scholarly journals Transport of α-Aminoisobutyric Acid across Brain Capillary and Cellular Membranes

1983 ◽  
Vol 3 (1) ◽  
pp. 8-32 ◽  
Author(s):  
Ronald G. Blasberg ◽  
Joseph D. Fenstermacher ◽  
Clifford S. Patlak
Keyword(s):  
Brain Tissue ◽  
Rate Constants ◽  
Transfer Rate ◽  
Brain Cell ◽  
Brain Cells ◽  
Normal Brain ◽  
Multiple Time ◽  
Brain Capillaries ◽  
Aminoisobutyric Acid ◽  
Blood Brain

The transport of α-aminoisobutyric acid (AIB), N-methyl-AIB (MeAIB), and diethylenetriaminepentaacetic acid (DTPA) from blood to brain was measured over different experimental periods in eight regions of the rat brain. Unidirectional transfer rate constants were determined from multiple-time/graphical and single-time analysis of the experimental data; values of 0,0018, 0,00057, and 0,000021 ml g−1 min−1, respectively, were obtained for the thalamus by graphical analysis, The initial distribution volume of AIB and MeAIB in brain tissue was several-fold greater than that of DTPA and the tissue plasma volume, and this difference was not accounted for by red blood cell uptake, This discrepancy could be due to rapid transport of AIB and MeAIB into brain endothelial cells in addition to the relatively rapid uptake by choroidal, meningeal, and ependymal associated tissues that was demonstrated by autoradiography. Thus, it may be misleading and erroneous to consider the blood–brain barrier (BBB) to be a simple, single-membrane structure when analyzing the blood–brain transfer data of solutes such as amino acids. The data from the ventriculocisternal perfusion experiments and previously published AIB uptake data in mouse brain slices were used to estimate the transfer rate constants across brain cell membranes. These studies indicated that the transport of AIB into brain cells was approximately 110 to 265 times greater than that across normal brain capillaries per unit mass of brain tissue, and that the BBB limits blood-to-brain cell transport of this amino acid. These observations (low rate of transport across normal brain capillaries and rapid concentrative uptake by brain cells) indicate that AIB is a good marker for measuring moderate to large increases in BBB permeability by experiments that require unidirectional flux of the tracer.

Intracarotid Infusion of Leukotriene C4 Selectively Increases Blood-Brain Barrier Permeability after Focal Ischemia in Rats

1991 ◽  
Vol 11 (4) ◽  
pp. 638-643 ◽  
Author(s):  
Takehiko Baba ◽  
Keith L. Black ◽  
Kiyonobu Ikezaki ◽  
Kangnian Chen ◽  
Donald P. Becker
Keyword(s):  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Normal Brain ◽  
Brain Capillaries ◽  
Transfer Constant ◽  
Blood Brain ◽  
Mca Occlusion ◽  
Ischemic Tissue ◽  
Bbb Permeability ◽  
Glutamyl Transpeptidase

Intracarotid infusions of leukotriene C4 (LTC4) were used to open selectively the blood–brain barrier (BBB) in ischemic tissue after middle cerebral artery (MCA) occlusion in rats. BBB permeability was determined by quantitative autoradiography using [14C]aminoisobutyric acid. Seventy-two hours after MCA occlusion, LTC4 (4 μg total dose) infused into the carotid artery ipsilateral to the MCA occlusion selectively increased the unidirectional transfer constant for permeability K1 approximately threefold within core ischemic tissue and tissue adjacent to the ischemic core. No effect on BBB permeability was seen within nonischemic brain tissue or in ischemic tissue after only 24 h after MCA occlusion. γ-Glutamyl transpeptidase (γ-GTP) activity was decreased in capillaries in ischemic tissue at 48 and 72 h after infarction, compared to high γ-GTP in normal brain capillaries and moderate γ-GTP in capillaries in the ischemic tissue at 24 h after infarction. These findings suggest that normal brain capillaries resist the vasogenic effects of LTC4. In contrast, LTC4 increases permeability in capillaries of ischemic tissue, where γ-GTP is decreased. γ-Glutamyl transpeptidase, an enzyme that inactivates LTC4 to LTD4 and LTE4 to LTF4, may act as an “enzymatic barrier” in normal brain capillaries to leukotrienes.

Abstract 19922: Phosphatidylserine Receptor Upregulation in Stroke for Targeted Carrier Delivery

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Tao Peng ◽  
David D McPherson ◽  
Shao-Ling Huang
Keyword(s):  
Endothelial Cells ◽  
Cerebral Ischemia ◽  
Brain Tissue ◽  
Umbilical Vein ◽  
Brain Cells ◽  
Normal Brain ◽  
Necrosis Factor Alpha ◽  
Stroke Treatment ◽  
Early Stages ◽  
Phosphatidylserine Receptor

Introduction: The phosphatidylserine receptor (PSR) plays an important role in apoptosis and inflammation. Whether this receptor is expressed in damaged brain cells in the early stages of stroke is unknown. Hypothesis: The purpose of this study was to determine if this receptor is expressed on ischemic neurovascular endothelial cells and if so, can it bind with PS-containing liposomes (PS-liposomes). The latter would allow for a neurovascular targeting strategy for therapeutic delivery to ischemic brain cells. Methods: Sprague Dawley rats (n=22) were randomly divided into sham, cerebral ischemia, and subarachnoid hemorrhage (SAH) injury groups. Cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 2 hours using an intraluminal suture method. SAH was induced by an endovascular perforation method. Brain tissue was harvested for immunofluorescence staining at day 2 after surgery. Cultured human umbilical vein endothelial cells (HUVEC) were pre-treated with tumor necrosis factor alpha (TNFα) to stimulated PSR. Fluorescence labeled PS-containing liposomes (FITC-PS-Liposomes) were used. The PSR expression and the adhesion of liposomes were observed using fluorescence microscopy. Results: Normal brain tissue did not demonstrate PSR expression (Fig.1A). After brain injury, PSR expression was observed in the injured area and colocalized with the endothelium both in the MCAO (Fig.1B) and SAH animals (Fig.1C). Normal HUVEC did not demonstrate fluorescence (Fig.1D). Weak fluorescence was observed in normal cells that incorporated the PS-liposomes (Fig.1E). Strong fluorescence was observed in the TNFα-activated HUVEC that incorporated the FITC-PS-Liposomes (Fig.1F). Conclusions: This is first report that PSR expression occurs in cerebral microvascular endothelial cells in the early stages of stroke and can be recognized by PS-liposomes. This may allow development of a targeted liposomal carrier for novel targeted stroke treatment.

scholarly journals Ultrastructural Analysis of ICP34.5− Herpes Simplex Virus 1 Replication in Mouse Brain Cells In Vivo

2010 ◽  
Vol 84 (21) ◽  
pp. 10982-10990 ◽  
Author(s):  
Hina Mehta ◽  
Jacqueline Muller ◽  
Nancy S. Markovitz
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Tumor Cells ◽  
Brain Cell ◽  
Brain Cells ◽  
Normal Brain ◽  
Herpes Simplex Virus 1 ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Replication-competent forms of herpes simplex virus 1 (HSV-1) defective in the viral neurovirulence factor infected cell protein 34.5 (ICP34.5) are under investigation for use in the therapeutic treatment of cancer. In mouse models, intratumoral injection of ICP34.5-defective oncolytic HSVs (oHSVs) has resulted in the infection and lysis of tumor cells, an associated decrease in tumor size, and increased survival times. The ability of these oHSVs to infect and lyse cells is frequently characterized as exclusive to or selective for tumor cells. However, the extent to which ICP34.5-deficient HSV-1 replicates in and may be neurotoxic to normal brain cell types in vivo is poorly understood. Here we report that HSV-1 defective in ICP34.5 expression is capable of establishing a productive infection in at least one normal mouse brain cell type. We show that γ34.5 deletion viruses replicate productively in and induce cellular damage in infected ependymal cells. Further evaluation of the effects of oHSVs on normal brain cells in animal models is needed to enhance our understanding of the risks associated with the use of current and future oHSVs in the brains of clinical trial subjects and to provide information that can be used to create improved oHSVs for future use.

scholarly journals Enzymatic Dissociation Induces Transcriptional and Proteotype Bias in Brain Cell Populations

2020 ◽  
Vol 21 (21) ◽  
pp. 7944
Author(s):  
Daniele Mattei ◽  
Andranik Ivanov ◽  
Marc van Oostrum ◽  
Stanislav Pantelyushin ◽  
Juliet Richetto ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Flow Cytometric Analysis ◽  
Enzymatic Digestion ◽  
Systematic Investigation ◽  
Cell Isolation ◽  
Brain Cell ◽  
Brain Cells ◽  
Enzymatic Dissociation ◽  
Isolation Techniques ◽  
Flow Cytometric

Different cell isolation techniques exist for transcriptomic and proteotype profiling of brain cells. Here, we provide a systematic investigation of the influence of different cell isolation protocols on transcriptional and proteotype profiles in mouse brain tissue by taking into account single-cell transcriptomics of brain cells, proteotypes of microglia and astrocytes, and flow cytometric analysis of microglia. We show that standard enzymatic digestion of brain tissue at 37 °C induces profound and consistent alterations in the transcriptome and proteotype of neuronal and glial cells, as compared to an optimized mechanical dissociation protocol at 4 °C. These findings emphasize the risk of introducing technical biases and biological artifacts when implementing enzymatic digestion-based isolation methods for brain cell analyses.

scholarly journals Enzymatic dissociation induces transcriptional and proteotype bias in brain cell populations

2020 ◽  
Author(s):  
Daniele Mattei ◽  
Andranik Ivanov ◽  
Marc van Oostrum ◽  
Stanislav Pantelyushin ◽  
Juliet Richetto ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Flow Cytometric Analysis ◽  
Enzymatic Digestion ◽  
Systematic Investigation ◽  
Cell Isolation ◽  
Brain Cell ◽  
Brain Cells ◽  
Enzymatic Dissociation ◽  
Isolation Techniques ◽  
Flow Cytometric

AbstractDifferent cell isolation techniques exist for transcriptomic and proteotype profiling of brain cells. Here, we provide a systematic investigation of the influence of different cell isolation protocols on transcriptional and proteotype profiles in mouse brain tissue by taking into account single-cell transcriptomics of brain cells, proteotypes of microglia and astrocytes, and flow cytometric analysis of microglia. We show that standard enzymatic digestion of brain tissue at 37°C induces profound and consistent alterations in the transcriptome and proteotype of neuronal and glial cells, as compared to an optimized mechanical dissociation protocol at 4°C. These findings emphasize the risk of introducing technical biases and biological artefacts when implementing enzymatic digestion-based isolation methods for brain cell analyses.

scholarly journals Thrombin Stimulates Activation of the Cerebral 5-Lipoxygenase Pathway during Blood-Brain Cell Contact

1996 ◽  
Vol 16 (4) ◽  
pp. 737-745 ◽  
Author(s):  
Michael Winking ◽  
Rochus M. Heldt ◽  
Thomas Simmet
Keyword(s):  
Rat Brain ◽  
Whole Blood ◽  
Brain Cell ◽  
Thrombin Inhibitors ◽  
Brain Cells ◽  
Cell Contact ◽  
Serum Samples ◽  
Lipoxygenase Pathway ◽  
Blood Brain ◽  
Stimulation Of

The purpose of this study was to identify the trigger mechanism activating the 5-lipoxygenase pathway during blood–brain cell contact and to estimate the contribution of blood and brain cells to the cysteinyl–leukotriene (LT) biosynthesis observed under these conditions. Incubation of dissociated rat brain cells in Krebs-Henseleit solution for up to 60 min did not stimulate any detectable cysteinyl–LT biosynthesis. Incubation of recalcified rat whole blood in vitro for up to 60 min led to release of only small amounts of cysteinyl–LT into the serum samples. However, coincubation of dissociated rat brain cells with physiologically recalcified autologous whole blood triggered a time-dependent release of large amounts of immunoreactive cysteinyl–LT into the serum samples. By reverse-phase HPLC, immunoreactive cysteinyl–LT was identified as a mixture of LTC4, LTD4, and LTE4. The extent of the 5-lipoxygenase stimulation depended on the amount of autologous blood coincubated with the dissociated brain cells. Activation of the 5-lipoxygenase pathway also occurred with coincubation of dissociated rat brain cells with recalcified autologous plasma. Stimulation of cysteinyl–LT biosynthesis during blood–brain cell contact remained unaffected by aprotinin, but concentration-dependent inhibition by the structurally and functionally unrelated thrombin inhibitors D-Phe-Pro-Arg-CH2Cl and recombinant hirudin was seen. Finally, when dissociated rat brain cells were incubated in Krebs-Henseleit solution in the presence of human α-thrombin, a concentration-dependent release of cysteinyl–LT into the buffer samples was observed. These data demonstrate that, in rats, during blood–brain cell contact, stimulation of the 5-lipoxygenase pathway in brain cells proceeds via α-thrombin as effector molecule.

scholarly journals Protein Kinase A Distribution in Meningioma

Cancers ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1686 ◽  
Author(s):  
Caretta ◽  
Denaro ◽  
D’Avella ◽  
Mucignat-Caretta
Keyword(s):  
Brain Tissue ◽  
Catalytic Subunit ◽  
Therapeutic Interventions ◽  
Normal Brain ◽  
Local Concentration ◽  
Correlation Pattern ◽  
Catalytic Subunits ◽  
Tissue Specimens ◽  
Pka Catalytic Subunit

Deregulation of intracellular signal transduction pathways is a hallmark of cancer cells, clearly differentiating them from healthy cells. Differential intracellular distribution of the cAMP-dependent protein kinases (PKA) was previously detected in cell cultures and in vivo in glioblastoma and medulloblastoma. Our goal is to extend this observation to meningioma, to explore possible differences among tumors of different origins and prospective outcomes. The distribution of regulatory and catalytic subunits of PKA has been examined in tissue specimens obtained during surgery from meningioma patients. PKA RI subunit appeared more evenly distributed throughout the cytoplasm, but it was clearly detectable only in some tumors. RII was present in discrete spots, presumably at high local concentration; these aggregates could also be visualized under equilibrium binding conditions with fluorescent 8-substituted cAMP analogues, at variance with normal brain tissue and other brain tumors. The PKA catalytic subunit showed exactly overlapping pattern to RII and in fixed sections could be visualized by fluorescent cAMP analogues. Gene expression analysis showed that the PKA catalytic subunit revealed a significant correlation pattern with genes involved in meningioma. Hence, meningioma patients show a distinctive distribution pattern of PKA regulatory and catalytic subunits, different from glioblastoma, medulloblastoma, and healthy brain tissue. These observations raise the possibility of exploiting the PKA intracellular pathway as a diagnostic tool and possible therapeutic interventions.

scholarly journals Neuroinflammatory changes of the normal brain tissue in cured mice following combined radiation and anti-PD-1 blockade therapy for glioma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mariano Guardia Clausi ◽  
Alexander M. Stessin ◽  
Zirun Zhao ◽  
Stella E. Tsirka ◽  
Samuel Ryu
Keyword(s):  
Brain Tissue ◽  
Cranial Irradiation ◽  
Subcortical White Matter ◽  
Glioma Cell Line ◽  
Hippocampal Neurogenesis ◽  
Normal Brain ◽  
Long Term Effects ◽  
Normal Brain Tissue ◽  
Molecule Inhibitor

AbstractThe efficacy of combining radiation therapy with immune checkpoint inhibitor blockade to treat brain tumors is currently the subject of multiple investigations and holds significant therapeutic promise. However, the long-term effects of this combination therapy on the normal brain tissue are unknown. Here, we examined mice that were intracranially implanted with murine glioma cell line and became long-term survivors after treatment with a combination of 10 Gy cranial irradiation (RT) and anti-PD-1 checkpoint blockade (aPD-1). Post-mortem analysis of the cerebral hemisphere contralateral to tumor implantation showed complete abolishment of hippocampal neurogenesis, but neural stem cells were well preserved in subventricular zone. In addition, we observed a drastic reduction in the number of mature oligodendrocytes in the subcortical white matter. Importantly, this observation was evident specifically in the combined (RT + aPD-1) treatment group but not in the single treatment arm of either RT alone or aPD-1 alone. Elimination of microglia with a small molecule inhibitor of colony stimulated factor-1 receptor (PLX5622) prevented the loss of mature oligodendrocytes. These results identify for the first time a unique pattern of normal tissue changes in the brain secondary to combination treatment with radiotherapy and immunotherapy. The results also suggest a role for microglia as key mediators of the adverse treatment effect.

Sign in / Sign up

Export Citation Format

Share Document