Innervation of the renal vasculature of the toad (Bufo marinus)

The innervation of the dorsal aorta and renal vasculature in the toad (Bufo marinus) has been studied with both fluorescence and ultrastructural histochemistry. The innervation consists primarily of a dense plexus of adrenergic nerves associated with all levels of the preglomerular vasculature. Non-adrenergic nerves are occasionally found in the renal artery, and even more rarely near the afferent arterioles. Many of the adrenergic nerve profiles in the dorsal aorta and renal vasculature are distinguished by high proportions of chromaffin-negative, large, filled vesicles. Close neuromuscular contacts are common in both the renal arteries and afferent arterioles. Possibly every smooth muscle cell in the afferent arterioles is multiply innervated. The glomerular capillaries and peritubular vessels are not innervated, and only 3-5% of efferent arterioles are accompanied by single adrenergic nerve fibres. Thus, nervous control of glomerular blood flow must be exerted primarily by adrenergic nerves acting on the preglomerular vasculature. The adrenergic innervation of the renal portal veins and efferent renal veins may play a role in regulating peritubular blood flow. In addition, glomerular and postglomerular control of renal blood flow could be achieved by circulating agents acting via contractile elements in the glomerular mesangial cells, and in the endothelial cells and pericytes of the efferent arterioles. Some adrenergic nerve profiles near afferent arterioles are as close as 70 nm to distal tubule cells, indicating that tubular function may be directly controlled by adrenergic nerves.     

The identification, purification and characterisation of an inhibitor of collagenase (20K) produced by neoplastic epithelial cells

A rat carcinoma cell line (T2/H7) constitutively synthesised interstitial collagenase. When these cells were incubated with 12-O-tetradecanoylphorbol 13-acetate (TPA) they secreted an inhibitor of collagenase, which resulted in a net decrease of collagenolytic activity being detected in conditioned medium. Using reverse zymography, the Mr of the inhibitor was found to be 20,000 which suggests that it may be the rat homologue of inhibitor of metalloproteinase 2 (IMP2; TIMP-2), as it inhibited both the gelatinolytic and collagenolytic activities of rat collagenase. The inhibitor was separated from collagenase by filtration through a YM30 membrane. The inhibitor was purified further by sequential chromatography on heparin-Sepharose and Con A-Sepharose. It bound to heparin-Sepharose in 75 mM NaCl and was eluted with 300 mM NaCl. It did not bind to Con A-Sepharose, suggesting that it was a non-glycosylated molecule. The inhibitor was resistant to treatment with either trypsin, APMA or heat.     

Ultrastructural and tissue-culture studies on the role of fibronectin,collagen and glycosaminoglycans in the migration of neural crest cells in the fowl embryo

Summary The initial migration of neural crest (NC) cells into cell-free space was studied by transmission electron microscopy at trunk levels of fowl embryos, some of which were fixed in the presence of ruthenium red. Migrating NC cells occurred in zones which contained fewer ruthenium-red stained 15–40 nm diameter granules than other regions. The ruthenium-red stained granules were linked by similarly stained thin ( 3 nm diameter) microfibrils. The granules resemble proteoglycan and the microfibrils may be hyaluronate. NC cells contacted thicker ( 10 nm diameter) fibrils and interstitial bodies, which did not require ruthenium red for visualization. Cytoplasmic microfilaments were sometimes aligned at the point of contact with the extracellular fibrils, which may be fibronectin and collagen.Phase-contrast time-lapse videotaping and scanning electron microscopy showed that NC cells of the fowl embryo in vitro migrated earlier and more extensively on glass coated with fibronectin-rich fibrous material and adsorbed fibronectin molecules than on glass coated with collagen type I (fibres and adsorbed molecules). NC cells became completely enmeshed in fibronectin-rich fibres, but generally remained on the surface of collagen-fibre gels. When given a choice, NC cells strongly preferred fibronectin coatings to plain glass, and plain glass to dried collagen gels. NC cells showed a slight preference for plain glass over glass to which collagen was adsorbed. Addition to the culture medium of hyaluronate (initial conc. 20 mg/ml), chondroitin (5 mg/ml) and fully sulphated chondroitin sulphate and dermatan sulphate (up to 10 mg/ml) did not drastically alter NC cell migration on fibronectin-rich fibrous substrates. However, partially desulphated chondroitin sulphate (5mg/ml) strongly retarded the migration of NC cells.The in vivo and in vitro studies suggest that fibronectin may dictate the pathways of NC cell migration by acting as a highly preferred physical substrate. However, the utilization of these pathways may be reduced by the presence of proteoglycans bearing undersulphated chondroitin sulphate.Abbreviations NC neural crest - ECM extracellular material - GAG glycosaminoglycan - FN fibronectin - CIG cold insoluble globulin - TEM transmission electron microscopy - SEM scanning electron microscopy - DMEM-H HEPES buffered Dulbecco's modified Eagle's medium - FCS foetal calf serum - CEE chick embryo extract - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - PBS phosphate-buffered saline     

Galanin is more common than NPY in vascular sympathetic neurons of the brush-tailed possum.

The distribution of galanin (Gal) in sympathetic vascular neurons of adult and juvenile brush-tailed possums (Trichosurus vulpecula), was examined using double-labelling immunohistochemistry. This was compared with the distribution of neuropeptide Y (NPY) in the same tissues. Immunoreactivity (IR) to galanin was present in the majority (64-99%) of nerve cell bodies in paravertebral sympathetic ganglia, where it mostly co-existed with IR to the catecholamine-synthesizing enzyme, tyrosine hydroxylase (TH). Gal-IR also was present in most, if not all, TH-IR perivascular axons supplying systemic arteries and veins. NPY-IR was less common than Gal-IR in all sympathetic ganglia and perivascular axons examined. Some sympathetic, TH-IR axons supplying the abdominal aorta and renal artery contained both Gal-IR and NPY-IR, while TH-IR axons supplying cephalic and thoracic vessels contained Gal-IR but not NPY-IR. Limited observations on sympathetic neurons in two species of wallabies indicated that Gal-IR also was more common than NPY-IR in other marsupial species, but the incidence of NPY-IR was higher in these wallabies than in the brush-tailed possum. Together with previous studies, this work suggests that the coexistence of galanin and NPY may be the primitive condition for sympathetic neurons in tetrapods. The differential expression of these peptides in specific populations of sympathetic neurons may have important functional consequences in the autonomic control of the circulation.     

Enzymes associated with metabolism of xylose and other pentoses by Prevotella (Bacteroides) ruminicola strains, Selenomonas ruminantium D, and Fibrobacter succinogenes S85.

Prevotella (Bacteroides) ruminicola strains B(1)4 and S23 and Selenomonas ruminantium strain D used xylose as the sole source of carbohydrate for growth, whereas Fibrobacter succinogenes was unable to metabolize xylose. Prevotella ruminicola strain B(1)4 exhibited transport activity for xylose. In contrast, F. succinogenes lacked typical xylose uptake activity but did exhibit low binding potential for the sugar. Prevotella ruminicola strains B(1)4 and S23 as well as S. ruminantium D showed low xylose isomerase activities but higher xylulokinase activities, using assays that gave high activities for these enzymes in Escherichia coli. Xylose isomerase appeared to be produced constitutively in these ruminal bacteria, but xylulokinase was induced to varying degrees with xylose as the source of carbohydrate. Fibrobacter succinogenes lacked xylose isomerase and xylulokinase. All three species of ruminal bacteria possessed transketolase, xylulose-5-phosphate epimerase, and ribose-5-phosphate isomerase activities. Neither P. ruminicola B(1)4 nor F. succinogenes S85 showed significant phosphoketolase activity. The data indicate that F. succinogenes is unable to either actively uptake or metabolize xylose as a result of the absence of functional xylose permease, xylose isomerase, and xylulokinase activities, although it and both P. ruminicola and S. ruminantium possess the essential enzymes of the nonoxidative branch of the pentose phosphate cycle.     

Jejunal malabsorption in the rat infected by the nematode Nippostrongylus brasiliensis

Symons L. E. A., Gibbins J. R. and Jones W. O., 1970. Jejunal malabsorption in the rat infected by the nematode Nippostrongylus brasiliensis. International Journal for Parasitology, 1: 179–187. The rate of jejunal absorption of a range of substances absorbed actively or by diffusion was depressed in the rat infected by the nematode Nippostrongylus brasiliensis. The degree of malabsorption of actively absorbed substances was directly related to the severity of the infection.     

Effects of long term denervation on smooth muscle of the chicken expansor secundariorum

Summary Denervation of the expansor secundariorum muscle of the adult and 2 week chicken, by sectioning the brachial plexus, resulted in an approximate twofold increase in dry weight over 8 weeks. Unlike skeletal muscle, no ultrastructural changes were exhibited by the smooth muscle cells for a period of up to 5 months post denervation. No evidence of hypertrophy of the individual muscle cells was observed, but following colchicine treatment a definite increase in the number of mitotic figures was noted within muscle bundles indicating that the increase in dry weight of the expansor muscle is due to hyperplasia of the smooth muscle cells. The results are discussed in relation to in vitro studies of the interaction of sympathetic nerves with smooth muscle.     

Neurochemically similar myenteric and submucous neurons directly traced to the mucosa of the small intestine

Summary Antisera to neuropeptide Y (NPY) gave an intense immunohistochemical reaction of certain nerve cells in the myenteric and submucous plexuses of the guinea-pig small intestine. Each nerve cell had up to 20 branching, tapering processes that were less than 50 m long and a long process that could be followed for a considerable distance. This morphology corresponds to that of the type-III cells of Dogiel. The long process of each myenteric cell ran through the circular muscle to the submucosa, and in most cases the process could be traced to the mucosa. The submucous nerve cell bodies also had processes that extended to the mucosa. These cell bodies, in both plexuses, also stained with antisera raised against calcitonin generelated peptide (CGRP), cholecystokinin (CCK), choline acetyltransferase (ChAT) and somatostatin (SOM), but did not stain with antibodies against enkephalin, substance P or vasoactive intestinal peptide. Thus, it has been possible for the first time to trace the processes of chemically specified neurons through the layers of the intestinal wall and to show by a direct method that CGRP/CCK/ChAT/NPY/ SOM myenteric and submucous nerves cells provide terminals in the mucosa.     

Differentiation of sympathetic and enteric neurons of the fowl embryo in grafts to the chorio-allantoic membrane

Summary Sympathetic cells (adrenergic neurons, SIF cells and chromaffin cells) and enteric neurons differentiate from migratory cells derived from the neural crest. The development of these cell types was studied in chorio-allantoic membrane (CAM) grafts, using combinations of tissues from domestic fowl embryos. Neural anlagen (neural tube and crest) of the vagal, cervico-thoracic and lumbo-sacral axial levels were equally capable of sympathetic differentiation, but this required somitic tissue for its significant expression. However, the vagal somites possessed only slight sympathogenic activity, thereby accounting for the negligible contribution of the vagal neural crest to the sympathetic nervous system.The same three levels of the neural anlage could furnish enteric neurons when combined directly with the aneuronal colo-rectum. However, the scale of this line of differentiation varied with the level of origin of the neural anlage, in contrast to the apparent equivalence in the ability to diffentiate as sympathetic cells. The density of enteric neurons in combinations with the vagal neural anlage was estimated as 60 times greater than the neuron density in combinations with the cervico-thoracic neural anlage. The lumbo-sacral neural anlage gave results similar to those of the cervico-thoracic level. Moreover, neural crest-derived pigment cells, positioned ectopically in the wall of the colo-rectum, were rare in combinations with the vagal neural anlage, but common in grafts with the other levels.When tested physiologically, the colo-rectum grown with the vagal neural anlage showed non-adrenergic, non-cholinergic inhibitory nervous activity in addition to the expected cholinergic excitatory responses. The neurons derived directly from vagal neural anlagen were similar to those that had reached the colo-rectum via their normal migratory pathways, when studied in terms of histological appearance, density of distribution and physiological responses.