Vascular pathology is a critical and more and more recognised component of AD, impairing blood-to-brain influx of nutrients essential for cerebral functioning and brain-to-blood efflux of cerebral waste products leading to cognitive deterioration [
10,
11,
64,
65]. By characterising the transgenic arcAβ mouse model of AD—which presents a strong vascular pathology—at early, mid and late disease stages we investigated what processes could possibly instigate the vascular and metabolic impairments early on in the disease and which mechanisms aggravate the vascular pathology at later stages in this AD mouse model. Decreased endothelial GLUT1 expression has been found in transgenic mouse models of AD and in human AD patients [
15,
22,
26,
47]. We observed lowered GLUT1 protein expression in TG animals starting at mid-stage disease pathology when CAA formation is apparent in these mice and diffuse Aβ plaques are becoming wide-spread. As GLUT1 is the sole glucose transporter expressed by brain endothelial cells and peripheral supply is the principal source of glucose for the brain [
46], it can be appreciated that a decrease in GLUT1 expression will have significant outcomes on cerebral glucose uptake. Indeed, cerebral imaging data from human subjects show a clear correlation between GLUT1 expression and blood-to-brain glucose uptake [
12]. Our findings of decreased brain glucose influx and concentration as measured by in vivo microdialysis in TG arcAβ mice, are in line with these data. The majority of glucose entering the brain via endothelial GLUT1 is subsequently taken up by astrocytes via astrocytic GLUT1 [
46]. Astrocyte pathology with altered expression of astrocyte-specific proteins essential for cerebral functioning and metabolism is recognised as a significant factor in AD and subject to intensive research [
3,
48]. To understand if a decreased astrocytic GLUT1 expression could also be an underlying factor in the reduced glucose uptake in our microdialysis studies, we cultured astrocytes from the same cohort of arcAβ mice ex vivo. We found that its expression was severely affected, starting at mid-stage disease pathology. Astrocytic glucose uptake is a vital step in the cerebral energy apparatus because astrocytes convert glucose into lactate which is used by neurons during neuronal stimulation [
14,
29,
38,
43,
44]. Abnormalities in this essential astrocytic contribution to neurometabolic coupling have detectable outcomes on cerebral performance and are known to occur in AD [
3,
50]. The impaired lactate release we observed upon in vivo neuronal stimulation during microdialysis in TG arcAβ mice, was confirmed by a decreased astrocytic MCT1 expression and lactate release deficiency ex vivo. The reciprocal character of the expression profiles of MCT1 and GLUT1 as shown by Maurer and colleagues [
33] makes plausible that the decreased astrocytic MCT1 protein levels could be related to lower astrocytic GLUT1. Astrocytic GLUT1 and MCT1 are mainly expressed on astrocyte endfeet which are crucial for neurovascular coupling [
1,
3,
65]. Impaired neurovascular coupling is common in neurodegenerative disorders including AD and AD mouse models with astrocyte endfeet retraction and swelling compromising the cross-talk between astrocytes, blood vessels and neurons and decreased cerebral metabolism as histological and functional hallmarks [
3,
37,
61]. We detected signs of neurovascular uncoupling represented by astrocyte endfeet retraction and swelling, possibly also (partly) explaining our findings on impaired metabolism. We observed this starting at an early pathological stage of the disease, at which extracellular Aβ plaque burden is small in the arcAβ mouse. Only astrocytes surrounding diffuse Aβ plaques and vascular Aβ deposits showed endfeet abnormalities even at the late-stage pathology, suggesting that Aβ instigates the loss of neurovascular coupling rather than the process of ageing or the mere presence of the APP transgene in these mice. The mechanism by which Aβ prevents contact between endfeet could be the formation of a physical barrier between blood vessels and astrocytes. Another likely mechanism underlying this impaired astrocyte-endothelium interaction could be the loss of expression of the astrocyte endfeet—VBM linker β dystroglycan as we observed in our TG arcAβ mice [
2,
53]. It is unclear whether Aβ toxicity is involved in the attenuated β dystroglycan expression or that the changes in the VBM we describe here and were described by others [
21] are culprit. Interestingly, however, the degree of β dystroglycan loss was paralleled by the presence of diffuse, parenchymal Aβ plaques and leakage of endogenous IgG into the brain. Astrocyte endfeet form an essential part of the BBB [
1] and the question is if detachment of astrocyte endfeet from the vasculature leading to disruption of the BBB and hence extravasation of serum proteins into the CNS or that entry of serum proteins into the brain is responsible for this aspect of neurovascular uncoupling. Brains of multiple sclerosis patients and its in vivo counterpart experimental autoimmune encephalomyelitis show that neuroinflammation and extravasation of components of the peripheral immune system induce astrocyte endfeet abnormalities including retraction and swelling [
2,
63]. It is well established that CNS entry of components of the peripheral immune system is part of the AD pathology [
13]. Taking into account our findings on the pathological cerebral extravasation of the BBB viability marker Trypan Blue and vessel wall instability as shown by leakage of vascular casting resin in the TG arcAβ mice, a role for components of the peripheral immune system in (further) triggering pathological astrocyte responses including endfeet retraction could indeed be possible [
1,
13,
65]. It is clear that with astrocyte endfeet abnormalities already present at the early-stage pathology, impaired BBB functioning is a premature process in the arcAβ mouse. An astrocytic involvement at premature time points in the pathology was further indicated by astrogliosis with paralleled increase in vascular laminin and astrocytic laminin secretion ex vivo in the early-stage mice. Increase in laminin VBM in these relatively young mice with little Aβ plaque burden seems to be contradictory with the unique CAA-related increase in laminin we found in mid-stage and late-stage mice. It is interesting, however, that mainly the incoming arteries from leptomeningeal origin showed this increased astrocyte activation in the early-stage TG mice with concomitant increase in VBM laminin. It is exactly these vessels which are the first to be affected by CAA at later disease stages in the arcAβ mouse and other transgenic AD mouse models [
58,
59]. The induction of signal transduction cascades within the endothelium can trigger astrocytic release of endothelium-protective molecules [
1]. This protective reaction could be elicited by soluble Aβ species which are known to induce astrocyte activation and to be toxic to endothelial cells and SMCs [
18,
51,
56,
60]. Furthermore, laminin expression is known to be up-regulated by damage to the vasculature [
4,
65]. Soluble Aβ species mainly as toxic protofibrils are abundantly present in arcAβ mice from 2 months of age onwards [
31,
35] and could be directed to the vasculature by the forces of the cerebrovascular drainage pathways in which especially the cerebral arteries are involved [
9,
20,
58], providing a possible explanation for the unique location of these reactive astrocytes around blood vessels. Seen in this light, laminin is known to interact with Aβ, inhibiting its fibrillisation and diluting its neurotoxicity [
27,
36], giving foundation to the idea that astrocytic laminin release as we detected could be a protective mechanism to both strengthen the vessel wall by
increasing VBM thickness and an attempt to reduce Aβ-induced neuro- and vascular toxicity by interfering with (further) Aβ fibrillisation. Although we found astrocytes to secrete laminin as was observed by others as well [
8,
45], it cannot be excluded that endothelial cells themselves are involved too as these cells can secrete laminin [
19]. Although possibly being an astrocytic protection attempt, an increase in laminin secretion paralleled by VBM thickening leads to vessel wall rigidity and hence reduced CBF, especially the leptomeningeal vessel wall thickening we observed in early-stage TG arcAβ mice which directly affects downstream perfusion of the arterioles and capillaries [
23,
30]. Cerebral hypoperfusion—and with that reduced oxygen and glucose supply and hindered cerebral drainage of cerebral waste products—starting at such early age could be at the base of the accumulation of Aβ and the severe CAA-related vascular pathology with vascular SMC loss and BBB impairments as we observed later on at mid- and late-stage pathology in the arcAβ mice.