This report describes a method for isolating single rabbit atrioventricular (AV) node myocytes which retain their normal morphology when exposed to millimolar levels of calcium. Previous attempts to isolate cells from the AV node have produced myocytes that "round up" (i.e., go into contracture) when exposed to calcium. We show that the cells isolated with our technique possess properties similar to those described for intact AV nodal tissue. We find that single AV node myocytes are shorter and thinner (mean dimension = 103.5 +/- 2.3 by 7.8 +/- 0.2 microns; mean +/- SE, n = 90) than atrial or ventricular cells. Many of the cells produced by this isolation procedure generate spontaneous action potentials (188 +/- 9 beats/min; n = 6), which resemble action potentials recorded previously from the intact AV node. Voltage-clamp recordings from spontaneously active cells revealed similar membrane currents to those seen in intact tissue: fast sodium current and a L-type calcium current, followed by a delayed outward current. However, we found little evidence for the hyperpolarization-activated current (I(f)). Because the cells responded normally to concentrations of acetylcholine and isoproterenol within the physiological range, their cholinergic and adrenergic receptors appear to be well preserved by the isolation procedure. The ability to isolate morphologically and functionally normal AV myocytes may represent a significant advance for the investigation of nodal physiology at the cellular level.
Electron-microscope autoradiographs have been prepared from the neural lobes of the pituitary glands of rats which had received intracisternal injections of [35S] cysteine at various times before gland removal. The rate of appearance and disappearance of autoradiographically demonstrable radioactivity in the neural lobe closely paralleled that previously determined, biochemically, for radioactive hormones and neurophysins. Radioactivity was appreciably associated with the undilated parts of neurosecretory axons only during the first few hours after injection of the label. The axonal dilations were subdivided into those in which small vesicles could be seen ("endings") and those in which no small vesicles could be seen ("swellings"). Radioactivity appeared first in "endings" and then in progressively larger and larger profiles of "swellings". It appeared that newly arrived granules were found close to the limiting membrane of the nerve swelling and that as time progressed they moved deeper and deeper into the swelling. On the basis of the results, suggestions were made for an anatomical explanation of the readily-releasable pool of hormone which has been demonstrated pharmacologically.
The sites and concentrations of Der p I within the house dust mite were determined. Highest concentrations were found in the epithelium of the posterior end of the mite stomach, implying that this is the site of Der p I synthesis and secretion.
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