Age-related changes in the photoreactivity of retinal lipofuscin granules

Role of chloroform-insoluble components

Małgorzata Rózanowska, Anna Pawlak, Bartosz Rózanowski, Christine Skumatz, Mariusz Zarȩba, Mike E. Boulton, Janice M. Burke, Tadeusz Sarna, John D. Simon

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

PURPOSE. Lipofuscin accumulates in human retinal pigment epithelium (RPE) cells with age and may be the main factor responsible for the increasing susceptibility of RPE to photo-oxidation with age. As the composition, absorption, and fluorescence of lipofuscin undergo age-related changes, the purpose of this study was to determine whether photoreactivity of lipofuscin granules also changes with the donor age. METHODS. To determine whether the photoreactivity of lipofuscin itself is age related, lipofuscin granules were isolated from human RPE and pooled into age groups. Photoreactivity was assessed by measuring action spectra of photo-induced oxygen uptake and photogeneration of reactive oxygen species. Separation of chloroform-soluble (ChS) and -insoluble (ChNS) components by Folch's extraction was used to determine the factors responsible for the age-related increase in lipofuscin photoreactivity. RESULTS. The observed rates of photo-induced oxygen uptake and photo-induced accumulation of superoxide-derived spin adducts indicated that when normalized to equal numbers of lipofuscin granules, aerobic photoreactivity of lipofuscin increased with age. Both ChS and ChNS mediated photogeneration of singlet oxygen, superoxide radical anion, and photo-oxidation of added lipids and proteins. Although both ChS and ChNS exhibited substantial photoreactivities, neither exhibited significant age-related changes when normalized to equal dry mass. In contrast, ChNS contents in lipofuscin granules significantly increased with aging. CONCLUSIONS. Aerobic photoreactivity of RPE lipofuscin substantially increases with aging. This effect may be ascribed to the increased content of insoluble components.

Original languageEnglish (US)
Pages (from-to)1052-1060
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume45
Issue number4
DOIs
StatePublished - Apr 2004
Externally publishedYes

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Lipofuscin
Chloroform
Retinal Pigment Epithelium
Superoxides
Reactive Oxygen Species
Oxygen
Singlet Oxygen
Age Factors
Age Groups
Fluorescence
Tissue Donors
Lipids

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Rózanowska, M., Pawlak, A., Rózanowski, B., Skumatz, C., Zarȩba, M., Boulton, M. E., ... Simon, J. D. (2004). Age-related changes in the photoreactivity of retinal lipofuscin granules: Role of chloroform-insoluble components. Investigative Ophthalmology and Visual Science, 45(4), 1052-1060. https://doi.org/10.1167/iovs.03-0277

Age-related changes in the photoreactivity of retinal lipofuscin granules : Role of chloroform-insoluble components. / Rózanowska, Małgorzata; Pawlak, Anna; Rózanowski, Bartosz; Skumatz, Christine; Zarȩba, Mariusz; Boulton, Mike E.; Burke, Janice M.; Sarna, Tadeusz; Simon, John D.

In: Investigative Ophthalmology and Visual Science, Vol. 45, No. 4, 04.2004, p. 1052-1060.

Research output: Contribution to journalArticle

Rózanowska, M, Pawlak, A, Rózanowski, B, Skumatz, C, Zarȩba, M, Boulton, ME, Burke, JM, Sarna, T & Simon, JD 2004, 'Age-related changes in the photoreactivity of retinal lipofuscin granules: Role of chloroform-insoluble components', Investigative Ophthalmology and Visual Science, vol. 45, no. 4, pp. 1052-1060. https://doi.org/10.1167/iovs.03-0277
Rózanowska, Małgorzata ; Pawlak, Anna ; Rózanowski, Bartosz ; Skumatz, Christine ; Zarȩba, Mariusz ; Boulton, Mike E. ; Burke, Janice M. ; Sarna, Tadeusz ; Simon, John D. / Age-related changes in the photoreactivity of retinal lipofuscin granules : Role of chloroform-insoluble components. In: Investigative Ophthalmology and Visual Science. 2004 ; Vol. 45, No. 4. pp. 1052-1060.
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AU - Pawlak, Anna

AU - Rózanowski, Bartosz

AU - Skumatz, Christine

AU - Zarȩba, Mariusz

AU - Boulton, Mike E.

AU - Burke, Janice M.

AU - Sarna, Tadeusz

AU - Simon, John D.

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N2 - PURPOSE. Lipofuscin accumulates in human retinal pigment epithelium (RPE) cells with age and may be the main factor responsible for the increasing susceptibility of RPE to photo-oxidation with age. As the composition, absorption, and fluorescence of lipofuscin undergo age-related changes, the purpose of this study was to determine whether photoreactivity of lipofuscin granules also changes with the donor age. METHODS. To determine whether the photoreactivity of lipofuscin itself is age related, lipofuscin granules were isolated from human RPE and pooled into age groups. Photoreactivity was assessed by measuring action spectra of photo-induced oxygen uptake and photogeneration of reactive oxygen species. Separation of chloroform-soluble (ChS) and -insoluble (ChNS) components by Folch's extraction was used to determine the factors responsible for the age-related increase in lipofuscin photoreactivity. RESULTS. The observed rates of photo-induced oxygen uptake and photo-induced accumulation of superoxide-derived spin adducts indicated that when normalized to equal numbers of lipofuscin granules, aerobic photoreactivity of lipofuscin increased with age. Both ChS and ChNS mediated photogeneration of singlet oxygen, superoxide radical anion, and photo-oxidation of added lipids and proteins. Although both ChS and ChNS exhibited substantial photoreactivities, neither exhibited significant age-related changes when normalized to equal dry mass. In contrast, ChNS contents in lipofuscin granules significantly increased with aging. CONCLUSIONS. Aerobic photoreactivity of RPE lipofuscin substantially increases with aging. This effect may be ascribed to the increased content of insoluble components.

AB - PURPOSE. Lipofuscin accumulates in human retinal pigment epithelium (RPE) cells with age and may be the main factor responsible for the increasing susceptibility of RPE to photo-oxidation with age. As the composition, absorption, and fluorescence of lipofuscin undergo age-related changes, the purpose of this study was to determine whether photoreactivity of lipofuscin granules also changes with the donor age. METHODS. To determine whether the photoreactivity of lipofuscin itself is age related, lipofuscin granules were isolated from human RPE and pooled into age groups. Photoreactivity was assessed by measuring action spectra of photo-induced oxygen uptake and photogeneration of reactive oxygen species. Separation of chloroform-soluble (ChS) and -insoluble (ChNS) components by Folch's extraction was used to determine the factors responsible for the age-related increase in lipofuscin photoreactivity. RESULTS. The observed rates of photo-induced oxygen uptake and photo-induced accumulation of superoxide-derived spin adducts indicated that when normalized to equal numbers of lipofuscin granules, aerobic photoreactivity of lipofuscin increased with age. Both ChS and ChNS mediated photogeneration of singlet oxygen, superoxide radical anion, and photo-oxidation of added lipids and proteins. Although both ChS and ChNS exhibited substantial photoreactivities, neither exhibited significant age-related changes when normalized to equal dry mass. In contrast, ChNS contents in lipofuscin granules significantly increased with aging. CONCLUSIONS. Aerobic photoreactivity of RPE lipofuscin substantially increases with aging. This effect may be ascribed to the increased content of insoluble components.

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