RESUMEN
During capillary network formation, ECs establish interconnecting tubes with defined lumens that reside within vascular guidance tunnels (physical spaces generated during EC tubulogenesis). Pericytes are recruited to EC tubes within these tunnels and capillary basement membrane deposition occurs to facilitate tube maturation. Here, we discuss molecular mechanisms controlling EC tubulogenesis demonstrating the involvement of integrins, MT1-MMP, extracellular matrix, Cdc42, Rac1, Rac2, k-Ras, Rap1b, and key downstream effectors including Pak2, Pak4, IQGAP1, MRCKß, and Rasip1. These molecules activate kinase cascades controlling EC tube formation, in conjunction with growth factor receptor signaling, which involve PKCÉ, Src family, Raf, Mek, and Erk kinases. These molecules and signaling cascades stimulate EC lumen and tube formation by: regulating MT-MMP-dependent lumen expansion and vascular guidance tunnel formation; generation of intracellular vacuoles/vesicles to create EC apical membranes; and establishing cytoskeletal polarity with acetylated tubulin distributed subapically (and F-actin basally) to facilitate vacuole trafficking/fusion in a polarized, perinuclear region. Using defined serum-free models, we have demonstrated that human EC tubulogenesis and EC-pericyte tube coassembly requires five exogenously applied growth factors which are SCF, IL-3, SDF-1α, FGF-2, and insulin (Factors). Also, we have demonstrated that EC-derived PDGF-BB and HB-EGF are necessary for pericytes to proliferate, recruit to tubes, and induce basement membrane assembly. Finally, we have shown that VEGF fails to directly stimulate EC tubulogenesis. In contrast, it acts as an upstream EC primer of downstream "Factor"-induced tubulogenic and EC-pericyte tube coassembly by upregulating c-Kit, IL-3Rα, and CXCR4 as well as PDGF-BB and HB-EGF expression.
Asunto(s)
Matriz Extracelular/metabolismo , Morfogénesis/fisiología , Pericitos/metabolismo , Animales , Becaplermina , Humanos , Proteínas Proto-Oncogénicas c-sis/metabolismo , Transducción de Señal/fisiologíaRESUMEN
Fever is initiated by activation of the arachidonic acid cascade and the biosynthesis of prostaglandins within the brain. Inducible cyclooxygenase (COX-2) is a rate-limiting enzyme in prostaglandin synthesis, and the number of blood vessels expressing COX-2 correlates with elevated body temperature following peripheral lipopolysaccharide (LPS). Despite its importance in host defense, fever is energetically expensive and we hypothesized that fever may be limited by available metabolic resources. During winter, when competing metabolic demands are constrained by low temperatures and food availability, it was predicted that fever duration would be reduced in seasonally breeding Siberian hamsters (Phodopus sungorus). We measured LPS-induced COX-2 expression in blood vessels of hamsters to test whether photoperiodic alterations in fever duration are centrally mediated, or whether they reflect changes in peripheral modulation of body temperature. Hamsters housed in long, 'summer-like' or short, 'winter-like' day lengths for 10 weeks were injected with LPS, and brains were collected 2, 4, or 8 h later. COX-2 expression was comparably increased in long- and short-day hamsters by 2 h and 4 h post-LPS; however, short-day hamsters exhibited significantly fewer COX-2-positive cells and blood vessels by 8 h post-LPS compared to long-day hamsters, corresponding with reduced fever duration in short-day hamsters. Cortisol concentrations increased more than two-fold in short-day compared to long-day hamsters by 4 h; this increase may have contributed to the decrease in COX-2 expression observed by 8 h in short days. We conclude that short photoperiods significantly altered the time course of central COX-2 protein expression in hamsters in a manner consistent with reduced fever duration.