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Notes: A theory for the order in σ phases based on the sphere-packing model of Wilson & Spooner [Acta Cryst. (1973), A29, 342-352] has been examined using a mathematical analysis. The analysis suggests that the prediction of order in these phases based on lattice-constant variations associated with variations in atomic diameter should be treated with caution, particularly where the results are at variance with experimentally determined results.

Notes: Abstract: Benzodiazepines bind to glial membranes on a single type of site, with a high affinity (KD= 5 × 10−9 M) on about 100 fmol of sites per mg protein. The number of binding sites is increased when the membranes are treated with Triton X-100. Antiepileptic drugs such as clonazepam and phenobarbital and hypnotic drugs such as Ro-11-3128 and Ro-11-6896 are able in pharmacological concentrations to displace [3H]flunitrazepam from its glial binding sites.

Notes: 1. We have studied the number and distribution of adrenaline synthesizing nerve cells in the medulla oblongata of the rat, using a combination of immunofluorescence to visualize the enzyme phenylethanolamine-N-methyltransferase (PNMT) and catecholamine fluorescence to detect central catecholamines.2. The distribution of adrenaline synthesizing nerve cells was similar in normo-tensive (Wistar Kyoto) rats, spontaneous hypertensive rats, and stroke-prone rats. Few of the cells visualized by PNMT immunofluorescence were detected by the Faglu fluorescence method for catecholamines. The CI (ventrolateral) and C2 (dorsomedial) groups of PNMT cells were anatomically distinct from the Al and A2 groups of catecholamine fluorescent cells and lay rostral to these cells within the medulla. There was a third group of adrenaline synthesizing cells close to the midline in the rostral medulla, and we have called this the C3 group.3. There was a 32% increase in the number of PNMT cells in the medulla of 4-week-old stroke-prone rats.4. PNMT enzyme activity in a cross-segment of the medulla containing the adrenaline synthesizing cells was also increased by 30% in both spontaneous hypertensive rats and stroke-prone rats.

Notes: 1. Concentrations of noradrenaline, adrenaline and dopamine were measured in the submucosa and myenteric plexus of innervated and extrinsically denervated guinea-pig ileum using a sensitive radioisotope enzymatic assay for catecholamines.2. Subcellular fractionation studies indicated that the microsomal fraction obtained from both layers of the normal ileum was greatly enriched with noradrenaline compared to the total homogenate. Low levels of adrenaline and dopamine were also detected in both layers of the ileum.3. After extrinsic denervation or pretreatment with reserpine, noradrenaline was reduced to less than 3% and could no longer be visualized histochemically. Small proportions of the adrenaline and dopamine also disappeared after extrinsic denervation.4. The residual amounts of noradrenaline, adrenaline and dopamine present after extrinsic denervation were not sensitive to reserpine and were not concentrated in microsomal fractions suggesting that these amines are not stored as neurotransmitters in intrinsic neurons of the intestine.

Notes: Summary In this paper a steady-state kinetic study on the system lactate dehydrogenase-β-thiopyruvate-β-thiolactate is presented and the possible mechanistic and physiological implications are discussed. At pH 7.4 the equilibrium between β-thiopyruvate and β-thiolactate, in the presence of NADH and lactate dehydrogenase is largerly shifted towards the formation of β-thiolactate as in the case of pyruvate and lactate. This can be relevant in connection with the mixed disulfide between cysteine and β-thiolactate that is observed to be present in the mammalian body fluids. The catalytic mechanism is of the bi-bi compulsory order type, and rapid equilibrium conditions for the binding of the first substrate (NADH) are shown to apply. A complex inhibition pattern of inhibitions by both substrates, however, prevents simple suggestions about the nature of the dead-end species involved.

Notes: Summary A method is described for the immunohistochemical localization of peptides in whole-mount preparations. Tissue is fixed as laminae with a picric acid/formaldehyde mixture and then dehydrated, cleared and rehydrated before exposure to antibodies. This procedure ensures adequate penetration of the antibody molecules without the need to freeze and thaw the tissue or to use detergents, preserves antigenicity and lowers non-specific background staining. The laminae are incubated with the primary antisera for 16 h at room temperature and, after washing, with a second, fluorescent tagged, antiserum. This can be followed by a peroxidase-anti-peroxidase localization of the second antiserum, which acts as a bridge. The method gives a precise and reproducible localization of immunoreactive peptides, with good penetration and low background even in thick preparations. Large areas can be scanned and neuroeffector relationships studied more easily than in sections.

Notes: Summary A fine structural study was made of the ganglia, neurons, Schwann cells and neuropil of the submucous plexus of the guinea-pig ileum. The arrangement of the plexus as seen by light microscopy is briefly described. Submucous ganglia are small, containing an average of eight neurons per ganglion (compared with 43 in myenteric ganglia) and are connected with each other by fine nerve strands. The cell bodies of neurons and Schwann cells and a neuropil consisting of neuronal and Schwann cell processes form the ganglia. No other cell types or blood vessels are found within the ganglia. Ganglia are surrounded by a continuous basal lamina but lack a well-defined connective tissue investment. The glial investment of neurons is incomplete: many neurons lie directly beneath the basal lamina with no intervening Schwann cell processes, and the plasma membranes of adjacent neurons are often directly apposed over large areas. Other areas of apposition occur between the cell bodies and processes of neurons and Schwann cells. Desmosome-like membrane specializations may be seen between neurons and other neurons or Schwann cells. Submucous neurons could not be categorized according to size, shape, organelle content or types of processes. Processes emerging from nerve-cell bodies were placed into four broad categories on the basis of shape and microtubule content. Many bundles of closely apposed small nerve profiles lacking intervening Schwann processes are found in the neuropil in addition to a large number of vesiculated varicosities, some of which are directly apposed to the plasma membranes of nerve-cell bodies. A small proportion of vesiculated profiles form synapses with nerve cell bodies, their processes and profiles in the neuropil. From their structure, submucous neurons appear to form a more homogeneous population than myenteric neurons. Because of their incomplete investment they are more likely to be freely exposed to substances diffusing in the extraganglionic tissue than are neurons of sympathetic ganglia.

Notes: Summary Noradrenergic axons in the enteric plexuses of the guinea-pig ileum have been identified at the ultrastructural level using three techniques: the chromaffin reaction, localization of dopamine-β-hydroxylase (DBH) with horseradish peroxidase-conjugated antibody, andin vivo andin vitro loading with 5-hydroxydopamine (5-OHDA). In the myenteric (Auerbach's) plexus from normal ileum all of these methods produced electron-dense deposits in a distinctive population of axonal varicosities that contained many flattened vesicles (usually more than 30% of the total number of vesicles), as well as oval or irregularly shaped vesicles. When noradrenergic axons to the small intestine had degenerated after surgical denervation, no profiles containing vesicles with electron-dense deposits were observed with the chromaffin reaction, DBH localization or loading with 5-OHDA. Pretreatment with 6-hydroxydopamine (6-OHDA) substantially reduced the number of noradrenergic axons identified by these three techniques. Axons with many flattened vesicles of similar dimensions but without dense cores were found in myenteric plexus from conventionally fixed intestine. These axons had the same distribution within the ganglia as cytochemically labelled noradrenergic terminals and disappeared after extrinsic denervation. In the normal submucous (Meissner's) plexus, both 5-OHDA loading and the chromaffin reaction produced electron-dense granules in small and large vesicles in some axon terminals. In ganglia labelled by these techniques, reactive terminals contained many small round vesicles and few flattened and large round vesicles as did a population of nonreactive terminals. In axon terminals of submucous plexus labelled with anti-DBH, flattened vesicles were found to be more numerous than with the other treatments. As in the myenteric plexus, all reactive axons disappeared from the submucous plexus after extrinsic denervation. In conventionally processed submucous ganglia, noradrenergic axon profiles could not be distinguished from some non-noradrenergic profiles on the basis of types and proportions of vesicles. In the myenteric plexus noradrenergic axon terminals were seen most often near the edges of ganglia. Noradrenergic varicosities also occurred near nerve cell bodies but were rarely found in internodal strands. In the submucous plexus noradrenergic terminals appeared to be randomly distributed throughout submucous ganglia. No axosomatic synapses formed by noradrenergic axons were found in either plexus, but synapses on nerve processes were occasionally encountered in submucous ganglia.

Notes: Summary A fine structural study has been made of the vesiculated nerve profiles of the submucous plexus of both normally innervated and extrinsically denervated segments of guinea-pig ileum. Two types of nerve profiles could be readily distinguished by their vesicular content after conventional fixation. The first type, comprising 5% of all intrinsic profiles, consisted of predominantly small vesicles containing electron dense material which usually formed a ring around the inner face of the vesicular membrane but sometimes partially or completely filled the vesicle. These profiles, termed ring-vesicle-containing profiles, remained after extrinsic denervation and their vesicular content did not change following injection of reserpine or 5-hydroxydopamine. Thus ring-vesicle-containing profiles are not noradrenergic. Profiles which were positive for the uranaffin method were similar in morphology and frequency of occurrence to ring-vesicle-containing profiles, although it is not possible to say that they are the same. The second type of profile, comprising 95% of all intrinsic profiles, contained varying proportions of large granular and small clear vesicles. These heterogeneous profiles were present in both normally innervated and extrinsically denervated tissue. Their vesicular content did not change following injection of reserpine, however, some profiles of this type in normally innervated, but not in extrinsically denervated, intestine contained electron dense deposits after injection of 5-hydroxydopamine. This means that noradrenergic profiles are a subpopulation of the heterogeneous profiles in normally innervated tissue. Analysis of intrinsic heterogeneous profiles showed that the proportion and packing density of large granular vesicles formed continuous distributions which did not provide any basis for further subdivision of this type of profile. Ring-vesicle-containing and heterogeneous profiles often formed synapses with neuronal cell bodies and processes. Two rarer types of profiles were also seen. The first type contained mainly small flattened vesicles which took up 5-hydroxydopamine and was not present in extrinsically denervated tissue. This type, like the group described above, is considered to be noradrenergic. The second rare type contained large numbers of lysosome-like dense bodies and vesicles of different sizes and content and was seen in both normally innervated and denervated tissue. This type probably represents spontaneously degenerating nerve profiles.