Scanning electron microscopy of microcorrosion casts of the vascular bed in the skin of the spotted salamander,Salamandra salamandra L.

Microcorrosion casts of blood vessels in the skin of the spotted salamander, Salamandra salamandra L., were studied using scanning eletron microscopy. The investigated vessels include a subepidermal network of respiratory capillaries and the vessels of poison glands. A hypothesis is proposed, according to which both types of vascular beds possess a common origin. Probable factors involved in the differentiation of the primary netword of the subepidermal vessels in larvae are indicated and speculation concerning the supposed mosaic type of circulation in the capillaries of the subepidermal respiratory bed is presented.     

The morphology of amphibian skin vascularization before and after metamorphosis

Summary The skin vascularization was investigated troughout the ontogenetic development and in adults of two anurans, Rana temporaria and Bufo bufo, and two urodeles, Triturus vulgaris and Triturus cristatus. It was found that, contrary to the urodele larvae, the anuran tadpoles have a very sparse skin vascularization. During the early stages of anuran metamorphosis the skin capillary network becomes dense; later, during skin metamorphosis, a second, venous, network is formed as anastomoses between the subcutaneous vein ramifications. In the urodeles, metamorphosis is not accompanied by any significant morphological changes in the skin vascularization, and a subcutaneous network is not formed. It is suggested that the reduced skin vascularization in anuran tadpoles is an advanced larval character relative to the rich vascularization of the skin in urodele larvae. It is further suggested that anuran tadpoles have a reduced ability to utilize gaseous exchange through the skin. The function of the subcutaneous venous network found in anurans after metamorphosis is obscure; experiments indicate a vasomotor regulation which is neither adrenergic nor cholinergic.Abbreviations a arteriole - A artery - an anastomosis between the subepidermal capillary network and the subcutaneous venous network - C stratum compactum - E epidermis - ec subepidermal capillaries - S stratum spongiosum - sv subcutaneous venous network - v venule - V vein - vc venae comitantes     

A systems biology view of blood vessel growth and remodelling

Blood travels throughout the body in an extensive network of vessels – arteries, veins and capillaries. This vascular network is not static, but instead dynamically remodels in response to stimuli from cells in the nearby tissue. In particular, the smallest vessels – arterioles, venules and capillaries – can be extended, expanded or pruned, in response to exercise, ischaemic events, pharmacological interventions, or other physiological and pathophysiological events. In this review, we describe the multi‐step morphogenic process of angiogenesis – the sprouting of new blood vessels – and the stability of vascular networks in vivo. In particular, we review the known interactions between endothelial cells and the various blood cells and plasma components they convey. We describe progress that has been made in applying computational modelling, quantitative biology and high‐throughput experimentation to the angiogenesis process.     

十一种无尾两栖类物种皮肤和肺显微结构

无尾两栖类动物的呼吸方式为肺呼吸和皮肤辅助呼吸,为探究两种呼吸器官的显微结构,本文采用解剖学和组织学的方法,对大蹼铃蟾(Bombina maxima)、腺角蟾(Megophrys glandulosa)、中华蟾蜍(Bufo gargarizans)、华西雨蛙(Hyla gongshanensis)、昭觉林蛙(Rana chaochiaoensis)、滇蛙(Dianrana pleuraden)、双团棘胸蛙(Gynandropaa yunnanensis)、贡山树蛙(Rhacophorus gongshanensis)、斑腿泛树蛙(Polypedates megacephalus)、饰纹姬蛙(Microhyla fissipes)、多疣狭口蛙(Kaloula verrucosa)的皮肤和肺的形态及显微结构进行观察。结果显示,背部和腹部皮下可见血管交错成网状,皮肤由表皮层和真皮层构成。除华西雨蛙外,其余10种均有分布于真皮疏松层与致密层间的钙化层;色素细胞位于疏松层上层,体背色素层较发达。肺囊状,中空密布血管,分为大小相当的左右肺叶,两肺叶相通并与心粘连,无气管和支气管。肺由肺囊壁、隔膜、毛细血管、肺泡细胞等结构组成。肺囊壁分为胸膜层、中间层和内层:胸膜层由扁平细胞构成,中间层由结缔组织构成,内层由肺细胞和毛细血管组成,隔膜由毛细血管和结缔组织构成,游离隔膜向中部靠拢可形成半封闭腔室,结合隔膜与肺囊壁形成封闭小腔室。在这11个物种中,肺较发达的个体,其皮肤结构中黏液腺的数量就会相对较少,组织形态学特征表现出与环境适应性关系较大,而受到系统发育关系影响较小。     

On the vascularization and structure of the skin of a Korean bullhead Pseudobagrus brevicorpus (Bagridae, Teleostei) based on its entire body and appendages

To investigate the vascularization and structure of the skin and its relationship to cutaneous respiration in Pseudobagrus brevicorpus , a histological study by light microscopy was carried out on 15 regions of the skin, including eight body regions, six fins and the barbel. The skin consisted of the epidermis, dermis and subcutis in all regions, except for the barbel that had a relatively thin dermis and subcutis. The epidermis was composed of the outermost layer, the middle layer and the stratum germinativum. There were two kinds of gland cells: the unicellular mucus cells and large club cells. The middle layer had a small number of fine blood capillaries accompanied by dermal collagen in all regions; the mean number of blood capillaries ranged from 0.9 to 5.9. The mean diffusion distance between the capillary endothelial cells and the surface of the epidermis ranged from 50.6 to 126.8 μm. Based on these intra-epithelial blood capillaries, the relative surface area of the respiratory epithelium ranged from 0.1 to a maximum value of 1.2%. The dermis lacking scales had collagen bundles arranged parallel to each other, but vertical fiber bundles around the dorso-lateral regions were seen at intervals. Sensory organs such as taste buds, pit organs and lateral canals were found whereby the taste buds in particular were more abundant in the epidermis of the barbel. The vascularization of the skin may be closely related to an additional respiratory system used to deal with an extreme hypoxic condition during dry seasons.     

Structure of the skin of an air-breathing mudskipper, Periophthalmus magnuspinnatus

The epidermis of the mudskipper Periophthalmus magnuspinnatus consisted of three layers: the outermost layer, middle layer and stratum germinativum. Extensive vascular capillary networks were present near the superficial layer of epidermis and outermost layer. The diffusion distance between the vascular capillaries and the surface of epidermis was c . 1.5 ± 0.9μm. The middle layer consisted of small or voluminous cells swollen by epidermal cells. Due to the swollen cells, the thickness of the epidermis increased and the epidermis appeared web-like. The swollen cells contained tonofilaments, lucent contents and desmosomes. Fine blood capillaries were also discernible in this layer. Well-developed lymphatic spaces containing lymphocytes existed in the stratum germinativum. Numerous blood capillaries were present under the basement membrane. The dermis consisted of a stratum laxum and stratum compactum, and there was a definite area with acid mucopolysaccharides and a small scale in the stratum laxum. The skin had an epidermal pigment cell, dendritic melanophores (-cytes) containing melanin granules within their cytoplasm, and two kinds of dermal pigment cells, melanophores and colourless pigments containing reflecting platelets.     

Structure and Histochemistry of the Skin of a Torrent Catfish, Liobagrus mediadiposalis

The epidermis of the torrent catfish, Liobagrus mediadiposalis, consists of three layers: the outermost layer, middle layer and stratum germinativum. The epidermis consists of two types of skin glands, small mucus cell and voluminous club cell. The unicellular mucus cell contains acid sulfomucins (some sialomucins) and the club cell, sometimes binucleate, is proteinaceous. Well-developed vascularization is one of the characteristics of epidermis of L. mediadiposalis. Well-developed lymphatic spaces contain lymphocytes in the epidermis. The dermis lacks scales and consists mostly of a thick, dense connective tissue; its superficial region just below the basal membrane is supplied with fine blood capillaries. These histological features of the skin in L. mediadiposalis are consistent with that required for cutaneous respiration.     

A study on the epidermal structure of <Emphasis Type="Italic">Periophthalmodon</Emphasis> and <Emphasis Type="Italic">Periophthalmus</Emphasis> mudskippers with reference to their terrestrial adaptation

Epidermal structures of three species of Periophthalmus (Ps.) and two species of Periophthalmodon (Pn.) were investigated in relation to their lifestyle. All species of both genera lack a dermal bulge, which species of other two oxudercine genera, Boleophthalmus and Scartelaos, have in their epidermis. In Periophthalmus and Periophthalmodon species, which are highly terrestrial, the middle cells are well developed in the epidermis and the capillaries are distributed in the surface of the epidermis on the head and dorsal body. In Periophthalmus species and Pn. septemradiatus, the capillaries and blood vessels are also distributed in the epidermis of the abdomen, superficially in Ps. modestus and deeply in other species. In Ps. modestus, the capillaries are also densely distributed on the surface of the epidermis in the caudal area, whereas in other species, the epidermal capillaries and blood vessels of this area are located deep with a very low density. In Pn. schlosseri, the epidermal capillaries are not found in either the abdominal area or caudal area. A comparison of the distribution of epidermal capillaries among Boleophthalmus, Periophthalmodon, Periophthalmus, and Scartelaos species revealed that the skin makes a larger contribution to respiration in the species having a more terrestrial lifestyle. Goblet mucous cells are completely lacking in Periophthalmus species, whereas slimelike materials were often found on the skin surface of Periophthalmus species. This finding suggests that Periophthalmus species have some unknown mechanism for producing mucus. In Pn. schlosseri, exposure of the dense capillary net on the surface of the head is likely to increase cutaneous respiration, but it also makes the fish an attractive target of bloodsucking insects.Supplementary material to this paper is available in electronic format at http://dx.doi.org/10.1007/s10228-003-00173-7     

Formation of the branching pattern of blood vessels in the wall of the avian yolk sac studied by a computer simulation

The present study was performed to provide data to support the notion previously believed but not proved experimentally or theoretically, that blood vessels are formed by the selection of capillaries in the network. In an attempt to understand the mechanism of formation of blood vessel branching structures, the transformation of a capillary network to a branching system in the wall of quail yolk sac was successively recorded by a series of photographs, and a computer simulation was carried out for the process of in vivo vascularization based on the photographs. The simulation demonstrated that a positive feedback system participated in the formation of a branching structure. That is, vessels which had been much used were enlarged, whereas less used vessels were reduced in their size and finally extinguished. The enlarged vessels became major components of the branching system. As the body of an embryo grew, it was observed that polygonal capillary networks enlarged, which led each polygon of the network to divide into a few finer polygons. Then, some of the capillary vessels were again selected and formed a branching system. This process repeated during the body growth, indicating that the vascular system developed adaptively to the body growth. A region where the growth was fast, received much blood flow and produced finer networks of capillaries. Thus, it was experimentally demonstrated for the first time that capillaries in the network are successively selected by a positive feedback mechanism and form blood vessels.     

Glomerular bypass shunts in the kidney of the Atlantic hagfish,Myxine glutinosa

Summary The vascular pathways associated with the glomerulus of the Atlantic hagfish, Myxine glutinosa have been studied by scanning electron microscopy of corroded resin casts of the vasculature. Although the overall pattern of the renal vasculature did not differ from earlier reports, a previously unreported vascular pathway which arose from the renal artery and bypassed the glomerular capillaries in 28% of glomeruli was clearly demonstrated. Glomerular bypass shunts either ran to join the loose capillary network around Bowman's capsule and thereby drain into the network of vessels associated with the mesonephric duct (ureter), or ran directly into the ureteral system of vessels and subsequently into the posterior cardinal veins. Glomerular bypass shunts which theoretically permit renal arterial blood to bypass the process of filtration may play a role in the regulation of body fluid volume.     

Vasculature of the head and respiratory organs in an obligate air-breathing fish,the swamp eel Monopterus (=Amphipnous) cuchia

Methyl methacrylate vascular corrosion replicas were used to examine the macrocirculation in the head region and the microcirculation of respiratory vessels in the air-breathing swamp eel Monopterus cuchia. Fixed respiratory tissue was also examined by SEM to verify capillary orientation. The respiratory and systemic circulations are only partially separated, presumably resulting in supply of mixed oxygenated and venous blood to the tissues. A long ventral aorta gives rise directly to the coronary and hypobranchial arteries. Two large shunt vessels connect the ventral aorta to the dorsal aorta, whereas the remaining ventral aortic flow goes to the respiratory islets and gills. Only two pairs of vestigial gill arches remain, equivalent to the second and third arches, yet five pairs of aortic arches were identified. Most aortic arches supply the respiratory islets. Respiratory islet capillaries are tightly coiled spirals with only a fraction of their total length in contact with the respiratory epithelium. Valve-like endothelial cells delimit the capillary spirals and are unlike endothelial cells in other vertebrates. The gills are highly modified in that the lamellae are reduced to a single-channel capillary with a characteristic three-dimensional zig-zag pathway. There are no arterio-arterial lamellar shunts, although the afferent branchial artery supplying the gill arches also supplies respiratory islets distally. A modified interlamellar filamental vasculature is present in gill tissue but absent or greatly reduced in the respiratory islets. The macro- and micro-circulatory systems of M. cuchia have been considerably modified presumably to accommodate aerial respiration. Some of these modifications involve retention of primitive vessel types, whereas others, especially in the microcirculation, incorporate new architectural designs some of whose functions are not readily apparent.     

Fine structure of the epidermis of the mudskipper,Periophthalmus modestus (Gobiidae)

The ultra-structure of the epidermis of the mudskipper,Periophthalmus modestus, was examined by both light and transmission electron microscopies. The epidermis is exceptionally not well endowed with mucous or granular cells. Filament-containing cells occur in three distinct layers of the surface, middle and basal epidermis. The surface layer is further subdivided into two layers, an outermost and less superficial one. Two different cell types were identified in the epidermis. Type I cells are fiat cells in a single stratum. Type II cells are enormous cells, characterized by having a large vacuole in the cytoplasm. The outermost layer is composed of a free surface of Type I cells and numerous microridges covered with a fuzzy, fibrillar substance. The “fuzz” forms a cuticule-like structure, but keratinization as found in terrestrial animals does not occur. The superficial layer contains Type I cells and intraepithelial blood capillaries. When Type I cells become senescent, numerous intercellular spaces are formed in the plasma membranes of adjacent cells, with the senescent cells finally falling off. Just beneath these cells, however, young cells of Type I are always found. The blood capillaries are usually reinforced with young Type I cells. A large volume of oxygen may be absorbed through the skin using the blood capillary network. The middle layer contains several strata of Type II cells. The special corky structure of these cells seems to play an important role in thermal insulation and protection against ultraviolet light in relation to life out of water. However, by comparison with terrestrial animals, the histological design of the epidermis of this goby appears incomplete, so as to reduce desiccation on land, owing to the epidermis lacking a keratinized stratum. The differentiation of the epidermis seems to be an adaptation for a terrestrial habit in this species.     

Skin structure and vascularization in the Antarctic notothenioid fish Gymnodraco acuticeps

The scaleless notothenioid Gymnodraco acuticeps is a bottom dweller beneath the sea ice of McMurdo Sound, Antarctica. Gymnodraco experience unusual environmental conditions, including highly oxygenated subzero water. Skin morphology is evaluated with reference to its potential as a barrier to ice propagation and as a surface for cutaneous respiration. Light and electron microscopy and histochemistry reveal skin structure that is generally similar to that of other teleosts. In the epidermis, epithelial cells are arranged in nine to fifteen layers, and two types of mucous cells are also present. Large mucous cells are most common on external epidermal surfaces, whereas small cells are more frequent on internal epithelial surfaces. Epithelial cell junctions have extensive areas of desmosomes as well as interdigitations of the cell membranes, especially in the basal and midepidermis. The dermis consists of an exceptionally dense stratum compactum. The skin is thicker than that of Bovichtus, a scaleless temperate notothenioid from New Zealand. Mean skin thicknesses at sites on the trunk are 371–711 μm. With the exception of fins that contact the substrate, epidermal thickness between rays of most fins is 70–118 μm. The epithelial surfaces of the oral and branchial cavities are 27–50 μm thick. An unusual type of connective tissue is present beneath the epidermis of the pelvic fin. It contains abundant ground substance and is similar to mucous connective tissue of the mammalian umbilical cord. Perfusions of a microvascular filling agent reveal a moderately developed cutaneous vasculature. These vessels have the dimensions of capillaries (mean external diameter 11 μm). They are confined to the dermis and are more prominent on the head than on the trunk. The skin is secondary to the gills as a respiratory surface in Gymnodraco.     

A study on the vascularization and structure of the epidermis of the air-breathing mudskipper, Periophthalmus magnuspinnatus (Gobiidae, Teleostei), along different parts of the body

A histological study on the epidermis of eight body regions, five fins and the sucking disc was performed on the mudskipper, Periophthalmus magnuspinnatus. The study aimed to determine the role of the skin in respiration and to assess which region of the skin was most effective. The structure of the epidermis, consisting of the superficial layer, middle layer and the stratum germinativum, was the same in all regions. Large numbers of blood capillaries were situated at the superficial layer and occasionally at the middle layer. The mean diffusion distance between the capillary endothelial cells and the surface of the epidermis ranged from 2.6 to 15.4 μm: the lowest value was on the back (mean 2.0 μm) and the highest value was at the base of the anal fin (mean 15.4 μm). Relative surface area of respiratory epithelium in 14 regions was highest in the 1st and 2nd dorsal fins with a thinner epidermis and a lower diffusion distance (mean value 3.2% and 2.5% respectively), whereas the lowest was found at the base of the anal fin (mean 0.7%). Among the 14 regions of the epidermis, it can be surmised that the two dorsal fins toward the upper region may often be more exposed to air and for longer time periods than the other body regions during the amphibious life phase of Periophthalmus magnuspinnatus.     

The tentacles of Ichthyophis (Amphibia: Caecilia) with special reference to the skin

The skin of the paired tentacles of Ichthyophis consists of a cornified epidermis of 5–7 layers of epidermal cells, and a glandular dermis of ducted mucous glands, in association with collagen, blood vessels, fibroblasts, granulocytes, sparse melanophores and characteristic laminophores of unknown function. The epidermis is highly innervated at all levels below the stratum corneum by naked neurites, which originate as branches from large unmyelinated nerve bundles (and associated Schwann cells), located sub-epidermally, and which are part of the trigeminal cranial nerve. Myelinated nerves are also present below the epidermis, spatially associated with capillaries and glands. The study of the ultrastructure of the tentacle supports a concept of a sensory function, possibly tactile, though until further information from experimentation is available, any ideas on the specific nature of these sensory activities must remain speculative.     

Basement membrane protein distribution in LYVE-1-immunoreactive lymphatic vessels of normal tissues and ovarian carcinomas

The endothelial cells of blood vessels assemble basement membranes that play a role in vessel formation, maintenance and function, and in the migration of inflammatory cells. However, little is known about the distribution of basement membrane constituents in lymphatic vessels. We studied the distribution of basement membrane proteins in lymphatic vessels of normal human skin, digestive tract, ovary and, as an example of tumours with abundant lymphatics, ovarian carcinomas. Basement membrane proteins were localized by immunohistochemistry with monoclonal antibodies, whereas lymphatic capillaries were detected with antibodies to the lymphatic vessel endothelial hyaluronan receptor-1, LYVE-1. In skin and ovary, fibrillar immunoreactivity for the laminin α4, β1, β2 and γ1 chains, type IV and XVIII collagens and nidogen-1 was found in the basement membrane region of the lymphatic endothelium, whereas also heterogeneous reactivity for the laminin α5 chain was detected in the digestive tract. Among ovarian carcinomas, intratumoural lymphatic vessels were found especially in endometrioid carcinomas. In addition to the laminin α4, β1, β2 and γ1 chains, type IV and XVIII collagens and nidogen-1, carcinoma lymphatics showed immunoreactivity for the laminin α5 chain and Lutheran glycoprotein, a receptor for the laminin α5 chain. In normal lymphatic capillaries, the presence of primarily α4 chain laminins may therefore compromise the formation of endothelial basement membrane, as these truncated laminins lack one of the three arms required for efficient network assembly. The localization of basement membrane proteins adjacent to lymphatic endothelia suggests a role for these proteins in lymphatic vessels. The distribution of the laminin α5 chain and Lutheran glycoprotein proposes a difference between normal and carcinoma lymphatic capillaries.     

Localized agglutinin staining in muscle capillaries from normal and very old atrophic human muscle using winged bean (Psophocarpus tetragonolobus) lectin

 The winged bean (Psophocarpus tetragonolobus) agglutinin (total lectin) and its basic (WBA I) and acidic isoform (WBA II) were used to analyze capillaries in sections from human muscle. The microvessels were clearly labeled after incubation with the lectins in both normal muscle and in old muscles with age-related type II atrophy or muscle fiber grouping. Muscle fibers, nerves, and connective tissue remained unstained. The total lectin detected muscle capillaries from all blood group AB0 individuals. The isoform WBA I reacted only with blood vessels in blood group A and B individuals, while the blood vessels in blood group 0 individuals were demonstrated with WBA II. WBA I staining was inhibited by p-nitrophenyl α-galactopyranoside and N-acetylgalactosamine, whereas 2′-fucosyllactose and preincubation with an antibody against type-1 chain H abolished capillary staining with WBA II. The study demonstrates the usefulness of WBA as a marker of capillaries in human muscle. Accepted: 2 September 1996     

Morphology and tyrosine-hydroxylase immunohistochemistry of the systemic secondary vessel system of the blue catfish,Arius graeffei

Fish have a secondary vessel system which emerges from the primary vasculature via large numbers of coiled origins. The precise role of this vessel system is unknown. Vascular casting techniques and scanning electron microscopy reveal that the secondary vessels of the blue catfish, Arius graeffei, originate from dorsal, lateral, and ventral segmental primary arteries and from the caudal dorsal aorta. These vessels anastomose with each other to form larger secondary arteries which parallel the primary vessels for their entire length. Secondary vessels do not appear to form a capillary bed in the skin in A. graeffei as they do in some fish species. Coiled secondary vessel origins are abundant within the tunica media and adventitia of the primary vessels from which they emerge. The origins of the secondary vessels are surrounded by the extensive cytoplasmic processes of specialized endothelial cells. These processes extend for up to 6 μm into the lumen of the primary vessel. Ultrastructurally the coiled secondary capillaries consist of an endothelial cell tube which is surrounded by a single layer of pericytes. These endothelial cells extend large numbers of microvilli into the lumen of the coiled secondary capillary. Nerve terminals are commonly associated with the coiled secondary capillaries. Immunohistochemistry has revealed the presence of tyrosine-hydroxylase, an enzyme involved in catecholamine synthesis in nerve varicosities close to secondary vessels in A. graeffei. This vessel system could therefore be regulated by adrenergic nerves. © 1996 Wiley-Liss, Inc.     

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.     

In vivo two-photon excited fluorescence microscopy reveals cardiac- and respiration-dependent pulsatile blood flow in cortical blood vessels in mice

Subtle alterations in cerebral blood flow can impact the health and function of brain cells and are linked to cognitive decline and dementia. To understand hemodynamics in the three-dimensional vascular network of the cerebral cortex, we applied two-photon excited fluorescence microscopy to measure the motion of red blood cells (RBCs) in individual microvessels throughout the vascular hierarchy in anesthetized mice. To resolve heartbeat- and respiration-dependent flow dynamics, we simultaneously recorded the electrocardiogram and respiratory waveform. We found that centerline RBC speed decreased with decreasing vessel diameter in arterioles, slowed further through the capillary bed, and then increased with increasing vessel diameter in venules. RBC flow was pulsatile in nearly all cortical vessels, including capillaries and venules. Heartbeat-induced speed modulation decreased through the vascular network, while the delay between heartbeat and the time of maximum speed increased. Capillary tube hematocrit was 0.21 and did not vary with centerline RBC speed or topological position. Spatial RBC flow profiles in surface vessels were blunted compared with a parabola and could be measured at vascular junctions. Finally, we observed a transient decrease in RBC speed in surface vessels before inspiration. In conclusion, we developed an approach to study detailed characteristics of RBC flow in the three-dimensional cortical vasculature, including quantification of fluctuations in centerline RBC speed due to cardiac and respiratory rhythms and flow profile measurements. These methods and the quantitative data on basal cerebral hemodynamics open the door to studies of the normal and diseased-state cerebral microcirculation.