Formation of insulin-secreting, Sertoli-enriched tissue constructs by microgravity coculture of isolated pig islets and rat Sertoli cells.

Pancreatic islets, isolated from neonatal pigs, and Sertoli cells, isolated from prepubertal rats, were cocultured in simulated microgravity utilizing the NASA-developed highly accelerating, rotating vessel (HARV) biochamber. Following 5 d of incubation, three-dimensional Sertoli-islet cell aggregates (SICA) retained the ability to secrete insulin when exposed to elevated glucose. SICA contained FasL-positive Sertoli cells and insulin-positive beta-cells randomly organized within the spherical construct. The addition of 1% Matrigel induced the reorganization of aggregates (SICAs formed in the presence of Matrigel [SICAmgs]) showing the peripherialization and epithelialization of Sertoli cells and the centralization of islets in association with lumen-like spaces. The Sertoli cells, but not Matrigel, aided in preserving the structural integrity of HARV-incubated islets. Neither Matrigel nor Sertoli cells appeared to interfere with the ability of SICA or SICA mg to secrete insulin and express FasL.

Leptin secretion from the epididymal fat pad is increased by the sexual maturation of the male rat.

Although leptin has been implicated as an important factor in triggering the onset of puberty in females, much less is known about the role of this adipose tissue hormone in the sexual maturation of males. Previous work in the rat has suggested that the peripubertal rise in testosterone precedes an increase in leptin secretion, and it has been suggested that the testosterone rise induces the leptin increase. These studies examined some of the interactions between leptin secretion and the peripubertal testosterone rise in male rats. Serum leptin concentrations were significantly elevated in young adult male rats compared with immature rats. Cultured epididymal fat pads obtained from adult animals secreted significantly more leptin than did those obtained from immature rats. Castration of immature rats with or without testosterone replacement for 1 week did not result in a significant change in either the serum leptin concentrations or the ability of the epididymal fat pad to secrete leptin. Exposure of epididymal fat to 5 ng/mL of testosterone in vitro resulted in a significantly enhanced secretion of leptin into the media compared with plain media controls. These results confirmed that there is an increase in serum leptin concentrations with sexual maturation in the male rat. They also suggest that this increase is due to an enhanced ability of adipose tissue to secrete leptin. Within a normal physiologic range, testosterone may play a role in inducing this increased ability to secrete leptin.

Corticosterone-attenuating and anxiolytic properties of mecamylamine in the rat.

1. The available evidence suggests that stress induced release of acetylcholine (ACh) in the brain has a significant role in mediating neuroendocrine, emotional, and physiological responses to stress. Recent findings also suggest that stress indirectly (via acetylcholine) and nicotine directly stimulates the HPA axis through activation of nAChRs. 2. Our working hypothesis is that under stressful conditions, nicotinic receptor antagonists, such as mecamylamine, should act to attenuate the activation of the HPA axis and exhibit anxiolytic behavioral effects. The purpose of this study was to determine whether or not mecamylamine would: a) produce anxiolytic effects in rats on the elevated plus maze and b) blunt the plasma corticosterone response to predator stress in rats. 3. Results suggested that mecamylamine has anxiolytic properties under stressful conditions. In the EPM experiment, mecamylamine (0.3 mg/kg) produced increased time spent in the open arms. Similarly, in the predator stressor experiment, mecamylamine blunted the stress-induced plasma corticosterone response, with the lowest dose of mecamylamine (0.1 mg/kg). 4. These findings may have important therapeutic implications since clinical observations have shown that low doses of mecamylamine reduce tension and anxiety in patients with Tourette syndrome.

Evidence supporting a physiological role for proANP-(1-30) in the regulation of renal excretion.

The experiments, performed in pentobarbital sodium-anesthetized rats, consisted of a 1-h equilibration period followed by two 30-min control periods. Subsequently, synthetic rat pro atrial natriuretic peptide (ANP) [proANP-(1-30)] (n = 8) was given as a bolus of 10 microg in 1 ml of 0.9% saline followed by an infusion at 30 ng/min (20 microl/min) for six additional periods. Control rats (n = 6) received only 0.45% saline in the appropriate volumes. Mean arterial pressure, renal blood flow, and glomerular filtration rate did not change significantly in either group during the proANP-(1-30) infusion. Urine flow and potassium excretion increased approximately 50% in the proANP-(1-30)-infused group only (P < 0.05). Sodium excretion and fractional excretion of sodium, expressed as the change from their own baselines, were significantly increased by the proANP-(1-30) infusion (P < 0.05), whereas cGMP excretion was similar in both groups. These results suggest that the rat sequence of proANP-(1-30) produces a natriuresis in the rat independent of changes in hemodynamics and renal cGMP production. In a second study, rats (n = 8) were prepared as above and pretreated with 0.4 ml iv of rabbit serum containing an antibody directed against proANP-(1-30) (anti-proANP group). The rats were volume expanded with 3 ml of 6% albumin in Krebs and observed for 3 h to determine if the anti-proANP would attenuate the responses to volume expansion. Control rats (n = 7) received 0.4 ml of normal rabbit serum. The elevation in potassium excretion in response to volume expansion was significantly attenuated in the anti-proANP group (P < 0.05). Sodium excretion and urine flow responses also tended to be reduced but not significantly. These results suggest that in the rat, proANP-(1-30) plays a physiological role in regulating renal excretion.

Formation of Sertoli cell-enriched tissue constructs utilizing simulated microgravity technology.

Cell transplantation therapy for diabetes and Parkinson's disease offers hope for long-term alleviation of symptoms. However, successful protocols remain elusive due to obstacles, including rejection and lack of tropic support for the graft. To enhance engraftment, testis-derived postmitotic Sertoli cells have been cotransplanted with islets in the diabetic rat (Db) and neurons in the Parkinsonian rat (PD). Sertoli cell tropic, regulatory, and nutritive factors that nourish and stimulate germ cells also support isolated neurons and islets in vitro. Likewise, immunosuppressive properties of Sertoli cells, extant in the testis, are expressed by extratesticular Sertoli cells evidenced by allo- and xenograft immunoprotection of grafts in both the CNS (in the PD model) and the periphery (in the Db model). On this basis, we have created Sertoli islet cell aggregates (SICA) and Sertoli neuron aggregated cells (SNAC) using simulated microgravity culture technology developed by NASA. Isolated rat and pig Sertoli cells were cocultured with neonatal pig islets (SICA) and with immortalized N-Terra-2 (NT2) neurons (SNAC) in the HARV biochamber. Formed aggregates were assayed for desirable functional and structural characteristics. Cell viability in SICA and SNAC exceeded 90% and FasL immunopositive Sertoli cells were present in both. Sertoli cells did not interfere with insulin secretion by SICA and promoted differentiation of NT2 cells to the dopaminergic hNT cell type in SNAC. Addition of Matrigel resulted in structural reorganization of the aggregates and enhanced insulin secretion. We conclude that SICA, SNAC, and Matrigel-induced islet- and neuron-filled "Sertoli cell biochambers" are suitable for long-term transplantation treatment of Db and PD.

Increased bioactivity of rat atrial extracts: relation to aging and blood pressure regulation.

The purpose of this study was to evaluate the possible role of atrial factor(s) in the regulation of cardiovascular homeostasis and their relationship to aging. Rats were anesthetized and received jugular vein, carotid artery, and bilateral ureteral catheterization. After a half-hour equilibration period, the rats received 0.5 ml of atrial extract with a concentration of proANP (atrial natriuretic peptide) of 150 microg/ml prepared from either aged (18-20 month, "aged extract group", n = 12) or young (2-3 month, "young extract group", n = 12) rats. Mean arterial pressure (MAP) and renal function were monitored over five 20-minute periods. The atrial extract caused MAP to fall significantly in the aged extract group (p < .05) but MAP was unchanged in young extract group. There was a significant difference in MAP between the two groups (p < .05). Urine output increased significantly in both groups after extract infusion (p < .05 in both cases). Sodium and potassium excretion showed similar responses. However, the diuresis, natriuresis, and kaliuresis after extract infusion would have been expected to be relatively lower in the aged extract group compared to the young extract group considering the significantly lower MAP in the aged extract group. High performance gel permeation chromatography (HP-GPC) analysis of the atrial extract showed an increased quantity of a large molecular weight C-terminal peptide in atrial extracts from aged rats compared to young rat atria. Plasma levels of ANP and proANP 1-30 both increased significantly after extract infusion in both aged and young groups, and there was no significant difference in ANP concentration between the two groups. However, the concentration of proANP 1-30 was significantly increased in the aged group compared to the young group after extract infusion. These results suggest that changes in the structure or processing of proANP in aging may contribute to the different hemodynamic responses.

Temporal relation between leptin and various indices of sexual maturation in the male rat.

Recent studies in humans and rhesus monkeys have suggested the possibility that the adipose tissue hormone leptin has a stimulatory and/or permissive effect on the onset of puberty in the male. We evaluated this hypothesis by measuring leptin in groups of male rats between the ages of 26 days and 96 days. A statistically significant positive correlation was present between serum leptin and age, body weight, prostate, seminal vesicle, and testes weight (both absolute and as a function of body weight). A statistically significant negative correlation was present between leptin and serum FSH and alpha-inhibin. There was not a statistically significant correlation between leptin and testosterone or LH. There was a statistically significant increase in the serum leptin concentrations at day 47. This rise was coincident with the peripubertal growth spurt in the secondary sexual organs and the peripubertal testosterone rise but occurred after the prepubertal rise in testicular weight, the appearance of elongating spermatids in the testes, and the start of the decline in FSH. In animals in which the peripubertal testosterone rise was delayed by the administration of EDS, serum leptin showed statistically significant differences from control. These data do not support the hypothesis that leptin provides a trigger for the onset of puberty in the male rat. They do suggest that leptin may be involved in the secondary sexual organ growth spurt and are consistent with the hypothesis that testosterone stimulates leptin synthesis during puberty.

Development of Sertoli cell binding competency in the peripubertal rat.

The peripheral distribution of Sertoli cell F-actin, a cytoskeletal protein found in Sertoli cell ectoplasmic specializations, is associated with enhanced spermatid binding to the Sertoli cell and, as such, serves as a functional marker for its acquisition of binding competency. Previous studies suggest that the peripheral distribution of actin is dependent on follicle-stimulating hormone (FSH). To investigate the developmental pattern of Sertoli cell actin distribution in relation to peripubertal FSH and testosterone levels, we examined epithelial sheets from 2-8 week-old-rats. Tissues were processed for light microscopy and for the visualization of rhodamine-labeled F-actin. At 2 weeks, actin staining was diffuse throughout most of the Sertoli cells and was similar to that observed in binding-incompetent Sertoli cells. By 4 weeks, actin distribution was peripheral, acquiring the same staining pattern as observed in binding-competent Sertoli cells. Serum levels of FSH peaked at 4 weeks and declined to adult levels thereafter. Testosterone levels did not increase significantly until 6 weeks. Results show that Sertoli cell actin undergoes peripheral reorganization concurrent with the peripubertal peak of FSH but prior to the peripubertal rise of testosterone. The study demonstrates a temporal correlation between the peripubertal FSH rise and the actin redistribution in Sertoli cells that is consistent with an induction of this redistribution by FSH. These results suggest that FSH induces binding competency in Sertoli cells.

Alterations in the opioid control of LHRH release from hypothalami isolated from aged male rats.

Several lines of evidence have suggested that the opioid control of gonadotropin secretion in the male rat is altered with aging. Because neural control of gonadotropins is mediated through luteinizing hormone releasing hormone (LHRH) secreting neurons, we examined the postulated changes in the opioid control of gonadotropins more directly by studying isolated hypothalamic fragments in vitro. Hypothalami from young (75-90 days) and old (18-20 months) males were examined for their ability to release LHRH when incubated with increasing doses of naloxone in a semi-static culture system. Serum concentrations of testosterone and luteinizing hormone (LH) in the donor animals were both significantly lower in old male rats compared with young males. Basal secretion of LHRH was similar in both age groups. Two-way repeated measures ANOVA indicated that naloxone stimulated a significant dose-dependent increase in the release of LHRH into the media. ANOVA also indicated a significant effect of age. We conclude that the changes in the endogenous opioid systems reported to occur with aging are, in fact, linked to differences in LHRH secretion and thus to differences in the dynamic relationship between testosterone and LH in older male rats.

Alterations in atrial natriuretic peptide (ANP) secretion and renal effects in aging.

Aging is associated with hypertension and electrolyte disturbances. The purpose of this study was to determine the effect of aging upon secretion and renal actions of atrial natriuretic peptide (ANP). Rats were anesthetized and received tracheal, jugular vein, carotid artery, and bilateral uretheral catheterization. One set of young (2-3 mo) rats (Group 2, n = 9) and one set of old (18-21 mo) rats (Group 4, n = 7) received bilateral atrial appendectomies. Control young (Group 1, n = 8) and old (Group 3, n = 8) rats received a sham appendectomy. All rats were infused (iv) with 6% albumin in Krebs buffer, sufficient to increase blood volume by 15%. Finally, each rat was injected with ANP (1 microgram/kg). Sodium excretion rate (U(Na+)V) in response to volume expansion was significantly decreased in all groups compared to Group 1 (young control, p < .05). All groups demonstrated a striking increase in U(Na+)V with the ANP injection, but the response was greatest in young control rats when factored by body weight (p < .05). There were no significant differences in MAP between the groups, suggesting that the differences in U(Na+)V observed were not the result of hemodynamic factors. Isolated perfused atria from young (n = 9) and old (n = 8) rats were subjected to stretch and endothelin stimulation (50 nM). Atria from young rats showed a dramatic increase in ANP secretion in response to atrial stretch and a further marked increase in secretion in response to endothelin, whereas both of these responses were markedly attenuated in old rats (p < .05). These results suggested that the secretion and renal effects of ANP are impaired in aging. Changes in secretion and actions of ANP in aging could contribute to the development of hypertension or heart failure.

Endothelin acts as a paracrine regulator of stretch-induced atrial natriuretic peptide release.

Several lines of evidence suggest a paracrine regulatory role for endothelin (ET) in the release of atrial natriuretic peptide (ANP). To investigate this possibility, we used the ET A-type receptor (ETA-R) competitive inhibitor cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123) in isolated perfused atria to determine the effect of endogenously produced ET on the release of ANP. Initially, we found that high pressure (8-10 mmHg) increased the mean ANP secretion rate by 117.3 +/- 21.2% (P < 0.05). Next, we found that at high pressure 50 nM of exogenously applied ET significantly augmented the stretch-induced release of ANP (P < 0.05) and that this response could be significantly attenuated in a dose-dependent manner by 1 and 3 microM BQ-123 (P < 0.05). These experiments proved the efficacy of the inhibitor in our model. Subsequently, we found that the stretch-induced release of ANP was significantly reduced to 51.5 +/- 13.0 and 22.3 +/- 11.8% by 1 and 3 microM BQ-123, respectively (P < 0.05). Because the perfused atria model eliminates systemic cardiovascular effects, allows control and direct recording of the intra-atrial pressure, and preserves the potential endothelium-myocyte control system, we conclude that the stretch-induced release of ANP is partially regulated by ET and that the ET is locally produced and constitutes a paracrine control system.

Secretion and renal effects of ANF prohormone peptides.

1. Atrial natriuretic factor (ANF) and pro ANF peptide appears to be secreted simultaneously from the atria in response to atrial stretch. 2. The major peptide forms secreted from rat atria appear to be ANF (pro ANF 99-126) as the primary C-terminal peptide and pro ANF 1-30 as the primary N-terminal peptide, as opposed to 1-67 or 1-98. 3. The plasma concentrations of ANF and pro ANF 1-30 are increased by acute stimulation with blood volume expansion and the plasma levels of ANF and N-terminal ANF prohormone peptides are chronically elevated by high salt diet. 4. Pro ANF 31-67 produces a natriuresis which is not dependent on an increase in renal cGMP excretion, decreases in plasma renin activity (PRA) or elevations in plasma ANF concentration.

Opioid inhibition of adrenergic and dopaminergic but not serotonergic stimulation of luteinizing hormone releasing hormone release from immortalized hypothalamic neurons.

Opioids are known to have an inhibitory effect on the secretion of luteinizing hormone releasing hormone (LHRH) when administered to whole animals in vivo or when applied to hypothalamic fragments in vitro. Whether opioids have this effect by acting directly on the LHRH secreting neurons or require the mediation of an interneuron is controversial. To examine this question, a clonal cell line derived from a hypothalamic neuron (GT1-7) was perfused and fractions collected every 6 min. Morphine treatment had no effect on basal secretion of LHRH, nor on the spontaneous, pulsatile release of LHRH. Isoproterenol, dopamine, and serotonin all produced significant increments in LHRH secretion. Pretreatment of GT1-7 cells for 2 h with morphine, suppressed the LHRH response to isoproterenol and dopamine but had no apparent effect on serotonin-induced LHRH release. These data indicate that morphine has a direct effect on GT1-7 cells that alters their responsiveness to some, but not all, LHRH secretagogues. These results suggest that, in vivo, the inhibitory effects that opioids have on LHRH release may not require an interneuron.

Possible mechanisms involved in the natriuretic response to atrial natriuretic factor (ANF) and proANF 31-67 in the rat.

1. The present study was conducted to compare the mechanisms involved in the natriuretic response to atrial natriuretic factor (ANF) and proANF 31-67. The peptides were infused intravenously into anaesthetized rats at 10 pmol/min for 40 min. 2. Only ANF produced a significant decrease in arterial pressure; the maximum decrease was 11 mmHg (P < 0.05). 3. Both peptides produced significant increases in sodium excretion (P < 0.05) but only ANF increased the cyclic GMP (cGMP) excretion rate (P < 0.01) and neither peptide had a significant effect on plasma renin activity or glomerular filtration rate (GFR). ProANF 31-67 did not increase the plasma levels of ANF. 4. These results demonstrate that both ANF and proANF 31-67 have natriuretic effects via a tubular mechanism and suggest that the natriuretic effects of ANF are mediated by cGMP while the excretion, changes in GFR or a reduction in renin secretion.

Effects of follicle-stimulating hormone on the junction-related Sertoli cell cytoskeleton and daily sperm production in testosterone-treated hypophysectomized rats.

Specialized junctional binding of step 8 spermatids to Sertoli cells is an important spermiogenic event. In the hypophysectomized (Hypox) rat, the daily sperm production (DSP) is reduced and Sertoli cells become binding incompetent. Delayed replacement of testosterone (DT-Hypox) does not restore the normal DSP and Sertoli cells remain binding incompetent. In this study, DT-Hypox rats received FSH daily for 2 days to 3 wk concurrent with delayed testosterone replacement and were killed 8 wk after the initiation of hormone treatment. The DT-Hypox rat treated with FSH for 2 days to 2 wk had a significantly reduced DSP. Sertoli cells remained binding incompetent as evidenced by structurally abnormal ectoplasmic specializations and an abnormal pattern of f-actin and vinculin immunostaining. The DT-Hypox rat treated with FSH for 3 wk had a normal DSP. Sertoli cells were binding-competent as evidenced by structurally intact ectoplasmic specializations and the normal pattern of f-actin and vinculin immunostaining. The results indicate that the "priming" effect of FSH necessary to restore normal spermiogenesis is associated with restoration of the junction-related Sertoli cell cytoskeleton (i.e., FSH induction of binding competency) expressed as intact ectoplasmic specializations and peripheral distribution of f-actin and vinculin.

Junction-related Sertoli cell cytoskeleton in testosterone-treated hypophysectomized rats.

Testosterone is the principal hormone necessary for insuring the completion of normal spermatogenesis. However, its precise role in spermatid maturation is not clear. In hypophysectomized rats, testosterone can maintain spermiogenesis if replaced soon after surgery. Delaying treatment results in a reduction in the number of mature spermatids. In culture, testosterone and FSH are required to maximize spermatid attachment to binding-competent Sertoli cells. This binding event is an essential step in the process of spermiogenesis and is dependent on components of the Sertoli cell cytoskeleton. The present study was undertaken to determine the binding competency of Sertoli cells and their junctional interaction with spermatids in the hypophysectomized rat after immediate testosterone replacement and after delayed testosterone replacement. Hypophysectomized rats treated with immediate testosterone replacement had peripheral distribution of Sertoli cell f-actin and vinculin, structurally intact Sertoli ectoplasmic specializations facing step 8 spermatids, and daily sperm production similar to these parameters as observed in intact controls. In the delayed treatment group, these parameters were abnormal and were similar to those observed in the untreated hypophysectomized animals. The results suggest that testosterone can maintain binding competency of the Sertoli cell and normal Sertoli-spermatid junctional interaction but cannot restore them.

Reduced testosterone during puberty results in a midspermiogenic lesion.

The aim of this study was to determine the role of testosterone, as reflected in the testicular interstitial fluid, in the completion of the first wave of spermatogenesis and to further elucidate its role in spermiogenesis. At weekly intervals beginning with 26-day-old rats, body and testis weights were obtained, testicular interstitial fluid testosterone (TIF-T) was assayed, daily sperm production (DSP) was determined, and testicular tissue was structurally analyzed by light and electron microscopy. At 40 days postpartum, half the rats were treated with ethane dimethanesulphonate (EDS) to temporarily reduce Leydig cells. The other half served as controls and were treated with the vehicle. The timing of EDS treatment was just prior to the elongation of spermatids. At Day 47 (1 week after EDS treatment), TIF-T, testis weight, DSP, and number of Leydig cells were significantly reduced. At Day 54 (2 weeks after treatment), TIF-T had returned to the normal adult level, Leydig cell repopulation was apparent, and testis weight was normal. The DSP returned to normal by Day 61 (3 weeks after treatment). At 1 and 2 weeks after treatment, Step 8-9 spermatids were partially or completely detached from Sertoli cells. Results indicate that a temporary reduction of testosterone during the peripubertal period leads to a temporary reduction of the DSP approximately 1 week later. It is suggested that reduced testosterone is associated with a mid-spermiogenic lesion interfering with stable attachment of Step 8-9 spermatids to Sertoli cells during Stage VIII-IX of the spermatogenic cycle.

Developmental expression of multiple forms of 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA in rat testes.

The developmental expression of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the enzyme that catalyzes the rate-limiting reaction of mevalonate formation, was investigated in rat testes. Adult testes were found to contain three distinct forms of HMG-CoA reductase mRNA of 4.8, 4.2, and 4.0 kilobases (kb) in length. In testes from 26-day-old rats the 4.8-kb species was the major form present. By 40 days of age, large increases in the levels of the testes-specific 4.2- and 4.0-kb forms occurred, with a small decrease in the amount of the 4.8-kb form. The level of the 4.8-kb form appeared to be associated with the level of testicular HMG-CoA reductase activity.

Development and control of the opioid inhibition of gonadotropin secretion during the sexual maturation of the male rat.

The endogenous opioid system tonically inhibits the LH-releasing apparatus in adult male rats but not in immature animals. To determine the relationship between the onset of this effect and the peripubertal testosterone rise, male rats were examined at 5-day intervals from day 25 through day 65. They were injected subcutaneously with saline, 0.25 or 1.0 mg/kg body weight naloxone and sacrificed 20 min later. Another group of immature males was castrated, implanted with testosterone-filled capsules and tested with naloxone 4 days later. The peripubertal increase in testosterone and the ability of naloxone to increase serum LH concentrations were both first statistically significant on day 45. Testosterone treatment of immature rats did not induce a naloxone effect. The ability of hypothalamic fragments to release LHRH in vitro in response to naloxone also appeared to occur at the same time as the peripubertal testosterone rise. Hypothalamic fragments obtained from immature male rats treated in vivo with testosterone were capable of responding to naloxone with LHRH release in vitro. These data suggest that in the rat the maturation of the endogenous opioid system is a component of male puberty that is induced by the peripubertal testosterone rise.

Effect of changes in sodium intake on atrial natriuretic factor (ANF) and peptides derived from the N terminus of the ANF prohormone in the rat.

The purpose of the present study was to determine whether variations in salt intake would alter the plasma concentrations of atrial natriuretic factor and the N-terminal atrial natriuretic factor prohormone peptides proANF 1-98 and proANF 31-67. Two groups of rats were placed on different salt intakes for 1 week. The low salt group of rats was fed a diet providing less than 0.1 mM NaCl/day and given deionized water to drink. The normal salt group of rats was fed regular rat chow with deionized water to drink, providing them with approximately 2 mM NaCl/day. Plasma atrial natriuretic factor was 204 +/- 60 pg/ml (mean +/- SE) in normal salt rats and was significantly lower in the low salt group (44 +/- 13 pg/ml, P less than 0.01). ProANF 1-98 was also significantly higher in the normal salt group (635 +/- 47 pg/ml) compared with the low salt group (353 +/- 33 pg/ml, P less than 0.01). ProANF 31-67 was 123 +/- 21 pg/ml in the normal salt group and 59 +/- 12 pg/ml in the low salt group (P less than 0.05). Plasma renin activity in ng angiotensin l/ml/hr averaged 1.80 +/- 0.15 in the normal salt group of rats and was significantly higher in the low salt group of rats (5.66 +/- 1.07, P less than 0.05). These results suggest that atrial natriuretic factor and the atrial natriuretic factor prohormones may play a role in the physiological adjustments to low salt intake.