PMID- 28622396
OWN - NLM
STAT- MEDLINE
DCOM- 20170719
LR  - 20181113
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 58
IP  - 7
DP  - 2017 Jun 1
TI  - Mutant Fibulin-3 Causes Proteoglycan Accumulation and Impaired Diffusion Across 
      Bruch's Membrane.
PG  - 3046-3054
LID - 10.1167/iovs.17-21720 [doi]
AB  - PURPOSE: The mutation R345W in EFEMP1 (fibulin-3) causes macular degeneration. 
      This study sought to determine whether proteoglycan content and diffusion across 
      Bruch's membrane are altered in Efemp1ki/ki mice carrying this mutation or in 
      Efemp1-/- mice. METHODS: Proteoglycans in mouse Bruch's membranes were stained 
      with Cupromeronic Blue (CB). Heparan sulfated proteoglycan (HSPG) and 
      chondroitin/dermatan sulfate proteoglycan (C/DSPG) distributions were visualized 
      following treatments with chondroitinase ABC (C-ABC) or nitrous acid. Total 
      sulfated glycosaminoglycans (sGAGs) in Bruch's membrane/choroid (BrM/Ch) were 
      measured with dimethylmethylene blue (DMMB). Matrix metalloprotease (MMP)-2, 
      MMP-9, and tissue inhibitor of metalloproteinase (TIMP)-3 were examined by 
      immunofluorescence and quantified using Image J. Molecules with different Stokes 
      radius (Rs) were allowed simultaneously to diffuse through mouse BrM/Ch mounted 
      in a modified Ussing chamber. Samples were quantified using gel exclusion 
      chromatography. RESULTS: HSPGs and C/DSPGs were markedly increased in Efemp1ki/ki 
      Bruch's membrane, and MMP-2 and MMP-9 were decreased, but TIMP-3 was increased. 
      Diffusion across Efemp1ki/ki Bruch's membrane was impaired. In contrast, the 
      proteoglycan amount in Efemp1-/- Bruch's membrane was not significantly 
      different, but the size of proteoglycans was much larger. MMP-2, MMP-3, and 
      TIMP-3 levels were similar to that of Efemp1+/+ mice, but they were localized 
      diffusely in retinal pigment epithelium (RPE) cells instead of Bruch's membrane. 
      Diffusion across Efemp1-/- Bruch's membrane was enhanced. CONCLUSIONS: Mutant 
      fibulin-3 causes proteoglycan accumulation, reduction of MMP-2 and MMP-9, but 
      increase of TIMP-3, and impairs diffusion across Bruch's membrane. Fibulin-3 
      ablation results in altered sizes of proteoglycans, altered distributions of 
      MMP-2, MMP-9, and TIMP-3, and enhances diffusion across Bruch's membrane.
FAU - Zayas-Santiago, Astrid
AU  - Zayas-Santiago A
AD  - Department of Pathology & Laboratory Medicine, Universidad Central del Caribe, 
      Bayamon, Puerto Rico.
FAU - Cross, Samuel D
AU  - Cross SD
AD  - Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota, United States.
FAU - Stanton, James B
AU  - Stanton JB
AD  - Department of Ophthalmology & Vision Science, University of Arizona, Tucson, 
      Arizona, United States.
FAU - Marmorstein, Alan D
AU  - Marmorstein AD
AD  - Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota, United States.
FAU - Marmorstein, Lihua Y
AU  - Marmorstein LY
AD  - Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota, United States.
LA  - eng
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Efemp1 protein, mouse)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Proteoglycans)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Aging/genetics
MH  - Animals
MH  - Bruch Membrane/*metabolism/pathology
MH  - DNA/*genetics
MH  - DNA Mutational Analysis
MH  - Disease Models, Animal
MH  - Extracellular Matrix Proteins/*genetics/metabolism
MH  - Macular Degeneration/diagnosis/*genetics/metabolism
MH  - Mice
MH  - Mice, Mutant Strains
MH  - *Mutation
MH  - Proteoglycans/*metabolism
PMC - PMC5477800
EDAT- 2017/06/18 06:00
MHDA- 2017/07/20 06:00
PMCR- 2017/07/01
CRDT- 2017/06/17 06:00
PHST- 2017/06/17 06:00 [entrez]
PHST- 2017/06/18 06:00 [pubmed]
PHST- 2017/07/20 06:00 [medline]
PHST- 2017/07/01 00:00 [pmc-release]
AID - 2633355 [pii]
AID - IOVS-17-21720 [pii]
AID - 10.1167/iovs.17-21720 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2017 Jun 1;58(7):3046-3054. doi: 
      10.1167/iovs.17-21720.