebellumGTP-Binding ProteinsMiceSerine-Type D-Ala-D-Ala CarboxypeptidaseAgtpbp1 protein, mouseAntibodiesGTP-Binding ProteinsSerine-Type D-Ala-D-Ala CarboxypeptidasecerebellumCP: NeuroscienceFANSisolationneurodegenerationnucleusphosphodiesterasePurkinje cellsRanBP2SCA7spinocerebellar ataxia

Identifiers

PMID38981474
PMCPMC11294835

What OpenQuestion holds

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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.