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Journals of Gerontology: Biological Sciences
cite as: J Gerontol A Biol Sci Med Sci, 2017, Vol. 72, No. 6, 740–746
doi:10.1093/gerona/glw213
Advance Access publication November 15, 2016
Original Article
Microvasculature of the Mouse Cerebral Cortex Exhibits
Increased Accumulation and Synthesis of Hyaluronan
With Aging
May J. Reed,
1
Robert B. Vernon,
2
Mamatha Damodarasamy,
1
Christina K. Chan,
2
Thomas N. Wight,
2
Itay Bentov,
3
and William A. Banks
1,4
1
Department of Medicine, University of Washington, Seattle.
2
Matrix Biology Program, Benaroya Research Institute at Virginia Mason,
Seattle, Washington.
3
Department of Anesthesia and Pain Medicine, University of Washington, Seattle.
4
VA Puget Sound Health Care
System, Geriatric Research Education and Clinical Center, Seattle, Washington.
Address correspondence to May J. Reed, MD, Department of Medicine, University of Washington, Harborview Medical Center, 325 9th Avenue,
Box 359625, Seattle, WA 98104. E-mail: mjr@uw.edu
Received May 18, 2016; Accepted October 10, 2016
Decision Editor: Rafael de Cabo, PhD
Abstract
The microvasculature of the aged brain is less dense and more vulnerable to dysfunction than that of the young brain. Brain microvasculature
is supported by its surrounding extracellular matrix, which is comprised largely of hyaluronan (HA). HA is continually degraded into lower
molecular weight forms that induce neuroinfammation. We examined HA associated with microvessels (MV) of the cerebral cortex of young
(4 months), middle-aged (14 months), and aged (24–26 months) mice. We confrmed that the density of cortical MV decreased with age.
Perivascular HA levels increased with age, but there was no age-associated change in HA molecular weight profle. MV isolated from aged
cortex had more HA than MV from young cortex. Examination of mechanisms that might account for elevated HA levels with aging showed
increased HA synthase 2 (HAS2) mRNA and protein in aged MV relative to young MV. In contrast, mRNAs for HA-degrading hyaluronidases
or hyaladherins that mitigate HA degradation showed no changes with age. Corresponding to increased HAS2, aged MV synthesized
signifcantly more HA (of all molecular weight classes) in vitro than young MV. We propose that increased HA synthesis and accumulation in
brain MV contributes to neuroinfammation and reduced MV density and function in aging.
Keywords: Aging—Brain—Microvasculature—Hyaluronan—Microvessels
It is increasingly appreciated that the microvasculature of the aged
brain is less dense and more vulnerable to injury than that of the
young brain (1–3). Changes in the brain microvasculature with aging
result in less effective perfusion and increased risk of exposure of
brain tissue to deleterious substances from the systemic circulation
(4–6). The resiliency of the microvasculature is determined, in part,
by the extracellular matrix (ECM) that surrounds it (7). This ECM
is comprised largely of hyaluronan (HA), an anionic, nonsulfated
glycosaminoglycan comprised of the repeating disaccharide unit
d-glucuronic acid and N-acetyl-D-glucosamine. In the brain, HA is
synthesized by multiple cell types, including cells of the microvessels
(MV), which consist of vascular endothelial cells and their associated
pericytes and astrocytes. Associated with the HA are hyaladherins:
a broad group of HA-binding molecules that include chondroitin
sulfate-rich proteoglycans (e.g., aggrecan, versican, neurocan, and
brevican), the glycoprotein tenascin-R, and tumor necrosis factor-
stimulated gene 6 protein (TSG-6) (8–11). HA linkage to these and
other components of brain ECM is mediated by proteins, such as
Bral1, which confer additional stability (12).
HA is produced by membrane-bound HA synthases (HASes)
which exist in at least three isoforms (HAS1, 2, and 3) that produce
HA of different chain lengths. HA is usually synthesized in forms
with high molecular weights (HMW) (e.g., >1,000 kDa) that are
subsequently cleaved by hyaluronidases (HYALs) into shorter chain
lengths with a wide range of MW. There are at least six HYAL iso-
forms with consensus that HYALs 1–3 could be detected in the set-
ting of brain injury (13). The biological activity of HA is determined
by its size, with many studies showing that HA of relatively low MW
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