The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Colak, Gozde; Filiano, Anthony J.; Johnson, Gail V.W.

doi:10.3791/3039

Cited by 21 publications

(16 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Construction of an "Ast-Tbr2DN" Mouse Model to Inhibit Astrocytic TGFb Signaling To investigate astrocytic TGFb signaling after stroke, we selected the model of distal middle cerebral artery occlusion (dMCAO) (Colak et al, 2011). The advantages of dMCAO are that it is directly caused by arterial occlusion and produces a highly consistent stroke with a clearly defined border.…”

Section: Resultsmentioning

confidence: 99%

Astrocytic transforming growth factor-beta signaling reduces subacute neuroinflammation after stroke in mice

Cekanaviciute

Fathali

Doyle

et al. 2014

Glia

175

110

View full text Add to dashboard Cite

Astrocytes limit inflammation after CNS injury, at least partially by physically containing it within an astrocytic scar at the injury border. We report here that astrocytic transforming growth factor-beta (TGFβ) signaling is a second, distinct mechanism that astrocytes utilize to limit neuroinflammation. TGFβs are anti-inflammatory and neuroprotective cytokines that are upregulated subacutely after stroke, during a clinically accessible time window. We have previously demonstrated that TGFβs signal to astrocytes, neurons, and microglia in the stroke border days after stroke. To investigate whether TGFβ affects astrocyte immunoregulatory functions, we engineered “Ast-Tbr2DN” mice where TGFβ signaling is inhibited specifically in astrocytes. Despite having a similar infarct size to wildtype controls, Ast-Tbr2DN mice exhibited significantly more neuroinflammation during the subacute period after distal middle cerebral occlusion (dMCAO) stroke. The peri-infarct cortex of Ast-Tbr2DN mice contained over 60% more activated CD11b+ monocytic cells and twice as much immunostaining for the activated microglia and macrophage marker CD68 than controls. Astrocytic scarring was not altered in Ast-Tbr2DN mice. However, Ast-Tbr2DN mice were unable to upregulate TGF-β1 and its activator thrombospondin-1 two days after dMCAO. As a result, the normal upregulation of peri-infarct TGFβ signaling was blunted in Ast-Tbr2DN mice. In this setting of lower TGFβ signaling and excessive neuroinflammation, we observed worse motor outcomes and late infarct expansion after photothrombotic motor cortex stroke. Taken together, these data demonstrate that TGFβ signaling is a molecular mechanism by which astrocytes limit neuroinflammation, activate TGFβ in the peri-infarct cortex, and preserve brain function during the subacute period after stroke.

show abstract

Section: Resultsmentioning

confidence: 99%

Astrocytic transforming growth factor-beta signaling reduces subacute neuroinflammation after stroke in mice

Cekanaviciute

Fathali

Doyle

et al. 2014

Glia

175

110

View full text Add to dashboard Cite

show abstract

“…Focal ischemic stroke was induced by pMCAO as previously referenced with slight modifications (Colak et al, 2011). Briefly, 8–12 week-old mice (25–35 g) were injected with analgesic Buprenorphine-SR (0.15 mg/kg), and anesthesia induced with 2% isoflurane-30% oxygen mixture.…”

Section: Methodsmentioning

confidence: 99%

Temporal remodeling of pial collaterals and functional deficits in a murine model of ischemic stroke

Okyere

Creasey

Lebovitz

et al. 2018

Journal of Neuroscience Methods

View full text Add to dashboard Cite

Background Leptomeningeal anastomoses play a critical role in regulating reperfusion following cerebrovascular obstruction; however, methods to evaluate their temporospatial remodeling remains under investigation. New method We combined arteriole-specific vessel painting with histological evaluation to assess the density and diameter of inter-collateral vessels between the middle cerebral artery and anterior cerebral artery (MCA-ACA) or posterior cerebral artery (MCA-PCA) in a murine model of permanent middle cerebral artery occlusion (pMCAO). Results While the overall density was not influenced by pMCAO, the size of MCA-ACA and MCA-PCA vessels had significantly increased 2 days post-pMCAO and peaked by 4 days compared to the un-injured hemisphere. Using a combination of vessel painting and immunofluorescence, we uniquely observed an induction of cellular division and a remodeling of the smooth muscle cells within the collateral niche following post-pMCAO on whole mount tissue sections. Vessel painting was also applied to pMCAO-injured Cx3cr1GFP mice, in order to identify the spatial relationship between Cx3cr1-positive peripheral-derived monocyte/macrophages and the vessel painted collaterals. Our histological findings were supplemented with analysis of cerebral blood flow using laser Doppler imaging and behavioral changes following pMCAO. Comparison with existing methods Compared to polyurethane and latex methods for collateral labeling, this new method provides detailed cell-type specific analysis within the collateral niche at the microscopic level, which has previously been unavailable. Conclusions This simple and reproducible combination of techniques is the first to dissect the temporospatial remodeling of pial collateral arterioles. The method will advance investigations into the underlying mechanisms governing the intricate processes of arteriogenesis.

show abstract

“…Body temperature was maintained at 37 °C with a heating pad during surgery (approx. 30 min) and recovery (Colak et al, 2011a). …”

Section: Methodsmentioning

confidence: 99%

Complete transglutaminase 2 ablation results in reduced stroke volumes and astrocytes that exhibit increased survival in response to ischemia

Colak

Johnson

2012

Neurobiology of Disease

Self Cite

View full text Add to dashboard Cite

Transglutaminase 2 (TG2) is a very multifunctional protein that is ubiquitously expressed in the body. It is a Ca2+-dependent transamidating enzyme, a GTPase, as well as a scaffolding protein. TG2 is the predominant form of transglutaminase expressed in the mammalian nervous system. Previously, it was shown that TG2 can affect both cell death and cell survival mechanisms depending on the cell type and the stressor. In the case of ischemic stress, TG2 was previously shown to play a protective role in the models used. For example in hTG2 transgenic mice, where TG2 is overexpressed only in neurons, middle cerebral artery ligation (MCAL) resulted in smaller infarct volumes compared to wild type mice. In this study TG2 knock out mice were used to determine how endogenous TG2 affected stroke volumes. Intriguingly, infarct volumes in TG2 knock out mice were significantly smaller compared to wild type mice. As expected, primary neurons isolated from TG2 knock out mice showed decreased viability in response to oxygen-glucose deprivation. However, primary astrocytes that were isolated from TG2 knock out mice were resistant to oxygen-glucose deprivation in situ. Both wild type and knock out neurons were protected against oxygen glucose deprivation when they were co-cultured with astrocytes from TG2 knockout mice. Therefore, the decreased stroke volumes observed in TG2 knock out mice after MCAL, can be correlated with the protective effects of TG2 knock out in astrocytes in response to oxygen glucose deprivation in situ. These findings suggest that neuron-astrocyte crosstalk plays a significant role in mediating ischemic cell death and that TG2 differentially impacts cell survival depending on cell context.

show abstract

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Cited by 21 publications

References 24 publications

Astrocytic transforming growth factor-beta signaling reduces subacute neuroinflammation after stroke in mice

Astrocytic transforming growth factor-beta signaling reduces subacute neuroinflammation after stroke in mice

Temporal remodeling of pial collaterals and functional deficits in a murine model of ischemic stroke

Complete transglutaminase 2 ablation results in reduced stroke volumes and astrocytes that exhibit increased survival in response to ischemia

Contact Info

Product

Resources

About