Laminin immunohistochemistry: a simple method to visualize and quantitate vascular structures in the mammalian brain

Eriksdotter, Maria; Björklund, Håkan; Olson, Lar̀s

doi:10.1016/0165-0270(86)90128-7

Cited by 74 publications

(31 citation statements)

References 25 publications

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“…One of the major functional components of the basement membrane of blood vessels is laminin, which is found underlying the endothelium, and is an excellent marker for brain blood vessels [37] . We used laminin immunohistochemistry to determine whether maternal MLT administration had an effect on the vascular structure or produced vascular expansion within the white matter of the FGR brain.…”

Section: Discussionmentioning

confidence: 99%

Effects of Antenatal Melatonin Treatment on the Cerebral Vasculature in an Ovine Model of Fetal Growth Restriction

et al. 2017

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Chronic moderate hypoxia, such as occurs in fetal growth restriction (FGR) during gestation, compromises the blood-brain barrier (BBB) and results in structural abnormalities of the cerebral vasculature. We have previously determined the neuroprotective and antioxidant effects of maternal administration of melatonin (MLT) on growth-restricted newborn lambs. The potential of maternal MLT therapy for the treatment of cerebrovascular dysfunction-associated developmental hypoxia has also been demonstrated in newborn lambs. We assessed whether MLT had an effect on the previously reported structural and cerebral vascular abnormalities in chronically hypoxic FGR lambs. Single umbilical-artery ligation surgery was performed in fetuses at approximately 105 days of gestation (term: 147 days) to induce placental insufficiency and FGR, and treatment with either saline or an MLT infusion (0.1 mg/kg) was started 4 h after surgery. Ewes delivered naturally at term and lambs were euthanased 24 h later. We found a significant reduction in the number of laminin-positive blood vessels within the subcortical and periventricular white matter (SCWM and PVWM) and the subventricular zone (SVZ) in FGR (p < 0.0005) and FGR + MLT brains (p < 0.0005 vs. controls), with no difference found between FGR and FGR + MLT animals. This was associated with a significant decrease in VEGF immunoreactivity in FGR and FGR + MLT brains versus controls (p < 0.0005; SCWM and PVWM) and in the SVZ in FGR brains versus controls (p < 0.005) and also with significantly lower levels of proliferating blood vessels versus controls (p < 0.0005). Glucose transporter-1 immunoreactivity (vascular endothelium) was decreased in FGR versus control lambs (p < 0.0005) in SCWM, PVWM, and the SVZ; it was significantly increased in FGR + MLT lambs compared with FGR lambs in SCWM and PVWM (p < 0.005) and even more markedly in the SVZ (p < 0.0005). FGR brains showed a 72% reduction in pericyte coverage versus control lambs and 68% versus FGR + MLT in PVWM. In SCWM, we found a 77 and 73% reduction compared with control and FGR + MLT lambs, respectively, while in the SVZ, we observed a 68% reduction versus controls and a 70% reduction in FGR versus FGR + MLT lambs. Astrocyte end-feet coverage in the SCWM showed a significant 24% reduction in FGR versus control levels, a 42% decrease within the PVWM, and a 35% decrease within the SVZ versus controls. MLT normalized astrocyte attachment to blood vessels, with no difference seen between controls and FGR + MLT animals in any of the brain regions examined. We also observed a decrease in albumin extravasation and microhemorrhage in controls and FGR + MLT brains versus FGR lambs. Our results demonstrate that umbilicoplacental insufficiency is associated with FGR-produced vascular changes in the white matter and SVZ of FGR newborn brains and that maternal MLT prevented disruption of the BBB by protecting perivascular cells essential for the maintenance of vascular homeostasis and stability.

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Section: Discussionmentioning

confidence: 99%

Effects of Antenatal Melatonin Treatment on the Cerebral Vasculature in an Ovine Model of Fetal Growth Restriction

et al. 2017

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“…For this study we performed experiments that demonstrated that it does not label rabbit blood vessels [data not shown]. In order to demon strate blood vessels independently from MESA-1 staining, we visu alized laminin (18]. Rabbit anti-laminin polyclonal antibodies (Gibco-BRL, Cergy, France) were used at a dilution of 1/100.…”

Section: A Te Ria Ls and M E Th O D Smentioning

confidence: 99%

Xenogenic Transplantation into Newborn Rodent Brain: Neovascularization of the Graft by the Host

Mrejen

Quinonero²,

Moinard³

et al. 1992

Dev Neurosci

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Neoangiogenesis of transplants implanted into the brains of newborn rodent hosts was evaluated by immunohistochemistry for 2 weeks after the operation. The use of species-specific antibodies directed against mouse endothelial cells demonstrated the respective participation of the host and the donor in the formation of new vessels in the graft after crossed rabbit into mouse and mouse into rat transplantation experiments. We show that blood vessels made by host endothelial cells begin to penetrate the transplant 24 h after grafting, and cross it completely by 72 h. Simultaneously, host astrocytes invade the transplant.

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“…A variety of antigens expressed on either endothelial cells or the vascular basement membrane have been used including laminin (Eriksdotter-Nilsson et al, 1986), fibronectin (Krum et al, 1991), von Willebrand factor (Theilen et al, 1992), collagen IV (Gidday et al, 2005) and the heparin sulfate proteoglycan (HSPG) perlecan (Chen et al, 2005;Gidday et al, 2005;Bailey et al, 2004;Buée et al, 1994). All work well in embryonic or neonatal mouse and rat brain.…”

Section: Introductionmentioning

confidence: 99%

Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain

Franciosi¹,

Gasperi²,

Dickstein³

et al. 2007

Journal of Neuroscience Methods

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While the brain vasculature can be imaged with many methods, immunohistochemistry has distinct advantages due to its simplicity and applicability to archival tissue. However, immunohistochemical staining of the murine brain vasculature in aldehyde fixed tissue has proven elusive and inconsistent using current protocols. Here we investigated whether antigen retrieval methods could improve vascular staining in the adult mouse brain. We found that pepsin digestion prior to immunostaining unmasked widespread collagen IV staining of the cerebrovasculature in the adult mouse brain. Pepsin treatment also unmasked widespread vascular staining with laminin, but only marginally improved

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Laminin immunohistochemistry: a simple method to visualize and quantitate vascular structures in the mammalian brain

Cited by 74 publications

References 25 publications

Effects of Antenatal Melatonin Treatment on the Cerebral Vasculature in an Ovine Model of Fetal Growth Restriction

Effects of Antenatal Melatonin Treatment on the Cerebral Vasculature in an Ovine Model of Fetal Growth Restriction

Xenogenic Transplantation into Newborn Rodent Brain: Neovascularization of the Graft by the Host

Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain

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