1887

Abstract

In Parkinson’s disease (PD), α-synuclein is a key protein in the process of neurodegeneration. Besides motor symptoms, most PD patients additionally suffer from gastrointestinal tract (GIT) dysfunctions, even several years before the onset of motor disabilities. Studies have reported a dysbiosis of gut bacteria in PD patients compared to healthy controls and have suggested that the enteric nervous system (ENS) can be involved in the development of the disease. As α-synuclein was found to be secreted by neurons of the ENS, we used RNA-based stable isotope probing (RNA-SIP) to identify gut bacteria that might be able to assimilate this protein. The gut contents of 24 mice were pooled and incubated with isotopically labelled (C) and unlabelled (C) α-synuclein. After incubation for 0, 4 and 24 h, RNA was extracted from the incubations and separated by density gradient centrifugation. However, RNA quantification of density-resolved fractions revealed no incorporation of the C isotope into the extracted RNA, suggesting that α-synuclein was not assimilated by the murine gut bacteria. Potential reasons and consequences for follow-up-studies are discussed.

Funding
This study was supported by the:
  • Bundesministerium für Bildung und Forschung (Award 03FH036PB5)
    • Principle Award Recipient: SeverinWeis
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/acmi/10.1099/acmi.0.000345
2022-05-04
2024-05-14
Loading full text...

Full text loading...

/deliver/fulltext/acmi/4/5/acmi000345.html?itemId=/content/journal/acmi/10.1099/acmi.0.000345&mimeType=html&fmt=ahah

References

  1. Perez-Pardo P, Kliest T, Dodiya HB, Broersen LM, Garssen J et al. The gut-brain axis in Parkinson’s disease: Possibilities for food-based therapies. Eur J Pharmacol 2017; 817:86–95 [View Article] [PubMed]
    [Google Scholar]
  2. Pfeiffer RF. Gastrointestinal dysfunction in Parkinson’s Disease. Curr Treat Options Neurol 2018; 20:54 [View Article] [PubMed]
    [Google Scholar]
  3. Lang AE. A critical appraisal of the premotor symptoms of Parkinson’s disease: potential usefulness in early diagnosis and design of neuroprotective trials. Mov Disord 2011; 26:775–783 [View Article] [PubMed]
    [Google Scholar]
  4. Savica R, Carlin JM, Grossardt BR, Bower JH, Ahlskog JE et al. Medical records documentation of constipation preceding Parkinson disease: A case-control study. Neurology 2009; 73:1752–1758 [View Article] [PubMed]
    [Google Scholar]
  5. Nair AT, Ramachandran V, Joghee NM, Antony S, Ramalingam G. Gut microbiota dysfunction as reliable non-invasive early diagnostic biomarkers in the pathophysiology of Parkinson’s disease: a critical review. J Neurogastroenterol Motil 2018; 24:30–42 [View Article] [PubMed]
    [Google Scholar]
  6. Rocha EM, De Miranda B, Sanders LH. Alpha-synuclein: Pathology, mitochondrial dysfunction and neuroinflammation in Parkinson’s disease. Neurobiol Dis 2018; 109:249–257 [View Article] [PubMed]
    [Google Scholar]
  7. Houser MC, Tansey MG. The gut-brain axis: is intestinal inflammation a silent driver of Parkinson’s disease pathogenesis?. NPJ Parkinsons Dis 2017; 3:3 [View Article] [PubMed]
    [Google Scholar]
  8. Gold A, Turkalp ZT, Munoz DG. Enteric alpha-synuclein expression is increased in Parkinson’s disease but not Alzheimer’s disease. Mov Disord 2013; 28:237–240 [View Article] [PubMed]
    [Google Scholar]
  9. Shannon KM, Keshavarzian A, Dodiya HB, Jakate S, Kordower JH. Is alpha-synuclein in the colon a biomarker for premotor Parkinson’s disease? Evidence from 3 cases. Mov Disord 2012; 27:716–719 [View Article] [PubMed]
    [Google Scholar]
  10. Hilton D, Stephens M, Kirk L, Edwards P, Potter R et al. Accumulation of α-synuclein in the bowel of patients in the pre-clinical phase of Parkinson’s disease. Acta Neuropathol 2014; 127:235–241 [View Article] [PubMed]
    [Google Scholar]
  11. Braak H, de Vos RAI, Bohl J, Del Tredici K. Gastric alpha-synuclein immunoreactive inclusions in Meissner’s and Auerbach’s plexuses in cases staged for Parkinson’s disease-related brain pathology. Neurosci Lett 2006; 396:67–72 [View Article] [PubMed]
    [Google Scholar]
  12. Gries M, Christmann A, Schulte S, Weyland M, Rommel S et al. Parkinson mice show functional and molecular changes in the gut long before motoric disease onset. Mol Neurodegener 2021; 16:34 [View Article] [PubMed]
    [Google Scholar]
  13. Paillusson S, Clairembault T, Biraud M, Neunlist M, Derkinderen P. Activity-dependent secretion of alpha-synuclein by enteric neurons. J Neurochem 2013; 125:512–517 [View Article] [PubMed]
    [Google Scholar]
  14. Weis S, Schwiertz A, Unger MM, Becker A, Faßbender K et al. Effect of Parkinson’s disease and related medications on the composition of the fecal bacterial microbiota. NPJ Parkinsons Dis 2019; 5:28 [View Article] [PubMed]
    [Google Scholar]
  15. Unger MM, Spiegel J, Dillmann K-U, Grundmann D, Philippeit H et al. Short chain fatty acids and gut microbiota differ between patients with Parkinson’s disease and age-matched controls. Parkinsonism Relat Disord 2016; 32:66–72 [View Article] [PubMed]
    [Google Scholar]
  16. Hill-Burns EM, Debelius JW, Morton JT, Wissemann WT, Lewis MR et al. Parkinson’s disease and Parkinson’s disease medications have distinct signatures of the gut microbiome. Mov Disord 2017; 32:739–749 [View Article] [PubMed]
    [Google Scholar]
  17. Scheperjans F, Aho V, Pereira PAB, Koskinen K, Paulin L et al. Gut microbiota are related to Parkinson’s disease and clinical phenotype. Mov Disord 2015; 30:350–358 [View Article] [PubMed]
    [Google Scholar]
  18. Li W, Wu X, Hu X, Wang T, Liang S et al. Structural changes of gut microbiota in Parkinson’s disease and its correlation with clinical features. Sci China Life Sci 2017; 60:1223–1233 [View Article] [PubMed]
    [Google Scholar]
  19. Hopfner F, Künstner A, Müller SH, Künzel S, Zeuner KE et al. Gut microbiota in Parkinson disease in a northern German cohort. Brain Res 2017; 1667:41–45 [View Article] [PubMed]
    [Google Scholar]
  20. Berry D, Loy A. Stable-isotope probing of human and animal microbiome function. Trends Microbiol 2018; 26:999–1007 [View Article] [PubMed]
    [Google Scholar]
  21. Lueders T, Dumont MG, Bradford L, Manefield M. RNA-stable isotope probing: from carbon flow within key microbiota to targeted transcriptomes. Curr Opin Biotechnol 2016; 41:83–89 [View Article]
    [Google Scholar]
  22. Weis S, Schnell S, Egert M. RNA-based stable isotope probing (RNA-SIP) in the gut environment. Methods Mol Biol 2019; 2046:221–231 [View Article]
    [Google Scholar]
  23. Egert M, Weis S, Schnell S. RNA-based stable isotope probing (RNA-SIP) to unravel intestinal host-microbe interactions. Methods 2018; 149:25–30 [View Article]
    [Google Scholar]
  24. Smith HO, Levine M. Two sequential repressions of DNA synthesis in the establishment of lysogeny by phage P22 and its mutants. Proc Natl Acad Sci U S A 1964; 52:356–363 [View Article]
    [Google Scholar]
  25. Herrmann E, Young W, Rosendale D, Conrad R, Riedel CU et al. Determination of resistant starch assimilating bacteria in fecal samples of mice by in vitro RNA-based stable isotope probing. Front Microbiol 2017; 8:1331 [View Article]
    [Google Scholar]
  26. Lane DJ. 16S/23S rRNA sequencing. In Stackebrandt E, Goodfellow M. eds Molecular Microbiological Methods: Nucleic Acid Techniques in Bacterial Systematics New York: Wiley; 1991 pp 115–175
    [Google Scholar]
  27. Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML et al. Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci U S A 1985; 82:6955–6959 [View Article] [PubMed]
    [Google Scholar]
  28. Herrmann E, Young W, Rosendale D, Reichert-Grimm V, Riedel CU et al. RNA-based stable isotope probing suggests Allobaculum spp. as particularly active glucose assimilators in a complex murine microbiota cultured in vitro. Biomed Res Int 2017; 2017:1829685 [View Article]
    [Google Scholar]
  29. Young W, Egert M, Bassett SA, Bibiloni R. Detection of sialic acid-utilising bacteria in a caecal community batch culture using RNA-based stable isotope probing. Nutrients 2015; 7:2109–2124 [View Article] [PubMed]
    [Google Scholar]
  30. Romano S, Savva GM, Bedarf JR, Charles IG, Hildebrand F et al. Meta-analysis of the Parkinson’s disease gut microbiome suggests alterations linked to intestinal inflammation. NPJ Parkinsons Dis 2021; 7:27 [View Article] [PubMed]
    [Google Scholar]
  31. Egert M, de Graaf AA, Maathuis A, de Waard P, Plugge CM et al. Identification of glucose-fermenting bacteria present in an in vitro model of the human intestine by RNA-stable isotope probing. FEMS Microbiol Ecol 2007; 60:126–135 [View Article] [PubMed]
    [Google Scholar]
  32. Whiteley AS, Thomson B, Lueders T, Manefield M. RNA stable-isotope probing. Nat Protoc 2007; 2:838–844 [View Article] [PubMed]
    [Google Scholar]
  33. Herrmann E, Young W, Reichert-Grimm V, Weis S, Riedel CU et al. In vivo assessment of resistant starch degradation by the caecal microbiota of mice using RNA-based stable isotope probing-a proof-of-principle study. Nutrients 2018; 10:E179 [View Article] [PubMed]
    [Google Scholar]
  34. Enggrob KL, Larsen T, Peixoto L, Rasmussen J. Gram-positive bacteria control the rapid anabolism of protein-sized soil organic nitrogen compounds questioning the present paradigm. Sci Rep 2020; 10:15840 [View Article] [PubMed]
    [Google Scholar]
  35. Jehmlich N, Vogt C, Lünsmann V, Richnow HH, von Bergen M. Protein-SIP in environmental studies. Curr Opin Biotechnol 2016; 41:26–33 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/acmi/10.1099/acmi.0.000345
Loading
/content/journal/acmi/10.1099/acmi.0.000345
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error