The Ulrastructure of the Gill of the Lugworm Arenicola marina (L.) (Annelida,Polychaeta)

Arenicola marina gills are hollow, branched, body outgrowths with a central coelomic cavity and afferent and efferent vessels. The gill surface area per unit body weight is about 4 cm²/g wet weight. The blood vascular system anatomy differs from the tip to the base of the gill. In the distal branches of the gill the superficial afferent and efferent vessels are joined by connecting vessels. All vessels arise as spacings between the basal laminae of the thin epidermis and of the coelomic myoepithelium. The contractile part of this epithelium mainly borders the afferent and efferent vessels, whereas pedicel-like cytoplasmic processes extend from the cell bodies and mainly line the connecting vessels. In the proximal branches of the gill the afferent and efferent vessels located in the coelomic cavity are surrounded by the coelomic myoepithelium, and a peripheral blood plexus is present below the epidermis. The gill epidermis is everywhere thin and does not exhibit the characters of a transporting epithelium. The gill coelomic myoepithelium has several functions: (i) periodic contractions of the gill, propelling blood and coelomic fluid toward the central vascular and coelomic compartments; (ii) blood ultrafilration toward the coelomic cavity; (iii) probably transport, suggested by the specialized structures of the lateral membranes of the cells.     

Indian-Ink Perfusion Based Method for Reconstructing Continuous Vascular Networks in Whole Mouse Brain

The topology of the cerebral vasculature, which is the energy transport corridor of the brain, can be used to study cerebral circulatory pathways. Limited by the restrictions of the vascular markers and imaging methods, studies on cerebral vascular structure now mainly focus on either observation of the macro vessels in a whole brain or imaging of the micro vessels in a small region. Simultaneous vascular studies of arteries, veins and capillaries have not been achieved in the whole brain of mammals. Here, we have combined the improved gelatin-Indian ink vessel perfusion process with Micro-Optical Sectioning Tomography for imaging the vessel network of an entire mouse brain. With 17 days of work, an integral dataset for the entire cerebral vessels was acquired. The voxel resolution is 0.35×0.4×2.0 µm³ for the whole brain. Besides the observations of fine and complex vascular networks in the reconstructed slices and entire brain views, a representative continuous vascular tracking has been demonstrated in the deep thalamus. This study provided an effective method for studying the entire macro and micro vascular networks of mouse brain simultaneously.     

3D-reconstruction of blood vessels by ultramicroscopy

As recently shown, ultramicroscopy (UM) allows 3D-visualization of even large microscopic structures with µm resolution. Thus, it can be applied to anatomical studies of numerous biological and medical specimens. We reconstructed the three-dimensional architecture of tomato-lectin (lycopersicon esculentum) stained vascular networks by UM in whole mouse organs. The topology of filigree branches of the microvasculature was visualized. Since tumors require an extensive growth of blood vessels to survive, this novel approach may open up new vistas in neurobiology and histology, particularly in cancer research.     

3D-reconstruction of blood vessels by ultramicroscopy

As recently shown, ultramicroscopy (UM) allows 3D-visualization of even large microscopic structures with µm resolution. Thus, it can be applied to anatomical studies of numerous biological and medical specimens. We reconstructed the three-dimensional architecture of tomato-lectin (Lycopersicon esculentum) stained vascular networks by UM in whole mouse organs. The topology of filigree branches of the microvasculature was visualized. Since tumors require an extensive growth of blood vessels to survive, this novel approach may open up new vistas in neurobiology and histology, particularly in cancer research.Key words: 3D-reconstruction, blood vessels, cancer, LEA, lectin, microvasculature, morphology, ultramicroscopy, whole mount     

A FERTILE AXIS OF TRILOBOXYLON ASHLANDICUM,A PROGYMNOSPERM FROM THE UPPER DEVONIAN OF NEW YORK

A partially petrified impression of Triloboxylon ashlandicum (Aneurophytales) is the first recognized fertile axis of the genus. Identification of the fertile axis rests on the similarity of its anatomy with that of previously described vegetative specimens. Fertile organs replace some vegetative branches along part of the main axis. Fertile organs are twice dichotomized in one plane and bear elongate sporangia arranged pinnately. Vegetative branches differ in that they bear the ultimate appendages of the plant helically. The latter organs dichotomize many times in one plane. Although similar in size and morphology to the ultimate appendages, the fertile organs are homologous by position and vascular supply to the vegetative branches which they replace. Sporangia of T. ashlandicum dehisce longitudinally and terminate in an apiculate tip. Spores are unknown. Fertile organs of T. ashlandicum resemble those of other Aneurophytales and support the earlier placement of Triloboxylon in the order on anatomical grounds. T. ashlandicum differs from other Aneurophytales, however, by bearing vegetative organs at the distal end of its fertile axis.     

Genetic investigation of equine recurrent uveitis in Appaloosa horses

Equine recurrent uveitis (ERU) is characterized by intraocular inflammation that often leads to blindness in horses. Appaloosas are more likely than any other breed to develop insidious ERU, distinguished by low-grade chronic intraocular inflammation, suggesting a genetic predisposition. Appaloosas are known for their white coat spotting patterns caused by the leopard complex spotting allele (LP) and the modifier PATN1. A marker linked to LP on ECA1 and markers near MHC on ECA20 were previously associated with increased ERU risk. This study aims to further investigate these loci and identify additional genetic risk factors. A GWAS was performed using the Illumina Equine SNP70 BeadChip in 91 horses. Additive mixed model approaches were used to correct for relatedness. Although they do not reach a strict Bonferroni genome-wide significance threshold, two SNPs on ECA1 and one SNP each on ECA12 and ECA29 were among the highest ranking SNPs and thus warranted further analysis (P = 1.20 × 10⁻⁵, P = 5.91 × 10⁻⁶, P = 4.91 × 10⁻⁵, P = 6.46 × 10⁻⁵). In a second cohort (n = 98), only an association with the LP allele on ECA1 was replicated (P = 5.33 × 10⁻⁵). Modeling disease risk with LP, age and additional depigmentation factors (PATN1 genotype and extent of roaning) supports an additive role for LP and suggests an additive role for PATN1. Genotyping for LP and PATN1 may help predict ERU risk (AUC = 0.83). The functional role of LP and PATN1 in ERU development requires further investigation. Testing samples across breeds with leopard complex spotting patterns and a denser set of markers is warranted to further refine the genetic components of ERU.     

Fluorescence decline as a function of redox potential and actinic light intensity in spinach thylakoids

Excitation of isolated thylakoids with sufficiently strong actinic light increases the fluorescence quantum yield up to a maximum level, F_max, followed by a slower decline under certain experimental conditions. In this study the latter effect was analyzed as a function of the ambient redox potential and the actinic light intensity. Two different types of fluorescence decrease were found. (a) In the presence of specific quinones widely used as redox mediators a fast and comparatively small decrease (30% of F_max), referred to as ΔF_SQ, was observed at moderate redox potentials (−300 <E_m < + 200 mV). ΔF_SQ disappears at positive values with E_{m, 7.5} = + 110 mV, whereas the decrease at negative redox potential depends on the midpoint potential of the quinone. (b) A more pronounced fluorescence decline was observed at redox potentials below −300 mV, which comprises 65–70% of the maximum fluorescence. The full expression of this effect, referred to as ΔF^max_LP, requires markedly higher actinic light intensities than ΔF^max_SQ. The extent of ΔF^max_LP as a function of the redox potential is dependent on the presence of redox mediators. In their absence the full expression of ΔF^max_LP can be only observed below −400 mV. Based on the hypothesis of Pheo⁻ photoaccumulation being responsible for the fluorescence decline at low redox potentials (Klimov, V.V., Klevanik, A.V. and Shuvalov, V.A. (1977) FEBS Lett. 82, 182–186), a reaction scheme is presented that qualitatively describes the time course of ΔF_LP at different actinic light intensities and redox potentials. Based on this analysis, the rate of Pheo^⨪ reoxidation is inferred to be limited by the reaction center apoprotein acting as a barrier to redox equilibration. The implications for the interpretations of redox titration curves are briefly discussed.     

Functional anatomy and xylem cavitation resistance of three species of monocotyledons grown on flooded substrates

There are few investigations that analyze the xylem functional anatomy of monocotyledons, as the methods have been developed for woody plants. This study describes the root, rhizome and aerial stem xylem anatomy and functional anatomy of Canna indica, Cyperus papyrus and Phragmites communis grown on flooded substrates; and it aims to evaluate the relationship between the xylem anatomy and its cavitation resistance. To calculate the indexes of vulnerability, mesomorphy, collapse and relative hydraulic conductivity in the three organs mentioned, the diameter, number of vessels per mm², thickness of the walls and the length of the tracheal elements were recorded. In addition, the xylem specific conductivity of the aerial stem was measured with the pipette method, and its resistance to cavitation was determined experimentally by the air injection technique. The protoxylem is xeromorphic, it has longer vessel elements, smaller diameters, thin walls and scalariform perforation plates, whereas the metaxylem is mesomorphic, with shorter vessel elements, larger diameters, thicker walls and simple perforation plates. Both present low collapse resistance but have a high relative hydraulic conductivity. P. communis recorded the highest cavitation resistance, and the number of vessels per mm² was related to xylem cavitation resistance in Canna indica and Cyperus papyrus. The experimental results of this investigation match partially the anatomical indexes and showed that the xylem of these species has a low specific conductivity and is more vulnerable to cavitation than that of other monocots.     

Blood Supply to the Chicken Femoral Head

The purpose of this study was to conduct a comprehensive evaluation of the vascular supply to the femoral head, including the vessels that give rise to the terminal perfusing branches. Using a casting agent, we highlighted the anatomy of the external iliac and ischiatic arteries with their associated branches after anatomic dissection of 24 hips from 12 Leghorn chickens. We confirmed published findings regarding perfusion of the femoral head and identified 3 previously undescribed arterial branches to this structure. The first branch (the acetabular branch of the femoralis artery) was supplied by the femoralis artery and directly perfused the acetabulum and femoral head. The second branch (the lateral retinacular artery) was a tributary of the femoralis artery that directly supplied the femoral head. Finally, we found that the middle femoral nutrient artery supplies a previously undescribed ascending intraosseous branch (the ascending branch of the middle femoral nutrient artery) that perfuses the femoral head. Precise understanding of the major vascular branches to the femoral head would allow for complete or selective ligation of its blood supply and enable the creation of a reproducible bipedal model of femoral head osteonecrosis.Like humans, chickens are bipedal animals that rely on the hip joint to absorb the majority of the body''s weight. This anatomy, in concert with their high activity level, makes chickens an attractive model for the study of osteonecrosis of the femoral head in humans. The vast majority of animal research on osteonecrosis of the femoral head has been performed on quadrupedal animals,^{3,4,10,19,25,26,28,29,31,36,37,41,51,52} thus limiting its application to bipedal species because most quadruped models fail to progress to end-stage mechanical collapse similar to that in humans.⁶Avascular necrosis is the death of bone that occurs from ischemia due to disruption of the vascular supply to bone through direct or indirect mechanisms.³⁸ Avascular necrosis should be differentiated from the broader term of osteonecrosis, which refers to bone death in general.³² Causes of femoral head osteonecrosis include direct and indirect disruption of vascular supply (traumatic injury, intravascular coagulation, extrinsic compression) as well as changes in cellular differentiation and cellular apoptosis.^{4,7,12,15,17,18,24,30-32,38,49,50} Accordingly, causes of osteonecrosis are both traumatic and nontraumatic.^16,31,32The arterial anatomy in the chicken hindlimb has been outlined by several authors.^{20,22,27,35,42,44,45} Briefly, the external iliac and ischiatic artery arise from the abdominal aorta to provide blood supply to the chicken hind limb. The external iliac artery has 2 main branches—the femoralis and femoral circumflex arteries—that distribute blood to the chicken hindlimb. The ischiatic artery provides 3 main branches: the trochanteric artery, superior femoral nutrient artery, and middle femoral nutrient artery. Although the terminal vascular supply to the femoral head of Leghorn and Broiler chickens has been described,^46,47 the origin of these terminal arteries with reference to the ischiatic and femoralis arteries and their respective branches has not been addressed. The current study will describe the blood vessels that feed these terminal branches to the chicken femoral head.     

Vascular Patterns in Iguanas and Other Squamates: Blood Vessels and Sites of Thermal Exchange

Squamates use the circulatory system to regulate body and head temperatures during both heating and cooling. The flexibility of this system, which possibly exceeds that of endotherms, offers a number of physiological mechanisms to gain or retain heat (e.g., increase peripheral blood flow and heart rate, cooling the head to prolong basking time for the body) as well as to shed heat (modulate peripheral blood flow, expose sites of thermal exchange). Squamates also have the ability to establish and maintain the same head-to-body temperature differential that birds, crocodilians, and mammals demonstrate, but without a discrete rete or other vascular physiological device. Squamates offer important anatomical and phylogenetic evidence for the inference of the blood vessels of dinosaurs and other extinct archosaurs in that they shed light on the basal diapsid condition. Given this basal positioning, squamates likewise inform and constrain the range of physiological thermoregulatory mechanisms that may have been found in Dinosauria. Unfortunately, the literature on squamate vascular anatomy is limited. Cephalic vascular anatomy of green iguanas (Iguana iguana) was investigated using a differential-contrast, dual-vascular injection (DCDVI) technique and high-resolution X-ray microcomputed tomography (μCT). Blood vessels were digitally segmented to create a surface representation of vascular pathways. Known sites of thermal exchange, consisting of the oral, nasal, and orbital regions, were given special attention due to their role in brain and cephalic thermoregulation. Blood vessels to and from sites of thermal exchange were investigated to detect conserved vascular patterns and to assess their ability to deliver cooled blood to the dural venous sinuses. Arteries within sites of thermal exchange were found to deliver blood directly and through collateral pathways. The venous drainage was found to have multiple pathways that could influence neurosensory tissue temperature, as well as pathways that would bypass neurosensory tissues. The orbital region houses a large venous sinus that receives cooled blood from the nasal region. Blood vessels from the nasal region and orbital sinus show anastomotic connections to the dural sinus system, allowing for the direct modulation of brain temperatures. The generality of the vascular patterns discovered in iguanas were assessed by firsthand comparison with other squamates taxa (e.g., via dissection and osteological study) as well as the literature. Similar to extant archosaurs, iguanas and other squamates have highly vascularized sites of thermal exchange that likely support physiological thermoregulation that “fine tunes” temperatures attained through behavioral thermoregulation.     

Engineering of In Vitro 3D Capillary Beds by Self-Directed Angiogenic Sprouting

In recent years, microfluidic systems have been used to study fundamental aspects of angiogenesis through the patterning of single-layered, linear or geometric vascular channels. In vivo, however, capillaries exist in complex, three-dimensional (3D) networks, and angiogenic sprouting occurs with a degree of unpredictability in all x,y,z planes. The ability to generate capillary beds in vitro that can support thick, biological tissues remains a key challenge to the regeneration of vital organs. Here, we report the engineering of 3D capillary beds in an in vitro microfluidic platform that is comprised of a biocompatible collagen I gel supported by a mechanical framework of alginate beads. The engineered vessels have patent lumens, form robust ∼1.5 mm capillary networks across the devices, and support the perfusion of 1 µm fluorescent beads through them. In addition, the alginate beads offer a modular method to encapsulate and co-culture cells that either promote angiogenesis or require perfusion for cell viability in engineered tissue constructs. This laboratory-constructed vascular supply may be clinically significant for the engineering of capillary beds and higher order biological tissues in a scalable and modular manner.     

Vascular and General Anatomy of the Rootstocks of Three Stemless Viola Species

The primary vascular system of the rootstock of Viola rotundifolia,V. odorata and V. cucullata consists of an open system of threesympodia, corresponding to three orthostichies (parastichies)of leaves in the 1 /3 phyllotaxy. Between the major, collateralbundles there are vascular strands consisting of only primaryphloem. The vascular supply to axillary buds developing in thefirst season of growth is the same in flowering and vegetativebuds and there is a homology between the bracts on the pedunclesof axillary flowers and the prophylls of the axillary vegetativebranches. The overwintering portions of the rootstocks are somewhatwoody. The uneven development of secondary xylem correspondsto the locations of the sympodia. Secondary xylem closes a leafgap of a median trace only slowly, but any gap in the phloicsystem is closed quickly by secondary growth. Viola rotundifolia, V. odorata, V. cucullata, rootstock, vascular anatomy     

Hypoxia Inducible Factor-2α Regulates the Development of Retinal Astrocytic Network by Maintaining Adequate Supply of Astrocyte Progenitors

Here we investigate the role of hypoxia inducible factor (HIF)-2α in coordinating the development of retinal astrocytic and vascular networks. Three Cre mouse lines were used to disrupt floxed Hif-2α, including Rosa26^CreERT2, Tie2^Cre, and GFAP^Cre. Global Hif-2α disruption by Rosa26^CreERT2 led to reduced astrocytic and vascular development in neonatal retinas, whereas endothelial disruption by Tie2^Cre had no apparent effects. Hif-2α deletion in astrocyte progenitors by GFAP^Cre significantly interfered with the development of astrocytic networks, which failed to reach the retinal periphery and were incapable of supporting vascular development. Perplexingly, the abundance of strongly GFAP⁺ mature astrocytes transiently increased at P0 before they began to lag behind the normal controls by P3. Pax2⁺ and PDGFRα⁺ astrocytic progenitors and immature astrocytes were dramatically diminished at all stages examined. Despite decreased number of astrocyte progenitors, their proliferation index or apoptosis was not altered. The above data can be reconciled by proposing that HIF-2α is required for maintaining the supply of astrocyte progenitors by slowing down their differentiation into non-proliferative mature astrocytes. HIF-2α deficiency in astrocyte progenitors may accelerate their differentiation into astrocytes, a change which greatly interferes with the replenishment of astrocyte progenitors due to insufficient time for proliferation. Rapidly declining progenitor supply may lead to premature cessation of astrocyte development. Given that HIF-2α protein undergoes oxygen dependent degradation, an interesting possibility is that retinal blood vessels may regulate astrocyte differentiation through their oxygen delivery function. While our findings support the consensus that retinal astrocytic template guides vascular development, they also raise the possibility that astrocytic and vascular networks may mutually regulate each other''s development, mediated at least in part by HIF-2α.     

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

In organs, the correct architecture of vascular and ductal structures is indispensable for proper physiological function, and the formation and maintenance of these structures is a highly regulated process. The analysis of these complex, 3-dimensional structures has greatly depended on either 2-dimensional examination in section or on dye injection studies. These techniques, however, are not able to provide a complete and quantifiable representation of the ductal or vascular structures they are intended to elucidate. Alternatively, the nature of 3-dimensional plastic resin casts generates a permanent snapshot of the system and is a novel and widely useful technique for visualizing and quantifying 3-dimensional structures and networks.A crucial advantage of the resin casting system is the ability to determine the intact and connected, or communicating, structure of a blood vessel or duct. The structure of vascular and ductal networks are crucial for organ function, and this technique has the potential to aid study of vascular and ductal networks in several ways. Resin casting may be used to analyze normal morphology and functional architecture of a luminal structure, identify developmental morphogenetic changes, and uncover morphological differences in tissue architecture between normal and disease states. Previous work has utilized resin casting to study, for example, architectural and functional defects within the mouse intrahepatic bile duct system that were not reflected in 2-dimensional analysis of the structure^1,2, alterations in brain vasculature of a Alzheimer''s disease mouse model³, portal vein abnormalities in portal hypertensive and cirrhotic mice⁴, developmental steps in rat lymphatic maturation between immature and adult lungs⁵, immediate microvascular changes in the rat liver, pancreas, and kidney in response in to chemical injury⁶.Here we present a method of generating a 3-dimensional resin cast of a mouse vascular or ductal network, focusing specifically on the portal vein and intrahepatic bile duct. These casts can be visualized by clearing or macerating the tissue and can then be analyzed. This technique can be applied to virtually any vascular or ductal system and would be directly applicable to any study inquiring into the development, function, maintenance, or injury of a 3-dimensional ductal or vascular structure.     

Productivity and mineral cycling in an oak coppice forest 2. Annual net production of the forest

Annual net production was estimated in the secondary coppice forest near Tokyo, which was dominated by a deciduous oak,Quercus serrata Thunb. Lateral growth of stems and old branches was directly estimated by examining the annual rings for 35 shoots in a clear-cut quadrat of 10m×10m. Phytomasses of current organs were also weighed in the quadrat. Preharvest losses of current organs were determined by twelve 0.5 m² litter traps for fine litter and twelve 6 m² quadrats for woody litter. Branch production was also assessed indirectly by use of the stem-branch allometry and death of branches. The results of the indirect method were in sufficient agreement with the result of the direct one. Grazing loss of leaves from the canopy was estimated directly from the loss in leaf area and indirectly from the animal faeces caught by the litter traps. Net production of the canopy trees was 149 kg a⁻¹ year⁻¹, in which leaf production was 36.9 kg. Animals grazed about 14% of the leaf area by the end of the growing season. True consumption of leaves by animals was 7.6% of leaf production or 10% of leaf mass. Production of undergrowth, mainly a dwarf bamboo,Pleioblastus chino Makino, was 28 kg a⁻¹ year⁻¹, being 15% of the total stand production. Productivity of this forest was significantly higher than that of cool-temperate deciduous broadleaf forests.     

Circulatory organs of Diplura (Hexapoda): the basic design in Hexapoda?

The circulatory systems of Campodea augens and Catajapyx aquilonaris (Hexapoda: Diplura) have been examined by means of light and electron microscopy. Hemolymph flow has also been investigated in vivo. Both species share features that deviate conspicuously from the common textbook design of the insect circulatory system: (i) antennal vessels connected to the anterior end of the dorsal vessel; (ii) presence of a circumoesophageal vessel ring in the head; (iii) a bidirectional flow within the dorsal vessel, made possible by intracardiac valves; (iv) posterior end of the dorsal vessel tube opens into a caudal chamber connected to cercal vessels (in Campodea) or to cercal channels (in Catajapyx); (v) dorsal diaphragm barely realized, ventral diaphragm absent altogether, and (vi) legs without specific organs serving hemolymph circulation. Comparative analysis has revealed that these characters in Diplura represent the most plesiomorphic condition in the circulatory organs of all extant Hexapoda. In the basic evolutionary lineages of insects, some organ components have been lost and the peripheral vessels decoupled from the dorsal vessel; as a result, autonomous accessory pulsatile organs have evolved to supply hemolymph to long body appendages and a unidirectional hemolymph flow mode prevailed within the dorsal vessel.     

Blood vascular system of the sea cucumber,Stichopus moebii

Stichopus moebii, a sea cucumber, has a closed circulatory system which is unique in its degree of development for the phylum Echinodermata. The gross anatomy, histology and fine structure of the system were studied. Blood vessels consist of a coelomic surface of ciliated epithelium, a layer of muscle and nerve cells, followed by connective tissue and luminal lining of endothelium. Basically the blood vascular system consists of two major vessels running parallel to the gut: the dorsal vessel pumps colorless blood via the vessels within the walls of the intestine into the ventral vessel. There are two specialized areas of the circulation: (1) At the upper small intestine 120 to 150 muscular single-chambered hearts pump blood from the dorsal vessel into a series of intestinal plates. (2) At the lower region of the small intestine the vasculature is associated with the left respiratory tree. Blood passing from the dorsal pulmonary vessel can take two routes to the gut, it either passes through myriads of minute respiratory shunt vessels entangled with the respiratory tree or it passes through a unique follicle network consisting of tiny channels periodically dilated into chambers filled with iron deposits, necrotic cells and developing coelomocytes.     

Branchial vascular pathways in two species of Tetraodontiformes and the concept of secondary vessels and nutrient arteries

The vascular organisation of the branchial basket was examined in two Tetraodontiform fishes; the three-barred porcupinefish, Dicotylichthys punctulatus and the banded toadfish, Marylina pleurosticta by scanning electron microscopy of vascular casts and standard histological approaches. In D. punctulatus, interarterial anastomoses (iaas) originated at high densities from the efferent filamental and branchial arteries, subsequently re-anastomosing to form progressively larger secondary vessels. Small branches of this system entered the filament body, where it was interspersed between the intrafilamental vessels. Large-bore secondary vessels ran parallel with the efferent branchial arteries, and were found to constitute an additional arterio–arterial pathway, in that these vessels exited the branchial basket in company with the mandibular, the carotid and the afferent and efferent branchial arteries, from where they gave rise to capillary beds after exit. Secondary vessels were not found to supply filament muscle; rather these tissues were supplied by single specialised vessels running in parallel between the efferent and afferent branchial arteries in both species examined. Although the branchial vascular anatomy was generally fairly similar for the two species examined, iaas were not found to originate from any branchial component in the banded toadfish, M. pleurosticta, which instead showed a moderate frequency of iaas on other vessels in the cephalic region. It is proposed that four independent vascular pathways may be present within the teleostean gill filament, the conventional arterio–arterial pathway across the respiratory lamellae; an arterio–arterial system of secondary vessels supplying the filament and non-branchial tissues; a system of vessels supplying the filament musculature; and the intrafilamental vessels (central venous sinus). The present study demonstrates that phylogenetic differences in the arrangement of the branchial vascular system occur between species of the same taxon.     

Vascularisation is not necessary for gut colonisation by enteric neural crest cells

The vasculature and nervous system share striking similarities in their networked, tree-like architecture and in the way they are super-imposed in mature organs. It has previously been suggested that the intestinal microvasculature network directs the migration of enteric neural crest cells (ENCC) along the gut to promote the formation of the enteric nervous system (ENS). To investigate the inter-relationship of migrating ENCC, ENS formation and gut vascular development we combined fate-mapping of ENCC with immunolabelling and intravascular dye injection to visualise nascent blood vessel networks. We found that the enteric and vascular networks initially had very distinct patterns of development. In the foregut, ENCC migrated through areas devoid of established vascular networks. In vessel-rich areas, such as the midgut and hindgut, the distribution of migrating ENCC did not support the idea that these cells followed a pre-established vascular network. Moreover, when gut vascular development was impaired, either genetically in Vegfa^120/120 or Tie2-Cre;Nrp1^fl/− mice or using an in vitro Wnt1-Cre;Rosa26^Yfp/+ mouse model of ENS development, ENCC still colonised the entire length of the gut, including the terminal hindgut. These results demonstrate that blood vessel networks are not necessary to guide migrating ENCC during ENS development. Conversely, in miRet⁵¹ mice, which lack ENS in the hindgut, the vascular network in this region appeared to be normal suggesting that in early development both networks form independently of each other.