Peptidergic innervation of mesenteric lymphatics in guinea pigs: an immunocytochemical and pharmacological study.

By immunocytochemistry, substance P immunoreactive (SP-IR) and vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibers were examined in guinea pig mesenteric lymph collectors. The immunoreactive nerve fibers, located in the adventitia of lymphatics, were few and were irregularly distributed along the vessel wall. These fibers appeared to be more numerous and more evenly distributed along the corresponding artery and vein walls within the same area. SP immunoreactivity in the vascular nerves was depleted in guinea pigs injected with capsaicin but was unaffected by the injection of 6-hydroxydopamine. By contrast, VIP-IR nerve fibers were unaffected by both treatments. It is concluded that SP-IR nerve fibers in the lymphatics are likely to be of sensory origin and that VIP containing nerves in the lymph collectors are distinct from SP-containing and noradrenergic nerves. It is also suggested that lymph collectors possess a complex although limited innervation pattern not only of autonomic nerve fibers containing classic neurotransmitters but also of peptidergic nerve fibers of a different origin with a vasomotor and/or sensory action.

Ultrastructural cytochemistry of anchoring filaments of human lymphatic capillaries and their relation to elastic fibers.

In previous studies "anchoring filaments" of human lymphatic capillaries have been shown to consist of microfibrils having histochemical and ultrastructural characteristics similar to elastin-associated microfibrils. When not associated with an elastin component, these microfibrils are referred to as "oxytalan microfibrils." In this study, alpha-glycol-containing carbohydrates and glycoconjugated sulfate groups, originating from sulphydryls and/or disulfide bridges, have been detected in anchoring filament microfibrils of human lymphatic capillaries by Thiery reaction (PA-TCH-SP) and "Hight Iron Diamine" cytochemical method (HID), respectively. Both of these chemical groups belong to the putative glycoprotein of which the microfibrils are constituted. Similar molecular characteristics have been demonstrated in elastic fiber microfibrils and oxytalan microfibrils of connective tissue. These findings suggest a close molecular similarity among these different types of microfibrils. Thus, whatever their individual location or denomination (anchoring filaments, oxytalan fibers, or elastin-associated microfibrils), these microfibrils form an uniform population of fibrous elements. These findings further support a structural (and functional) continuity between the lymphatic capillary wall and the elastic network of adjacent connective tissues previously described and termed "Fibrillar Elastic Apparatus" (FEA). Of interest, endothelial cells also selectively react positively to the PA-TCH-SP and HID methods.

A fibrillar elastic apparatus around human lymph capillaries.

A fibrillar elastic apparatus around the wall of human lymph capillaries is demonstrated by means of histochemical and ultrastructural techniques. This apparatus consists of three interlinked components listed here in order of increasing distance from the capillary wall: 1) oxytalan fibres connected to the abluminal surface of the endothelial cells, known also as "anchoring filaments" and consisting of bundles of microfibrils; 2) elaunin fibres consisting of microfibrils and a small amount of elastin; and 3) typical elastic fibres consisting of microfibrils and abundant elastin. The microfibrillar constituent has similar ultrastructural features in the three components of the elastic apparatus. Microfibrils have a diameter of 12-14 nm, an electron-transparent core and a wall with 3-5 electron-dense subunits and oblique cross striations with a period of 15-17 nm. Microfibrils are the common element of the three components of the elastic apparatus and they link them to one another and to the elastic network of the perivascular connective tissue. An elastic apparatus was not found around blood capillaries and it can thus provide a histological marker to identify lymph capillaries. The possible role of the lymphatic elastic apparatus in the physiological activity of the lymphatic absorbing network is discussed and it is proposed that its disconnection from the elastic network of the tissue may promote pathological conditions such as lymphoedema or diseases related to impaired immune responses.

Morphometric analysis of elastic fibers in human skin lymphatic capillaries.

In contrast to their absence near dermal blood capillaries, elastic fibers are commonly seen adjacent to dermal lymphatic capillaries under light microscopy. Based on morphometric analysis, the elastic fiber network that surrounds these skin lymphatic capillaries is predominantly oriented longitudinally to the lymphatic vessel wall. Quantitative analysis reveals that the density of these pericapillary elastic fibers are almost twice that of the intercapillary elastic fibers but only about one-half as thick. These data suggest that dermal lymph capillaries are surrounded by a specific elastic network of functional significance, morphologically distinct from that seen in the intercapillary dermis. Because lymphatic capillaries are often difficult to identify especially when collapsed, this elastic network may facilitate the positive identification of dermal lymphatic capillaries by light microscopy and thereby help differentiate them from blood capillaries. The possible role of this lymphatic elastic network in the absorptive activity of the dermal lymphatic system is also discussed.

Type 3 (chronic) GM1 gangliosidosis presenting as infanto-choreo-athetotic dementia, without epilepsy, in three sisters.

Three sisters (ages 27, 24, and 17 years) presented with slowly progressing dystonic dementia and spastic tetraparesis with infantile onset. CSF, bone marrow, and conjunctival cells showed storage vacuoles. Biochemical analysis revealed increased urinary oligosaccharide excretion and decreased activity of acid beta-D-galactosidase and beta-D-fucosidase in serum, leukocytes, and cultured fibroblasts. The parents' enzyme values were in the heterozygous range. This is the only case in the literature of severe dementia associated with the clinical symptoms of type 3 GM1 gangliosidosis. The clinical heterogeneity of GM1 gangliosidosis and the significance of the combination of beta-D-galactosidase and beta-D-fucosidase defects in this syndrome are discussed.

Identification of beta-adrenergic sensitive adenylate cyclase in rat thoracic duct.

The effect of L-isoproterenol on the 3',5'-cyclic adenosine monophosphate (cAMP) generating system in rat thoracic duct membranes was investigated in order to identify beta-adrenergic receptors. L-Isoproterenol elicited a dose-dependent stimulation of cAMP formation; L-noradrenaline was less effective than L-isoproterenol in stimulating cAMP increase, whereas L-phenylephrine was without important effects on cAMP levels. L-Propranolol, a selective antagonist of beta-adrenergic receptors, caused a dose-dependent decrease of the effects of L-isoproterenol. In contrast, the L-isoproterenol-elicited increase of cAMP was unaffected by the alpha-adrenergic and dopamine receptor-blocking agents phentolamine and haloperidol. These data indicate that L-isoproterenol stimulates cAMP formation in the rat thoracic duct by a specific interaction with beta-adrenergic receptors positively coupled to adenylate cyclase.

Increased density of perivascular nerves to the major cerebral vessels of the spontaneously hypertensive rat: differential changes in noradrenaline and neuropeptide Y during development.

Fluorescence and immunohistochemical techniques were used to study the pattern and density of perivascular nerves containing noradrenaline (NA) and neuropeptide Y (NPY) supplying the major cerebral arteries of 4-, 6-, 8- and 12-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar (WIS) controls. Levels of NA and NPY in the superior cervical ganglia were measured. The density of nerves containing NA and NPY was greater in the hypertensive animals at all ages studied. However, the developmental changes in the density of innervation showed similar trends in both SHR and WIS groups. With few exceptions, there was a significant increase in the density of nerves containing NA from 4 to 6 weeks and from 8 to 12 weeks of age. This was in contrast to a low expression, and in some vessels a significant decrease in the number of NPY-containing nerves from 4 to 6 weeks. The density of nerve fibres containing NPY increased significantly in almost all vessels between 6 and 8 weeks of age and then stabilized. Thus there is a differential time course for the appearance of NA and NPY during development. Furthermore, the hyperinnervation of cerebral vessels in SHR by nerves containing NA and NPY precedes the onset of hypertension and associated medial hypertrophy. High-performance liquid chromatography and enzyme-linked immunosorbant assays show that the NA and NPY contents of the superior cervical ganglion do not reflect the changes in innervation pattern seen in the terminal fibres in the cerebral arteries. This tends to support the view that a local neurovascular mechanism is involved in the maintenance of hypertension. The possibility that increase in NPY as well as NA in cerebral perivascular nerves of hypertensive animals is involved in the protection of the blood-brain barrier against oedema and cerebral haemorrhage is raised.

[Morphological study by scanning electron microscopy of the lymphatic vessels of the guinea pig]. / Studio morfologico al microscopio elettronico a scansione (SEM) dei vasi linfatici di cavia.

The purpose of this study was to describe the morphology of the whole lymphatic way: from capillaries to thoracic duct including cisterna chili using scanning electron microscopy and Evan's technique. We observed the lymph vascular wall that is: the endothelial surface, the muscular layer and the adventitial one. All these vessels were covered by an endothelial surface, with raised nuclei and long cell axes oriented parallel to the direction of flow. The borders between adjacent endothelial cell were often seen and open junctions were noted in lymphatic capillaries. The technique we used, permitted the removal of connective tissue by HC1 hydrolysis, so that smooth muscle cells could be examined. The latter showed a great variety of aspects and a very irregular course. The adventitial layer was thin in capillaries and became complex in thoracic duct where collagen fibers and connective elements were seen.

[Mesentery collector valves in the guinea pig: morpho-functional aspects]. / Le valvole dei collecttori mesenteriali di cavia: aspetti morfo-funzionali.

In guinea pig mesenterial lymph vessels there are many bicuspid valves, which determine the flow centralwards of lymph. Our observations, based on the study of 52 guinea pig lymph collectors, demonstrated a different number of valves on different parts of each vessel. In fact we found more valves in the part near the intestinal wall and the mesenterial lymph node than in the middle part of the vessel. Besides we measured the time of flow in these different portions by Indian ink injection and we found a correlation between valve number and flow. In fact the time of flow decreases with the increase of the number of valves the shortest being near the intestinal wall and the mesenterial lymph node and the longest being in the middle part of the collectors. The experimental data indicate that the valves have an important role in the lymph circulation because they favour the flow lymph velocity.

Localization of dopamine receptors in the rabbit renal artery: a histoautoradiographic study.

The localization of dopamine receptors within rabbit renal artery was studied using 3H-spiroperidol as a label for dopamine receptors and a histoautoradiographic technique. Preliminary radioreceptor binding studies showed that 3H-spiroperidol was bound to sections of renal artery in a manner consistent with the existence of dopamine receptors. In fact the binding was found to be saturable, stereospecific and of high affinity, with a Bmax approximately of 158.3 fmol/mg protein and a Kd of 13.5 nmol/l. The microscopic examination of sections processed for the histoautoradiographic demonstration of 3'-spiroperidol binding sites showed that the distribution of dopamine receptors in the renal artery was widespread. The highest concentration of dopamine receptors was found primarily in the smooth muscle cells of the media and then, in the following order, in endothelial cells of the intima and in fibrous and connective cells of the adventitia. The direct demonstration of dopamine receptors in the media of rabbit renal artery strongly supports the hypothesis that these receptors may be involved in the relaxation of the artery caused by infusion or application of dopamine.

[Histochemical and ultrastructural studies of the innervation of the lymph-vessel and blood-vessel wall. 1. Adrenergic innervation]. / Richerche istochimiche ed ultrastrutturali sull'innervazione della parete vascolare linfatica e sanguifera. 1) L'innervazione adrenergica.

The adrenergic innervation of the lymph vascular wall was studied by means of the Falck fluorescence histochemical tecnique and electron microscopy with Tranzer and Richards' histochemical tecnique. The lymph vessels wall, compared with that of blood vessels, shows very few adrenergic nerve fibers located in the adventitia outside the smooth muscle cells. The possible role of the nervous system in the motor control of the lymph vessels is discussed.

[Histochemical and ultrastructural studies of the innervation of the lymph-vessel and blood-vessel wall. II. Cholinergic innervation]. / Richerche istochimiche e ultrastrutturali sull'innervazione della parete vascolare linfatica e sanguifera. II. L'innervazione colinergica.

Using the Acetylcholinesterase (AChE) tecnique applied to light and electron microscopy, was observed that the lymph vascular wall shows very few and inconstant AChE-positive fibers. The cholinergic fibers run prevalently longitudinal in the perivascular connective tissue, only brief segments show a loose network. The results are discussed and compared with blood vessels innervation.

Ultrastructural modifications of the testes and accessory sex glands of genetically dwarf mice after treatment with human chorionic somatomammotropin.

We have studied the effects of treatment with human chorionic somatommatropin (hCS) in the dwarf mouse on the weight and ultrastructure of the testes and accessory sex glands. We used animals of the Snell-Bagg (dw/dw) strain, whose hypophyses secrete neither prolactin nor growth hormone. The effects found were compared with those produced by a similar treatment with ovine prolactin. The administration of hCS causes a significant increase in the wet weight of the testes and accessory sex glands. Ultrastructural observations have demonstrated that hCS is able to stimulate sperminogenesis and the secretory function of the accessory sex glands. Prolactin, as shown in previous reports in the literature, has a similar action on the male reproductive system. However, contrary to the observation after prolactin therapy, no matings were observed after treatment with hCS.