Time Course Of Regulation Research Articles

Influence of olive (cv Grignano) fruit ripening and oil extraction under different nitrogen regimes on volatile organic compound emissions studied by PTR-MS technique

Volatile organic compounds of extra virgin olive oils obtained from the local Italian cultivar Grignano were measured by proton transfer reaction-mass spectrometry (PTR-MS). Oils were extracted by olives harvested at different ripening stages across veraison, performing each extraction step and the whole extraction process in nitrogen atmosphere to observe the changes in the volatile profiles of the oils. Principal component analysis carried out on the full spectral signature of the PTR-MS measurements showed that the stage of ripening has a stronger effect on the global definition of volatile profiles than the use of nitrogen during oil extraction. The fingerprint-like chemical information provided by the spectra were used to construct a heat map, which allowed the dynamical representation of the multivariate nature of mass evolution during the ripening process. This provided the first evidence that some groups of volatile organic compounds displayed a time course of regulation with coordinated increasing or decreasing trends in association with specific stages of fruit ripening.

Both synthesis and reuptake are critical for replenishing the releasable serotonin pool in Drosophila

The two main sources of serotonin available for release are expected to be newly synthesized serotonin and serotonin recycled after reuptake by the serotonin transporter. However, their relative importance for maintaining release and the time course of regulation are unknown. We studied serotonin signaling in the ventral nerve cord of the larval Drosophila CNS. Fast-scan cyclic voltammetry at implanted microelectrodes was used to detect serotonin elicited by channelrhodopsin2-mediated depolarization. The effects of reuptake were probed by incubating in cocaine, which is selective for the serotonin transporter in Drosophila. p-chlorophenylalanine, an inhibitor of tryptophan hydroxylase2, was used to investigate the effects of synthesis. Stimulations were repeated at various intervals to assess the time course of recovery of the releasable pool. Reuptake is important for the rapid replenishment of the releasable pool, on the 1 min time scale. Synthesis is critical to the longer-term replenishment (10 min) of the releasable pool, especially when reuptake is also inhibited. Concurrent synthesis and reuptake inhibition decreased both serotonin tissue content measured by immunohistochemistry (by 50%) and the initial amount of evoked serotonin (by 65%). Decreases in evoked serotonin are rescued by inhibiting action potential propagation with tetrodotoxin, implicating endogenous activity in the depletion. These results show synthesis is necessary to replenish part of the releasable serotonin pool that is depleted after reuptake inhibition, suggesting that regulation of synthesis may modulate the effects of serotonin reuptake inhibitors.

Modulation of calcium currents by electrical activity.

Electrical activation of mouse dorsal root ganglion (DRG) neurons in cultures for 1-2 days produced a downregulation of voltage sensitive calcium currents, which persisted for > or = 24 h after stimulation was terminated. This regulation varied with different patterns of activation. Both the magnitude and time course of regulation of the low-threshold voltage-activated (LVA) and high-threshold voltage-activated (HVA) currents were differentially sensitive to neural impulse activity. Tonic stimulation at 0.5 Hz did not affect the HVA currents, but 2.5 Hz did produce a significant decrease. Phasic stimulation (10 Hz for 0.5 s every 2 s) with an average frequency of 2.5 Hz produced significantly more downregulation of HVA currents than did the tonic 2.5-Hz stimulation. The efficacy of phasic stimulation varied inversely with the interval between bursts. Thus phasic stimulation of 10 Hz for 0.5 s but delivered every 4 s produced no effects on HVA currents. Stimulation optimal for downregulation of Ca2+ currents also produced a decreased binding by the DRG neurons of an L-type Ca2+ channel antagonist. This suggests a downregulation by electrical activity of the number of Ca2+ channels, rather than an alteration in a constant number of channels. Depression of LVA currents was produced by all stimulus patterns tested, including 0.5-Hz tonic stimulation. Chronic stimulation with a stimulation pattern that downregulated Ca2+ currents also produced a slowing of the increase in intracellular Ca2+ (as measured by Fura-2/AM) that is produced acutely by repetitive stimulation. This is consonant with earlier studies of intracellular Ca2+ concentration kinetics in growth cones.

Regulation of the adrenomedullary catecholaminergic system after mild, acute stress.

The time course of regulation of rat adrenomedullary phenylethanolamine N-methyltransferase (PNMT) and tyrosine hydroxylase (TH) activity was studied after a single episode of 20-min restraint stress. Significant increases in PNMT and TH activity were observed 18 h after the beginning of the stress. The time course of acute stress-induced regulation of PNMT and TH was examined for the influence of neural and hormonal input. Unilateral denervation was performed, and the animals were exposed to a single episode of restraint. PNMT activity increased similarly in both the innervated and denervated adrenals, with a significant increase observed at 36 h after the stress. TH activity was similar in both denervated and innervated adrenals, with a significant increase observed at 24 and 36 h after stress. Finally, suppression of endogenous corticosterone with dexamethasone delayed the stress-induced increase in activity of PNMT but not TH. The present study indicates that increases in catecholamine biosynthetic enzymes can be observed after a single episode of mild, acute stress. In addition, glucocorticoids appear to be important in the time course of the stress-induced increase in PNMT but not TH activity, whereas neural input does not seem to affect the time course of these increases.

Brief cortisol exposure elevates adrenal phenylethanolamine N-methyltransferase after a necessary lag period

The present study, using bovine adrenal medullary cells, characterized in detail the time course of regulation of phenylethanolamine N-methyltransferase activity following brief glucocorticoid exposure. Cortisol pulses (10 −4 and 10 −5M), as short as 15 min, increased phenylethanolamine N-methyltransferase activity measured 2 days following cortisol exposure, with a required lag period of 18 h or more. Phenylethanolamine N-methyltransferase activity was increased 2 days following brief (2 h) exposure to cortisol in concentrations that reach the medulla in vivo (10 −6 to 10 −4 M). Phenylethanolamine N-methyltransferase activity following both continuous and 2 h pulses of 10 −5 M cortisol were reduced by the glucocorticoid receptor antagonist, RU 38486. A 2 h pulse of nicotine (10 −5 M) increased phenylethanolamine N-methyltransferase activity with a lag period of at least 18 h, while combination treatment of nicotine and cortisol (10 −4 M) produced significantly higher increases in phenylethanolamine N-methyltransferase compared to either treatment alone. Therefore, this study provides novel in vitro evidence for the regulation of adrenomedullary phenylethanolamine N-methyltransferase activity, following a necessary lag period, by acute changes in both cortisol and nicotine.

Differential Regulation by 1,25-Dihydroxyvitamin D3of Calbindin-D9kand Calbindin-D28kGene Expression in Mouse Kidney*

The mouse kidney is a unique tissue since both vitamin D-dependent calcium binding proteins (calbindin-D9k and calbindin-D28k) are present in the same cells of the distal convoluted tubule. We have used specific complementary DNAs to mouse calbindin-D9k and mouse calbindin-D28k and Northern and slot blot analyses in order to obtain a better understanding of the regulation of two different molecular expressions of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] action in the same cells. Both calbindins were found to be regulated developmentally in a similar manner (an increase in gene expression between birth and 1 week of age, coinciding with nephron differentiation, and a peak at 3 weeks of age). However, the time course of response of the messenger RNA of each calbindin to 1,25(OH)2D3 was markedly different. The peak of induction of renal calbindin-D28k mRNA was at 12 h after a single injection of 1,25(OH)2D3 (200 ng/100 g body wt) to vitamin D-deficient mice, and a decrease was observed at 24 h (similar to the time course of response of other steroid-regulated genes). Interestingly, unlike calbindin-D28k, a delayed response of renal calbindin-D9k mRNA to 1,25(OH)2D3 was observed (the peak of induction was at 24 h after 1,25(OH)2D3 administration). Both genes in mouse kidney did not respond to glucocorticoids, although a dose-dependent decrease (12-86%) of mouse intestinal calbindin-D9k mRNA was observed after dexamethasone treatment, suggesting tissue-specific multiple steroid interactions in the regulation of calbindin gene expression. The finding of a different time course of regulation of each calbindin by 1,25(OH)2D3 suggests that different factors may be regulating the expression of the two different calbindins in mouse kidney and that elucidation of these control mechanisms should provide new insight concerning 1,25(OH)2D3-regulated gene expression.

Characteristics of pressureflow regulation by the kidney.

Pressure-flow relationships in the dog kidney were studied by means of a square wave, noncannulating electromagnetic flowmeter probe placed around the renal artery by way of a retroperitoneal approach. Renal artery pressure was varied by a clamp on the artery and recorded via a needle inserted distal to the probe and clamp. Reductions in artery pressure from control levels demonstrated complete regulation of flow from control level down to 70 mm Hg, incomplete regulation from 50–70 mm Hg, and inability to compensate further below 50 mm Hg. Insignificant regulation was found in the iliac vascular bed by this technique. Recovery time of flow occurred within 5–60 sec following occlusion and was more prolonged at lower levels of artery pressure. The findings of a sharp inflection in the pressure-flow plot at approximately 70 mm Hg and a time course of regulation and recovery pattern varying with the reduced artery pressure suggest an active mechanism controlled by a sensitive feedback system. Submitted on September 18, 1961