A specialized vascular niche for adult neural stem cells

Stem cells reside in specialized niches that regulate their self-renewal and differentiation. The vasculature is emerging as an important component of stem cell niches. Here, we show that the adult subventricular zone (SVZ) neural stem cell niche contains an extensive planar vascular plexus that has specialized properties. Dividing stem cells and their transit-amplifying progeny are tightly apposed to SVZ blood vessels both during homeostasis and regeneration. They frequently contact the vasculature at sites that lack astrocyte endfeet and pericyte coverage, a modification of the blood-brain barrier unique to the SVZ. Moreover, regeneration often occurs at these sites. Finally, we find that circulating small molecules in the blood enter the SVZ. Thus, the vasculature is a key component of the adult SVZ neural stem cell niche, with SVZ stem cells and transit-amplifying cells uniquely poised to receive spatial cues and regulatory signals from diverse elements of the vascular system.     

Genetic determinants of hyaloid and retinal vasculature in zebrafish

Background

The retinal vasculature is a capillary network of blood vessels that nourishes the inner retina of most mammals. Developmental abnormalities or microvascular complications in the retinal vasculature result in severe human eye diseases that lead to blindness. To exploit the advantages of zebrafish for genetic, developmental and pharmacological studies of retinal vasculature, we characterised the intraocular vasculature in zebrafish.

Results

We show a detailed morphological and developmental analysis of the retinal blood supply in zebrafish. Similar to the transient hyaloid vasculature in mammalian embryos, vessels are first found attached to the zebrafish lens at 2.5 days post fertilisation. These vessels progressively lose contact with the lens and by 30 days post fertilisation adhere to the inner limiting membrane of the juvenile retina. Ultrastructure analysis shows these vessels to exhibit distinctive hallmarks of mammalian retinal vasculature. For example, smooth muscle actin-expressing pericytes are ensheathed by the basal lamina of the blood vessel, and vesicle vacuolar organelles (VVO), subcellular mediators of vessel-retinal nourishment, are present. Finally, we identify 9 genes with cell membrane, extracellular matrix and unknown identity that are necessary for zebrafish hyaloid and retinal vasculature development.

Conclusion

Zebrafish have a retinal blood supply with a characteristic developmental and adult morphology. Abnormalities of these intraocular vessels are easily observed, enabling application of genetic and chemical approaches in zebrafish to identify molecular regulators of hyaloid and retinal vasculature in development and disease.     

The Ocular Lens Epithelium

An adult lens contains two easily discernible, morphologically distinct compartments, the epithelium and the fiber-cell mass. The fiber-cell mass provides the lens with its functional phenotype, transparency. Metabolically, in comparison to the fiber cells the epithelium is the more active compartment of the ocular lens. For the purposes of this review we will only discuss the surface epithelium that covers the anterior face of the adult ocular lens. This single layer of cells, in addition to acting as a metabolic engine that sustains the physiological health of this tissue, also works as a source of stem cells, providing precursor cells, which through molecular and morphological differentiation give rise to fiber cells. Morphological simplicity, defined developmental history and easy access to the experimenter make this epithelium a choice starting material for investigations that seek to address universal questions of cell growth, development, epithelial function, cancer and aging. There are two important aspects of the lens epithelium that make it highly relevant to the modern biologist. Firstly, there are no known clinically recognizable cancers of the ocular lens. Considering that most of the known malignancies are epithelial in origin this observation is more than an academic curiosity. The lack of vasculature in the lens may explain the absence of tumors in this tissue, but this provides only a teleological basis to a very important question for which the answers must reside in the molecular make-up and physiology of the lens epithelial cells. Secondly, lens epithelium as a morphological entity in the human lens is first recognizable in the 5th–6th week of gestation. It stays in this morphological state as the anterior epithelium of the lens for the rest of the life, making it an attractive paradigm for the study of the effects of aging on epithelial function. What follows is a brief overview of the present status and lacunae in our understanding of the biology of the lens epithelium.     

Lymphatic vasculature development

Although the process of blood vasculature formation has been well documented, little is known about lymphatic vasculature development, despite its importance in normal and pathological conditions. The lack of specific lymphatic markers has hampered progress in this field. However, the recent identification of genes that participate in the formation of the lymphatic vasculature denotes the beginning of a new era in which better diagnoses and therapeutic treatment(s) of lymphatic disorders could become a reachable goal.     

Lama1 mutations lead to vitreoretinal blood vessel formation, persistence of fetal vasculature, and epiretinal membrane formation in mice

Background

Valuable insights into the complex process of retinal vascular development can be gained using models with abnormal retinal vasculature. Two such models are the recently described mouse lines with mutations in Lama1, an important component of the retinal internal limiting membrane (ILM). These mutants have a persistence of the fetal vasculature of vitreous (FVV) but lack a primary retinal vascular plexus. The present study provides a detailed analysis of astrocyte and vascular development in these Lama1 mutants.

Results

Although astrocytes and blood vessels initially migrate into Lama1 mutant retinas, both traverse the peripapillary ILM into the vitreous by P3. Once in the vitreous, blood vessels anastomose with vessels of the vasa hyaloidea propria, part of the FVV, and eventually re-enter the retina where they dive to form the inner and outer retinal capillary networks. Astrocytes continue proliferating within the vitreous to form a dense mesh that resembles epiretinal membranes associated with persistent fetal vasculature and proliferative vitreoretinopathy.

Conclusions

Lama1 and a fully intact ILM are required for normal retinal vascular development. Mutations in Lama1 allow developing retinal vessels to enter the vitreous where they anastomose with vessels of the hyaloid system which persist and expand. Together, these vessels branch into the retina to form fairly normal inner retinal vascular capillary plexi. The Lama1 mutants described in this report are potential models for studying the human conditions persistent fetal vasculature and proliferative vitreoretinopathy.     

Can the circadian system of a diurnal and a nocturnal rodent entrain to ultraviolet light?

Spectral measurements of sunlight throughout the day show close correspondence between the timing of above ground activity of the European ground squirrel and the presence of ultraviolet light in the solar spectrum. However, in a standard entrainment experiment ground squirrels show no entrainment to ultraviolet light, while Syrian hamsters do entrain under the same protocol. Presented transmittance spectra for lenses, corneas, and vitreous bodies may explain the different results of the entrainment experiment. We found ultraviolet light transmittance in the colourless hamster lens (50% cut-off at 341 nm), but not in the yellow ground squirrel lens (50% cut-off around 493 nm). Ultraviolet sensitivity in the ground squirrels based upon possible fluorescence mechanisms was not evident. Possible functions of ultraviolet lens filters in diurnal mammals are discussed, and compared with nocturnal mammals and diurnal birds. Species of the latter two groups lack ultraviolet filtering properties of their lenses and their circadian system is known to respond to ultraviolet light, a feature that does not necessarily has to depend on ultraviolet photoreceptors. Although the circadian system of several species responds to ultraviolet light, we argue that the role of ultraviolet light as a natural Zeitgeber is probably limited.     

Leaky livers, portal shunting and immunity to schistosomes

In mice, concomitant immunity to schistosomes seems to be largely dependent on factors that lack immunological specificity. In this review Alan Wilson describes the anatomical changes stimulated in the host's vasculature after the formation of granulomas around schistosome eggs in the liver. The blockage of portal venules by the granulomas leads to portal hypertension and the development of anastomoses that direct blood flow, schistosomula and eggs from the liver. An intact portal vasculature is required for schistosome maturation; some strains of mice appear to be resistant to schistosome infection because they have a predisposition to form anastomoses in the absence of infection.     

Photoacoustic imaging of the human placental vasculature

Minimally invasive fetal interventions require accurate imaging from inside the uterine cavity. Twin‐to‐twin transfusion syndrome (TTTS), a condition considered in this study, occurs from abnormal vascular anastomoses in the placenta that allow blood to flow unevenly between the fetuses. Currently, TTTS is treated fetoscopically by identifying the anastomosing vessels, and then performing laser photocoagulation. However, white light fetoscopy provides limited visibility of placental vasculature, which can lead to missed anastomoses or incomplete photocoagulation. Photoacoustic (PA) imaging is an alternative imaging method that provides contrast for hemoglobin, and in this study, two PA systems were used to visualize chorionic (fetal) superficial and subsurface vasculature in human placentas. The first system comprised an optical parametric oscillator for PA excitation and a 2D Fabry‐Pérot cavity ultrasound sensor; the second, light emitting diode arrays and a 1D clinical linear‐array ultrasound imaging probe. Volumetric photoacoustic images were acquired from ex vivo normal term and TTTS‐treated placentas. It was shown that superficial and subsurface branching blood vessels could be visualized to depths of approximately 7 mm, and that ablated tissue yielded negative image contrast. This study demonstrated the strong potential of PA imaging to guide minimally invasive fetal therapies.      

Renal connexins and blood pressure

The kidneys are centrally involved in the regulation of blood pressure. Kidney function requires the coordinated actions of a number of different vascular and tubular cell types in the renal vasculature and in the renal tubular system. The intrarenal coordination of these actions is not well understood. Since gap junctions have been identified in the kidneys, possible pathways involved in this context could be direct intercellular communication via gap junctions or via connexin hemichannels. In this context nine different connexins have been found to be expressed in the kidney, either localized to the vasculature or to the tubular system. Evidence is arising that malfunctions of certain connexins have an impact on the capability of the kidney to maintain blood pressure homeostasis. Findings reported in this context will be outlined and discussed in this review. This article is part of a Special Issue entitled: The Communicating junctions, composition, structure and characteristics.     

Autonomic nervous control of the circulatory system in the air-breathing fish Channa argus

The control of the cardiovascular system with particular emphasis on the regulation of blood distribution in the gills and air-breathing organ was studied in the air-breathing teleost Channa argus. Perfused head preparations were used in addition to experiments with isolated strip preparations of arteries and heart chambers. The distribution of adrenergic nerves was investigated using Falck-Hillarp fluorescence histochemistry. This preliminary study shows an adrenergic control system composed of chromaffin cells and adrenergic nerves similar to that found in other teleosts investigated, although the systemic arteries (coeliac artery, dorsal aorta and the vasculature of the air-breathing organ) appear to lack an adrenergic innervation. The reactions of isolated artery strip preparations to acetylcholine and adrenaline resemble those seen in other teleosts, and there is a prominent inhibitory effect of L-isoprenaline suggestive of arterial beta-adrenoceptors. The general vascular resistance of the gill apparatus-air-breathing organ increases in response to acetylcholine or adrenaline, and there is a redistribution of perfusion flow from the air-breathing organ circuit (anterior venous outflow from the first and second pair of gills and the air-breathing organ) to the general systemic circuit (dorsal aortic outflow from the third and fourth pair of gills). Stimulation of the vagal branch entering the air-breathing organ mimics the effects of acetylcholine or adrenaline. This innervation is probably non-adrenergic since no adrenergic nerve fibres could be demonstrated in the vasculature of the air-breathing organ using the histochemical technique. An adrenergic control of the vasculature of the air-breathing organ is not likely, since the concentration of adrenaline needed to affect the vasculature is not reached in the plasma even during "stress".     

The enigmatic role of angiopoietin-1 in tumor angiogenesis

A tumor vasculature is highly unstable and immature, characterized by a high proliferation rate of endothelial cells, hyper-permeability, and chaotic blood flow. The dysfunctional vasculature gives rise to continual plasma leakage and hypoxia in the tumor, resulting in constant on-sets of inflammation and angiogenesis. Tumors are thus likened to wounds that will not heal. The lack of functional mural cells, including pericytes and vascular smooth muscle cells, in tumor vascular structure contributes significantly to the abnormality of tumor vessels. Angiopoietin-1 (Ang 1) is aphysiological angiogenesis promoter during embryonic development. The function of Angl is essential to endothelial cell survival, vascular branching, and pericyte recruitment. However, an increasing amount of experimental data suggest that Angl-stimulated association of mural cells with endothelial cells lead to stabilization of newly formed blood vessels. This in turn may limit the otherwise continuous angiogenesis in the tumor, and consequently give riseto inhibition of tumor growth. We discuss the enigmatic role of Angl in tumor angiogenesis in this review.     

Influence of small molecules on the photo-stability of water soluble porcine lens proteins

The eye lens is a biconvex structure composed of lens fibres, cells that lack of blood and nerve supply and of any organelle, allowing for a high concentration of water soluble proteins that determine the lens transparency and refractive index. The lens water soluble protein pool in mammals is composed of α-, β-, and γ-crystallins, the latter being involved in calcium homeostasis and having structural importance, the first playing a crucial role in preventing protein aggregation and the consequent lens obfuscation, which leads to the clinical outcome of cataract. Among different factors, oxidative stress, free radicals, and reactive oxygen species (ROSs) generated by the exposure to UV light are widely recognized to cause cataract formation. Taking advantage of synchrotron radiation circular dichroism, fluorescence, and circular dichroism spectroscopies, in the present study we investigate the influence of different small molecules with the potential to either quench ROS generation or to stabilize protein conformation. Therefore, ascorbic acid, an excellent antioxidant agent already present in the eye aqueous humour, has been tested along with ceftriaxone, mannitol and trehalose, which osmolyte activity was demonstrated interfering with model proteins misfolding. Our results showed that ascorbic acid strongly inhibits the ROS production without, however, preserving the native protein structure, whereas mannitol had no effect on the ROS production but retained better the secondary structure of WS proteins. Collectively, the use of a mixture of ascorbic acid and mannitol could be used to better protect eye lens proteins from ROS damage preventing the cataract onset.     

北京萤火虫复眼的模型小眼设计与光线追迹

以实际的北京萤火虫复眼结构与生理参数与依据,基于已知的ＧＲＩＮ透镜特征,把小眼的整个光学系统分成几个部分被模型化。模型中,在晶体柱部分使用了一个径向梯度折射率透镜柱具有的抛物线轮廓折射率公式;在晶锥末端所接的半椭球ＧＲＩＮ透镜部分,使用了一个径向梯度和纵向梯度分布都存在的折射率公式．模型的尺寸与折射率的大小根据实际晶锥的测量值．通过对模型小眼的光线追迹,证明模型小眼基本上是一个平行光入射,年行光出射的望远镜系统．小眼光学系统的前半部分把入射的平行光束聚焦到中间焦点,而这个点的位置恰好位于实际晶锥的腰的部位,而光线再次经过晶锥近端以后又以大致平行光束从晶锥末端出射。此模型小眼的最大入射角为３０度,在这个入射角内其角放大率基本上是相同的。随着对平行入射光束与小眼光轴的角度的增加,所产生的中间像越来越远离晶锥光轴。     

Development of a 3D finite element model of lens microcirculation

Background

It has been proposed that in the absence of a blood supply, the ocular lens operates an internal microcirculation system. This system delivers nutrients, removes waste products and maintains ionic homeostasis in the lens. The microcirculation is generated by spatial differences in membrane transport properties; and previously has been modelled by an equivalent electrical circuit and solved analytically. While effective, this approach did not fully account for all the anatomical and functional complexities of the lens. To encapsulate these complexities we have created a 3D finite element computer model of the lens.

Methods

Initially, we created an anatomically-correct representative mesh of the lens. We then implemented the Stokes and advective Nernst-Plank equations, in order to model the water and ion fluxes respectively. Next we complemented the model with experimentally-measured surface ionic concentrations as boundary conditions and solved it.

Results

Our model calculated the standing ionic concentrations and electrical potential gradients in the lens. Furthermore, it generated vector maps of intra- and extracellular space ion and water fluxes that are proposed to circulate throughout the lens. These fields have only been measured on the surface of the lens and our calculations are the first 3D representation of their direction and magnitude in the lens.

Conclusion

Values for steady state standing fields for concentration and electrical potential plus ionic and fluid fluxes calculated by our model exhibited broad agreement with observed experimental values. Our model of lens function represents a platform to integrate new experimental data as they emerge and assist us to understand how the integrated structure and function of the lens contributes to the maintenance of its transparency.     

The circadian clock of fruit flies is blind after elimination of all known photoreceptors

Circadian rhythms are entrained by light to follow the daily solar cycle. We show that Drosophila uses at least three light input pathways for this entrainment: (1) cryptochrome, acting in the pacemaker cells themselves, (2) the compound eyes, and (3) extraocular photoreception, possibly involving an internal structure known as the Hofbauer-Buchner eyelet, which is located underneath the compound eye and projects to the pacemaker center in the brain. Although influencing the circadian system in different ways, each input pathway appears capable of entraining circadian rhythms at the molecular and behavioral level. This entrainment is completely abolished in glass(60j) cry(b) double mutants, which lack all known external and internal eye structures in addition to being devoid of cryptochrome.     

The initiation of blood flow and flow induced events in early vascular development

Within a day of gastrulation, the embryonic heart begins to beat and creates blood flow in the developing cardiovascular system. The onset of blood flow completely changes the environment in which the cardiovascular system is forming. Flow provides physiological feedback such that the developing network adapts to cue provided by the flow. Targeted inactivation of genes that alter early blood fluid dynamics induce secondary defects in the heart and vasculature and therefore proper blood flow is known to be essential for vascular development. Though hemodynamics, or blood fluid dynamics, are known to activate signaling pathways in the mature cardiovascular system in pathologies ranging from artherosclerosis to angiogenesis, the role in development has not been as intensively studied. The question arises how blood vessels in the embryos, which initially lack cells types such as smooth muscle cells, differ in their response to mechanical signals from blood flow as compared to the more mature cardiovascular system. Many genes known to be regulated by hemodynamics in the adult are important for developmental angiogenesis. Therefore the onset of blood flow is of primary importance to vascular development. This review will focus on how blood flow initiates and the effects of the mechanical signals created by blood flow on cardiovascular development.     

Prospects for Vasculature Reorganization in Sentinel Lymph Nodes

Primary tumors can induce vasculature and lymph channel reorganizations within sentinel lymph nodes before the arrival of cancer cells. The key blood vessels in such nodes that are remodeled are high endothelial venules (HEVs). The morphological alteration of HEVs in the presence of a cancer, coupled with the increased proliferation rate of the endothelial cells, results in a functional shifting of HEVs from immune response to blood-flow carrier. This tumor-induced reorganization is quite different from an endotoxin-induced inflammatory vasculature alteration. We review some of the accepted doctrines on lymph flow and lymphatic metastasis in light of this reorganization. More investigations are needed to elucidate the molecular mechanisms underlying the morphological and functional alteration of HEVs, the fluid exchanges between lymph and blood, the microenvironmental preparation for cancer cells to survive and expand in the lymph node, and the detail process of further distant dissemination of cancer cells from the lymph nodes. Further study of this pathological process may help to explain some clinical phenomena, and should aid in developing novel therapeutics and prevention strategies against cancer metastasis.     

Embryonic stem cell-derived cystic embryoid bodies form vascular channels: an in vitro model of blood vessel development.

Murine embryonic stem cells can differentiate in vitro to form cystic embryoid bodies (CEB) that contain different structures and cell types. The blood islands are one such structure that consist of immature hematopoietic cells surrounded by endothelial cells, the first identifiable vascular cells. CEBs differentiated in vitro developed blood islands initially, and subsequently these blood islands matured to form vascular channels containing hematopoietic cells. Phase contrast microscopy demonstrated the presence of channels in mature CEBs grown in suspension culture, and high resolution light and electron microscopy showed that the cells lining these channels were endothelial cells. The channels appeared less organized than the vasculature of the mature yolk sac. The hematopoietic cells were occasionally seen 'flowing' through the CEB channels, although their numbers were reduced relative to the yolk sac. Analysis of primary CEB cultures showed the presence of cells with two characteristics of endothelial cells: approximately 30% of the cells labelled with fluorescent acetylated low density lipoprotein and a small number of cells were positive for von Willebrand's factor by immunostaining. Thus we conclude that a primitive vasculature forms in CEBs differentiated in vitro, and that not only primary differentiation of endothelial cells but also some aspects of vascular maturation are intrinsic to this cell culture system. CEBs are therefore a useful model for the study of developmental blood vessel formation.     

Hypothalamohypophysial vascular and neurosecretory link in the teleost Bagarius bagarius (Ham.).

A three-dimensional picture of the hypothalamohypophysial neurosecretory system of B. bargarius could be obtained by adopting in situ staining techniques. The paired NPO give rise to left and right neurosecretory tracts which, although they approximate as a common tract, maintain their individuality till entering the pars intermedia. The ventral hypothalamic and the hypophysial arteries take their origin from the internal carotid artery. The former contributes to the formation of the primary plexus of the median eminence and the latter enters the pituitary directly, giving rise to the neuroadreno-interface vasculature. The vasculature of the median eminence lies in close contact with the long common neurosecretory tract. Morphological evidence suggests that in B. bagarius there are three pathways of hypothalamic control of the hypophysis: (1) A direct neuroglandular pathway, where the neurosecretory axons come directly in contact with the adrenohypophysial cells. (2) An indirect neurovasculoglandular pathway, where the blood exposed to the NSM at the median eminence comes in contact with gland cells. (3) Another indirect neurovasculoglandular pathway, where the blood is exposed to NSM at the neurohypophysis prior coming in contact with the gland cells. Thus, B. bagarius has the 'median eminence equivalent' neuroadeno-interface vasculature typical of the teleosts and the median eminence comparable to the Elasmobranchs, Holocephali and primitive Actinopterygians. This shows that the pituitary portal system of teleosts is in general agreement with that of other fish groups, except that in some it is restricted to the neuroadreno-interface, whereas in others like B. bagarius it extends to the hypothalamus also. These may be termed anterior and posterior median eminence.