Inhibition of sodium transport by angiotensin II in the main duct of the rabbit mandibular gland isolated and perfused in vitro

Summary The effect of angiotensin II on nett electrolyte transport by the main duct of the rabbit mandibular gland was investigated in vitro using a perfused duct preparation bathed in a Haemaccel^®-nutrient salt solution. In a bath concentration of 4×10^–10 M, angiotensin reduced nett absorption of Na⁺ and Cl^– by about 8% and depolarized the transepithelial electrical potential difference (P.D.) by about 13%; the drug had no effect on ductal transport of K⁺ and HCO ₃ ^– . In both lower (4×10^–11 M) and higher (4×10^–9 M) concentrations, angiotensin had qualitatively similar effects. After exposure to the hormone for about 30 min, Na⁺ transport and P.D. became unstable and gradually fell away towards zero. It is concluded that angiotensin in physiological concentrations has a specific inhibitory effect on Na⁺ absorption by salivary duct cells which could arise either from a change in the Na⁺ pump rate or from a conductance change in the apical or basal membrane of the epithelial cell.     

Effects of saline loading on jejunal absorption of calcium,sodium, and water,and on parathyroid hormone secretion in the rat

Summary To investigate whether intestinal calcium absorption parallels that of sodium following extracellular fluid volume expansion, the effects of saline loading on intestinal transport of calcium. sodium and water were studied in rats by perfusing jejunal loops in situ.After calcium-free saline infusion net calcium absorption was reversed similar to that of sodium and water and net secretion occurred. Concurrently, blood-to-lumen (b-l) calcium flux, measured using⁴⁵Ca, increased significantly (P<0.001). Following expansion with calcium-containing Ringer a similar reversal of net calcium, sodium and water flux was also observed. Again, the b-l calcium flux increased but to a significantly lesser extent (P<0.05). Plasma ionized calcium remained unchanged after calcium-rich Ringer loading, but decreased significantly (P<0.001) when calcium was omitted from the solution. Plasma immunoreactive parathyroid hormone was unchanged after expansion with the calcium containing solution but increased following calcium-free infusion.It is concluded that after extracellular fluid volume expansion: 1. net jejunal calcium absorption is decreased; 2. the decrease parallels that of sodium and water; 3. b-l calcium transport is enhanced to a greater degree by calcium-free Ringer infusion than by a calcium-rich solution. This difference could be the result of increased parathyroid hormone secretion.     

Intracellular potentials of microperfused human sweat duct cells

Intracellular potentials of cells from isolated segments of microperfused human sweat ducts were measured in order to determine the electrical profiles of these cells under resting, transporting, and inhibited conditions. Even though the cells are relatively small (ca. 6–8 m), continuous recordings of intracellular potentials from the same impalement were stable for up to 2 h. In the resting condition in normal Ringer's solution when the lumen of the duct was collapsed and not perfused, the intracellular potential measured across the basal membrane was 34.6±1.5 mV (n=31; mean±SE). In the same bathing medium, when the duct lumen was also perfused with normal Ringer's solution, the basolateral membrane potential (V _b), the apical membrane potential (V _a) and transepithelial potential (V _t) was –33.8±0.47 mV, –23.7±0.48 mV and –9.6±0.9 mV (n=73), respectively. The average input impedence (R _i) of these cells was 19.6±0.4 M (n=36). The frequency distribution ofV _b was unimodal suggesting that only one functional cell type exists in this tissue. Amiloride (0.1 mM) in the lumen hyperpolarized bothV _a andV _b by –40.5±3.6 mV and –33.2±3.7 mV (n=15), respectively, with a slight but significant increase inR _i (15%), while abolishingV _t. Removing luminal Cl^– depolarizedV _a by +37.0±4.2 mV and hyperpolarizedV _b by –19.0±4.2 mV (n=11). Removing Cl^– from the bath hyperpolarizedV _a by –3.3±2.3 mV and depolarizedV _b by +24.3±2.7 mV (n=15). Ouabain caused an initial fast depolarization (+8 mV) followed by a prolonged slow depolarization ofV _b, and an increase inR _i of about 84%. These results not only provide the first electrical profile of the human sweat duct tissue, but they also show that its cell membrane potentials are unusually low. This unusual property of this epithelium appears to be due to the combination of a significant Na⁺ conductance at the apical membrane and a remarkably high tissue Cl^– conductance.     

A long term study of axonal transport in the central visual system following eye enucleation in the adult cat

Summary The effect of the enucleation of one eye on anterograde and retrograde labelling in geniculo-cortical, cortico-geniculate and commissural projections was investigated in adult cats by means of horseradish peroxidase (HRP) and tritiated aminoacids. It was found that in addition to the immediate decrease of retrograde labelling with HRP in the cortical projections from the deafferented A-laminae of the dorsal part of the lateral geniculate nucleus (Singer et al. 1977) there is a further reduction which lasts up to 75 days after enucleation. At 146 and 363 days after enucleation a slight increase in the number of labelled neurones was noted in the deafferented lamina. Qualitative assessment did not reveal any changes of anterograde labelling with tritiated amino acids in geniculo-cortical, cortico-geniculate and commissural axones. In addition, the retrograde labelling with HRP in cortico-geniculate and commissural projections seemed to be unaffected by eye enucleation.Deceased     

Ionic requirements for in vitro retrograde axonal transport of acetylcholinesterase

The ionic requirements for retrograde axonal transport of acetylcholinesterase were studied in vitro in desheathed spinal nerves of the bullfrog; the accumulation of enzyme activity distal to a ligature served to evaluate the status of retrograde transport. After an 18 h incubation in control medium, the 2 mm segment of nerve immediately distal to the ligature contained approximately twice as much acetylcholinesterase activity as other more distal segment. Accumulation of acetylcholinesterase distal to the ligature was reduced during incubation in Ca2+-free medium plus 1 mM EGTA; retrograde transport was not diminished by the substitution of sucrose for the NaCl of the medium. In comparison with ionic requirements for orthograde fast axonal transport of protein, retrograde transport of acetylcholinesterase thus appears to share a Ca2+ requirement, but not a requirement for NaCl.     

Effect of vinpocetine on retrograde axoplasmic transport

Vinpocetine, a derivate of vincamine, is widely used in the clinical pharmacotherapy of cerebral circulatory diseases. Herewith we report on a novel effect of vinpocetine: inhibition of retrograde axoplasmic transport of nerve growth factor (NGF) in the peripheral nerve. Blockade of retrograde transport of NGF results in transganglionic degenerative atrophy (TDA) in the segmentally related ipsilateral superficial spinal dorsal horn, which is characterized by depletion of the marker enzymes fluoride-resistant acid phosphatase (FRAP) and thiamine monophosphatase (TMP). At the same time, pain-related neuropeptides such as substance P (SP) and calcitonin gene-related peptide (CGRP), are depleted from lamina I-III from the segmentally related, ipsitateral Rolando substance of the spinal cord. On the basis of these experiments it is suggested that vinpocetine may result in a locally restricted decrease of nociception, that might be useful in clinical treatment of intractable pain. Pilot self-experiments support this assumption.     

The somatotopic organization of primary afferent terminals in the superficial laminae of the dorsal horn of the rat spinal cord

Transganglionic transport of wheatgerm agglutinin conjugated horse-radish peroxidase (WGA-HRP) was used to reveal the central distribution of terminals of primary afferent fibers from peripheral nerves innervating the hind leg of the rat. In separate experiments the sizes and locations of cutaneous peripheral receptive fields were determined by electrophysiological recording techniques for each of the nerves that had been labeled with WGA-HRP. By using digital image analysis, the sizes and positions of the peripheral receptive fields were correlated with the areas of superficial dorsal horn occupied by terminals of primary afferents from each of these receptive fields. Data were obtained from the posterior cutaneous nerve of the thigh, lateral sural, sural, saphenous, superficial peroneal, and tibial nerves. The subdivisions of the sciatic nerve, the sural, lateral sural, superficial peroneal, and tibial nerves each projected to a separate and distinct region of the superficial dorsal horn and collectively formed a "U"-shaped zone of terminal labeling extending from lumbar spinal segments L2 to the caudal portions of L5. The gap in the "U" extended from L2 to the L3-4 boundary and was occupied by terminals from the saphenous nerve. Collectively, all primary afferents supplying the hindlimb occupied the medial 3/4 of the superficial dorsal horn with terminals from the tibial nerve lying most medially and occupying the largest of all the terminal fields. Afferents from the superficial peroneal lay in a zone between the medially situated tibial zone and the more laterally placed sural zone. Afferents from the posterior cutaneous nerve were located most caudally and laterally. Terminal fields from the posterior cutaneous and saphenous nerves differed from the others in having split representations caused presumably by their proximity to the mid-axial line of the limb. Comparisons between the peripheral and the central representations of each nerve revealed that 1 mm2 of surface area of the superficial dorsal horn serves approximately 600-900 mm2 of hairy skin and roughly 300 mm2 of glabrous skin. The vast majority of terminal labeling observed in the dorsal horn was found in the marginal layer and substantia gelatinosa, suggesting that small diameter afferents have an orderly somatotopic arrangement in which each portion of the skin surface is innervated by afferent fibers that terminate in preferred localities within the dorsal horn.     

Strontium transport in the rat colon

Summary Concentration dependence of strontium (Sr) fluxes across the colon ascendens and descendens of the rat were measured in a modified Ussing-chamber. Mucosa (m) to serosa (s) and s to m Sr fluxes across both colonic segments were linearly related to the Sr concentration from 0.125 mmol/l to 10 mmol/l. In the colon ascendens m to s Sr fluxes were slightly higher than the fluxes in the opposite direction, resulting in net Sr absorption. In the colon descendens s to m fluxes were higher than the ms fluxes, resulting in net Sr secretion. Neither Sr nor calcium (Ca) showed a concentration dependent interaction with respect to their unidirectional fluxes in both parts of the colon. Only in the colon ascendens Sr at the highest concentration (10 mmol/l) inhibited m to s calcium transport.Experiments, in which the voltage dependence of the unidirectional Sr fluxes was measured confirmed the results obtained from the concentration dependence: (1) The unidirectional fluxes of Sr across the colon ascendens and descendens were totally voltage dependent, i.e. diffusive. (2) In the colon descendens the voltage dependence of the s to m flux was steeper than the flux from m to s. It is hypothesized that this prevalence is caused by an anomalous solvent drag effect. 1.25-Dihydroxyvitamin D₃ [1.25 (OH)₂D₃] stimulated m to s calcium flux in the colon descendens but had no effect on Sr flux.The results demonstrate that Sr and Ca in the rat colon are transported by different mechanisms. In contrast to the Ca transport the Sr flux is only diffusive and insensitive to 1.25 (OH)₂D₃.Supported by the Deutsche Forschungsgemeinschaft SFB 38 Membranforschung Send offprint requests to U. Karbach at the above address