Abstract:
:Alterations in the retinal microvessel (RMV) compartment occurring in systemic disease states such as diabetes may eventually contribute to blindness. To specifically address the pathophysiological role of the microvasculature we developed a new method for RMV bulk isolation from individual rats. The extraction procedure performed in the cold throughout takes less than one hour. Slight modifications enable isolation of brain microvessels (BMVs) for comparison. Microscopically, RMVs and BMVs consisted mainly of capillaries of good structural integrity. The endothelial cell/pericyte ratio was approximately 1.8 in RMVs and 2.7 in BMVs, well in agreement with data from intact vascular beds. Total RNA extracted from individual rats amounted to approximately 7 ng in RMVs, 50 ng in BMVs, and 155 ng in pial arteries (which were also isolated) with highly preserved integrity throughout. Measurements using microfluidic card methodology revealed segregation of RMVs, BMVs, and pial arteries in distinct clusters based on principal component analysis. In all three vascular compartments endothelial cell-specific markers were significantly enriched. Similarly, pericyte-specific markers displayed accumulation in RMVs, BMVs, and pial arteries, the latter probably reflecting the common ontogenetic origin of pericytes and smooth muscle cells. Isolation of RMVs, BMVs, and pial arteries from rats suffering from 8-weeks hyperglycemia yielded expression patterns of endothelial cell- and pericyte-specific marker genes largely comparable to those obtained in control rats. Our newly developed protocols allow for selective studies of RMVs from individual rats to characterize reactive pathways, in comparison with the ontogenetically closely related BMVs. Moreover, our protocols with inclusion of pial arteries enable comparative studies of the macro- and microvasculature from the same organ.
journal_name
Vascul Pharmacoljournal_title
Vascular pharmacologyauthors
Li Y,Lapina N,Weinzierl N,Bonde L,Boedtkjer E,Schubert R,Moshage H,Wohlfart P,Schilling Ldoi
10.1016/j.vph.2018.07.001subject
Has Abstractpub_date
2018-11-01 00:00:00pages
24-30eissn
1537-1891issn
1879-3649pii
S1537-1891(18)30086-7journal_volume
110pub_type
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journal_title:Vascular pharmacology
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journal_title:Vascular pharmacology
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journal_title:Vascular pharmacology
pub_type: 杂志文章
doi:10.1016/j.vph.2008.03.004
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journal_title:Vascular pharmacology
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doi:10.1016/j.vph.2008.11.005
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journal_title:Vascular pharmacology
pub_type: 杂志文章
doi:10.1016/j.vph.2006.01.002
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journal_title:Vascular pharmacology
pub_type: 杂志文章
doi:10.1016/j.vph.2004.10.002
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journal_title:Vascular pharmacology
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journal_title:Vascular pharmacology
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journal_title:Vascular pharmacology
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journal_title:Vascular pharmacology
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doi:10.1016/j.vph.2007.08.001
更新日期:2007-11-01 00:00:00
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journal_title:Vascular pharmacology
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doi:10.1016/j.vph.2008.02.002
更新日期:2008-04-01 00:00:00
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journal_title:Vascular pharmacology
pub_type: 杂志文章
doi:10.1016/j.vph.2006.01.004
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journal_title:Vascular pharmacology
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doi:10.1016/j.vph.2016.08.001
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journal_title:Vascular pharmacology
pub_type: 杂志文章
doi:10.1016/j.vph.2015.07.011
更新日期:2016-01-01 00:00:00
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journal_title:Vascular pharmacology
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