A Search for Discrete Cholinergic Nuclei in the Human Ventral Forebrain

Abstract: Slices cut from five frozen human brains were dissected into 2-mm cubes and assayed for choline acetyltransferase (ChAT) activity and protein content. A pattern of enrichment of ChAT activity was found ventral to the anterior commissure; this finding is consistent with the location of the enzyme in the cells of the nucleus basalis of Meynert. The region beneath the anterior commissure was the only place a discrete enrichment of activity could be found, and the precise topography of the enrichment was somewhat variable from brain to brain. The results are discussed in the light of recent knowledge concerning the source of the cortical cholinergic innervation.     

Muscarinic Agonists Evoke Neurotransmitter Release: Possible Roles for Phosphatidyl Inositol Bisphosphate Breakdown Products in Neuromodulation

Carbachol (CCh), a muscarinic agonist that elicits the formation of inositol trisphosphate (IP3) and diacylglycerol (DG), induces a calcium-dependent [3H]norepinephrine ([3H]NE) release [IC50 = (2.7 +/- 0.5) X 10(-4) M] in rat brain slices. Similarly, other muscarinic agonists evoke [3H]NE release which is specifically inhibited by muscarinic antagonists such as 3-quinuclidinyl benzilate, atropine, and N-methyl-4-piperidyl benzilate. The atropine-sensitive evoked release is effectively inhibited by neomycin (IC50 = 50 microM), a phospholipase C inhibitor that interferes with IP3-dependent cellular processes. In addition, polymyxin B, a rather selective inhibitor of protein kinase C (PK-C), abolishes the agonist-mediated release with a half-maximal effective concentration of 0.53 microM (750 ng/ml). These results have a significant implication for the mechanism by which agonists generating IP3 and DG act as inducers of neurotransmitter release in the CNS. However, since both neomycin and polymyxin B act also as N-calcium-channel blockers, other possible mechanisms are discussed. The CCh-induced release suggests that in the CNS an agonist-receptor interaction leads to a calcium-dependent neurotransmitter release, most likely via promoting the IP3/DG as second messengers followed by activation of PK-C.     

Aortic input impedance during Mueller maneuver: an evaluation of "effective length"

Aortic input impedance was calculated in seven subjects in the control state (normal reflection) and during the Mueller maneuver (increased reflection) to evaluate "effective arterial length" under altered physiological conditions. Regional foot-to-foot pulse wave velocities and pressure waveforms along the aorta were used to define an "apparent anatomic length" or distance to a dominant discrete site of reflection "seen" by the ejecting ventricle. Time of wave travel was taken to be one-half the interval from the foot of the incident wave to the midsystolic inflection point. Knowing the time of travel from the returning reflection and velocity, distances calculated to the "apparent anatomic length" were 35 +/- 2 and 34 +/- 2 during control and Mueller maneuver, respectively (P = NS). The frequency of the first minimum of the modulus (fmin) and the first zero crossing of the phase angle (f phi) were determined from the input impedance spectra. During baseline conditions, fmin (3.9 +/- 0.2 Hz) approximately equaled f phi (4.2 +/- 0.2 Hz), and the resulting "effective lengths" calculated using the quarter-wavelength formula were similar to the apparent anatomic length. These data suggested that the aortic region incorporating the renal arterial branches as a site of discrete reflection and that terminal load was not significantly frequency dependent. During Mueller maneuver, however, f min (3.3 +/- 0.2 Hz) and f phi (5.1 +/- 0.2 Hz) were significantly discordant, the terminal load became strongly frequency dependent, and effective length calculated from f min was dissimilar (P less than 0.05) from the unchanged apparent anatomic length.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro OKT3-induced mitogenesis in selenium-deficient patients on a diet for phenylketonuria

Patients with phenylketonuria (PKU) are frequently deficient in the essential trace element selenium (Se), because of their very low protein diet. Using two approaches to investigate T-cell response to proliferative signaling, viz, mitogenesis caused by the monoclonal antibody OKT3 and the plant lectin phytohaemagglutinin (PHA), we demonstrated significantly reduced responses to optimal concentrations of OKT3 in a group of PKU patients with reduced serum Se compared with a normal group (p = 0.0005) and with a group of PKU patients whose serum Se was normal (p = 0.0023). The response of the Se-deficient group to optimal levels of PHA did not differ from that of the normal controls or from that of Se-normal PKU patients. A dose-dependent relationship between serum Se levels and mitogenic response was evident for OKT3 (r = 0.34, p = 0.0154), but not for PHA (r = -0.02, p = 0.9086). We suggest that the reduced response to OKT3 mitogenesis in Se-deficient PKU patients is possibly the consequence of impaired Se-dependent metabolic activity, which affects mitogenic signaling via the T cell antigen receptor (TCR/CD3) complex.     

Effects of Neuronal Activity on Inositol Phospholipid Metabolism in the Rat Autonomic Nervous System

The effect of nerve stimulation on inositol phospholipid hydrolysis in autonomic tissue was assessed by direct measurement of [3H]inositol phosphate production in ganglia that had been preincubated with [3H]inositol. Within minutes, stimulation of the preganglionic nerve increased the [3H]inositol phosphate content of the superior cervical sympathetic ganglion indicating increased hydrolysis of inositol phospholipids. This effect was blocked in a low Ca2+, high Mg2+ medium. It was also greatly reduced when nicotinic and muscarinic antagonists were present together in normal medium. However, neither the nicotinic antagonist nor the muscarinic antagonist alone appeared to be as effective as both in combination. In other experiments, stimulation of the vagus nerve caused dramatic increases in [3H]inositol phosphate in the nodose ganglion but did not increase [3H]inositol phosphate in the nerve itself. This effect was insensitive to the cholinergic antagonists. Thus, neuronal activity increased inositol phospholipid hydrolysis in a sympathetic ganglion rich in synapses, as well as in a sensory ganglion that contains few synapses. In the sympathetic ganglion, synaptic stimulation activated inositol phospholipid hydrolysis and this was primarily due to cholinergic transmission; both nicotinic and muscarinic pathways appeared to be involved.     

Comparison of direct and indirect measurements of pulmonary capillary transit times

Using in vivo microscopy, we made direct measurements of pulmonary capillary transit time by determining the time required for fluorescent dye to pass from an arteriole to a venule on the dependent surface of the dog lung. Concurrently, in the same animals, pulmonary capillary transit time was measured indirectly in the entire lung using the diffusing capacity method (capillary blood volume divided by cardiac output). Transit times by each method were the same in a group of five dogs [direct: 1.75 +/- 0.27 (SE) s; indirect: 1.85 +/- 0.33 s; P = 0.7]. The similarity of these transit times is important, because the widely used indirect determinations based on diffusing capacity are now shown to coincide with direct measurements and also because it demonstrates that measurements of capillary transit times on the surface of the dependent lung bear a useful relationship to measurements on the capillaries in the rest of the lung.     

G-suit inflation to 50 mmHg alters the cardiovascular transients when entering micro-G in parabolic flight

In the present experiments it was decided to have each test-subject serve as his own control by fitting the test-subjects with a G-suit and comparing the condition of inflated G-suit to the normal situation. G-suit inflation was intended to only displace blood on the venous side of the circulation, not to increase total peripheral resistance. Therefore, a very modest inflation of 50 mmHg was applied. This was considered sufficient to expel most of the blood from the venous pool in abdomen and legs, even under the condition of increased G-loading in the pull-up phase. The parabolas were to be undergone in three body positions: standing upright, sitting and supine. The prediction of the experimental outcome was that we would find no difference between transients with and without G-suit inflation in the supine position, that an initial overshoot in pressure and stroke volume in the upright position would be very much damped by the G-suit, even more in the standing than in the sitting position. Studies were performed in 5 flights of NASA's KC-135, in January 1993. Per flight 40 parabolas were flown in an adapted 'roller coaster profile', i.e. 0-G phases were followed by a 2-G pull-out phase, after a very brief 1-G phase again followed by the next 2-G pull-up phase. This sequence was flown for 10 parabolas, then a 1-G horizontal flight period was inserted. The first 3 parabolas of each set of 10 the subjects were sitting upright, seat belt fastened. The next three they were standing, feet stuck under a load strap on the floor, stabilizing themselves by a grip on the ceiling. Then three parabolas were flown with the test-subject supine, loosely attached to the floor by a load strap and further aided by a grip to another strap on the floor. The last parabola of a set was used as 'spare' to repeat any failed maneuver.