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1.
Glia ; 59(10): 1489-502, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21638341

RESUMEN

Activation of the unfolded protein response (UPR) is involved in the pathogenesis of numerous CNS myelin abnormalities; yet, its direct role in traumatic spinal cord injury (SCI)-induced demyelination is not known. The UPR is an evolutionarily conserved cell defense mechanism initiated to restore endoplasmic reticulum homeostasis in response to various cellular stresses including infection, trauma, and oxidative damage. However, if uncompensated, the UPR triggers apoptotic cell death. We demonstrate that the three signaling branches of UPR including the PERK, ATF6, and IRE1α are rapidly initiated in a mouse model of contusive SCI specifically at the injury epicenter. Immunohistochemical analyses of the various UPR markers revealed that in neurons, the UPR appeared at 6 and 24-h post-SCI. In contrast, in oligodendrocytes and astroglia, UPR persisted at least for up to 3 days post-SCI. The UPR-associated proapoptotic transcriptional regulator CHOP was among the UPR markers upregulated in neurons and oligodendrocytes, but not in astrocytes, of traumatized mouse spinal cords. To directly analyze its role in SCI, WT and CHOP null mice received a moderate T9 contusive injury. Deletion of CHOP led to an overall attenuation of the UPR after contusive SCI. Furthermore, analyses of hindlimb locomotion demonstrated a significant functional recovery that correlated with an increase in white-matter sparing, transcript levels of myelin basic protein, and Claudin 11 and decreased oligodendrocyte apoptosis in CHOP null mice in contrast to WT animals. Thus, our study provides evidence that the UPR contributes to oligodendrocyte loss after traumatic SCI.


Asunto(s)
Estrés del Retículo Endoplásmico/fisiología , Recuperación de la Función/fisiología , Traumatismos de la Médula Espinal/fisiopatología , Regulación hacia Arriba/fisiología , Factor de Transcripción Activador 4/metabolismo , Análisis de Varianza , Animales , Apoptosis/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Caspasa 3/genética , Caspasa 3/metabolismo , Proteínas de Unión al ADN/metabolismo , Modelos Animales de Enfermedad , Estrés del Retículo Endoplásmico/genética , Factor 2 Eucariótico de Iniciación/metabolismo , Femenino , Locomoción/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína Básica de Mielina/genética , Proteína Básica de Mielina/metabolismo , Fibras Nerviosas Mielínicas/metabolismo , Fibras Nerviosas Mielínicas/patología , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Factor de Transcripción 2 de los Oligodendrocitos , Oligodendroglía/metabolismo , Oligodendroglía/patología , ARN Mensajero/metabolismo , Factores de Transcripción del Factor Regulador X , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/patología , Factores de Tiempo , Factor de Transcripción CHOP/deficiencia , Factores de Transcripción/metabolismo , Respuesta de Proteína Desplegada/genética , Respuesta de Proteína Desplegada/fisiología , Regulación hacia Arriba/genética
2.
Brain ; 133(Pt 4): 1026-42, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20375135

RESUMEN

Blood vessel loss and inflammation cause secondary degeneration following spinal cord injury. Angiopoietin-1 through the Tie2 receptor, and other ligands through alphavbeta3 integrin, promote endothelial cell survival during developmental or tumour angiogenesis. Here, daily intravenous injections with an alphavbeta3-binding peptide named C16 or an angiopoietin-1 mimetic following a spinal cord contusion at thoracic level 9 in mice rescued epicentre blood vessels, white matter and locomotor function, and reduced detrimental inflammation. Preserved vascularity and reduced inflammation correlated with improved outcomes. C16 and angiopoietin-1 reduced leukocyte transmigration in vitro. Growth factor receptors and integrins facilitate each others' function. Therefore, angiopoietin-1 and C16 were combined and the effects were additive, resulting in almost complete functional recovery. The treatment had lasting effects when started 4 h following injury and terminated after one week. These results identify alphavbeta3 integrin and the endothelial-selective angiopoietin-1 as vascular and inflammatory regulators that can be targeted in a clinically relevant manner for neuroprotection after central nervous system trauma.


Asunto(s)
Angiopoyetina 1/administración & dosificación , Integrina alfaVbeta3/administración & dosificación , Fármacos Neuroprotectores/administración & dosificación , Fragmentos de Péptidos/administración & dosificación , Traumatismos de la Médula Espinal/prevención & control , Médula Espinal/irrigación sanguínea , Médula Espinal/efectos de los fármacos , Secuencia de Aminoácidos , Animales , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Quimioterapia Combinada , Femenino , Humanos , Inyecciones Intravenosas , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Recuperación de la Función/efectos de los fármacos , Recuperación de la Función/fisiología , Traumatismos de la Médula Espinal/fisiopatología , Vértebras Torácicas , Factores de Tiempo
3.
Microcirculation ; 17(7): 557-67, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21040121

RESUMEN

UNLABELLED: We have demonstrated that MFs isolated from adipose retain angiogenic potential in vitro and form a mature, perfused network when implanted. However, adipose-derived microvessels are rich in provascularizing cells that could uniquely drive neovascularization in adipose-derived MFs implants. OBJECTIVE: Investigate the ability of MFs from a different vascular bed to recapitulate adipose-derived microvessel angiogenesis and network formation and analyze adipose-derived vessel plasticity by assessing whether vessel function could be modulated by astrocyte-like cells. METHODS: MFs were isolated by limited collagenase digestion from rodent brain or adipose and assembled into 3D collagen gels in the presence or absence of GRPs. The resulting neovasculatures that formed following implantation were assessed by measuring 3D vascularity and vessel permeability to small and large molecular tracers. RESULTS: Similar to adipose-derived MFs, brain-derived MFs can sprout and form a perfused neovascular network when implanted. Furthermore, when co-implanted in the constructs, GRPs caused adipose-derived vessels to express the brain endothelial marker glucose transporter-1 and to significantly reduce microvessel permeability. CONCLUSION: Neovascularization involving isolated microvessel elements is independent of the tissue origin and degree of vessel specialization. In addition, adipose-derived vessels have the ability to respond to environmental signals and change vessel characteristics.


Asunto(s)
Microvasos/crecimiento & desarrollo , Microvasos/trasplante , Neovascularización Fisiológica , Adipocitos/citología , Adipocitos/trasplante , Animales , Astrocitos/citología , Permeabilidad Capilar , Separación Celular , Corteza Cerebral/irrigación sanguínea , Células Madre Embrionarias/citología , Células Madre Embrionarias/trasplante , Epidídimo/irrigación sanguínea , Epidídimo/citología , Técnicas In Vitro , Masculino , Microvasos/citología , Microvasos/fisiología , Neuroglía/citología , Neuroglía/trasplante , Ratas
4.
J Med Chem ; 61(21): 9691-9721, 2018 11 08.
Artículo en Inglés | MEDLINE | ID: mdl-30289716

RESUMEN

HIF-2α, a member of the HIF family of transcription factors, is a key oncogenic driver in cancers such as clear cell renal cell carcinoma (ccRCC). A signature feature of these cancers is the overaccumulation of HIF-2α protein, often by inactivation of the E3 ligase VHL (von Hippel-Lindau). Herein we disclose our structure based drug design (SBDD) approach that culminated in the identification of PT2385, the first HIF-2α antagonist to enter clinical trials. Highlights include the use of a putative n → π*Ar interaction to guide early analog design, the conformational restriction of an essential hydroxyl moiety, and the remarkable impact of fluorination near the hydroxyl group. Evaluation of select compounds from two structural classes in a sequence of PK/PD, efficacy, PK, and metabolite profiling identified 10i (PT2385, luciferase EC50 = 27 nM) as the clinical candidate. Finally, a retrospective crystallographic analysis describes the structural perturbations necessary for efficient antagonism.


Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/antagonistas & inhibidores , Carcinoma de Células Renales/patología , Diseño de Fármacos , Indanos/química , Indanos/farmacología , Neoplasias Renales/patología , Sulfonas/química , Sulfonas/farmacología , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/química , Línea Celular Tumoral , Perros , Indanos/farmacocinética , Ratones , Modelos Moleculares , Conformación Proteica , Ratas , Relación Estructura-Actividad , Sulfonas/farmacocinética , Distribución Tisular
5.
Cancer Res ; 76(18): 5491-500, 2016 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-27635045

RESUMEN

More than 90% of clear cell renal cell carcinomas (ccRCC) exhibit inactivation of the von Hippel-Lindau (pVHL) tumor suppressor, establishing it as the major underlying cause of this malignancy. pVHL inactivation results in stabilization of the hypoxia-inducible transcription factors, HIF1α and HIF2α, leading to expression of a genetic program essential for the initiation and progression of ccRCC. Herein, we describe the potent, selective, and orally active small-molecule inhibitor PT2385 as a specific antagonist of HIF2α that allosterically blocks its dimerization with the HIF1α/2α transcriptional dimerization partner ARNT/HIF1ß. PT2385 inhibited the expression of HIF2α-dependent genes, including VEGF-A, PAI-1, and cyclin D1 in ccRCC cell lines and tumor xenografts. Treatment of tumor-bearing mice with PT2385 caused dramatic tumor regressions, validating HIF2α as a pivotal oncogenic driver in ccRCC. Notably, unlike other anticancer agents that inhibit VEGF receptor signaling, PT2385 exhibited no adverse effect on cardiovascular performance. Thus, PT2385 represents a novel class of therapeutics for the treatment of RCC with potent preclincal efficacy as well as improved tolerability relative to current agents that target the VEGF pathway. Cancer Res; 76(18); 5491-500. ©2016 AACR.


Asunto(s)
Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/antagonistas & inhibidores , Carcinoma de Células Renales/patología , Neoplasias Renales/patología , Animales , Antineoplásicos/química , Calorimetría , Línea Celular Tumoral , Cristalografía por Rayos X , Humanos , Inmunohistoquímica , Inmunoprecipitación , Ratones , Ratones SCID , Reacción en Cadena de la Polimerasa , Ensayos Antitumor por Modelo de Xenoinjerto
6.
J Neurotrauma ; 29(3): 579-88, 2012 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-21933012

RESUMEN

Manipulation of various components of the endoplasmic reticulum (ER) stress response (ERSR) has led to functional recovery in diabetes, cancer, and several neurodegenerative diseases, indicating its use as a potential therapeutic intervention. One of the downstream pro-apoptotic transcription factors activated by the ERSR is CCAAT enhancer binding protein (C/EBP) homologous protein (CHOP). Recently, we showed significant recovery in hindlimb locomotion function after moderate contusive spinal cord injury (SCI) in mice null for CHOP. However, more than 40% of human SCI are complete. Thus the present study examined the potential therapeutic modulation of CHOP in a more severe SCI injury. Contused wild-type spinal cords showed a rapid activation of PERK, ATF6, and IRE-1, the three arms of the ERSR signaling pathway, specifically at the injury epicenter. Confocal images of phosphorylated EIF2α, GRP78, CHOP, ATF4, and GADD34 localized the activation of the ERSR in neurons and oligodendrocytes at the injury epicenter. To directly determine the role of CHOP, wild-type and CHOP-null mice with severe contusive SCI were analyzed for improvement in hindlimb locomotion. Despite the loss of CHOP, the other effectors in the ERSR pathway were significantly increased beyond that observed previously with moderate injury. Concomitantly, Basso Mouse Scale (BMS) scores and white matter sparing between the wild-type and CHOP-null mice revealed no significant differences. Given the complex pathophysiology of severe SCI, ablation of CHOP alone is not sufficient to rescue functional deficits. These data raise the caution that injury severity may be a key variable in attempting to translate preclinical therapies to clinical practice.


Asunto(s)
Apoptosis/fisiología , Retículo Endoplásmico/fisiología , Locomoción/fisiología , Traumatismos de la Médula Espinal/fisiopatología , Factor de Transcripción CHOP/fisiología , Animales , Conducta Animal/fisiología , Western Blotting , Claudinas , Contusiones/patología , Contusiones/fisiopatología , Chaperón BiP del Retículo Endoplásmico , Femenino , Proteínas de Choque Térmico/metabolismo , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína Básica de Mielina/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteína Fosfatasa 1/metabolismo , ARN/biosíntesis , ARN/aislamiento & purificación , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Médula Espinal/patología , Traumatismos de la Médula Espinal/patología , Factor de Transcripción CHOP/genética
7.
Cell Metab ; 15(6): 827-37, 2012 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-22682223

RESUMEN

Dysregulated metabolism is a hallmark of cancer cell lines, but little is known about the fate of glucose and other nutrients in tumors growing in their native microenvironment. To study tumor metabolism in vivo, we used an orthotopic mouse model of primary human glioblastoma (GBM). We infused (13)C-labeled nutrients into mice bearing three independent GBM lines, each with a distinct set of mutations. All three lines displayed glycolysis, as expected for aggressive tumors. They also displayed unexpected metabolic complexity, oxidizing glucose via pyruvate dehydrogenase and the citric acid cycle, and using glucose to supply anaplerosis and other biosynthetic activities. Comparing the tumors to surrounding brain revealed obvious metabolic differences, notably the accumulation of a large glutamine pool within the tumors. Many of these same activities were conserved in cells cultured ex vivo from the tumors. Thus GBM cells utilize mitochondrial glucose oxidation during aggressive tumor growth in vivo.


Asunto(s)
Neoplasias Encefálicas/metabolismo , Glioblastoma/metabolismo , Glucosa/metabolismo , Mitocondrias/metabolismo , Animales , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Glioblastoma/genética , Glioblastoma/patología , Gluconeogénesis , Glutamato-Amoníaco Ligasa/metabolismo , Ácido Glutámico/metabolismo , Glutaminasa/metabolismo , Glutamina/metabolismo , Glucólisis , Humanos , Ratones , Ratones Endogámicos NOD , Ratones SCID , Trasplante de Neoplasias , Neostriado/metabolismo , Neostriado/patología , Oxidación-Reducción , Fenotipo , Piruvato Carboxilasa/metabolismo , Estadísticas no Paramétricas , Células Tumorales Cultivadas
8.
J Comp Neurol ; 512(2): 243-55, 2009 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-19003792

RESUMEN

Endothelial cell (EC) loss and subsequent angiogenesis occur over the first week after spinal cord injury (SCI). To identify molecular mechanisms that could be targeted with intravenous (i.v.) treatments, we determined whether transmembrane "a disintegrin and metalloprotease" (ADAM) proteins are expressed in ECs of the injured spinal cord. ADAMs bind to integrins, which are important for EC survival and angiogenesis. Female adult C57Bl/6 mice with a spinal cord contusion had progressively more ADAM8 (CD156) immunostaining in blood vessels and individual ECs between 1 and 28 days following injury. Uninjured spinal cords had little ADAM8 staining. The increase in ADAM8 mRNA and protein was confirmed in spinal cord lysates, and ADAM8 mRNA was present in FACS-enriched ECs. ADAM8 colocalized extensively and exclusively with the EC marker PECAM and also with i.v.-injected lectins. Intravenous isolectin B4 (IB4) labels a subpopulation of blood vessels at and within the injury epicenter 3-7 days after injury, coincident with angiogenesis. Both ADAM8 and the proliferation marker Ki-67 were present in IB4-positive microvessels. ADAM8-positive proliferating cells were seen at the leading end of IB4-positive blood vessels. Angiogenesis was confirmed by BrdU incorporation, binding of i.v.-injected nucleolin antibodies, and MT1-MMP immunostaining in a subset of blood vessels. These data suggest that ADAM8 is vascular selective and plays a role in proliferation and/or migration of ECs during angiogenesis following SCI.


Asunto(s)
Proteínas ADAM/metabolismo , Antígenos CD/metabolismo , Células Endoteliales/metabolismo , Proteínas de la Membrana/metabolismo , Neovascularización Fisiológica/fisiología , Traumatismos de la Médula Espinal/metabolismo , Adulto , Animales , Biomarcadores/metabolismo , Vasos Sanguíneos/anatomía & histología , Vasos Sanguíneos/metabolismo , Células Endoteliales/citología , Femenino , Humanos , Ratones , Ratones Endogámicos C57BL , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Médula Espinal/citología , Médula Espinal/metabolismo , Médula Espinal/patología , Traumatismos de la Médula Espinal/patología
9.
Curr Neurovasc Res ; 6(2): 124-31, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-19442162

RESUMEN

Acute loss of spinal cord vascularity followed by an endogenous adaptive angiogenic response with concomitant microvascular dysfunction is a hallmark of traumatic spinal cord injury (SCI). Recently, the potent vasoactive factor vascular endothelial growth factor (VEGF) has received much attention as a putative therapeutic for the treatment of various neurodegenerative disorders, including SCI. Exogenous VEGF exerts both protective and destabilizing effects on microvascular elements and tissue following SCI but the role of endogenous VEGF is unclear. In the present study, we systemically applied a potent and well characterized soluble VEGF antagonist to adult C57Bl/6 mice post-SCI to elucidate the relative contribution of VEGF on the acute evolving microvascular response and its impact on functional recovery. While the VEGF Trap did not alter vascular density in the injury epicenter or penumbra, an overall increase in the number of Griffonia simplicifolia isolectin-B4 bound microvessels was observed, suggesting a VEGF-dependency to more subtle aspects of endothelial plasticity post-SCI. Neutralizing endogenous VEGF neither attenuated nor exacerbated chronic histopathology or functional recovery. These results support the idea that overall, endogenous VEGF is not neuroprotective or detrimental following traumatic SCI. Furthermore, they suggest that angiogenesis in traumatically injured spinal tissue is regulated by multiple effectors and is not limited by endogenous VEGF activation of affected spinal microvessels.


Asunto(s)
Microvasos/efectos de los fármacos , Regeneración Nerviosa/efectos de los fármacos , Recuperación de la Función/efectos de los fármacos , Traumatismos de la Médula Espinal/patología , Traumatismos de la Médula Espinal/fisiopatología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Análisis de Varianza , Animales , Modelos Animales de Enfermedad , Femenino , Inmunoglobulina G/uso terapéutico , Lectinas/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Microvasos/patología , Receptores de Factores de Crecimiento Endotelial Vascular , Proteínas Recombinantes de Fusión/uso terapéutico , Recuperación de la Función/fisiología , Traumatismos de la Médula Espinal/tratamiento farmacológico , Factores de Tiempo , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Factor A de Crecimiento Endotelial Vascular/inmunología , Receptor 2 de Factores de Crecimiento Endotelial Vascular/uso terapéutico
10.
ASN Neuro ; 1(3)2009 Aug 26.
Artículo en Inglés | MEDLINE | ID: mdl-19663807

RESUMEN

Microvascular dysfunction, loss of vascular support, ischaemia and sub-acute vascular instability in surviving blood vessels contribute to secondary injury following SCI (spinal cord injury). Neither the precise temporal profile of the cellular dynamics of spinal microvasculature nor the potential molecular effectors regulating this plasticity are well understood. TGFß (transforming growth factor ß) isoforms have been shown to be rapidly increased in response to SCI and CNS (central nervous system) ischaemia, but no data exist regarding their contribution to microvascular dysfunction following SCI. To examine these issues, in the present study we used a model of focal spinal cord ischaemia/reperfusion SCI to examine the cellular response(s) of affected microvessels from 30 min to 14 days post-ischaemia. Spinal endothelial cells were isolated from affected tissue and subjected to focused microarray analysis of TGFß-responsive/related mRNAs 6 and 24 h post-SCI. Immunohistochemical analyses of histopathology show neuronal disruption/loss and astroglial regression from spinal microvessels by 3 h post-ischaemia, with complete dissolution of functional endfeet (loss of aquaporin-4) by 12 h post-ischaemia. Coincident with this microvascular plasticity, results from microarray analyses show 9 out of 22 TGFß-responsive mRNAs significantly up-regulated by 6 h post-ischaemia. Of these, serpine 1/PAI-1 (plasminogen-activator inhibitor 1) demonstrated the greatest increase (>40-fold). Furthermore, uPA (urokinase-type plasminogen activator), another member of the PAS (plasminogen activator system), was also significantly increased (>7.5-fold). These results, along with other select up-regulated mRNAs, were confirmed biochemically or immunohistochemically. Taken together, these results implicate TGFß as a potential molecular effector of the anatomical and functional plasticity of microvessels following SCI.


Asunto(s)
Células Endoteliales/metabolismo , Microvasos/fisiología , Daño por Reperfusión/metabolismo , Isquemia de la Médula Espinal/metabolismo , Activación Transcripcional/fisiología , Factor de Crecimiento Transformador beta/biosíntesis , Animales , Células Endoteliales/patología , Femenino , Microvasos/patología , Plasticidad Neuronal/fisiología , ARN Mensajero/biosíntesis , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/patología , Isquemia de la Médula Espinal/patología
11.
J Cereb Blood Flow Metab ; 28(11): 1771-85, 2008 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-18612314

RESUMEN

Microvascular dysfunction is a critical pathology that underlies the evolution of secondary injury mechanisms after traumatic spinal cord injury (SCI). However, little is known of the molecular regulation of endothelial cell (EC) plasticity observed acutely after injury. One reason for this is the relative lack of methods to quickly and efficiently obtain highly enriched spinal microvascular ECs for high-throughput molecular and biochemical analyses. Adult C57Bl/6 mice received an intravenous injection of fluorescein isothiocyanate (FITC)-conjugated Lycopersicon esculentum lectin, and FITC-lectin-bound spinal microvessels were greatly enriched by fluorescence-activated cell sorter (FACS) purification. This technique allows for rapid (<1.5 h postmortem) isolation of spinal cord microvascular ECs (smvECs). The results from cell counting, reverse-transcription polymerase chain reaction (RT-PCR), and western blot analyses show a high degree of EC enrichment at mRNA and protein levels. Furthermore, a focused EC biology microarray analysis identified multiple mRNAs dramatically increased in the EC compartment 24 h after SCI, which is a time point associated with the pathologic loss of spinal vasculature. These included thrombospondin-1, CCL5/RANTES, and urokinase plasminogen activator, suggesting they may represent targets for therapeutic intervention. Furthermore, these novel methodologic approaches will likely facilitate the discovery of molecular regulators of endothelial dysfunction in a variety of central nervous system (CNS) disorders including stroke and other neurodegenerative diseases having a vascular component.


Asunto(s)
Endotelio Vascular/fisiopatología , Perfilación de la Expresión Génica , Microcirculación/fisiología , ARN Mensajero/genética , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/fisiopatología , Médula Espinal/irrigación sanguínea , Transcripción Genética , Animales , Anexinas/genética , Endotelio Vascular/patología , Femenino , Fibrinolisina/genética , Citometría de Flujo , Regulación de la Expresión Génica , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Microcirculación/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Lectinas de Plantas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Trombospondinas/genética , Activador de Plasminógeno de Tipo Uroquinasa/genética
12.
J Comp Neurol ; 507(1): 1031-52, 2008 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-18092342

RESUMEN

After traumatic spinal cord injury (SCI), disruption and plasticity of the microvasculature within injured spinal tissue contribute to the pathological cascades associated with the evolution of both primary and secondary injury. Conversely, preserved vascular function most likely results in tissue sparing and subsequent functional recovery. It has been difficult to identify subclasses of damaged or regenerating blood vessels at the cellular level. Here, adult mice received a single intravenous injection of the Griffonia simplicifolia isolectin B4 (IB4) at 1-28 days following a moderate thoracic (T9) contusion. Vascular binding of IB4 was maximally observed 7 days following injury, a time associated with multiple pathologic aspects of the intrinsic adaptive angiogenesis, with numbers of IB4 vascular profiles decreasing by 21 days postinjury. Quantitative assessment of IB4 binding shows that it occurs within the evolving lesion epicenter, with affected vessels expressing a temporally specific dysfunctional tight junctional phenotype as assessed by occludin, claudin-5, and ZO-1 immunoreactivities. Taken together, these results demonstrate that intravascular lectin delivery following SCI is a useful approach not only for observing the functional status of neovascular formation but also for definitively identifying specific subpopulations of reactive spinal microvascular elements.


Asunto(s)
Ratones Endogámicos C57BL/fisiología , Neovascularización Fisiológica/fisiología , Lectinas de Plantas , Traumatismos de la Médula Espinal/fisiopatología , Columna Vertebral/irrigación sanguínea , Adaptación Fisiológica/fisiología , Factores de Edad , Animales , Capilares/citología , Capilares/fisiología , Capilares/ultraestructura , División Celular/fisiología , Endotelio Vascular/citología , Endotelio Vascular/fisiología , Endotelio Vascular/ultraestructura , Femenino , Ratones , Microscopía Inmunoelectrónica , Traumatismos de la Médula Espinal/patología , Columna Vertebral/patología , Uniones Estrechas/fisiología
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