SEM observations of the elastic networks in canine femoral artery

Scanning electron microscopy was used to study the normal architectural arrangement of elastic tissue in a medium-sized muscular artery. Selective NaOH sonication digestion or formic acid digestion was used to expose and isolate the elastic networks in the femoral arteries of four healthy dogs. The digested segments were neutralized and freeze-dried before mounting for scanning electron microscopy (SEM) observation. The fenestrated internal elastic lamina (IEL) had a smooth surface with scattered regions of the fine elastic fibers that made up lacy networks protruding from the luminal surface. Prominent ellipsoid fenestrae, randomly scattered across the surface, were grouped into small and large sizes based on their mean diameter. The openings of most fenestrae were bridged by elastic fibers to give the fenestrae a sieve-like appearance. Large, transversely oriented, fusiform gaps were randomly scattered along the length of the IEL. These gaps, filled in by an elastic fiber network, sometimes spanned as much as a quarter of the vessel circumference. It is suggested that these gaps represent splits in the IEL that have been repaired. The tunica media contained a complex network of anastomosing elastic fibers and lamellae that were primarily circumferential in orientation. A well-defined external elastic lamina formed a solid sheet at the junction of the tunica media and the tunica adventitia. The tunica adventitia contained 8-10 incomplete lamellae of large, interconnecting, longitudinally oriented fibers. The architecture of the elastic network in canine femoral artery was compared with that previously described in medium-sized canine veins and in the rat femoral artery.     

Electron microscopic structure and innervation of the carotid baroreceptor region in the rock hyrax (Procavia capensis).

Semi-thin plastic sections reveal that the carotid baroreceptor region in the rock hyrax comprising the origin of the internal carotid artery has a preponderantly elastic structure and a thick tunica adventitia. In contrast, the common carotid artery has a musculoelastic structure, whereas the cranial segment of the internal carotid artery (immediately distal to the baroreceptor areas) shows the features of a muscular artery. Electron microscopy discloses the presence of sensory nerve endings within the parts of the tunica adventitia adjoining the preponderantly elastic zone of the internal carotid artery. These nerve endings are characterized by varicose regions containing a large quantity of mitochondria. Bundles of collagen fibers in the tunica adventitia form convolutions or whorls around the nerve terminals and often terminate on the surface of the elastic fibers or into the basement membranes of the neuronal profiles. The large content of elastic tissue in the tunica media of the baroreceptor region may render the vessel wall highly distensible to intraluminal pressure changes. This, in turn, would facilitate the transmission of the stimulus intensity to the sensory nerve terminals located in the tunica adventitia. It is suggested that the stretching of elastic fibers may form the main mechanical event leading to the distortion of the associated nerve terminals. However, a change in the geometrical configuration of the bundles of collagen under the influence of the elastic fibers may provide a better insight into the mechanisms of distortion of the baroreceptors related to and/or in contact with collagen fibers.     

Microscopic studies on the transition between the sigmoid sinus, the superior bulb of the jugular vein and the first portion of the internal jugular vein

The author studied the structure of the tissue components of the tunicae of the terminal segment of the sigmoid sinus, particularly at the level of the transition between the sigmoid sinus, the superior bulb of the jugular vein and the first portion of the human internal jugular vein; it was established that the transition between the sigmoid sinus and the first portion of the internal jugular vein occupies the whole extension of the superior bulb of the jugular vein up to the inferior third of the first portion of this vessel. These vascular walls exhibit a structure similar to that of the dura, i.e. the tunica adventitia is formed by fascicles of collagenic fibers which describe discontinuous spirals, more open proximal to the beginning of the first portion of the internal jugular vein. Approximately in the inferior third of the first portion of the internal jugular vein, there appear fascicles of smooth muscle fibers which are arranged similarly to those of the venous walls. The tunica intima of these vascular segments exhibits an endothelium resting on a network of elastic fibers which may play the role of an internal elastic lamina. From the bony border of the jugular foramen there originates a connective system whose fascicles of collagenic and elastic fibers incorporate to the wall of the internal jugular vein after describing a stretch in spiral around the vascular lumen.     

Neuronal systems immunoreactive with antiserum to lamprey gonadotropin-releasing hormone in the brain of Petromyzon marinus

Summary The wall structure of arteriovenous anastomoses in the rabbit ear was investigated. (1) Clusters of epithelioid smooth muscle cells form 3–4 longitudinally oriented plicae. The channel shows a single, irregularly outlined lumen, and its wall is very thin between adjacent plicae. (2) Endothelial cells covering the plicae protrude into the lumen, thus suggesting active contraction or shortening of the plicae. (3) The tunica adventitia is composed of 4–6 sheaths of flat fibroblasts, which may serve as a barrier to prevent loss of neurotransmitters. Processes of some of the fibroblasts also extend into the tunica media. (4) The tunica media is composed of an outer circular layer of typical smooth muscle cells, and an inner longitudinally running plica of ramified smooth muscle cells. Wide intercellular spaces between these ramified cells are filled with collagen fibrils, microfibrils, amorphous intercellular substances, and fibroblasts. Fibroblasts form close membrane contacts with each other, and with the smooth muscle cells. (5) Fibroblasts and other connective tissue components may function as an elastic support during active motility of the anastomotic channel.     

Interstitial flow through the internal elastic lamina affects shear stress on arterial smooth muscle cells

Interstitial flow through the tunica media of an artery wall in the presence of the internal elastic lamina (IEL), which separates it from the subendothelial intima, has been studied numerically. A two-dimensional analysis applying the Brinkman model as the governing equation for the porous media flow field was performed. In the numerical simulation, the IEL was modeled as an impermeable barrier to water flux, except for the fenestral pores, which were uniformly distributed over the IEL. The tunica media was modeled as a heterogeneous medium composed of a periodic array of cylindrical smooth muscle cells (SMCs) embedded in a fiber matrix simulating the interstitial proteoglycan and collagen fibers. A series of calculations was conducted by varying the physical parameters describing the problem: the area fraction of the fenestral pore (0. 001-0.036), the diameter of the fenestral pore (0.4-4.0 microm), and the distance between the IEL and the nearest SMC (0.2-0.8 microm). The results indicate that the value of the average shear stress around the circumference of the SMC in the immediate vicinity of the fenestral pore could be as much as 100 times greater than that around an SMC in the fully developed interstitial flow region away from the IEL. These high shear stresses can affect SMC physiological function.     

Histoarchitecture of the human parotid duct. Light-microscopic study

A light-microscopical study of the histoarchitecture of the human parotid duct was carried out. The parotid duct, much like other excretory passages, possessed three histological coats: mucosa, muscular layer and adventitia. The mucous epithelium was composed of the innermost tall columnar cells and the basal cubical cells. Numerous, circular elastic fibers were in close vicinity to the epithelium. The muscular layer consisted of smooth muscular fibers running in longitudinal direction. The adventitia consisted of lipofibrous connective tissue and contained many vessels. The result of the present study suggests that the parotid duct contributes to control of salivary secretion, since its architecture is basically similar to that of other excretory passages which have peristaltic activity.     

Observations on the fine structure of the baroreceptors and adrenergic innervation of the guinea-pig carotid sinus

We examined the fine structure of the baroreceptors and the adrenergic innervation of the guinea-pig carotid sinus. The tunica adventitia contained many nerve bundles whose perineuria enclosed unmyelinated nerve fibers, alone or together with myelinated nerve fibers. Baroreceptors, which lay close to elastic and collagen fibers in the adventitia and media, were surrounded by “terminal” cells with ultrastructural features characteristic of Schwann cells and contained inclusions of various types. Morphologic features of the baroreceptors included densely packed mitochondria, osmiophilic lamellated and homogeneous bodies, clear and granular vesicles, lamellar systems, glycogen granules, neurofilaments, neurotubuli, and vacuolated mitochondria. In animals that had been treated with 6-hydroxydopamine, occasional electrondense endings (or fibers) were observed in the adventitial layer. The baroreceptors in the guinea-pig carotid sinus appear to have most of the morphologic features reported for other species.     

The mammalian testicular capsule and its muscle elements

This study of the testicular capsule of rat, dog, cat and human has confirmed the presence of three layers, viz., the tunica vaginalis, the tunica albuginea proper and an innermost tunica vasculosa. Smooth muscle cells are present in the tunica albuginea of all four species and are more prominent at the posterior pole of the testis where the capsule merges with the mediastinum testis. In the rat and the dog, a few striated muscle fibers also are present. While the tunica albuginea is to be considered as a dense connective tissue, the arrangement of the collagen bundles and the presence of a relatively high content of elastic fibers probably permits changes in size of the testis following spontaneous contractions of the muscle elements, which are known to occur. The role of the testicular capsule in sperm transport is discussed in relation to other factors, the spontaneous contractions of the capsule presumably having a “pumping” action and aiding the movement of non-motile spermatozoa from the testis to the epididymis. The presence of striated muscle fibers in two species is of interest and, while these may function in a similar manner to the smooth muscle, they may represent simply an unusual differentiation of embryonic myoblasts.     

Fluid-structure Interaction within a Layered Aortic Arch Model

The response of wall stress to the elasticity of each layer in the aorta wall was investigated to understand the role of the different elastic properties of layers in the aortic dissection. The complex mechanical interaction between blood flow and wall dynamics in a three-dimensional arch model of an aorta was studied by means of computational coupled fluid-structure interaction analysis. The results show that stresses in the media layer are highest in three layers and that shear stress is concentrated in the media layer near to the adventitia layer. Hence, the difference in the elastic properties of the layers could be responsible for the pathological state in which a tear splits across the tunica media to near to the tunica adventitia and the dissection spreads along the laminar planes of the media layer where it is near the adventitia layer.     

Innervation of the urinary bladder of Ovis aries L

The authors have extended a preliminary study about the innervation of urinary bladder, confirming the previous results pointing out the presence of metasympathetic ganglions in the wall of urinary bladder. Therefore nine urinary bladders of Ovis aries of different age and both sexes have been studied by Ruffini, Bodian and Bielschowsky's staining methods. It's possible summarize the data on the innervation of urinary bladder in the following way: in the tunica adventitia there are motor and sensitive bundles of myelinated nervous fiber. The formers, after many divisions, penetrate into the tunica muscularis contacting bundles of smooth muscle fibers, while the latters after several divisions after giving rise to thinner bundles, produce Pacini-like and Ruffini-like sense corpuscles and free nervous terminations. Furthermore, some metasympathetic ganglions of different size have been detected throughout the running of the bundles. In the tunica submucosa a diffuse and peculiar non myelinated network is observed, arising from the vegetative nervous fibers.     

A histological study on the coronary artery of the indigenous black Bengal goat in Bangladesh.

The coronary artery of the black Bengal goat was studied by light microscopy. The wall of the coronary artery consisted of the tunica intima, tunica media and tunica externa. The tunica intima consisted of a single layer of flattened endothelium. The tunica media was well-developed and composed of mainly of smooth muscle cells together with some fine elastic fibers. The tunica externa consisted of predominant collagen fibers, and some elastic fibers and smooth muscle cells. Elastic fibers in the tunica externa formed a circular arrangement around the tunica media. Sex differences were not observed. The media with well-developed smooth muscle cells may be responsible for changes in functional physiological conditions of the heart.     

Thromboxane A2 analogue contracts predominantly the hepatic veins in isolated canine liver

Thromboxane A₂ (TxA₂) is a potent vasoconstrictor and has been implicated as a mediator of liver diseases such as ischemic-reperfusion injury. We determined the effects of TxA₂ and the well-known hepatic venoconstrictor histamine, on the vascular resistance distribution and liver weight in isolated canine livers perfused with blood via the portal vein. The stable TxA₂ (STA₂; 20 μg, n=5) and histamine (5 μg, n=6) similarly increased the hepatic total vascular resistance, 2.5- and 2.4-fold, respectively. The increase in the hepatic venous resistance was significantly greater than that of the portal resistance (threefold vs. 1.9-fold for STA2; threefold vs. 1.8-fold for histamine). Predominant hepatic venoconstriction induced by both agents was confirmed in livers perfused in a reverse direction from the hepatic vein to the portal vein, as shown by marked precapillary vasoconstriction. STA2 transiently increased liver weight loss (−3.6 g/100g liver weight), followed by a gradual weight gain (9.0 g/100 g). Histamine caused a progressive weight gain (9.1 g/100 g). In conclusion, similar to histamine, TxA₂ constricts predominantly the hepatic vein in isolated canine livers.     

The association of medial collagenous tissue with atheroma formation in the aging human aorta as revealed by a special technique

A new technique which brilliantly colors collagen fibers in a field of polarized light reveals that during mid-life the smooth muscle cells in the tunica media of the human aorta begin to disappear. The connective tissue is divided between two regions; one below the subintimal layer and the other under the adventitia. Fine collagen fibers extend upward from the former into the subintima and beyond into the intima and the overlying atheromatous plaques of the aging aorta. Thus, the source of fibrous thickening of the vessel is not confined solely to the intimal layer; at least, a portion of the total collagen content arises deep within the aortic wall.     

The elastic fibre system in the veins of the human lower oesophagus

The elastic, elaunin and oxytalan fibres in the tunica mucosa and tela submucosa of veins of the human lower oesophagus were studied in 30 necropsy and biopsy specimens using appropriate histological and ultrastructural methodologies. Elaunin fibres predominate in the veins endowed with muscle cells. Scattered oxytalan and elastic fibres were also observed, the latter being more numerous at the vein periphery. No noteworthy fibre arrangement was encountered in veins lacking muscle cells. Those fibres disposed around the veins were considered to belong to the connective tissue of the tunica mucosa and tela submucosa in which they are embedded. The veins of the lower oesophagus seem to be of low elasticity, which may be related to a blockage mechanism described for the gastro-oesophageal blood stream in cases of portal hypertension.     

Identification and characterisation of regional variations in the material properties of ureter according to microstructure

There are few previous studies on the elastic properties of ureter and most have been limited to essentially one-dimensional deformation measurements. The object of this study was, therefore, to identify regional variations in the multiaxial behaviour of rabbit ureter, subjected to in vitro inflation/extension testing under a physiological range of intraluminal pressures and longitudinal forces. A microstructure-motivated material model (via two- and four-fibre families in turn for elastin and collagen) was implemented and its capacity to mathematically characterise the experimental data contrasted favourably with that of the well-established phenomenological models, but it was compromised by parameter covariance. Extensive optimisation studies confirmed that the reduced model without contribution to the elastin and circumferential-fibre (collagen) families characterised the data equally well without over-parameterisation. In view of the fitted parameters, the ureteral tissue was stiffer longitudinally, justified by the preferential alignment of collagen along that axis and the lower ureter was stiffer than the upper ureter, justified by the histological observation of a thickest lamina propria, i.e. of highest collagen content, there. The lower ureter was less anisotropic than the upper ureter, possessing a comparatively larger amount of diagonally arranged collagen fibres in tunica mucosa, while having the usual amounts of longitudinally arranged fibres in tunica adventitia and of circumferentially arranged fibres in tunica muscularis. The present data may be used as inputs to mathematical models of the ureter, assessing regional and intramural stress distributions, through which it is hoped that an improved appreciation of ureteral function may be attained in both health and disease.     

Elastic fibers in the tunica propria of the seminiferous tubules. Light and electron microscopic investigations.

Histological examination of numerous biopsies from mature testes, repeatedly showed irregularities of the elastin staining in the tunica propria of the seminiferous tubules. Even when abundant elastic fibers were visible by light microscopy, no elastic fibers were demonstrable in the electron microscope in ultrathin sections of testicular tubules embedded in Epon and contrasted with phosphotungstic acid (PTA). In 43 biopsies from 22 men aged 17-39 years (19 investigations of sterility and three patients with hypogonadotropic hypogonadism) we therefore checked for the occurrence of elastic fibers in the wall of the seminiferous tubules. Matrix loci of elastic fibers could indeed by demonstrated by electron microscopy using PTA and potassium permanganate (KMnO4), but only after embedding in araldite. Under these conditions, light and electron microscopic findings agreed with each other. The appearances of moderate and severe testicular tubular atrophy differed slightly from one another with regard to the amount of elastin. In the "Sertoli cells only syndrome", elastic fibers were demonstrable only outside the hyalinized inner layer. In the Klinefelter syndrome, only "uncertain" elastin loci were present, but greatly increased microfibrils were to be seen using the electron microscope. No elastic elements and only very sparse microfibrils were present in the tunica propria of the tubules of young men with hypogonadotropic hypogonadism.     

Potential role of the neuropeptide CGRP in the induction of differentiation of rat hepatic portal vein wall

The media of the rat hepatic portal vein is composed of an internal circular muscular layer (CL) and an external longitudinal muscular layer (LL). These two perpendicular layers differentiate progressively from mesenchymal cells within the first month after birth. In this paper, we studied the development of calcitonin gene-related peptide (CGRP) innervation during post-natal differentiation of the vessel. We show that CGRP innervation is already present around the vessel at birth in the future adventitia but far from the lumen of the vessel. Progressively, CGRP immunoreactive fibers reached first LL then CL. CL by itself become only innervated at day 14 after birth. This corresponds to the time at which thick filaments (myosin) are visible in electron microscopy and desmin visualisable by immunocytochemistry. Furthermore, we provide evidence by autoradiography, that binding sites for CGRP are transiently expressed on the portal vein media at day 1 and 14 after birth. Vascular smooth muscle cells were transfected with constructs containing promoters for desmin or smooth muscle myosin heavy chain (smMHC). CGRP treatment of the cells significantly increased the expression of smMHC. Overall these results suggest that CGRP can potentially influence the differentiation of smooth muscle cells from the vessel wall.     

Localization of dopamine D1 and D2 receptor mRNAs in the rat systemic and pulmonary vasculatures.

The present study was designed to evaluate the expression of dopamine D1 and D2 receptor mRNAs in systemic and pulmonary vasculatures. Using specific antisense riboprobes for dopamine D1 and D2 receptor cDNAs, in situ hybridization histochemistry was performed in the aorta, common carotid artery, vertebral artery, pulmonary artery, and superior vena cava of the adult male Sprague Dawley rat. In the case of the aorta, common carotid artery, and vertebral artery, dopamine D1 receptor mRNAs localized mainly in the smooth muscle cells of the tunica media. However, the signals of dopamine D2 receptor mRNAs were found in the endothelium and subendothelial layer of tunica intima, and interstitial cells of tunica adventitia. In the case of the pulmonary artery, signals of dopamine D1 receptor mRNAs were detected within the tunica intima, media, and adventitia. Expression of D2 receptor mRNAs was detected in the walls of small blood vessels within the tunica adventitia of the pulmonary artery. There were no detectable signals of dopamine D1 and D2 receptor mRNAs in the vein. The uneven distribution of dopamine D1 and D2 receptor mRNAs in the rat systemic vasculatures and pulmonary artery suggests that dopamine differentially regulates the vasodilation of the systemic and pulmonary arteries through the differential stimulation of dopamine D1 and D2 receptor.     

Electron microscopic observations on the formation of elastic fibres in cultures of aortic medial cells and adventitial cells

The formation of elastic fibres was observed in the cultured cells derived from the tunica media and the tunica adventitia of mouse aorta. Bundles of myofilaments with dense bodies were abundantly observed in the cytoplasm of the cultured medial cells, and numerous bundles of microfibrillar components were present in the intercellular spaces. Fine granules of approximately 50 nm in diameter were observed in the bundles of microfibrillar components. It was supposed that these fine granules of elastin fused with each other and formed elastic aggregates and then formed large elastic clumps. Numerous bundles of microfibrillar components were also present in the intercellular spaces of the cultured adventitial cells. Elastic aggregates were scarcely observed in the bundles of microfibrillar components. However, large elastic clumps as observed in the medial cell culture could not be found in the adventitial cell culture. It is suggested that the formation of large elastic clumps might be related to the sheet structures or lamellae of elastic fibres in the tunica media.     

Predictive Factors of Late Venous Aortocoronary Graft Failure: Ultrastructural Studies

Background

Venous aortocoronary graft arterialization may precede a preterm occlusion in some coronary artery bypass grafting (CABG) patients. The aim of the present study was to identify ultrastructural variations in the saphenous vein wall that may have an impact on the development of venous graft disease in CABG patients.

Methods

The study involved 365 consecutive patients with a mean age of 62.9±9.4 years who underwent isolated CABG. The thickness and area of the whole venous wall, the tunica intima, the tunica media and the adventitia and the number and shape (length, thickness and length/thickness ratio) of the nuclei in the medial smooth muscle cells nuclei in the distal saphenous vein segments were evaluated by ultrastructural studies. Patients were followed up for 41 to 50 months (mean 45.1±5.1). Saphenous vein graft patency was assessed by follow-up coronary angiography. Logistic regression models were used to identify independent risk factors for late graft failure.

Results

In 71 patients significant lesions in the saphenous vein grafts were observed. The whole venous wall thickness (437.5 µm vs. 405.5 µm), tunica media thickness (257.2 µm vs. 211.5 µm), whole venous wall area (2.23 mm² vs. 2.02 mm²) and tunica media area (1.09 mm² vs. 0.93 mm²) were significantly larger for this group of patients than for those without graft disease. In the latter group more elongated smooth muscle cell nuclei (higher length/thickness ratio) were found in the tunica media of the saphenous vein segments. Thickening of the saphenous vein tunica media and chunky smooth muscle cell nuclei were identified as independent risk factors for graft disease development.

Conclusions

Saphenous vein tunica media hypertrophy (resulting in wall thickening) and chunky smooth muscle cell nuclei might predict the development of venous graft disease.