Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells

Lisa Ann Brennan, Wanda Lee Kantorow, Daniel Chauss, Rebecca S. Estrada, Shuying He, Lyndzie Mattucci, Jianning Wei, S. Amer Riazuddin, Ales Cvekl, J. Fielding Hejtmancik, Marc Kantorow

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Purpose: Mutation of the autophagy gene FYVE (named after the four cysteine-rich proteins: Fab 1 [yeast orthologue of PIKfyve], YOTB, Vac 1 [vesicle transport protein], and EEA1) and coiled coil containing 1 (fyco1) causes human cataract suggesting a role for autophagy in lens function. Here, we analyzed the range and spatial expression patterns of lens autophagy genes and we evaluated whether autophagy could be induced in lens cells exposed to stress. Methods: Autophagy gene expression levels and their spatial distribution patterns were evaluated between microdissected human lens epithelium and fibers at the mRNA and protein levels by microarray data analysis, real-time PCR and western blot analysis. Selected autophagy protein spatial expression patterns were also examined in newborn mouse lenses by immunohistochemistry. The autophagosomal content of cultured human lens epithelial cells was determined by counting the number of microtubule-associated protein 1 light chain 3B (LC3B)-positive puncta in cells cultured in the presence or absence of serum. Results: A total of 42 autophagy genes were detected as being expressed by human lens epithelium and fibers. The autophagosomal markers LC3B and FYCO1 were detected throughout the newborn mouse lens. Consistently, the autophagy active form of LC3B (LC3B II) was detected in microdissected human lens fibers. An increased number of LC3B-positive puncta was detected in cultured lens cells upon serum starvation suggesting induction of autophagy in lens cells under stress conditions. Conclusions: The data provide evidence that autophagy is an important component for the function of lens epithelial and fiber cells. The data are consistent with the notion that disruption of lens autophagy through mutation or inactivation of specific autophagy proteins could lead to loss of lens resistance to stress and/or loss of lens differentiation resulting in cataract formation.

Original languageEnglish (US)
Pages (from-to)1773-1786
Number of pages14
JournalMolecular Vision
Volume18
StatePublished - Jun 30 2012

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Crystalline Lens
Autophagy
Lenses
Genes
Light
Cataract
Cultured Cells
Epithelium
Epithelial Cells
Protein Array Analysis
Mutation
Proteins
Microtubule-Associated Proteins
Microarray Analysis
Starvation
Serum

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Brennan, L. A., Kantorow, W. L., Chauss, D., Estrada, R. S., He, S., Mattucci, L., ... Kantorow, M. (2012). Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells. Molecular Vision, 18, 1773-1786.

Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells. / Brennan, Lisa Ann; Kantorow, Wanda Lee; Chauss, Daniel; Estrada, Rebecca S.; He, Shuying; Mattucci, Lyndzie; Wei, Jianning; Amer Riazuddin, S.; Cvekl, Ales; Fielding Hejtmancik, J.; Kantorow, Marc.

In: Molecular Vision, Vol. 18, 30.06.2012, p. 1773-1786.

Research output: Contribution to journalArticle

Brennan, LA, Kantorow, WL, Chauss, D, Estrada, RS, He, S, Mattucci, L, Wei, J, Amer Riazuddin, S, Cvekl, A, Fielding Hejtmancik, J & Kantorow, M 2012, 'Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells', Molecular Vision, vol. 18, pp. 1773-1786.
Brennan LA, Kantorow WL, Chauss D, Estrada RS, He S, Mattucci L et al. Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells. Molecular Vision. 2012 Jun 30;18:1773-1786.
Brennan, Lisa Ann ; Kantorow, Wanda Lee ; Chauss, Daniel ; Estrada, Rebecca S. ; He, Shuying ; Mattucci, Lyndzie ; Wei, Jianning ; Amer Riazuddin, S. ; Cvekl, Ales ; Fielding Hejtmancik, J. ; Kantorow, Marc. / Spatial expression patterns of autophagy genes in the eye lens and induction of autophagy in lens cells. In: Molecular Vision. 2012 ; Vol. 18. pp. 1773-1786.
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abstract = "Purpose: Mutation of the autophagy gene FYVE (named after the four cysteine-rich proteins: Fab 1 [yeast orthologue of PIKfyve], YOTB, Vac 1 [vesicle transport protein], and EEA1) and coiled coil containing 1 (fyco1) causes human cataract suggesting a role for autophagy in lens function. Here, we analyzed the range and spatial expression patterns of lens autophagy genes and we evaluated whether autophagy could be induced in lens cells exposed to stress. Methods: Autophagy gene expression levels and their spatial distribution patterns were evaluated between microdissected human lens epithelium and fibers at the mRNA and protein levels by microarray data analysis, real-time PCR and western blot analysis. Selected autophagy protein spatial expression patterns were also examined in newborn mouse lenses by immunohistochemistry. The autophagosomal content of cultured human lens epithelial cells was determined by counting the number of microtubule-associated protein 1 light chain 3B (LC3B)-positive puncta in cells cultured in the presence or absence of serum. Results: A total of 42 autophagy genes were detected as being expressed by human lens epithelium and fibers. The autophagosomal markers LC3B and FYCO1 were detected throughout the newborn mouse lens. Consistently, the autophagy active form of LC3B (LC3B II) was detected in microdissected human lens fibers. An increased number of LC3B-positive puncta was detected in cultured lens cells upon serum starvation suggesting induction of autophagy in lens cells under stress conditions. Conclusions: The data provide evidence that autophagy is an important component for the function of lens epithelial and fiber cells. The data are consistent with the notion that disruption of lens autophagy through mutation or inactivation of specific autophagy proteins could lead to loss of lens resistance to stress and/or loss of lens differentiation resulting in cataract formation.",
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AU - Kantorow, Wanda Lee

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AU - He, Shuying

AU - Mattucci, Lyndzie

AU - Wei, Jianning

AU - Amer Riazuddin, S.

AU - Cvekl, Ales

AU - Fielding Hejtmancik, J.

AU - Kantorow, Marc

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N2 - Purpose: Mutation of the autophagy gene FYVE (named after the four cysteine-rich proteins: Fab 1 [yeast orthologue of PIKfyve], YOTB, Vac 1 [vesicle transport protein], and EEA1) and coiled coil containing 1 (fyco1) causes human cataract suggesting a role for autophagy in lens function. Here, we analyzed the range and spatial expression patterns of lens autophagy genes and we evaluated whether autophagy could be induced in lens cells exposed to stress. Methods: Autophagy gene expression levels and their spatial distribution patterns were evaluated between microdissected human lens epithelium and fibers at the mRNA and protein levels by microarray data analysis, real-time PCR and western blot analysis. Selected autophagy protein spatial expression patterns were also examined in newborn mouse lenses by immunohistochemistry. The autophagosomal content of cultured human lens epithelial cells was determined by counting the number of microtubule-associated protein 1 light chain 3B (LC3B)-positive puncta in cells cultured in the presence or absence of serum. Results: A total of 42 autophagy genes were detected as being expressed by human lens epithelium and fibers. The autophagosomal markers LC3B and FYCO1 were detected throughout the newborn mouse lens. Consistently, the autophagy active form of LC3B (LC3B II) was detected in microdissected human lens fibers. An increased number of LC3B-positive puncta was detected in cultured lens cells upon serum starvation suggesting induction of autophagy in lens cells under stress conditions. Conclusions: The data provide evidence that autophagy is an important component for the function of lens epithelial and fiber cells. The data are consistent with the notion that disruption of lens autophagy through mutation or inactivation of specific autophagy proteins could lead to loss of lens resistance to stress and/or loss of lens differentiation resulting in cataract formation.

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