A correlative lipid and lipolytic study of the pancreas and small intestine of the guinea-pig

With the aid of a variety of TLC preparations the phospholipid and neutral lipid profiles of guinea-pig pancreas and small intestine has been established. Changes in these profiles, subsequent to incubation of these tissues and prior to lipid extraction, has provided evidence relating to their respective lipolytic enzyme capabilities, mainly in the phospholipase group.     

Lysine transport in the guinea-pig small intestine

Interactions between cationic and neutral amino acids in transport across the brush-border membrane, Jmc, of the small intestine have been examined using preparations from the distal rabbit ileum and the rat and guinea-pig mid-small intestine. (1) In the guinea pig, the dependence of Jmc Lys on the concentration of lysine is best described in terms of two saturable transport mechanism in addition to free diffusion. (2) It is shown that the discrepancy between cis-effects of low concentrations of neutral amino acids on the Jmc of cationic amino acids, cis-stimulation in the guinea pig contra cis-inhibition in the rabbit and rat, represents species differences. In the guinea pig, imposing sodium-free conditions turns cis-stimulation into cis-inhibition. (3) It is demonstrated that in rat and guinea pig, leucine is transported both by the transport system(s) for cationic amino acids and by transport system(s) which cannot be inhibited by cationic amino acids.     

Catalase particles in the epithelial cells of the guinea-pig small intestine

Synopsis Purified preparations of epithelial cells have been made from the guinea-pig small intestine. Homogenates of these preparations have been analysed by centrifugation in a zonal rotor. The results confirm the presence of lysosomes in these cells and indicate the existence of catalase particles which equilibrate in a sucrose gradient at a density of between 1.21 and 1.23 and which have a different distribution from other subcellular particles except lysosomes. Injection of Triton WR-1339 into fasting animals enables the separation of lysosomes and catalase particles.     

Carboxylic ester hydrolases in the thyroid gland of the guinea-pig. A light microscopic study

Synopsis The location of cholinesterase and non-specific esterase in the thyroid gland of the guinea-pig was studied with the light microscope. It was found that the inxodyl method for non-specific esterase activity under special conditions is superior to the cholinesterase method in a number of respects for the demonstration of the intra-, inter-and parafollicular cells. When using the indoxyl method the incubation period can be reduced from 2.5–3 hr to 40–50 min. Further, the reaction can be followed during the incubation. False localization of the reaction products is avoided, and nerves and erythrocytes are not stained.By varying the fixation time and the time of storage in gum arabic-sucrose, it was found that the miscellaneous activity of non-specific esterase in APUD cells (C-cells) and follicle cells may be due to both factors. In fresh tissue the activity of the enzyme was equal in follicle and C cells.Special cyst-like structures containing an esterase which is NaF-resistant whenn-naphthyl acetate is employed as a substrate and which gives a strong reaction at low pH values when 5-bromo indoxyl acetate is the substrate, are described, and their nature and possible origin are discussed.     

Zonal rotor analysis of the subcellular localization of alpha-glycerphosphate dehydrogenase, alpha-naphthyl palmitate and beta-naphthyl laurate hydrolases in the mucosa of the guinea-pig small intestine.

Whole homogenate of guinea-pig small intestine mucosa was analysed by centrifugation in a zonal rotor. The results indicate that FAD-linked alpha-glycerophosphate dehydrogenase is localized in the mitochondria and that NAD-linked alpha-glycerophosphate dehydrogeanse is a soluble phase enzyme. An enzyme hydrolysing alpha-naphthyl palmitate at an acid pH was localized in the lysosomes and was activated by 0.1% Triton X-100 and by freezing and thawing. An alkaline hydrolase acting on beta-naphthyl laurate was localized in the 'microsomes'. The possibility of this enzyme being different from alpha-naphthyl acetate hydrolase is discussed.     

A revised nomenclature for mammalian acyl-CoA thioesterases/hydrolases

Acyl-CoA thioesterases, also known as acyl-CoA hydrolases, are a group of enzymes that hydrolyze CoA esters such as acyl-CoAs (saturated, unsaturated, branched-chain), bile acid-CoAs, CoA esters of prostaglandins, etc., to the corresponding free acid and CoA. However, there is significant confusion regarding the nomenclature of these genes. In agreement with the HUGO Gene Nomenclature Committee and the Mouse Genomic Nomenclature Committee, a revised nomenclature for mammalian acyl-CoA thioesterases/hydrolases has been suggested for the 12 member family. The family root symbol is ACOT, with human genes named ACOT1-ACOT12, and rat and mouse genes named Acot1-Acot12. Several of the ACOT genes are the result of splicing events, and these splice variants are cataloged.     

Shapes and projections of tertiary plexus neurons of the guinea-pig small intestine

The axons of neurons that innervate the longitudinal muscle of the small intestine in small mammals such as rabbit, rat, guinea pig and mouse form a network, the tertiary plexus, against the inner surface of the muscle. In general, because of their substantial overlap, it has not been possible to follow the ramifications of individual axons in the tertiary plexus. In the present work, the longitudinal muscle motor neurons were filled with marker dyes through an intracellular microelectrode, and their morphologies and projections were examined in whole-mount preparations of longitudinal muscle and myenteric plexus. Most neurons that were examined were in the small intestine (ileum and duodenum), but a few were examined in the distal colon. Neurons in all regions had similar morphologies and projections. The cell bodies were amongst the smallest in myenteric ganglia, with major and minor axes of 14 microns and 25 microns (mean, n = 40) in the plane of the myenteric plexus. Each neuron had a single axon that branched extensively in the tertiary plexus, most had multiple lamellar dendrites and a few had filamentous dendrites or a mixture of filamentous and lamellar dendrites. The mean area of muscle covered by an axon and its branches extended 1.6 mm orally to anally and 1.7 mm circumferentially. The area covered was 2.8 +/- 1.9 mm2 (mean +/- SD, n = 23). From the density of occurrence of cell bodies, it can be calculated that each point in the longitudinal muscle is innervated by the processes of about 100 motor neurons and is influenced by electrotonic conduction of signals through the muscle by about 300 motor neurons.     

Total numbers of neurons in myenteric ganglia of the guinea-pig small intestine

Two techniques that are thought to stain all of the neurons in the myenteric ganglia of the intestine are NADH diaphorase histochemistry and immunhistochemistry using a nerve cell body antiserum. However, this assumption has never been directly verified. In the present study myenteric ganglia of the guinea-pig ileum were prepared as whole-mounts and stained with either of these techniques. All nerve cells that could be identified in the whole-mounts were counted. The whole-mounts were then embedded flat in resin and serially sectioned at 1 m. Nerve cells were identified and counted from the serial sections, and the data compared to those obtained from the whole-mounts. NADH diaphorase histochemistry did not reveal all the neurons at incubation times that gave selective staining. In contrast, nerve cell body antiserum stained the entire neuronal population. To determine the total number of nerve cell bodies/ganglion and the proportion of nerve cell bodies with calbindin immunoreactivity, whole-mounts that had been processed for calbindin immunohistochemistry were serially sectioned and reconstructed. The total number of neurons per myenteric ganglion was 105±10 (SE). Calbindin-immunoreactive neurons comprised about 20% of the myenteric neurons, which is considerably less than previous estimates, because previously the total population has been underestimated. The spatial density of myenteric neurons in the undistended ileum of the guinea-pig is 17300 nerve cells/cm².     

Phosphodiesterase II in epithelial cells from guinea-pig and rat small intestine

Phosphodiesterase II activity was determined by using a synthetic substrate, the 2,4-dinitrophenyl ester of thymidine 3'-phosphate. The enzyme activity was determined in fractions obtained by differential centrifugation of homogenates of epithelial cells from the small intestinal mucosa of guinea pigs and rats. In guinea-pig preparations phosphodiesterase II occurred with highest specific activity in those fractions rich in succinate dehydrogenase and acid phosphatase. A lysosomal location for the guinea-pig enzyme was indicated by its structure-linked latency and by its association with particles that under-went a characteristic decrease in equilibrium density when Triton WR-1339 was injected into the animals. With rat preparations a much greater proportion of the phosphodiesterase II activity was found in the soluble fraction after ultracentrifugation. The rat enzyme exhibited a lower degree of latency and administration of Triton WR-1339 had no effect. The rat enzyme further differed from that of the guinea pig in other respects; it was more labile at 60 degrees C, it exhibited a lower pH optimum and it had a higher molecular weight as determined by gel-filtration chromatography.     

Acid hydrolases in the suckling rat small intestine I. Fractionation of mucosal homogenates

Synopsis Small intestine mucosal homogenates of suckling rats have been fractionated by centrifugation and analyzed for acid hydrolases and for biochemical markers of subcellular organelles. The results indicate that the acid hydrolases are associated with particles having sedimentation properties similar to those of mitochrondria. The acid hydrolases exhibited latent activity. Subfractionation on a continuous density gradient of sucrose in deuterium oxide demonstrated that these enzymes are associated with particles distinct from other subcellular organelles. Electron micrographs of the acid hydrolase-rich region of the gradient show the presence of numerous small electron dense bodies bounded by a unit membrane.     

Scanning electron microscopy of the muscle coat of the guinea-pig small intestine

Summary We have studied the layers of the muscular coat of the guinea-pig small intestine after enzymatic and chemical removal of extracellular connective tissue. The cells of the longitudinal muscle layer are wider, have rougher surfaces, more finger-like processes and more complex terminations, but fewer intercellular junctions than cells in the circular muscle layer. A special layer of wide, flat cells with a dense innervation exists at the inner margin of the circular muscle layer, facing the submucosa. The ganglia of the myenteric and submucosal plexuses are covered by a smooth basal lamina, a delicate feltwork of collagen fibrils, and innumerable connective tissue cells. The neuronal and glial cell processes at the surface of ganglia form an interlocking mosaic, which is loosely packed in newborn and young animals, but becomes tightly packed in adults. The arrangement of glial cells becomes progressively looser along finer nerve bundles. Single varicose nerve fibres are rarely exposed, but multiaxonal bundles are common. Fibroblast-like cells of characteristic shape and orientation are found in the serosa; around nerve ganglia; in the intermuscular connective tissue layer and in the circular muscle, where they bridge nerve bundles and muscle cells; at the submucosal face of the special, flattened inner circular muscle layer; and in the submucosa. Some of these fibroblast like cells correspond to interstitial cells of Cajal. Other structures readily visualized by scanning electron microscopy are blood and lymphatic vessels and their periendothelial cells. The relationship of cellular elements to connective tissue was studied with three different preparative procedures: (1) freeze-cracked specimens of intact, undigested intestine; (2) stretch preparations of longitudinal muscle with adhering myenteric plexus; (3) sheets of submucosal collagen bundles from which all cellular elements had been removed by prolonged detergent extraction.