The β2-agonist salbutamol affects the expression of phospholamban and both isoforms of SERCA in canine skeletal muscle and blocks changes in these induced by neuromuscular stimulation

 Chronic administration of salbutamol induced expression of hybrid fibers in canine skeletal muscles. Fast-twitch fibers expressed SERCA2a (the slow-twitch isoform of sarcoplasmic reticulum Ca²⁺-ATPase) and slow-twitch fibers expressed SERCA1 (the fast-twitch isoform of the Ca²⁺-ATPase). The proportion of fibers that became hybrid increased from a small percentage in the control muscles to 30% in the predominantly fast-twitch latissimus dorsi and to 45% in the predominantly slow-twitch vastus intermedius. In contrast to this response by the SERCA genes the phospholamban gene response was muscle specific. The fraction of fibers that expressed phospholamban decreased slightly in the latissimus dorsi while increasing moderately in the vastus intermedius. The effects of chronic neurostimulation of the latissimus dorsi on SERCA1, SERCA2a and phospholamban levels were mostly blocked by salbutamol. While 100% of fibers from neurostimulated muscles expressed phospholamban, only 51% of the fibers from the neurostimulated and salbutamol-treated muscles expressed it. In the neurostimulated muscle, very few muscle fibers expressed SERCA1a while 61% of the fibers that received salbutamol expressed it, albeit as hybrid fibers. The levels of SERCA2a in response to these interventions were just the opposite. In the neurostimulated muscle 37.5% of fibers were hybrid and 62.5% expressed SERCA2a only. With co-administration of neurostimulation and salbutamol, 61.3% of fibers were hybrid and 38.7% expressed SERCA2a only. Received: 4 July 1997 / Received after revision: 16 October 1997 / Accepted: 30 October 1997     

The early phase of experimental acute renal failure

Experiments were conducted to establish whether diminished solute reabsorption in the loop of Henle during acute renal failure could explain the loss of urinary concentration and participate in generating a tubuloglomerular feedback-mediated reduction in filtration rate. The electrolyte content of the fluid in the ascending limb of the loop of Henle was determined in situ by monitoring its electrical conductivity after propulsion into the distal tubule with a sudden burst perfusion. The value of the minimum electrolyte concentration decreased exponentially with increasing equilibration time, reaching a steady-state value equivalent to 27±9 mM NaCl in normal kidneys, 34±15 mM in mercuric chloride kidneys and 53±22 mM following ischaemia. A mathematical model was derived to describe the process of sodium chloride dilution from which it was possible to calculate both the permeability and transport velocity of the cortical thick ascending limb. In the normal kidney, the transport velocity was calculated to be 4.65±0.92 ·10^–5 cm/s, a value not significantly different from that of the mercuric chloride or ischaemic kidneys, and the estimated permeability was 1.13±0.52·10^–5 cm/s, not different from that of the mercuric chloride kidneys but significantly lower than that calculated for the ischaemic kidneys. It is concluded that for the more severely damaged ischaemic model, the loss of urinary concentrating ability was accompanied by a reduction in diluting ability of the ascending limb of the short loop of Henle, which appears to be due, at least in part, to an elevation of the passive permeability to sodium chloride in this segment.     

A proposed autacoid mechanism controlling mastocyte behaviour

Evidence is provided here supporting the existence of a novel autacoid mechanism negatively modulating mast cell behaviour in response to noxious stimuliin vivo; hence, the denomination “autacoid local inflammation antagonism” (ALIA). In particular, as lipid amides of theN-acylethanolamine type have been reported to accumulate in tissues in degenerative inflammatory conditions, we examined whether theseN-acylated lipids could exert regulatory effects on mast cell activationin vivo. The results reported show that both long- and short-chainN-acylethanolamines, when systemically administered, are effective in reducing mast cell degranulation induced by local injection of substance P in the earpinna of developing rats. These and other data suggest that the endogenous production ofN-acylethanolamines may constitute a local autocrine/paracrine response for the negative feedback control of mast cell responses to various activating signals. Such a process may be of physio-pathological relevance in the regulation of functional neuroimmune-mast cell interactions.

     

Electron microscopy of the red pulp of human spleen

Terminating arterial vessels, the structure of sinuses and cords, and the passage of cells through the sinus wall in the red pulp of human spleen were studied. All terminating arterial capillaries arterial capillaries observed opened into cords. The distance between terminating arterial capillaries and sinuses varied. Macrophages were commonly present at arterial terminations. Arterial capillary endothelium contained filaments about 80 Å in diameter. Blood cells were frequently present in the capillary lumen or in passage through the capillary wall into cords. Endothelial cells of sinuses contained three distinctive structures: loosely organized cytoplasmic filaments, tightly organized finer filaments, and micropinocytotic vesicles. Many micropinocytotic vesicles about 0.1 μ in diameter were present just beneath the plasma membrane of the lateral and luminal sides of sinus endothelial cells and a few at the basal aspect. Loosely organized filaments about 80 Å in diameter ran parallel to the longitudinal axis of the sinus endothelium. The finer filaments about 30–50 Å in diameter were tightly organized as filamentous bands and present basally. The filaments of the bands appeared inserted upon the plasma membrane. They were also present in the cordal reticular cells and terminating arterial capillaries. Free cells were frequently present in passage through the slits of the sinus wall. There were no preformed or fixed apertures in the sinus wall. The basement membrane and reticular fibers were completely covered by the endothelial cells and/or cordal reticular cells. It is likely that those slits between endothelial cells in the sinus wall not covered by the basement membrane are potential passageways for cells moving from the cords into the sinus. The larger cytoplasmic filaments are likely contractile. The filamentous bands appear to maintain cell shape, stabilize the wall in relation to the basement membrane, and are probably operative in the control of cellular passage through the slits of sinus wall.     

Cellular localization of nerve growth factor-like immunoreactivity in hippocampus and septum of adult rat brain.

The cellular localization of the nerve growth factor-like immunoreactivity (NGF-LIR) has been studied in the intact adult rat brain at the level of the hippocampus and the septum. Immunolabelling for NGF combined with counterstaining with cresyl violet and double immunostaining technique, which allowed simultaneous localization of NGF-LIR and that of astroglial marker -GFAP, were used. The data indicate neuronal localization of NGF-like immunoreactivity and a lack of colocalization of NGF-LIR with the immunoreactivity of GFAP in the hippocampus. These data are consistent with in situ hybridization results for NGF and immunocytochemical results for pro-NGF localization obtained by others. At the septal level, apart from neuronal localization of NGF-LIR, single NGF-like immunoreactive astrocytes have been observed. This suggests that, although to a very small extent, in vivo intact brain astrocytes may, just as astrocytes growing in vitro, synthesize NGF-like molecules. This finding may be of importance in better understanding the trophic support for NGF responsive cholinergic neurones in the brain.