Perlecan Heparan Sulfate Proteoglycan Is a Critical Determinant of Angiogenesis in Response to Mouse Hind-Limb Ischemia
(2014) In Canadian Journal of Cardiology 30(11). p.1444-1451- Abstract
Background: Perlecan is a heparan sulfate proteoglycan (HSPG) constituent of the extracellular matrix with roles in cell growth, differentiation, and angiogenesis. The role of the HS side chains in regulating invivo angiogenesis after hind-limb ischemia is unknown. Methods: Heparan sulfate (HS)-deficient perlecan (Hspg2
δ3/δ3
) mice (n= 35), containing normal perlecan core protein but deficient in HS side chains, and wild-type (n= 33) littermates underwent surgical induction of hind-limb ischemia. Laser Doppler perfusion imaging (LDPI) and contrast-enhanced ultrasonography (CEU) provided serial assessment of hind-limb perfusion.... (More)
(Less)
Background: Perlecan is a heparan sulfate proteoglycan (HSPG) constituent of the extracellular matrix with roles in cell growth, differentiation, and angiogenesis. The role of the HS side chains in regulating invivo angiogenesis after hind-limb ischemia is unknown. Methods: Heparan sulfate (HS)-deficient perlecan (Hspg2
δ3/δ3
) mice (n= 35), containing normal perlecan core protein but deficient in HS side chains, and wild-type (n= 33) littermates underwent surgical induction of hind-limb ischemia. Laser Doppler perfusion imaging (LDPI) and contrast-enhanced ultrasonography (CEU) provided serial assessment of hind-limb perfusion. Harvested muscles underwent immunostaining for endothelial cell density (CD31), real-time reverse transcription polymerase chain reaction RT-PCR for vascular endothelial growth factor (VEGF) mRNA expression and western blot analysis for VEGF and fibroblast growth factor (FGF)2 protein expression at days 2 and28. Results: Serial LDPI showed significantly greater perfusion recovery in ischemic limbs of wild-type compared with Hspg2
δ3/δ3
mice. CEU showed that normalized microvascular perfusion was increased in wild-type compared with Hspg2
δ3/δ3
mice at day 28 (0.67 ± 0.12 vs 0.26 ± 0.08; P= 0.001). CD31-positive cell counts were significantly higher in wild-type compared with Hspg2
δ3/δ3
mice on day 28 (122 ± 30 cells vs 84 ± 34 cells per high-power field [HPF]; P < 0.05). Endogenous VEGF mRNA expression (P < 0.05) and VEGF protein expression (P < 0.002) were significantly decreased in the ischemic limbs of Hspg2
δ3/δ3
mice compared with wild-type mice at day 2 and day 28, respectively. FGF2 protein expression showed no significant differences. Conclusions: These results suggest that the HS side chains in perlecan are important mediators of the angiogenic response to ischemia through a mechanism that involves upregulation of VEGF expression.
- author
- publishing date
- 2014-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Canadian Journal of Cardiology
- volume
- 30
- issue
- 11
- pages
- 8 pages
- publisher
- Elsevier
- external identifiers
-
- pmid:25249499
- scopus:84908478240
- ISSN
- 0828-282X
- DOI
- 10.1016/j.cjca.2014.06.003
- language
- English
- LU publication?
- no
- id
- d8712bc8-9907-4ae2-9fa3-33d27393ad9d
- date added to LUP
- 2019-07-01 22:15:19
- date last changed
- 2024-04-30 17:04:37
@article{d8712bc8-9907-4ae2-9fa3-33d27393ad9d,
abstract = {{<p><br>
Background: Perlecan is a heparan sulfate proteoglycan (HSPG) constituent of the extracellular matrix with roles in cell growth, differentiation, and angiogenesis. The role of the HS side chains in regulating invivo angiogenesis after hind-limb ischemia is unknown. Methods: Heparan sulfate (HS)-deficient perlecan (Hspg2<br>
<sup>δ3/δ3</sup><br>
) mice (n= 35), containing normal perlecan core protein but deficient in HS side chains, and wild-type (n= 33) littermates underwent surgical induction of hind-limb ischemia. Laser Doppler perfusion imaging (LDPI) and contrast-enhanced ultrasonography (CEU) provided serial assessment of hind-limb perfusion. Harvested muscles underwent immunostaining for endothelial cell density (CD31), real-time reverse transcription polymerase chain reaction RT-PCR for vascular endothelial growth factor (VEGF) mRNA expression and western blot analysis for VEGF and fibroblast growth factor (FGF)2 protein expression at days 2 and28. Results: Serial LDPI showed significantly greater perfusion recovery in ischemic limbs of wild-type compared with Hspg2<br>
<sup>δ3/δ3</sup><br>
mice. CEU showed that normalized microvascular perfusion was increased in wild-type compared with Hspg2<br>
<sup>δ3/δ3</sup><br>
mice at day 28 (0.67 ± 0.12 vs 0.26 ± 0.08; P= 0.001). CD31-positive cell counts were significantly higher in wild-type compared with Hspg2<br>
<sup>δ3/δ3</sup><br>
mice on day 28 (122 ± 30 cells vs 84 ± 34 cells per high-power field [HPF]; P < 0.05). Endogenous VEGF mRNA expression (P < 0.05) and VEGF protein expression (P < 0.002) were significantly decreased in the ischemic limbs of Hspg2<br>
<sup>δ3/δ3</sup><br>
mice compared with wild-type mice at day 2 and day 28, respectively. FGF2 protein expression showed no significant differences. Conclusions: These results suggest that the HS side chains in perlecan are important mediators of the angiogenic response to ischemia through a mechanism that involves upregulation of VEGF expression.<br>
</p>}},
author = {{Qiang, Beiping and Lim, Sang Yup and Lekas, Michael and Kuliszewski, Michael A. and Wolff, Rafael and Osherov, Azriel B. and Rudenko, Dmitriy and Leong-Poi, Howard and Noyan, Hossein and Husain, Mansoor and Tran, Kiet and Tryggvason, Karl and Hedin, Ulf and Tran-Lundmark, Karin and Strauss, Bradley H.}},
issn = {{0828-282X}},
language = {{eng}},
month = {{01}},
number = {{11}},
pages = {{1444--1451}},
publisher = {{Elsevier}},
series = {{Canadian Journal of Cardiology}},
title = {{Perlecan Heparan Sulfate Proteoglycan Is a Critical Determinant of Angiogenesis in Response to Mouse Hind-Limb Ischemia}},
url = {{http://dx.doi.org/10.1016/j.cjca.2014.06.003}},
doi = {{10.1016/j.cjca.2014.06.003}},
volume = {{30}},
year = {{2014}},
}