title: A ratiometric two-photon fluorescent probe reveals reduction in mitochondrial H2S production in Parkinson's disease gene knockout astrocytes.
abstract: Hydrogen sulfide (H2S) is a multifunctional signaling molecule that exerts neuroprotective effects in oxidative stress. In this article, we report a mitochondria-localized two-photon probe, SHS-M2, that can be excited by 750 nm femtosecond pulses and employed for ratiometric detection of H2S in live astrocytes and living brain slices using two-photon microscopy (TPM). SHS-M2 shows bright two-photon-excited fluorescence and a marked change in emission color from blue to yellow in response to H2S, low cytotoxicity, easy loading, and minimum interference from other biologically relevant species including reactive sulfur, oxygen, and nitrogen species, thereby allowing quantitative analysis of H2S levels. Molecular TPM imaging with SHS-M2 in astrocytes revealed that there is a correlation between the ratiometric analysis and expression levels of cystathionine β-synthase (CBS), the major enzyme that catalyzes H2S production. In studies involving DJ-1, a Parkinson's disease (PD) gene, attenuated H2S production in comparison with wild-type controls was observed in DJ-1-knockout astrocytes and brain slices, where CBS expression was decreased. These findings demonstrate that reduced H2S levels in astrocytes may contribute to the development of PD and that SHS-M2 may be useful as a marker to detect a risk of neurodegenerative diseases, including PD.
authors.affiliation: Division of Energy Systems Research, Ajou University, Suwon 443-749, Korea.
authors.forename: Sung Keun
authors.initials: SK
authors.lastname: Bae
authors.forename: Cheol Ho
authors.initials: CH
authors.lastname: Heo
authors.forename: Dong Joo
authors.initials: DJ
authors.lastname: Choi
authors.forename: Debabrata
authors.initials: D
authors.lastname: Sen
authors.forename: Eun-Hye
authors.initials: EH
authors.lastname: Joe
authors.forename: Bong Rae
authors.initials: BR
authors.lastname: Cho
authors.forename: Hwan Myung
authors.initials: HM
authors.lastname: Kim
mesh_terms: D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D006862:Hydrogen Sulfide / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D015394:Molecular Structure; D015513:Oncogene Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism*; D000071617:Protein Deglycase DJ-1; D011522:Protons
chemical_list: D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D011522:Protons; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D006862:Hydrogen Sulfide