NPY Y1 receptors exert opposite effects on corticotropin releasing factor and noradrenaline overflow from the rat hypothalamus in vitro.

Neuropeptide Y (NPY), corticotropin releasing factor (CRF) and noradrenaline play important roles in the regulation of a number of endocrine and autonomic functions. NPY is co-localised with noradrenaline in the central nervous system and has been observed to modulate noradrenaline release. Recent morphological and physiological studies also support co-modulatory interactions between NPY and CRF. Earlier in vivo studies in our laboratory showed a potentiation of K(+)-stimulated noradrenaline release following NPY administration, possibly due to an NPY Y1 receptor mechanism. In this study, in vitro superfusion techniques were established to simultaneously monitor the release of endogenous noradrenaline and CRF from the hypothalamus of adult rats and to examine the direct neuromodulatory action of NPY on the overflow of CRF and noradrenaline. Administration of 0.10 microM NPY significantly increased CRF overflow to 395% basal levels and reduced hypothalamic noradrenaline overflow to 61% of basal levels. These effects were blocked by prior administration of the NPY Y1 receptor antagonist GR231118. Thus, this study suggests that NPY, working through a Y1 receptor, has dual and opposing effects on CRF and noradrenaline overflow in vitro.

Phenotypic evidence of faulty neuronal norepinephrine reuptake in essential hypertension.

Previous reports suggest that neuronal norepinephrine (NE) reuptake may be impaired in essential hypertension, perhaps because of dysfunction of the NE transporter, although the evidence is inconclusive. To further test this proposition, we applied phenotypically relevant radiotracer methodology, infusion of tritiated NE and quantification of NE metabolites, to 34 healthy lean subjects (body mass index <27.0 kg/m(2)), 19 overweight (body mass index >28.0 kg/m(2)) but otherwise healthy normotensive subjects, 13 untreated lean patients with essential hypertension, and 14 obesity-related hypertensives. Spillover of NE from the heart was increased in lean hypertensives only (mean+/-SD 33.4+/-20.6 versus 16.1+/-11.7 ng/min in lean normotensives, P<0.05), but this could have resulted from high cardiac sympathetic nerve firing rates, faulty NE reuptake, or both. The arterial plasma concentration of 3-methoxy-4-hydroxylphenylglycol, an extraneuronal metabolite of NE, was elevated in lean hypertensives only (3942+/-1068 versus 3055+/-888 pg/mL in healthy subjects, P:<0.05). The fractional extraction of plasma tritiated NE in passage through the heart, determined on the basis of neuronal NE uptake, was reduced in lean essential hypertensives (0.65+/-0.19 versus 0.81+/-0.11 in healthy subjects, P<0.05). Cardiac release of the tritiated NE metabolite [(3)H]dihydroxylphenylglycol, produced intraneuronally by monoamine oxidase after uptake of [(3)H]NE by the transporter, was reduced in lean hypertensives only (992+/-1435 versus 4588+/-3189 dpm/min in healthy subjects, P<0.01) These findings suggest that neuronal reuptake of NE is impaired in essential hypertension. Through amplification of the neural signal, such a defect could constitute a neurogenic variant of essential hypertension. In obesity-related hypertension, there was no phenotypic evidence of NE transporter dysfunction.

Performance of digital filtering methods in orthotic and prosthetic CAD/CAM.

This report characterizes the performance of three digital filters, when applied to residual limb shape maps. The three filters were an averaging filter, a uniform window Fourier filter, and a Hamming window Fourier filter. The frequency responses of the three filters were calculated from theory, and experimentally observed. Experimental observations consisted of responses on single-frequency lobed shapes, and on residual limb shapes. Seven trans-tibial limb molds were digitized, three times each. Each resulting shape was then passed through each of the three filters. The before and after shapes were then compared. A Hamming window filter (low-pass frequency of 10 cycles per revolution, 24 coefficients, Hamming number of 0.25) achieved the best performance based on maintained amount of residual limb frequencies, and on visual observation.

A method of residual limb stiffness distribution measurement.

A method of recording a residual limb indentation stiffness map was developed for possible use as an aid in calculating prosthetic socket rectifications. The method was tested to determine the level of repeatability attainable. A hand-held, pencil-like device was used, with an air-driven piston that indented the tissue 10 times per second. The indentor tip contained an electromagnetic digitizer element that sensed position and orientation 120 times per second. The examiner moved the device around the limb; sampling was variable in density, and typically concentrated on critical areas. An interactive visual display of sampled data quality was used to guide sampling. The indentation maps typically contained approximately 4,000 locations, in a cylindrical coordinate system, with sampling locations spaced every 3.2 mm vertically, and every 0.087 radians tangentially. The behavior of the system was characterized using six test subjects on whom recorded indentations ranged from 1.5 to 21 mm. The largest range of indentations (i.e., worst disagreement) recorded at a single location was 5.4 mm. The average standard deviation on repeated measurement ranged from 7 to 15%, and averaged 0.67 mm in absolute terms. Many of the structurally significant anatomical features of the limbs were visible, including the patella and patellar tendon, fibular head, shin, biceps femoris tendon, semitendinosus, and popliteal area.

In vitro studies of endogenous noradrenaline and NPY overflow from the rat hypothalamus during maturation and ageing.

Marked changes in brain monoamine content and NPY content occur during maturation and ageing. Earlier in vivo studies in our laboratory have reported blunted K+ stimulated noradrenaline release and reduced NPY overflow in aged animals using microdialysis and push pull techniques. In this study, in vitro superfusion techniques were established to measure endogenous noradrenaline, NPY, DOPAC and 5-HIAA overflow from the hypothalamus of 1, 5 and 16 month old Sprague-Dawley rats. A period of high K+ (56 mM) stimulation was carried out to elicit maximal release. Basal noradrenaline overflow was similar in all age groups of rats and during K+-induced depolarisation similar 3-4 fold increases were observed. On the other hand, basal and K+ stimulated NPY overflow were significantly greater in the adult rats compared to 1 month and 16 month old rats. Despite differences in absolute NPY overflow, the relative increase over resting was not significantly different across age groups. The molar quantities of hypothalamic NPY overflow at rest and under K+ stimulated conditions were three orders of magnitude lower than noradrenaline. Results of these studies suggest that both NPY and noradrenaline can be released from a similar hypothalamic pool. Basal and K+-evoked DOPAC and 5-HIAA overflow were similar between the 3 age groups. Thus the overflow of hypothalamic noradrenaline, DOPAC and 5-HIAA under in vitro conditions was not altered from 1 to 16 months. In contrast, 5 month old rats had significantly higher NPY overflow than the other age groups (P < 0.05), consistent with a reported decline in NPY content with advanced age. Hypothalamic noradrenaline overflow was not affected by ageing, suggesting that a selective loss of NPY in the arcuo-PVN projection, or other projections to the hypothalamus with ageing may contribute to the reduction in NPY overflow in aged rats.

Central interactions between noradrenaline and neuropeptide Y in the rat: implications for blood pressure control.

Neuropeptide Y (NPY) and noradrenaline are co-localised in central neurones and both transmitters exert cardiovascular effects. Using microdialysis and push-pull techniques to measure transmitter release in vivo, and microinjection studies, we examined the role(s) of central noradrenaline and NPY in blood pressure regulation in the hypothalamus and nucleus tractus solitarius (NTS) of the rat. Hypothalamic noradrenaline release was increased following haemorrhage and reduced after phenylephrine infusion. Ageing is associated with markedly reduced NPY concentrations in the hypothalamus. 18-month old animals showed a reduced ability to release both NPY and noradrenaline to a potassium depolarisation stimulus. NTS administration of NPY induced dose-dependent decreases in blood pressure and heart rate. The depressor but not the bradycardic response was attenuated by prior administration of yohimbine. NTS microinjection of 23 pmol NPY induced similar cardiovascular effects in spontaneously hypertensive and Wistar Kyoto rats. NPY and noradrenaline appear to interact at several sites in the brain known to be important for blood pressure control.

Neuropeptide Y and [Leu31,Pro34]neuropeptide Y potentiate potassium-induced noradrenaline release in the paraventricular nucleus of the aged rat.

This microdialysis study investigated the effects of NPY and the Y1 selective agonist [Leu31, Pro34]NPY on basal and potassium-stimulated noradrenaline release in the PVN of 18-month-old anaesthetised male Sprague-Dawley rats. Microdialysate noradrenaline, DOPAC and HVA concentrations were measured by HPLC after i.c.v. administration of 2 nmol NPY, [Leu31, Pro34]NPY or vehicle. [Leu31, Pro34]NPY produced a significant 40% reduction in basal noradrenaline concentration (P < 0.05). Aged rats had blunted noradrenaline response to potassium stimulation, however stimulated noradrenaline release was similar in 18-month-old NPY-treated animals and 3-month-old saline treated age controls (2.8 and 3.2 times resting, respectively). [Leu31, Pro34]NPY induced a significantly greater release of noradrenaline in response to KC1 (5.0 times resting, P < 0.05). Thus, in 18-month-old animals with reduced endogenous hypothalamic NPY content, administration of NPY or [Leu31, Pro34]NPY increased potassium-induced noradrenaline release to levels seen in 3-month-old rats. This effect may be mediated by an NPY Y1 receptor.

Altered in vivo catecholamine release in the hypothalamic paraventricular nucleus of the aged rat.

The effect of age on basal and stimulated noradrenaline release in the hypothalamic paraventricular nucleus (PVN) of the rat was examined by in vivo microdialysis. Microdialysis probes were inserted into the PVN of 3 and 18 month old anaesthetised Sprague Dawley rats and perfused with a modified Ringer solution. Following four basal 30-min collections, transmitter release was stimulated by perfusion with 100 mM potassium for one collection. After re-equilibration, blood pressure was raised 60 mmHg for 30 min by phenylephrine infusion (1-1.3 mg/kg) then a 2-h recovery period followed. Dialysate collections were injected directly onto a reverse phase HPLC-ECD (HPLC with electrochemical detection). Basal extracellular noradrenaline concentrations were found to be similar in adult and old animals. Basal dihydroxyphenylacetic acid (DOPAC) concentrations were significantly greater in old compared to adult rats (P < 0.05). Potassium depolarisation induced a significant increase in noradrenaline concentrations in both age groups (P < 0.001), however the noradrenaline response to potassium stimulation was significantly reduced in the aged rats (P < 0.05). Potassium-induced decreases in DOPAC and homovanillic acid (HVA) concentrations were seen in both age groups. Following phenylephrine infusion, a modest delayed reduction in noradrenaline levels, which failed to reach statistical significance, was seen. Phenylephrine-induced hypertension was associated with decreased DOPAC and HVA concentrations in adult (P < 0.05) and old (P < 0.05) rats, respectively. These results indicate that ageing is associated with changes in dopaminergic and noradrenergic activity in the PVN of the rat. A reduction in noradrenaline response to maximal stimulation induced by potassium depolarisation was observed with ageing. The alteration in the activity of the catecholaminergic pathways to the PVN induced by phenylephrine infusion appears to be age dependent.

Neuropeptide Y potentiation of potassium-induced noradrenaline release in the hypothalamic paraventricular nucleus of the rat in vivo.

NPY is co-localised with catecholamines in the brain and periphery. Noradrenaline and NPY are present in high concentrations in the PVN of the hypothalamus, an area implicated in autonomic regulation. This microdialysis study examined whether NPY can modulate rat PVN noradrenaline release in vivo, as has been shown in vitro. Basal and K(+)-stimulated noradrenaline release was measured after i.c.v. administration of 2 nmol NPY or vehicle. No effect of NPY was observed on basal release, however a significant doubling of K(+)-induced release was observed, both 60 and 150 min following i.c.v. NPY. This raises the possibility that NPY may potentiate rather than inhibit brain noradrenaline release in vivo.

Catecholamine release in the rat hypothalamic paraventricular nucleus in response to haemorrhage, desipramine and potassium.

In vivo microdialysis and HPLC were used to measure catecholamine release in the rat hypothalamic paraventricular nucleus (PVN) during haemorrhage. The effects of noradrenaline uptake blockade with 1 microM desipramine (DMI) and a depolarising concentration of potassium (100 mM) through the probe were also examined. Dialysis probes implanted in the PVN of urethane anesthetised rats were perfused with modified Ringer solution at 1.1 microliter/min. Thirty minute collections were analysed for DOPA, noradrenaline, DOPAC, HVA and 5-HIAA. Basal concentrations, in the absence of DMI, were: DOPA 203.6 +/- 44.0 pg/ml, noradrenaline 128.0 +/- 20.4 pg/ml; DOPAC 5.6 +/- 0.7, HVA 5.1 +/- 2.2 and 5-HIAA 87.2 +/- 17.8 ng/ml. Basal noradrenaline was doubled in the presence of DMI while basal and stimulated DOPA, DOPAC, HVA and 5-HIAA were not affected by DMI. Haemorrhage resulted in a significant noradrenaline release (48% over resting levels) in the presence of DMI (n = 10, P < 0.05); in the absence of DMI, a smaller and non-significant increase (30% over basal levels) was observed. Potassium-induced depolarisation caused a significant two- and four-fold increase in noradrenaline release (P < 0.001), with decreases in the dopamine metabolites DOPAC (31%, 44%) and HVA (35%, 28%), and the serotonin metabolite, 5-HIAA (41%, 33%), in the presence and absence of DMI, respectively. The catecholamine precursor DOPA did not vary throughout either experiment. The results indicate that haemorrhage induces a 48% increase in noradrenaline release in the rat PVN which provides evidence for a role of noradrenergic projections to the PVN in cardiovascular control.