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Multiparity is associated with increased risk of cardiovascular disease. We tested whether multiparity induces oxidative stress in rat vascular tissue. Coronary arteries and thoracic aorta were isolated from multiparous and age-matched virgin rats. Relaxation to ACh and sodium nitroprusside (SNP) was measured by wire myography. We also tested the effect of the superoxide dismutase mimetic MnTE2PyP (30 μM), the NADPH oxidase inhibitor apocynin (10 μM), and the peroxynitrite scavenger FeTPPs (10 μM) on ACh-mediated relaxation in coronary arteries. Vascular superoxide anion was measured using the luminol derivative L-012 and nitric oxide (NO) generation by the Griess reaction. Multiparity reduced maximal response and sensitivity to ACh in coronary arteries [maximal relaxation (Emax): multiparous 49 ± 3% vs. virgins 95% ± 3%; EC50: multiparous 135 ± 1 nM vs. virgins 60 ± 1 nM], and in aortic rings (Emax: multiparous 38 ± 3% vs. virgins 79 ± 4%; EC50: multiparous 160 ± 2 nM vs. virgins 90 ± 3 nM). Coronary arteries from the two groups relaxed similarly to SNP. Superoxide anions formation was significantly higher in both coronary arteries (2.8-fold increase) and aorta (4.1-fold increase) from multiparous rats compared with virgins. In multiparous rats, incubation with MnTE2PyP, apocynin, and FeTPPs improved maximal relaxation to ACh (MnTE2PyP: 74 ± 5%; vehicle: 41 ± 5%; apocynin: 73 ± 3% vs. vehicle: 41 ± 3%; FeTPPs: 72 ± 3% vs. vehicle: 46 ± 3%) and increased sensitivity (EC50: MnTE2PyP: 61 ± 0.5 nM vs. vehicle: 91 ± 1 nM; apocynin: 45 ± 3 nM vs. vehicle: 91 ± 6 nM; FeTPP: 131 ± 2 nM vs. vehicle: 185 ± 1 nM). Multiparity also reduced total nitrate/nitrite levels (multiparous: 2.5 ± 2 μmol/mg protein vs. virgins: 7 ± 1 μmol/mg protein) and endothelial nitric oxide synthase protein levels (multiparous: 0.53 ± 0.1 protein/actin vs. virgins: 1.0 ± 0.14 protein/actin). These data suggest that multiparity induces endothelial dysfunction through decreased NO bioavailability and increased reactive oxygen species formation.

The mechanisms whereby maternal nutritional manipulation through pregnancy result in altered blood pressure in the offspring may include changes in fetal and newborn and adult renal prostaglandin (PG) synthesis, metabolism, and receptor expression. Since the postnatal effects of nutrient restriction on the renal PG synthesis and receptor system during nephrogenesis in conjunction with nephron numbers and blood pressure have not been evaluated in the rat, the present study examined the effect of reducing maternal food intake by 50% of ad libitum through pregnancy on young male rats. Six control-fed mothers and eight nutrient-restricted pregnant rats with single litter mates were used at each sampling time point, most of which occurred during nephrogenesis. Offspring of nutrient-restricted dams were lighter from birth to 3 days. This was accompanied by reduced PGE2, with smaller kidneys up to 14 days. Nutrient restriction also decreased mRNA expression of the PG synthesis enzyme, had little effect on the PG receptors, and increased mRNA expression of the degradation enzyme during nephrogenesis and the glucocorticoid receptor in the adult kidney. These mRNA changes were normally accompanied by similar changes in protein. Nephron number was also reduced from 7 days up to adulthood when blood pressure (measured by telemetry) did not increase as much as in control offspring during the dark, active period. In conclusion, maternal nutrient restriction suppressed renal PG concentrations in the offspring, and this was associated with suppressed kidney growth and development and decreased blood pressure.

The in vivo cardiovascular effects of acutely administered neurokinin B (NKB) have been attributed both to direct effects on vascular tone and to indirect effects on central neuroendocrine control of the circulation. We proposed: 1) that a modest long-term increase in plasma NKB levels would decrease mean arterial pressure (MAP) due to attenuated peripheral vascular tone, and 2) that chronic high-dose NKB would increase MAP, due to increased sympathetic outflow which would override the peripheral vasodilation. We examined the in vivo and in vitro cardiovascular effects of chronic peripheral NKB. Low- (1.8 nmol/h) or high- (20 nmol/h) dose NKB was infused into conscious female rats bearing telemetric pressure transducers. MAP, heart rate (HR) and the pressor responses to I.V. phenylephrine (PE, 8 microg) and angiotensin II (Ang II, 150 ng) were measured. Concentration-response curves of small mesenteric arteries were constructed to PE using wire myography. Low-dose NKB reduced basal MAP (88+/-2 mm Hg to 83+/-2 mm Hg), did not affect resting HR, reduced the pressor responses to PE, and attenuated the maximal constriction of mesenteric arteries to PE and KCl. By contrast, high-dose NKB increased basal MAP (86+/-1 mm Hg to 89+/-1 mm Hg), increased HR (350+/-3 beats/min to 371+/-3 beats/min), increased the pressor responses to Ang II and, contrary to our hypothesis, increased the maximum contractile responses of mesenteric arteries to PE and KCl. The cardiovascular effects of NKB are thus dose-dependent: whereas chronic low-dose NKB directly modulates vascular tone to reduce blood pressure, chronic high-dose NKB induces an increase in blood pressure through both central (indirect) and peripheral (direct) pathways.

We investigated the effect of relaxin on vasodilation, myogenic reactivity, and compliance of small mesenteric arteries and veins. In acute experiments, small (second order) mesenteric arteries and veins from female rats were mounted in a pressure myograph, perfused intraluminally with relaxin, and exposed to incremental increases in intraluminal pressure (20-120 mm Hg for arteries, 2-12 mm Hg for veins). We expressed myogenic reactivity as the ratio of active to passive diameter at each pressure step. In chronic experiments, relaxin was administered to rats (4 microg/h) for 3 days prior to isolating the vessels and measuring myogenic reactivity. Arteries were more sensitive than were veins to the acute vasodilatory activity of relaxin (EC50: arteries=1.32+/-0.18x10(-8) M; veins=3.19+/-0.88x10(-8) M, P0.05). Acute relaxin reduced myogenic reactivity of mesenteric arteries, but not veins. Chronic pretreatment with relaxin did alter the pressure/diameter relationship in Ca(2+)-containing medium, but this was due to increased passive compliance (control: 2.96+/-0.14 microm mm Hg(-1), n=5; relaxin: 3.72+/-0.16 microm mm Hg(-1), n=5) rather than to reduced myogenic reactivity. Chronic relaxin did not alter myogenic reactivity or compliance (control: 43.8+/-1.4 microm mm Hg(-1), n=5; relaxin: 46.1+/-2.3 microm mm Hg(-1), n=5) of veins. Thus, although relaxin reduces total peripheral resistance, it does not affect splanchnic venous capacitance or tone. In the face of elevated plasma relaxin levels, such as during pregnancy, cardiac preload may thus be maintained, concurrent with a reduction in cardiac afterload and blood pressure. We caution that, if an experimental treatment alters compliance, myogenic reactivity must be expressed as the ratio of active:passive diameter.

In humans, multiparity (repeated pregnancy) is associated with increased risk of cardiovascular disease. In rats, multiparity increases the pressor response to phenylephrine and to acute stress, due in part to changes in tone of the splanchnic arterial vasculature. Given that the venous system also changes during pregnancy, we studied the effects of multiparity on venous tone and compliance. Cardiovascular responses to volume loading (2 ml/100 g body wt), and mean circulatory filling pressure (MCFP, an index of venomotor tone) were measured in conscious, repeatedly bred (RB), and age-matched virgin rats. In addition, passive compliance and venous reactivity of isolated mesenteric veins were measured by pressure myography. There was a greater increase in mean arterial pressure after volume loading in RB rats (+7.2 ± 2.5 mmHg, n = 8) than virgin rats (−1.4 ± 1.7 mmHg, n = 7) ( P < 0.05). The increase in MCFP in response to norepinephrine (NE) was also greater in RB rats [half maximal effective dose (ED50) 3.1 ± 0.5 nmol·kg−1·min−1, n = 6] than virgins (ED50: 12.1 ± 2.7 nmol·kg−1·min−1, n = 6) ( P < 0.05). Pressure-induced changes in passive diameter were lower in isolated mesenteric veins from RB rats (29.3 ± 1.8 μm/mmHg, n = 6) than from virgins (36.9 ± 1.3 μm/mmHg, n = 6) ( P < 0.05). Venous reactivity to NE in isolated veins was also greater in RB rats (EC50: 2.68 ± 0.37×10−8 M, n = 5) than virgins (EC50: 4.67 ± 0.93 × 10−8 M, n = 8). We conclude that repeated pregnancy induces a long-term reduction in splanchnic venous compliance and augments splanchnic venous reactivity and sympathetic tonic control of total body venous tone. This compromises the ability of the capacitance (venous) system to accommodate volume overloads and to buffer changes in cardiac preload.

We developed a novel model using fluorescent intravital microscopy to study the effect of atrial natriuretic peptide (ANP) on the extrasplenic microcirculation. Continuous infusion of ANP into the splenic artery (10 ng min−1 for 60 min) of male Long–Evans rats (220–250 g, n= 24) induced constriction of the splenic arterioles after 15 min (−7.2 ± 6.6% from baseline diameter of 96 ± 18.3 μm, mean ±s.e.m.) and venules (−14.4 ± 4.0% from 249 ± 25.8 μm; P < 0.05). At the same time flow did not change in the arterioles (from 1.58 ± 0.34 to 1.27 ± 0.27 ml min−1), although it decreased in venules (from 1.67 ± 0.23 to 1.15 ± 0.20 ml min−1) and increased in the lymphatics (from 0.007 ± 0.001 to 0.034 ± 0.008 ml min−1; P < 0.05). There was no significant change in mean arterial pressure (from 118 ± 5 to 112 ± 5 mmHg). After continuous ANP infusion for 60 min, the arterioles were dilated (108 ± 16 μm, P < 0.05) but the venules remained constricted (223 ± 24 μm). Blood flow decreased in both arterioles (0.76 ± 0.12 ml min−1) and venules (1.03 ± 0.18 ml min−1; P < 0.05), but was now unchanged from baseline in the lymphatics (0.01 ± 0.001 ml min−1). This was accompanied by a significant decrease in MAP (104 ± 5 mmHg; P < 0.05). At 60 min, there was macromolecular leak from the lymphatics, as indicated by increased interstitial fluorescein isothiocyanate–bovine serum albumin fluorescence (grey level: 0 = black; 255 = white; from 55.8 ± 7.6 to 71.8 ± 5.9, P < 0.05). This study confirms our previous proposition that, in the extrasplenic microcirculation, ANP causes greater increases in post- than precapillary resistance, thus increasing intrasplenic capillary hydrostatic pressure (Pc) and fluid efflux into the lymphatic system. Longer-term infusion of ANP also increases Pc, but this is accompanied by increased ‘permeability’ of the extrasplenic lymphatics, such that fluid is lost to perivascular third spaces.

We have previously shown that the splenorenal reflex controls renin release through splenic afferent and renal sympathetic nerves. We proposed that this reflex would also affect renal blood flow (RBF). RBF was measured in male Long Evans rats using transit-time flow probes. There were no significant differences between any of the experimental groups with respect to baseline values of RBF (8.9 +/- 0.4 ml min(-1), n= 25) or mean arterial pressure (MAP, 98.7 +/- 2.5 mmHg, n= 25). Splenic venous pressure was selectively raised (from 7.9 +/- 0.6 to 21.6 +/- 0.3 mmHg, n= 25) in anaesthetized rats by partial ligation of the splenic vein. This caused an immediate fall in RBF (-2.1 +/- 0.2 ml min(-1), n= 7) and in MAP (-12.4 +/- 2.8 mmHg, n= 7). The fall in RBF, but not the fall in blood pressure, was attenuated by renal denervation (DeltaRBF: - 0.7 +/- 0.1 ml min(-1), n= 6), splenic denervation (DeltaRBF: -0.8 +/- 0.1 ml min(-1), n= 6) and close renal arterial injection of the alpha1-adrenergic blocker phenoxybenzamine (12.5 microg; DeltaRBF: -0.8 +/- 0.1 ml min(-1), n= 6). Renal conductance fell only in the intact control group, i.e. the residual fall in RBF in the denervated and phenoxybenzamine-treated animals could be attributed to the fall in MAP. We also showed that splenic vein occlusion increased both splenic afferent (from 3.0 +/- 0.3 to 6.6 +/- 0.6 spikes s(-1), n= 5) and renal efferent (from 24.8 +/- 2.0 to 50.2 +/- 4.9 spikes s(-1), n= 9) nerve activity. We conclude that obstruction to splenic venous outflow, such as would occur in portal hypertension, initiates increased splenic afferent nerve activity and renal vasoconstriction through the splenorenal reflex, as well as a fall in blood pressure. We propose that this contributes to the renal and cardiovascular dysfunction observed in portal hypertension.

Increases in intravascular volume are detected by mechanoreceptors situated at the junctions of the great veins with the atria. We had previously shown that localized distension of the superior vena caval/right atrial junction, simulating increased cardiac preload, elicits release of ANF remotely from the atrial appendage. We proposed that ANF secretion is stimulated via intrinsic neural pathways running from the venoatrial junctions to the appendage. We developed a technique whereby non-adrenergic, non-cholinergic sensory nerves could be selectively destroyed in the heart of adult rats by instilling capsaicin into the pericardial space. Four days later, the animals were killed, and isolated perfused atria were prepared with small balloons positioned so that the superior vena caval/right atrial junction could be discretely stretched. Immunoreactive ANF secretion into the perfusate was measured. Although distension of the venoatrial junction increased ANF secretion from the control atria, there was no such response in the denervated atria. We conclude (A) that local application of capsaicin to the heart of adult rats induces selective functional neural deficits and (B) that information regarding distension of the junction of the great veins and the atria is normally transmitted across the atrium via these nerves to stimulate ANF secretion from peptide stores located in the atrial appendage. We propose that these pathways are crucial to ensure appropriate ANF secretion in response to an increase in circulating blood volume.

We have previously shown that intrasplenic fluid extravasation is important in controlling blood volume. We proposed that, because the splenic vein flows in the portal vein, portal hypertension would increase splenic venous pressure and thus increase intrasplenic microvascular pressure and fluid extravasation. Given that the rat spleen has no capacity to store/release blood, intrasplenic fluid extravasation can be estimated by measuring the difference between splenic arterial inflow and venous outflow. In anesthetized rats, partial ligation of the portal vein rostral to the junction with the splenic vein caused portal venous pressure to rise from 4.5 ± 0.5 to 12.0 ± 0.9 mmHg ( n = 6); there was no change in portal venous pressure downstream of the ligation, although blood flow in the liver fell. Splenic arterial flow did not change, but the arteriovenous flow differential increased from 0.8 ± 0.3 to 1.2 ± 0.1 ml/min ( n = 6), and splenic venous hematocrit rose. Mean arterial pressure fell (101 ± 5.5 to 95 ± 4 mmHg). Splenic afferent nerve activity increased (5.6 ± 0.9 to 16.2 ± 0.7 spikes/s, n = 5). Contrary to our hypothesis, partial ligation of the portal vein caudal to the junction with the splenic vein (same increase in portal venous pressure but no increase in splenic venous pressure) also caused the splenic arteriovenous flow differential to increase (0.6 ± 0.1 to 1.0 ± 0.2 ml/min; n = 8). The increase in intrasplenic fluid efflux and the fall in mean arterial pressure after rostral portal vein ligation were abolished by splenic denervation. We propose there to be an intestinal/hepatic/splenic reflex pathway, through which is mediated the changes in intrasplenic extravasation and systemic blood pressure observed during portal hypertension.

In the rat, the spleen is a major site of fluid efflux out of the blood. By contrast, the mesenteric vasculature serves as a blood reservoir. We proposed that the compliance and myogenic responses of these vascular beds would reflect their different functional demands. Mesenteric and splenic arterioles ( approximately 150-200 microm) and venules (

Background We have previously characterized several antihypertensive peptides in simulated digests of cooked eggs and showed blood pressure lowering property of fried whole egg digest. However, the long-term effects of this hydrolysate and its fractions on blood pressure are not known. Therefore, the objectives of the study were to determine the effects of long term administration of fried whole egg hydrolysate and its fractions (i.e. egg white and egg yolk) on regulation of blood pressure and associated factors in cardiovascular disease such as plasma lipid profile and tissue oxidative stress. Methods and Results We used spontaneously hypertensive rats (SHR), an animal model of essential hypertension. Hydrolysates of fried egg and its fractions were prepared by simulated gastro-intestinal digestion with pepsin and pancreatin. 16–17 week old male SHRs were orally administered fried whole egg hydrolysate, non-hydrolyzed fried whole egg, egg white hydrolysate or egg yolk hydrolysates (either defatted, or not) daily for 18 days. Blood pressure (BP) and heart rate were monitored by telemetry. Animals were sacrificed at the end of the treatment for vascular function studies and evaluating plasma lipid profile and tissue oxidative stress. BP was reduced by feeding fried whole egg hydrolysate but not by the non-hydrolyzed product suggesting a critical role for in vitro digestion in releasing anti-hypertensive peptides. Egg white hydrolysate and defatted egg yolk hydrolysate (but not egg yolk hydrolysate) also had similar effects. Reduction in BP was accompanied by the restoration of nitric oxide (NO) dependent vasorelaxation and reduction of plasma angiotensin II. Fried whole egg hydrolysate also reduced plasma levels of triglyceride although it was increased by the non-hydrolyzed sample. Additionally the hydrolyzed preparations attenuated tissue oxidative stress. Conclusion Our results demonstrate that fried egg hydrolysates exert anti-hypertensive effects, improve plasma lipid profile and attenuate tissue oxidative stress in vivo.

Plasma 5α-pregnan-3α-ol-20-one (pregnan) levels and nitric oxide (NO) biosynthesis increase during pregnancy. These factors have independently been implicated in the control of blood pressure and volume. We wished to determine whether pregnan might be responsible both for the increase in NO biosynthesis and for the increase in plasma volume observed during pregnancy. Virgin female Long-Evans rats were implanted with indwelling cannulas and maintained on a low nitrate/nitrite diet. After the rats recovered from surgery, 500 μg of pregnan or vehicle were given daily for 2 days. NO biosynthesis and plasma volume were measured in conscious animals before and after treatment. Pregnan caused a significant increase in NO biosynthesis (1.9 ± 0.8 μmol/24 h, n = 10) compared with the vehicle-treated control group (0.3 ± 0.4 μmol/24 h, n = 10, P < 0.05). Similarly, there was a significant increase in plasma volume in the pregnan-treated group (0.7 ± 0.2 ml/100 g, n = 11) compared with the vehicle-treated control group (0.2 ± 0.1 ml/100 g, n = 11, P < 0.05). These results confirm that pregnan can mimic pregnancy by its ability to increase both NO biosynthesis and plasma volume.

Atrial natriuretic factor (ANF(1-28); ANF) is important in fluid volume homeostasis. It is released in response to atrial distention, as would normally occur during hypervolaemia (Levin et al. 1998). Exogenously administered ANF causes an increase in haematocrit and a decrease in plasma volume, which cannot be accounted for by urinary losses (De Bold et al. 1981). We have shown that these responses are abolished by splenectomy (Kaufman, 1992) and that ANF causes a sustained increase in the haematocrit of blood as it passes through the spleen (Deng & Kaufman, 1996). Since the rat spleen is non-contractile and has virtually no blood storage capacity (Reilly, 1985), the additional red cell mass cannot originate from splenic reservoirs. The increase in haematocrit must therefore derive from a reduction in plasma volume. Indeed, further investigations have revealed that, under normovolaemic conditions, about 25 % of fluid volume flowing into the spleen is removed from the circulating blood and drains into the systemic lymphatic system (Chen & Kaufman, 1996). These findings have led us to believe that the spleen is a major site of ANF-induced translocation of fluid out of the intravascular space. ANF could potentially cause this increase in fluid extravasation by altering capillary permeability within the spleen, as it has been shown to do this at high doses in a variety of tissues (Trippodo & Barbee, 1987). However, splenic capillary beds have been proven to have a discontinuous endothelium (Takubo et al. 1999), which makes it unlikely that fluid extravasation results from increased capillary permeability. Furthermore, we have shown that there is no change in plasma protein concentration of blood flowing through the spleen, and that lymph draining from the spleen is isoncotic to the plasma; this indicates that the capillary beds are indeed freely permeable to plasma proteins (Kaufman & Deng, 1993). Therefore, we have proposed that changes in splenic fluid extravasation in response to physiological levels of ANF occur, not through changes in capillary permeability, but via alterations in splenic haemodynamics to increase intrasplenic filtration pressure. Such a mechanism would be analogous to the role of ANF in the kidney, where it raises glomerular filtration pressure, thereby increasing glomerular filtration rate (Marin Grez et al. 1986). The present study was designed to determine the mechanism by which ANF increases fluid efflux from the spleen. The experiments were done in a blood-perfused rat spleen, which enabled us to measure changes in arterial and venous blood flow, intrasplenic microvascular pressure, and pre-/post-capillary resistance in response to close-arterial infusion of ANF.

We investigated the effect of the NO donor SNAP (6.7 nM) on basal and stretch-induced ANF release from isolated perfused rat atria. There was no significant difference in basal ANF secretion between the vehicle- and SNAP-infused atria (SNAP: 388+/-63 pg. 100 microl(-1), n = 13 vs. vehicle: 349+/-26 pg. 100 microl(-1), n = 5). Atrial distention caused an increase in ANF secretion in both the buffer- and SNAP-treated groups. SNAP greatly attenuated the stretch-induced increase in ANF (SNAP: 225+/-7 pg. 100 microl(-1), n = 5 vs. vehicle: 448+/-72 pg. 100 microl(-1), n = 13, P < 0.05). The compliance of atria treated with SNAP was lower than that of the vehicle-perfused atria (P < 0.05). Thus, although SNAP appeared to attenuate stretch-induced ANF secretion, there was in fact no significant difference in the ratio of Delta[ANF] to Deltaintraluminal volume (SNAP: 5.8+/-1.3 pg. 100 microl(-1). microl(-1) vs. vehicle: 8.2+/-1.4 pg. 100 microl(-1). microl(-1).). In conclusion, we found no evidence that NO alters the control of basal or stretch-induced ANF secretion. NO can however reduce ANF release by shifting the pressure-volume curve, so that a given increase in atrial pressure is associated with a smaller increase in intraluminal volume and reduced atrial distention.

We hypothesized that nitric oxide (NO) contributes to intrasplenic fluid extravasation by inducing greater relaxation in splenic resistance arteries than veins such that intrasplenic microvascular pressure (PC) rises. Fluid efflux was estimated by measuring the difference between splenic blood inflow and outflow. Intrasplenic infusion of the NO donor S-nitroso- N-acetylpenicillamine (SNAP) (0.3 μg · 10 μl−1 · min−1) caused a significant increase in intrasplenic fluid efflux (baseline: 0.8 ± 0.4 ml/min, n = 10 vs. peak rise during SNAP infusion: 1.3 ± 0.4 ml/min, n = 10; P < 0.05). Intrasplenic PC was measured in the isolated, blood-perfused rat spleen. Intrasplenic infusion of SNAP (0.1 μg · 10 μl−1 · min−1) caused a significant increase in PC (saline: 10.9 ± 0.2 mmHg, n = 3 vs. SNAP: 12.2 ± 0.2 mmHg, n = 3; P < 0.05). Vasoreactivity of preconstricted splenic resistance vessels to sodium nitroprusside (SNP) (1 × 10−12-1 × 10−4 M) and SNAP (1 × 10−10-3 × 10−4 M) was investigated with the use of a wire myograph system. Significantly greater relaxation of arterioles than of venules occurred with both SNP (%maximal vasorelaxation: artery 96 ± 2.3, n = 9 vs. vein 26 ± 1.9, n = 10) and SNAP (%maximal vasorelaxation: artery 50 ± 3.5, n = 11 vs. vein 32 ± 1.7, n = 8). These results are consistent with our proposal that differential vasoreactivity of splenic resistance arteries and veins to NO elevates intrasplenic PC and increases fluid extravasation into the systemic lymphatic system.

Atrial natriuretic factor (ANF) release was studied in isolated perfused atria prepared from rats. When the vein-atrial junction (VAJ) was distended with an inflatable balloon, ANF release into the perfusate was greater in intact atria than in appendectomized atria. It was concluded that distention of the VAJ causes ANF release from the atrial appendage. A cascade experiment was then prepared whereby buffer from one isolated atrium perfused a second atrium. Although the VAJ of the first atrium could be distended by balloon, the atrial appendage was ligated so ANF was not secreted into the perfusate. The second atrium was intact, but no balloon was inserted. Despite the fact that there were no changes in intraluminal pressure, ANF secretion from the second atrium increased when the VAJ of the first atrium was distended. This response was blocked by the endothelin (ET) A receptor antagonist BQ-123. However, no distention-induced changes in ET-1 levels could be found in the perfusate from the first atrium. It is proposed that, in response to changes in distention of the VAJ, ANF is released remotely from the atrial appendage. The mediator does not appear to be ET-1 itself, but rather some factor that stimulates ET-1-induced ANF release within the tissue of the atrial appendage.Key words: atrial natriuretic factor (ANF) secretion, endothelin, atrial distention, BQ-123, atrial appendectomy.

Objective: We examined the response to hypertonic saline challenge (SC) as a potential predictor of fluid retention during heart failure induced by rapid ventricular pacing. Methods: Twelve dogs (22 ± 4 kg) were given an intra-arterial bolus of 30 ml of 20% saline after establishing baseline fluid intake and urine output (24 h). Dogs were classified according to whether they drank more (Group A) or less (Group B) than the amount required to dilute the SC to isotonicity. Fluid retention was then assessed during heart failure after rapid ventricular pacing according to a graded ordinal scale and correlated with the responses to SC. Results: No difference was noted in baseline fluid intake (1112 ± 236 ml in Group A vs. 809 ± 129 ml in Group B). Five hours after SC cumulative water intake was significantly greater in Group A than in Group B (1018 ± 136 vs. 591 ± 17 ml) ( P < 0.01). Urine sodium concentration was 113 ± 11 and 124 ± 28 mmol/l at baseline in Group A and B, respectively; increased to 190 ± 21 and 295 ± 59 mmol/l at 5 h and remained elevated 24 h after SC, 177 ± 60 and 274 ± 55 mmol/l (both P < 0.01 for within-group comparisons vs. baseline). Urine sodium concentration was less in Group A than in Group B at 5 and 24 h ( P < 0.05). The fluid retention score was greater in Group A (3.6 ± 0.5) than in Group B (0.8 ± 0.4) ( P < 0.01). Fluid retention in heart failure correlated with water intake after the pre-pacing SC ( r = 0.68, P < 0.025) and inversely with urine concentrating ability ( r = −0.58, P < 0.02). Furthermore, water intake and urine concentrating ability following the SC were inversely related ( r = −0.67, P < 0.02). Conclusions: We conclude that normal dogs may be classified according to their fluid intake after SC. Those dogs that drank excessively and produced a dilute urine were more likely to retain fluid during pacing-induced heart failure. Hence, fluid intake and the ability to excrete a concentrated urine after a saline challenge may be useful variables to predict fluid retention in pacing-induced heart failure.

1. Atrial natriuretic factor (ANF) causes a reduction in plasma volume that is abolished by splenectomy. Experiments were conceived to investigate whether ANF acts within the spleen to increase efflux of fluid from the intravascular to the extravascular space. 2. ANF, infused into the splenic artery of anaesthetized rats at rates of 1, 5 and 20 ng min -1, caused a dose-dependent increase in the arteriovenous difference in haematocrit as blood flowed through the spleen (basal difference, 0.18 +/- 0.10%; difference after 10 min at 20 ng min -1 ANF, 1.5 +/- 0.18%; n = 6). There was no such change in plasma protein concentration. 3. ANF (20 ng min -1) did not alter splenic arterial blood flow. However, splenic venous blood flow fell so that the arteriovenous difference increased significantly (basal difference, 0.34 +/- 0.19 ml min -1; difference at 60 min, 1.1 +/- 0.20 ml min-1, n = 7). There was no change in mean arterial pressure. 4. These data confirm our hypothesis that ANF acts within the spleen to increase fluid efflux from the intravascular to the extravascular space. Since there is no change in total splenic blood flow, we propose that the effects of ANF are mediated by dilatation of the splenic afferent arterioles and constriction of the efferent venules, thus increasing filtration pressure.

1. Previous evidence has suggested that fluid is extracted from the blood during its passage through the splenic circulation. In order to further investigate this phenomenon, flow probes were placed around the splenic artery and vein of rats. Splenic blood flow and mean arterial blood pressure were measured immediately after surgery, while the rats were still anaesthetized. Five days later, the measurements were repeated first in conscious rats, and then after administering sodium pentobarbitone. Final measurements were made in the conscious animals at day 10 post surgery. 2. Splenic arterial blood flow was lowest at the time of surgery. It had doubled by day 5, and remained stable thereafter at about 8 ml min-1. Splenic venous blood flow also increased after surgery, but less so than the arterial flow, so that the arteriovenous difference had increased to 2.0 +/- 0.4 ml min-1 by day 5. 3. In response to sodium pentobarbitone (20 mg I.V.), there was a transient fall in splenic arterial blood flow from 6.7 +/- 0.9 to 4.9 +/- 0.7 ml min-1. There was no significant change in the arteriovenous difference of blood flow. 4. In conclusion, we have shown that splenic blood flow is considerably higher than previously reported. We have also revealed a significant new path for fluid efflux from the intravascular space; at least 25% of fluid volume flowing into the splen is removed from the circulating blood. 5. Although anaesthesia modulates splenic blood flow, the changes are not as pronounced as those observed in the acute surgically prepared animal.

1. We investigated the control of atrial natriuretic factor (ANF) secretion during pregnancy. 2. Plasma ANF levels were measured in conscious virgin female rats under basal conditions, and after atrial distension with an indwelling balloon catheter. The rats were then mated, and the measurements repeated at 7, 14 and 21 days of pregnancy, and at 1 week postpartum. Plasma ANF levels were also measured in ovariectomized rats injected with progesterone, oestradiol, or oestradiol plus progesterone. 3. Basal plasma ANF levels were elevated at 7 and 14 days of pregnancy, but returned to prepregnant levels by 21 days. At 1 week postpartum, they were again elevated. 4. In response to atrial stretch, plasma ANF increased significantly in virgin rats (from 100 +/- 10 to 148 +/- 13 pg ml-1, P < 0.001, n = 20). In contrast, there was no such secretory response observed in the pregnant and postpartum animals i.e. stretch-induced secretion of ANF was markedly attenuated. 5. Treatment with exogenous oestradiol caused a significant increase in plasma ANF levels in acyclic rats. However, neither progesterone nor a combination of oestradiol plus progesterone had any effect. 6. It is concluded that basal and stretch-induced ANF secretion are differentially influenced by pregnancy; oestradiol is identified as a potential stimulatory factor.

Isolated perfused right atria were prepared from virgin cycling rats and from rats at 7, 14, and 21 days of pregnancy. Intraluminal pressure was raised from the basal control level (atmospheric) to 4, 6, and 10 cmH2O. Basal and stretch-induced release of atrial natriuretic factor (ANF) into the perfusate was measured by radioimmunoassay. When atria derived from unmated and from 7-day pregnant rats were distended, mean ANF secretion into the perfusion medium increased by 64 +/- 16 and 89 +/- 31 pg/ml at the highest distending pressure of 10 mmHg. (Mean basal secretion for the two groups was 196 +/- 104 and 181 +/- 63 pg/ml, respectively.) However, at 14 and 21 days, distension failed to elicit any significant increase in ANF release. There was no significant difference between the groups with respect to basal secretion of ANF, nor did pregnancy influence atrial compliance or volume. The slopes of the pressure-volume curves for atria derived from virgin, 7-, 14-, and 21-day-pregnant rats were 0.76 +/- 0.17, 0.623 +/- 0.178, 0.811 +/- 0.177, and 1.050 +/- 0.173, respectively. These results are in agreement with our findings in vivo that plasma ANF levels are elevated at 7 days of pregnancy but that, despite the progressive increase in intravascular volume, they decrease to control levels at 14 and 21 days. Our latest data suggest that this fall is a result of changes in the secretory characteristics of the atrial tissue itself and not necessarily of pregnancy-induced changes in atrial filling.

Chronic indwelling balloons were placed at the junction of the right atrium and the right superior vena cava of virgin female rats. The renal response to discrete standardized atrial stretch was tested in the conscious animals. The rats were then mated, and the renal response was retested at days 7, 14, and 20 of pregnancy. On day 21, the rats were killed, and the pressure-volume relationship of the right atria was measured. In response to atrial stretch, there was an increase in urine volume, urine sodium output, and urine potassium output and a decrease in free water clearance in the virgin rats. Subsequent to mating, these responses were completely abolished at all stages of pregnancy. The renal responses to atrial stretch were also abolished during pseudopregnancy. Central venous pressure did not change when the intracardiac balloons were inflated; there were also no changes in resting central venous pressure during pregnancy. There were no significant changes in unstressed atrial volume or in atrial compliance during pregnancy.

1. We tested the hypothesis that hypervolaemia causes an increase in intrasplenic filtration of cell-free fluid out of the vasculature. To this end we developed a preparation in the anaesthetized rat whereby the splenic vein could be non-occlusively cannulated. 2. Haematocrit and plasma protein concentrations were measured in the splenic afferent and efferent blood supplies. 3. In response to volume loading with saline (1% body weight), there was a sustained increase in the arterial-venous differential of haematocrit, i.e. there was a relative increase in the haematocrit of the blood draining from the spleen. There was no such change in plasma protein concentration. By contrast, this degree of volume loading had no effect on the haematocrit of blood passing through the hindquarters of the animal. 4. Following volume expansion, there was no significant difference in the protein concentration of the plasma and the lymph fluid collected from the splenic lymphatic duct. 5. Distension of the superior vena caval-right atrial junction by means of a small inflatable balloon, caused a similar increase in the splenic venous haematocrit, and again, no change in plasma protein concentration. 6. We interpret these results to mean that, in response to expansion of the intravascular space, there is increased intrasplenic filtration of plasma out of the blood and into the lymphatic system.

Atrial natriuretic factor (ANF) causes an increase in hematocrit that cannot be accounted for by urinary losses. The mechanism behind this phenomenon was studied in intact and splenectomized rats. Rat ANF 99–126 was infused i.v. for 30 min into conscious rats at rates of 0 (saline control), 0.05, or 0.1 μg/min. Plasma volume was then determined by dilution of the dye, Evan's Blue. In one group of rats, red cell volume was determined using 51Cr-labelled erythrocytes. ANF infusion was continued uninterrupted throughout the experiments. In the intact rats, ANF (0.10 μg/min) caused hematocrit to increase from 38.9 ± 0.5 to 41.2 ± 0.4% (p

Adrenomedullin is synthesized and secreted by fetoplacental tissues. Given that the placenta lacks autonomic innervation, we proposed that adrenomedullin acts locally to control blood flow in the placental vasculature through a balance of dilatory and constrictive pathways. Placental stem villous arteries (200 microm) from normotensive human pregnancies were dissected and mounted on a wire myograph. The vessels were preconstricted with the thromboxane A(2) mimetic U46619 (EC(80) concentration), and exposed to cumulative concentrations of adrenomedullin (1 x 10(-9) to 3 x 10(-7) mol/L). Adrenomedullin caused concentration-dependent vasorelaxation which, in endothelium-intact vessels, was attenuated in the presence of the nitric oxide synthase inhibitor L-NMMA. This suggested that the vasodilation was mediated, at least in part, through nitric oxide. However, removal of the endothelium did not similarly alter the response. Nor did L-NMMA have any effect in endothelium-denuded vessels. We hypothesized that adrenomedullin must induce release of both endothelium-derived relaxing (nitric oxide) and constricting factors. When we blocked the two major pathways through which adrenomedullin is known to induce vasodilation, by incubating the vessels with L-NMMA (nitric oxide synthase inhibitor) and Rp-cAMPS (cAMP-dependent protein kinase inhibitor), adrenomedullin induced concentration-dependent vasoconstriction. This was not mediated through endothelin, since addition of the non-specific endothelin receptor antagonist PD142893 failed to alter the response to adrenomedullin. We conclude that, in addition to increasing endothelial nitric oxide biosynthesis in placental stem villous arteries, adrenomedullin induces release of an endothelium-derived constricting factor.

The splenorenal reflex induces changes in mean arterial pressure (MAP) and renal function. We hypothesized that, in addition to spinal pathways previously identified, these effects are also mediated through central pathways. We investigated the effect of elevated splenic venous pressure on central neural activation in intact, renal-denervated, and renal + splenic-denervated rats. Fos-labeled neurons were quantified in the nucleus of the tractus solitarius (NTS), paraventricular nucleus (PVN), supraoptic nucleus (SON), and subfornical organ (SFO) after 1-h partial splenic vein occlusion (SVO) in conscious rats bearing balloon occluders around the splenic vein, telemetric pressure transducers in the gastric vein (splenic venous pressure), and abdominal aorta catheters (MAP). SVO stimulated Fos expression in the PVN and SON, but not NTS or SFO of intact rats. Renal denervation abolished this response in the parvocellular PVN, while renal + splenic denervation abolished activation in the magnocellular PVN and the SON. In renal-denervated animals, SVO depressed Fos expression in the NTS and increased expression in the SFO, responses that were abolished by renal + splenic denervation. In intact rats, SVO also induced a fall in right atrial pressure, an increase in renal afferent nerve activity, and an increase in MAP. We conclude that elevated splenic venous pressure does induce hypothalamic activation and that this is mediated through both splenic and renal afferent nerves. However, in the absence of renal afferent input, SVO depressed NTS activation, probably as a result of the accompanying fall in cardiac preload and reduced afferent signaling from the cardiopulmonary receptors.

Stenosis of either the portal or splenic vein increases splenic afferent nerve activity (SANA), which, through the splenorenal reflex, reduces renal blood flow. Because these maneuvers not only raise splenic venous pressure but also reduce splenic venous outflow, the question remained as to whether it is increased intrasplenic postcapillary pressure and/or reduced intrasplenic blood flow, which stimulates SANA. In anesthetized rats, we measured the changes in SANA in response to partial occlusion of either the splenic artery or vein. Splenic venous and arterial pressures and flows were simultaneously monitored. Splenic vein occlusion increased splenic venous pressure (9.5 +/- 0.5 to 22.9 +/- 0.8 mmHg, n = 6), reduced splenic arterial blood flow (1.7 +/- 0.1 to 0.9 +/- 0.1 ml/min, n = 6) and splenic venous blood flow (1.3 +/- 0.1 to 0.6 +/- 0.1 ml/min, n = 6), and increased SANA (1.7 +/- 0.4 to 2.2 +/- 0.5 spikes/s, n = 6). During splenic artery occlusion, we matched the reduction in either splenic arterial blood flow (1.7 +/- 0.1 to 0.7 +/- 0.05, n = 6) or splenic venous blood flow (1.2 +/- 0.1 to 0.5 +/- 0.04, n = 5) with that seen during splenic vein occlusion. In neither case was there any change in either splenic venous pressure (-0.4 +/- 0.9 mmHg, n = 6 and +0.1 +/- 0.3 mmHg, n = 5) or SANA (-0.11 +/- 0.15 spikes/s, n = 6 and -0.05 +/- 0.08 spikes/s, n = 5), respectively. Furthermore, there was a linear relationship between SANA and splenic venous pressure (r = 0.619, P = 0.008, n = 17). There was no such relationship with splenic venous (r = 0.371, P = 0.236, n = 12) or arterial (r = 0.275, P = 0.413, n = 11) blood flow. We conclude that it is splenic venous pressure, not flow, which stimulates splenic afferent nerve activity and activates the splenorenal reflex in portal and splenic venous hypertension.

The atrial volume reflex is attenuated in pregnancy. This may be mimicked by chronic administration of 5α-pregnan-3α-ol-20-one (pregnan). We investigated whether afferent output from sensory receptors may be suppressed at this time. Vagal afferent nerve activity was measured during discrete localized stimulation of the atrial volume receptors by inflation of a balloon at the superior vena caval-right atrial junction. The receptors were classified as high- (HF) or low- (LF) frequency subtypes on the basis of their response to graded atrial distension. Although both HF (regression coefficient = 0.50 ± 0.11 Hz/μl, r2 = 0.47, P < 0.001) and LF (regression coefficient = 0.03 ± 0.05 Hz/μl, r2 = 0.009, P = 0.613) subtypes could be identified in virgin rats, only LF (regression coefficient = 0.09 ± 0.05 Hz/μl, r2 = 0.044, P = 0.099) receptors were found in late-pregnant animals. Similarly, in virgin rats treated chronically with pregnan (500 μg/24 h for 2 days), only LF receptors were identified (regression coefficient = −0.004 ± 0.078 Hz/μl, r2 = 0.000, P = 0.962), whereas both subtypes were present in the vehicle-treated animals (HF regression coefficient = 0.626 ± 0.255 Hz/μl, r2 = 0.317, P = 0.029; LF regression coefficient = −0.012 ± 0.071 Hz/μl, r2 = 0.002, P = 0.866). By contrast, acute intracardiac pregnan (2.6 μg/kg) did not alter vagal afferent nerve activity. In conclusion, stretch-induced discharge of high-frequency atrial receptors is suppressed during pregnancy, whereas that of low-frequency receptors is preserved. This effect may be mimicked by chronic, but not acute, pregnan. We propose that, during pregnancy, pregnan alters the transducer properties of the atrial volume receptors, thus allowing blood volume to increase.

Placental neurokinin B (NKB) was recently identified as the causative agent in preeclampsia, a condition characterized by increased maternal and feto-placental vascular resistance. We hypothesized that NKB should constrict placental resistance vessels. Placentas were obtained from normotensive pregnancies. Immediately after delivery, stem villous arteries (300 microm diameter, 1.2 mm long) were dissected from macroscopically normal tissue in cold HEPES-physiological salt solution (PSS), mounted on a wire myograph system, and bathed in HEPES-PSS at 37 degrees C. After determination of the passive-tension internal circumference characteristics, the arteries were set to 90% of the internal circumference they would have under a normal physiological transmural pressure. Cumulative concentration-response curves were constructed for NKB (1 x 10(-12) to 1 x 10(-5) mol/l). Since there was no constrictive response to NKB, cumulative constrictive concentration-response curves were constructed to the thromboxane A(2) mimetic U46619 (1 x 10(-9) to 1 x 10(-5) mol/l). The vessels were then pre-constricted to 80% of maximal response and exposed to cumulative concentrations of NKB (1 x 10(-12) to 1 x 10(-6) mol/l). NKB caused a concentration-dependent relaxation (Maximal response NKB, 51+/-5%, n=5; time control, 12+/-6%, n=4; P0.05). Removal of the endothelium did not alter the vasodilatory response to NKB. We conclude that, contrary to our hypothesis, NKB causes an endothelium-independent relaxation of the placental resistance vessels. We propose that NKB plays a role in the maintenance of high placental blood flow in normal pregnancy.

Close-arterial infusion of test substances allows one to study the responses of a selected vascular bed without inducing confounding systemic effects. Unfortunately, laminar flow patterns within the artery cause streaming of the injected factor, so that distribution within the target organ is not homogeneous. We describe a reliable method of overcoming these problems. Specifically, we attach a vibrator (i-Vibe egg) to the syringe containing the test substance. We showed that, without vibration, infusion of a solution of Evans blue (0.5% wt/vol) results in uneven distribution of the dye in the kidney. Vibration of the syringe during infusion allows for uniform coloration of the kidney surface. There is also functional improvement of drug distribution during vibration. Renal blood flow was measured during intrarenal infusion of phenylephrine (150 μl, 0.05–0.5 μg). Vibration caused a significant leftward shift in the dose-response curve, i.e., the phenylephrine-induced reduction in renal blood flow was enhanced by vibration. This cheap, simple method for ensuring adequate mixing of intra-arterially infused substances will facilitate not only the study of renal function in the rat but also infusion of test and therapeutic substances into other organs.

We have previously shown that ANP causes differential constriction of the splenic vasculature of the rat (veins greater than arteries), which may be inhibited by blocking the production of cGMP with A7195. In this paper, we report experiments done on vessels derived from guanylyl cyclase (GC)-A knockout mice. Small splenic arteries (∼150-μm diameter) and veins (∼250-μm diameter) were dissected from male GC-A-deficient 129sv mice or age-matched wild-type controls and mounted in a wire myograph. In the wild-type mice, ANP exhibited higher potency in the veins than in the arteries (EC50 values wild-type mice: artery, 8 ± 3 × 10−9 M, n = 5 vs. vein, 6 ± 4 × 10−10 M, n = 5; P < 0.05). The concentration-response curve for ANP-induced vasoconstriction was also shifted leftward in denuded compared with intact arteries (EC50 values: denuded artery: 5 ± 3 × 10−10 M, n = 5 vs. intact artery, 8 ± 3 × 10−9 M, n = 5; P < 0.05), i.e., the denuded vessels were more reactive. By contrast, ANP caused no significant change in tension from baseline in intact splenic arteries, intact splenic veins, or denuded splenic arteries derived from the GC-A-deficient mice, although these vessels did show normal concentration-dependent increases in tension to phenylephrine. We conclude that ANP causes vasoconstriction in the splenic vasculature by an endothelium-independent mechanism, mediated via guanylyl cyclase.

Atrial distension increases c- fosexpression in the paraventricular nucleus of virgin, but not pregnant, rats. We proposed that nitric oxide (NO), biosynthesis of which increases during pregnancy, blunts this reflex and that blocking NO biosynthesis would restore the response. Female rats were implanted with indwelling intracardiac balloons. On day 14 of pregnancy, osmotic minipumps containing either d- or N G-nitro-l-arginine methyl ester (l-NAME) (120 mg/2 ml at 10 μg/min) were implanted. On day 20, the rats were infused with saline (3 ml/h) with or without atrial balloon inflation (1 h). The brains were then processed for quantitation of c- fos expression. In the virgin rats, and in the pregnant rats treated with l-NAME, atrial distension significantly increased hypothalamic c- fosexpression. In the pregnant animals treated with d-NAME, the response was greatly attenuated. NO had no effect on the increase in atrial receptor afferent discharge (single-fiber recordings) elicited by atrial distension. We conclude that, during pregnancy, NO attenuates central processing of the reflex response to atrial distension but does not alter the transducer properties of the volume receptors.

Nitric oxide (NO) biosynthesis increases during pregnancy and has been shown to suppress baroreceptor activity. The renal response to a simulated increase in circulating blood volume (atrial distension) is also attenuated at this time. We hypothesized that blocking NO biosynthesis during pregnancy would restore the renal response. Female rats were implanted with indwelling intracardiac balloons and central venous cannulas. After recovery, they were mated, and on day 14 of pregnancy, osmotic minipumps containing the NO synthase inhibitor N G-nitro-l-arginine methyl ester (l-NAME) or its inactive enantiomer N G-nitro-d-arginine methyl ester (d-NAME) (120 mg/2 ml at 10 μg/min) were implanted. In response to atrial distension (1 h), urine output increased in the d- and l-NAME-treated virgin rats. During pregnancy ( day 20), this response was attenuated in the d-NAME-treated, but not thel-NAME-treated, animals, i.e., after a simulated increase in circulating blood volume, inhibition of NO biosynthesis restored the renal response of pregnant rats to that seen in virgin animals. We conclude that, during normal pregnancy, increased NO biosynthesis blunts the reflex renal response to atrial distension.

During lipopolysaccharide (LPS)-induced endotoxemia, increased intrasplenic fluid efflux contributes to a reduction in plasma volume. We hypothesized that splenic sympathetic nerve activity (SSNA), which increases during endotoxemia, limits intrasplenic fluid efflux. We reasoned that splenic denervation would exaggerate LPS-induced intrasplenic fluid efflux and worsen the hypotension, hemoconcentration, and hypovolemia. A nonlethal dose of LPS (150 μg · kg−1 · h−1 for 18 h) was infused into conscious male rats bearing transit time flow probes on the splenic artery and vein. Fluid efflux was estimated from the difference in splenic arterial inflow and venous outflow (A-V). LPS significantly increased the (A-V) flow differential (fluid efflux) in intact rats (saline −0.01 ± 0.02 ml/min, n = 8 vs. LPS +0.21 ± 0.06 ml/min, n = 8); this was exaggerated in splenic denervated rats (saline −0.03 ± 0.01 ml/min, n = 7 vs. LPS +0.41 ± 0.08 ml/min, n = 8). Splenic denervation also exacerbated the LPS-induced hypotension, hemoconcentration, and hypovolemia (peak fall in mean arterial pressure: denervated 19 ± 3 mmHg, n = 10 vs. intact 12 ± 1 mmHg, n= 8; peak rise in hematocrit: denervated 6.7 ± 0.3%, n = 8 vs. intact 5.0 ± 0.3%, n = 8; decrease in plasma volume at 90-min post-LPS infusion: denervated 1.08 ± 0.15 ml/100 g body wt, n = 7 vs. intact 0.54 ± 0.08 ml/100 g body wt, n = 8). The exaggerated LPS-induced hypovolemia associated with splenic denervation was mirrored in the rise in plasma renin activity (90 min post-LPS: denervated 11.5 ± 0.8 ng · ml−1 · h−1, n = 9 vs. intact 6.6 ± 0.7 ng · ml−1 · h−1, n = 8). These results are consistent with our proposal that SSNA normally limits LPS-induced intrasplenic fluid efflux.

We recently demonstrated that fluid is filtered out of the splenic circulation and into the lymphatic system. The current experiments were designed to investigate the importance of this route of fluid extravasation in endotoxemia. Lipopolysaccharide (LPS) was infused into conscious intact and splenectomized rats (150 μg ⋅ kg−1⋅ h−1iv for 18 h). In the intact rats, mean arterial pressure (MAP) fell from 101 ± 2.4 to 88 ± 3.9 mmHg ( n = 7) and then stabilized at about 90 mmHg. Hematocrit rose from 41 ± 0.9 to 45 ± 0.4% at 40 min, at which time plasma volume had fallen from 4.7 ± 0.12 to 4.0 ± 0.05 ml/100 g body wt. In the splenectomized rats MAP did not fall and hematocrit did not rise. There also was no change in plasma volume, i.e., splenectomy prevented the hypotension and hemoconcentration customarily induced by LPS. In a second series of experiments, splenic arterial and venous blood flows were simultaneously measured in anesthetized rats infused with LPS (150 μg ⋅ kg−1⋅ h−1). LPS increased splenic fluid efflux. We conclude that during endotoxemia the initial fall in circulating blood volume may be attributed to fluid extravasation from the splenic vasculature.

In virgin female rats, atrial distension (an index of blood volume expansion) causes an increase in c- fos expression in the paraventricular nucleus of the lateral hypothalamus. During pregnancy, this response is markedly attenuated. We tested the effects of 3α-hydroxy-5α-pregnan-20-one (3α-OH-DHP) on activation of central pathways following stimulation of the atrial volume receptors. Not only does this progesterone metabolite increase during pregnancy, but it has already been implicated in pregnancy-induced changes in the baroreflex. Female rats were prepared with indwelling venous cannulas and intracardiac balloons that, when inflated, caused a discrete localized stimulation of the atrial volume receptors in the absence of changes in cardiac hemodynamics. Seven days later, the rats were infused with 3α-OH-DHP dissolved in cyclodextrin and the intracardiac balloons were inflated. One hour later, the rats were killed and fixed by perfusion and the brains were prepared for visualization of c- fos activity. Infusion with 3α-OH-DHP significantly reduced the central response to atrial distension, i.e., it mimicked pregnancy. These results are consistent with the suggestion that this metabolite of progesterone may be an important factor in cardiovascular adaptation to pregnancy.

On sait que les humains et les rats âges possedent de hauts taux plasmatiques de facteur natriuretique auriculaire (FNA), ce qui pourrait etre explique en partie par une reduction de clairance de la circulation. Dans cette etude, nous avons voulu verifier si la nature secretrice du tissu auriculaire change aussi avec l'âge. Pour ce faire, nous avons examine la liberation du FNA en reponse a une distension auriculaire, chez des rats mâles au debut de l'âge adulte (2-3 mois) et des rats mâles reproducteurs inactifs plus âges (4-6 mois), tant in vivo (animaux instrumentes, conscients) que in vitro (oreillettes perfusees, isolees). Une augmentation de la pression intraluminale a induit une augmentation de la secretion de FNA dans les oreillettes des jeunes rats, mais non pas dans celles des rats plus âges, c.-a-d. que, dans ce cas, la secretion induite par l'etirement a ete alteree. Une reduction de la compliance auriculaire serait responsable de l'effet observe chez les rats plus âges. Les taux de FNA plasmatiques de base de nos rats reproducteurs inactifs etaient plus faibles que ceux signales dans les etudes anterieures, bien que la reponse a la charge de volume ait ete maintenue. Ces resultats suggerent que, vers l'âge de 6 mois, le tissu auriculaire est deja moins sensible aux variations de pression auriculaire. Ainsi, l'augmentation des taux plasmatiques de FNA, observee anterieurement chez les rats vieillissants, resulte probablement d'une diminution de clairance plutot que d'une augmentation de secretion.

Stimulation of the atrial volume receptors increases neural traffic to the ventrolateral medulla, which in turn sends output to, and receives input from, the lateral hypothalamic area. An integrated reflex and hormonal response is thus initiated. We wished to investigate first whether atrial distension results in activation of selected nuclei in the forebrain and, second, whether pregnancy modifies this response. Rats were implanted with indwelling intracardiac balloons positioned at the superior vena caval/right atrial junction. One week later, the balloons were inflated. The animals were then anesthetized, their brains fixed by perfusion, and the tissue prepared for visualization of c-fos activity. Atrial distension caused a significant increase in c-fos expression in the paraventricular nucleus, the medial preoptic area, and the lateral septum. This response was markedly attenuated in the pregnant animals. In conclusion, during pregnancy central pathways that are normally activated in responses to volume expansion, fail to respond to atrial distension. We propose that this allows blood volume to increase in the pregnant animal, without triggering homeostatic mechanisms.

SUMMARY 1. We wished to determine whether, during pregnancy, there is reduced renal response to atrial distension, whether secretion of atrial natriuretic factor (ANF) is suppressed and whether neural input from the atrial volume receptors to the central nervous system is altered. 2. Conscious, chronically instrumented female rats were used. Atrial distension was achieved by implanting small balloons positioned at the superior vena caval/right atrial junction; inflation of the balloon did not impede blood flow through the heart. 3. In virgin rats, atrial stretch caused increased urine volume, urine sodium and potassium output and decreased free water clearance. These responses were abolished during pregnancy. 4. In response to atrial stretch, plasma ANF levels increased significantly in virgin rats. No such secretory response was observed in the pregnant animals. 5. Distension of isolated atria derived from unmated and 7 day pregnant rats resulted in an increase in secretion of ANF into the perfusate. Atria from 14 and 21 day pregnant rats were unresponsive to distension. 6. Pretreatment with oestradiol (50 μg daily for 10 days) caused plasma ANF levels and ANF secretion by isolated perfused atria to increase. By contrast, testosterone pretreatment (15 mg twice weekly for 2 weeks) abolished stretch induced secretion of ANF by isolated atria. 7. C-fos activity in the paraventricular nucleus of virgin rats increased in response to atrial distension. This response was reduced in the 7 day pregnant rats and abolished at 21 days. 8. We conclude that there is attenuation of both hormonal and neural responses to atrial distension in the pregnant animal. This allows blood volume to increase without eliciting homeostatic mechanisms to eliminate the extra fluid.

The pressor and heart rate responses to infused angiotensin (ANG) II were measured in conscious pregnant Long-Evans rats. Responses were recorded 7 days after chronic indwelling venous and arterial cannulas were implanted (virgin rats) and again at days 7, 14, and 21 of pregnancy. A significant reduction in the pressor response was noted as early as day 7 of pregnancy; this is comparable to the human condition. It was also noted that the reflex bradycardia associated with the pressor response was entirely absent at 21 days. To determine a possible cause of these pregnancy-induced changes, ovariectomized rats were injected subcutaneously for 10 days with estradiol, progesterone, or a combination of both. They were then tested for pressor responses to ANG II. None of the hormone-injected groups showed any significant deviation in their pressor response compared with saline-injected controls. Similar results were obtained in acutely prepared rats under chloroform anesthesia. However, the magnitude of the pressor response in the chloroformed animals was significantly reduced compared with the conscious unrestrained animals. It is concluded that 1) the rat is an appropriate model of human pregnancy in which to study changes in ANG responsiveness in pregnancy and 2) neither estradiol nor progesterone is responsible for the reduced pressor response.

The relationship between prolonged (6 h) atrial stretch, urine output, and plasma atrial natriuretic factor (ANF) was investigated in conscious rats. On inflation of a balloon at the superior venal caval-right atrial junction, urine volume and sodium output increased. However, this renal response was no longer apparent after the first hour, and there were no further changes in urine output when the balloon was deflated. This transient renal response was most marked during the first 30 min of balloon inflation, at which time plasma ANF was also elevated. At 6 h, plasma ANF was still elevated, although urine output had, by then, returned to normal. On deflation of the balloon, plasma ANF levels returned to normal. It is clear that the renal and hormonal responses to atrial stretch may be temporally uncoupled. This probably reflects adaptation of the atrial volume receptors in the absence of such an effect on ANF release. Although this might be interpreted to mean that ANF is not of physiological importance in fluid and electrolyte balance, a more plausible explanation is that ANF needs the backing of the reflex pathways arising from stimulation of the atrial receptors to express its natriuretic activity.

1. Extracellular fluid deficits of 33% were produced in male Long-Evans rats by peritoneal dialysis. The conscious, unrestrained animals were then infused I.V. for 30 min with atriopeptin III at doses of 0.01, 0.1, 0.5 and 1.0 micrograms/min. At 5 min into the infusion, the rats were offered water and subsequent intakes were monitored. Since atrial natriuretic peptide (ANP) causes hypotension, one group of control animals was given an injection of diazoxide sufficient to match this fall in blood pressure. 2. A similar group of rats was prepared for measurement of plasma ANP achieved by infusion. 3. Relative to the saline-infused controls, atriopeptin III did not reduce water intake. Indeed, intake was increased at the highest dose of 1.0 micrograms/min. 4. Relative to the diazoxide controls, water intake was influenced by atriopeptin III in a dose-dependent manner, the greatest attenuation being observed at infusions of 0.1 microgram/min. 5. Infusion of atriopeptin III at 0.1 microgram/min caused plasma ANP levels to rise from 252 +/- 21 to 532 +/- 136 pg/ml (n = 9, P less than 0.05) at 15 min. The lowest dose (0.01 microgram/min) caused no detectable increase in plasma levels. 6. It is concluded that, in groups of hypovolaemic rats matched for blood pressure, atriopeptin III caused a dose-related reduction in water intake.

Female rats subjected to 48 h of water deprivation drank more than similarly deprived males. There was no sexual difference in the dipsogenic responses to intracellular dehydration produced by intravenous hypertonic NaCl but females drank considerably more than males in response to the extracellular stimuli of hyperoncotic polyethylene glycol and angiotensin II. It is concluded that intact adult female rats are dipsogenically more responsive than adult male rats to stimuli acting through the pathways of extracellularly induced thirst but not to those arising from the intracellular fluid compartment.

Two groups of male Wistar rats were chronically implanted with small inflatable balloons at the right superior vena caval (SVC)-atrial junction. The balloons were inflated for 60 min in one-third of the rats in each group; the remainder served as noninflated controls. Extract was prepared from the right atria of the first and from the left atria of the second group and bioassayed in male Wistars to determine the content of atrial natriuretic factor (ANF). The bioassay rats received either two injections of sham extract (control) or an initial injection (I1) of sham extract followed by an injection (I2) of balloon-inflated extract (experimental). For right atrial extract, the incremental response to the experimental injection (I2/I1) was significantly less than the response to the control injection (experimental, 1.15 +/- 0.09 mueq/min, n = 14; control, 1.59 +/- 0.15 mueq/min, n = 13; P less than 0.01). However, in the case of left atrial extract, there was no such difference between the control group (1.35 +/- 0.13 mueq/min, n = 19) and the experimental group (1.25 +/- 0.11 mueq/min, n = 11; Student's t test). We conclude that right atrial distension causes release of ANF from the right atrium, but not the left, and that this release is probably locally mediated.

Rats were prepared with inflatable balloons at the superior vena cava – right atrium junction. After recovery 1 week later, when blood was taken from conscious, normovolaemic animals plasma renin activity was found not to be influenced by right atrial stretch. Plasma renin activity was then measured in rats in which an extracellular fluid deficit had been produced by peritoneal dialysis against a hyperoncotic, isotonic solution. Although basal plasma renin activity was elevated (6.8 ± 0.9 from 1.5 ± 0.2 ng∙mL∙h, n = 19), no depression was observed in the experimental group after 15 or 90 min of balloon inflation. In rats pretreated with isoprenaline (10 μg/kg body wt.) plasma renin activity was also increased over basal levels, but again balloon inflation caused no reduction in plasma renin activity. It would appear that right atrial stretch has little, if any, influence on renin release in the conscious rat.

1. Studies were made on the effects of right atrial stretch and atrial natriuretic factor (ANF) infusion on fluid movement into the intestinal tract. 2. Stimulation of the atrial volume receptors by inflation of an intracardiac balloon in the conscious, unrestrained rat did not change intestinal fluid content under normovolaemic conditions. 3. When the rat was rendered hypovolaemic by peritoneal dialysis (34% deficit in extracellular fluid volume), right atrial stretch significantly increased intestinal fluid content. Under these conditions, inflation of the balloon restored large intestinal fluid content to the pre-dialysis state, i.e. right atrial stretch completely abolished that component of fluid absorption attributable to the extracellular fluid volume deficit. 4. These data suggest that stimulation of the right atrial receptors inhibits intestinal fluid reabsorption but probably does not initiate fluid secretion. 5. There was no evidence that this might be mediated by ANF since rat ANF (twenty-eight residue peptide, Ser-99-Tyr-126), infused for 1 h at doses of 0, 0.05, 0.1 and 0.5 microgram/min, did not cause any detectable changes in the fluid content of the large or small intestine of similarly prepared hypovolaemic rats.