Functional development of grafted vasopressin neurons.

Vasopressin neurons, transplanted from normal rat fetuses into the third ventricle of adult Brattleboro rats, alleviate the polydipsia and polyuria of the hosts. Determination of the antidiuretic activity of grafted neurons in hosts with congenital diabetes insipidus provides a convenient model for analyzing the development, plasticity, and function of transplanted central nervous system neurons in mammals.

Cryopreservation, culture, and transplantation of human fetal mesencephalic tissue into monkeys.

Studies in animals suggest that fetal neural grafts might restore lost neurological function in Parkinson's disease. In monkeys, such grafts survive for many months and reverse signs of parkinsonism, without attendant graft rejection. The successful and reliable application of a similar transplantation procedure to human patients, however, will require neural tissue obtained from human fetal cadavers, with demonstrated cellular identity, viability, and biological safety. In this report, human fetal neural tissue was successfully grafted into the brains of monkeys. Neural tissue was collected from human fetal cadavers after 9 to 12 weeks of gestation and cryopreserved in liquid nitrogen. Viability after up to 2 months of storage was demonstrated by cell culture and by transplantation into monkeys. Cryopreservation and storage of human fetal neural tissue would allow formation of a tissue bank. The stored cells could then be specifically tested to assure their cellular identity, viability, and bacteriological and virological safety before clinical use. The capacity to collect and maintain viable human fetal neural tissue would also facilitate research efforts to understand the development and function of the human brain and provide opportunities to study neurological diseases.

Site of ATP and phenylephrine synergistic stimulation of vasopressin release from the hypothalamo-neurohypophyseal system.

ATP and norepinephrine are neurotransmitters carrying cardiovascular information to vasopressin (AVP) neurones. As shown previously, exposure of hypothalamo-neurohypophyseal system explants to ATP and phenylephrine (PE) (alpha(1)-adrenergic agonist) causes a significantly larger increase in AVP release than with either agent alone and converts the response from a transient to a sustained stimulation of AVP release. Potential mechanisms for this synergism include presynaptic stimulation of excitatory afferent input (i.e. glutamate release), postsynaptic activation of receptors on AVP neurones, modulation of stimulus-secretion coupling in the neural lobe and/or involvement of glial/neuronal interactions. The response to ATP + PE (100 microM each) was not altered in the presence of either a cocktail of ionotropic glutamate receptor antagonists (CNQX + AP5) or a nonselective metabotropic glutamate receptor antagonist [(RS)-alpha-methyl-4-carboxyphenylglycine]. Thus, it is not dependent on activation of glutamate receptors. Treatment with tetrodotoxin (3 microM) eliminated the response to ATP + PE. Because this could reflect blockade of action potentials propagated from the AVP perikarya to the nerve terminals in the neural lobe or action potentials generated in the neural lobe directly, synergism in the neural lobe was addressed by perifusing isolated neural lobes with ATP and PE alone or together. Synergistic stimulation of AVP release by ATP + PE was not observed in isolated, perifused neural lobes. Thus, the synergistic effect persists in the absence of glutamate transmission, is not due to synergistic actions of P(2) and alpha(1)-adrenergic receptors occurring at the level of the neural lobe and requires action potentials in either the hypothalamus or neural lobe.

Purinergic and adrenergic agonists synergize in stimulating vasopressin and oxytocin release.

The A1 catecholamine neurons of the caudal ventrolateral medulla transmit hemodynamic information to the vasopressin (VP) neurons in the hypothalamus. These neurons corelease ATP with norepinephrine. Perifused explants of the hypothalamoneurohypophyseal system were used to investigate the role of these substances on VP release. ATP (100 micrometer) increased VP release 1.5-fold (p = 0.027). The response was rapid but unsustained. It was blocked by the P(2) receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS). The alpha(1)-adrenergic agonist phenylephrine (PE; 100 micrometer) also increased VP release by 1.5-fold (p = 0.014). Again, the response was rapid and unsustained. However, simultaneous perifusion of explants with ATP (100 micrometer) and PE (100 micrometer) resulted in a threefold to fourfold increase in VP release, which was sustained for as long as 4 hr. There was a similar synergistic effect of ATP and PE on oxytocin release. Interestingly, the synergistic response was delayed approximately 40 min relative to the response to either agent alone. Several experiments were performed to elucidate the cellular mechanisms of this synergism. The effect was blocked by PPADS, a protein kinase C inhibitor (bisindolylmaleimide I HCl), and actinomycin, an inhibitor of gene transcription. These data suggest that P(2X) receptor activation, PKC-mediated phosphorylation, and gene transcription are required for the synergistic response. The marked synergism of these coreleased agents is probably important to achieve sustained increases in plasma VP in response to prolonged hypotension. These observations may also have broad applications to CNS function, because ATP may be coreleased at noradrenergic synapses throughout the CNS.

Gonadal steroid modulation of vasopressin secretion in response to osmotic stimulation.

As deficiencies in osmotic stimulation of vasopressin (VP) messenger RNA (mRNA) content in castrated rats have been reported, experiments were performed to determine whether castration altered osmotically stimulated VP release in vitro. Perifused explants of the hypothalamo-neurohypophyseal system were obtained from sham and gonadectomized male rats. There were no significant differences in VP release stimulated by a ramp increase in the osmolality of the culture medium between the two groups. As testosterone was undetectable in the perifusion medium, the effect of addition of testosterone on osmotically stimulated VP release was evaluated. Testosterone (3 ng/ml) and its metabolites, estradiol (50 pg/ml) and dihydrotestosterone (DHT; 3 ng/ml), inhibited osmotically stimulated VP release in hypothalamo-neurohypophyseal system explants. The osmotically induced increase in VP mRNA content was also inhibited by testosterone and estradiol, but not by DHT. Neither estradiol nor DHT affected stimulus-secretion coupling of hormone secretion, because they did not inhibit KCl (25 mM)-stimulated VP release. BSA conjugates of estradiol (200 nM) and DHT (10 mM) also inhibited osmotically stimulated VP release, and VP mRNA content was inhibited by BSA-estradiol, but not by BSA-DHT, suggesting nongenomic actions of the steroids. The differential effects of estradiol and DHT on VP mRNA imply distinct actions for these steroids, and the DHT mechanism uncouples regulation of VP release from VP mRNA content.

Characterization of cholinergic control of vasopressin release by the organ-cultured rat hypothalamo-neurohypophyseal system.

Acetylcholine and nicotine stimulated vasopressin (VP) release from the organ-cultured rat hypothalamo-neurohypophyseal system (HNS). Nicotinic antagonists, hexamethonium, tetraethylammonium chloride, and trimethaphan blocked VP release in response to acetylcholine and nicotine. A muscarinic agonist, methacholine, was ineffective in eliciting VP release from HNS explants at a molar concentration equal to the maximally effective concentration of acetylcholine (10(-5) M). Atropine, a muscarinic antagonist, was an ineffective blocking agent for acetylcholine. These data indicate that the cholinergic receptor in the HNS explant is nicotinic rather than muscarinic in character.

Cholinergic involvement in osmotic control of vasopressin release by the organ-cultured rat hypothalamo-neurohypophyseal system.

A nicotinic-cholinergic receptor appears to mediate osmotic stimulation of vasopressin (VP) release by the hypothalamo-neurohypophyseal explant. Nicotinic blocking agents, hexamethonium, tetraethylammonium chloride, and trimethaphan, blocked VP release in response to the addition of sufficient NaCl to yield a 10 mosm/kg H2O increase in culture medium osmolality. Atropine at a similar molar concentration was ineffective in blocking VP release in response to the same osmotic stimulus. Tetraethylammonium chloride and trimethaphan also blocked acetylcholine-stimulated VP release. These findings support the hypothesis that the osmoreceptive element responsible for controlling VP release resides in a separate cell and communicates with the VP cell by way of a nicotiniccholinergic receptor.

Angiotensin stimulation of vasopressin release from the rat hypothalamo-neurohypophyseal system in organ culture.

Angiotensin II (AII) stimulated vasopressin (VP) release from the rat hypothalamo-neurohypophyseal system (HNS) in organ culture in a concentration-dependent manner. Exposure to AII at 10(-8) M for 1 hr yielded a 1.8-fold increase in VP release over control release (P less than 0.01), while a 1-h exposure to 10(-5) M AII resulted in a 4-fold increment over control VP release by HNS explants maintained in organ culture for 3 days (P less than 0.01). Saralasin, an AII antagonist, blocked AII stimulation of VP release without significantly altering basal VP release by the HNS explants. Saralasin did not interfere with stimulation of VP release by acetylcholine or nicotine. Tetrodotoxin (10(-7) g/ml) also blocked AII stimulation of VP release. These findings suggest that action potentials are generated in response to AII stimulation of specific receptors in the HNS and are requisite for VP release in response to this stimulus.

Role of angiotensin in the osmotic control of vasopressin release by the organ-cultured rat hypothalamo-neurohypophyseal system.

Angiotensin II (AII) appears to either mediate or modulate osmotically stimulated vasopressin (VP) release by the organ-cultured rat hypothalamo-neurohypophyseal system. Saralasin, an AII antagonist, blocked VP release in response to a 10-mosmol increment in culture medium osmolality achieved by the addition of NaCl. This was observed at all concentrations tested (10(-7), 10(-6), 10(-4) M). Saralasin (10(-4) M) also blocked VP release in response to a comparable mannitol-induced increase in osmolality. Since nicotinic-cholinergic antagonists previously were shown to inhibit osmotically stimulated VP release, the effect of hexamthonium, a nicotinic, a nicotinic-cholinergic antagonist, on AII-stimulated VP release was examined. Hexamethonium (10(-5)-10(-3) M) was ineffective in blocking AII stimulation of VP release. This finding coupled with the previous observation that saralasin does not block acetylcholine stimulation of VP release suggests independent AII and cholinergic mechanisms controlling VP release; however, the effectiveness of both types of antagonists in blocking osmotically stimulated VP release indicates some interaction between these regulators of VP release.

The stimulation of vasopressin gene expression in cultured hypothalamic neurons by cyclic adenosine 3',5'-monophosphate is reversible.

Previous studies have demonstrated that the number of vasopressin (VP) neurons present in primary diencephalic cultures can be markedly augmented by treatment with drugs that elevate intracellular cAMP. To evaluate the effect of this drug treatment on VP secretion by hypothalamic cultures and to determine if this represents a developmental phenomenon or a mechanism involved in the continuing dynamic regulation of the VP gene, we have exposed primary dispersed hypothalamic cultures derived from 14-day-old fetal Sprague-Dawley rats to forskolin (25 microM) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX; 500 microM), either continually or intermittently, for up to 39 days. Culture medium was collected throughout the culture period for VP RIA, and at the end of the culture period, cultures were stained immunocytochemically for neurophysin (NP). As reported by previous investigators, exposure to the drugs for 11 days resulted in an increase in the number of NP-positive neurons. The increase was sustained with longer periods of exposure up to 39 days. IBMX and forskolin treatment also resulted in detectable release of VP into the culture medium, which increased from 1.4 +/- 0.15 pg/ml at 11 days to 8.4 +/- 0.6 pg/ml after 32 days of drug treatment. The VP concentration remained undetectable (< 1.25 pg/ml) in nontreated cultures throughout this period. The effect on VP expression did not require immediate exposure to the drugs in culture, but did require the continuous presence of the drugs. Removal of the drugs from days 11-18 of culture resulted in an almost complete loss of NP-positive cells; however, reexposure to the drugs reinstated NP expression in a time-dependent fashion. The effect of IBMX/forskolin treatment on the expression of other neuronal markers was also evaluated. The treatment did not alter the total number of neurons, and there was no evidence of stimulation of oxytocin expression. There was a marked increase in the number and size of neurons stained immunocytochemically for tyrosine hydroxylase and a small increase in the number of cells staining for somatostatin. These results demonstrate that treatment with cAMP-elevating drugs markedly and selectively elevates VP secretion from dispersed hypothalamic cultures, but continuous exposure to the drugs is necessary to sustain the effect. These findings suggest that although cAMP is required in hypothalamic cultures for VP gene expression, it may also participate in the dynamic regulation of VP gene transcription in response to physiological challenges.

Effect of extended exposure to hypertonicity on vasopressin messenger ribonucleic acid content in hypothalamo-neurohypophyseal explants.

The feasibility of using organ-cultured explants of the rat hypothalamo-neurohypophyseal system (HNS) to study the mechanisms regulating the vasopressin (VP) mRNA content of the HNS was examined by evaluating the effect of exposure to hypertonicity on the VP mRNA content of these explants. Different effects were observed after a step increase in osmolality and a gradual increase in the same amount over 24 h. The VP mRNA content of control HNS explants determined from a RNA protection assay was 22 +/- 6 pg. It gradually decreased to 23% and 9% of the control value during 24 and 48 h in culture, respectively. Northern blot analysis revealed a single band of VP mRNA approximately 700 bases long in explants cultured for 36 h. Explants exposed to the step increase in osmolality were maintained in static culture. The control explants were placed directly into isotonic medium (299 mosmol/kg H2O). The explants exposed to the step increase were placed directly into hypertonic medium (greater than 304 mosmol/kg H2O). After 24 h in culture, basal VP release was measured, and all explants were then exposed to a further acute 15 mosm/kg H2O increase in osmolality. The highest basal release of VP was observed in the explants maintained under isotonic conditions (299 mosm/kg H2O). These explants significantly increased VP release in response to the acute increase in osmolality. Basal VP release was lower in explants maintained in hypertonic medium (greater than 304 mosmol/kg H2O), and these explants did not respond to the acute hypertonic pulse. VP mRNA content was significantly decreased in explants maintained for 24 or 48 h in hypertonic medium compared to that in explants maintained in isotonic medium (47 +/- 10% and 57 +/- 6%, respectively; P less than 0.01). No significant difference existed in the VP content of the posterior pituitary between the groups. To achieve a slow increase in osmolality, explants were perifused in individual chambers with medium at 2.1 ml/h. A gradual increase in osmolality (16 mosmol/kg H2O medium) was achieved by increasing the NaCl concentration in the perifusion medium. In response to this stimulus there was a significant increase in VP release, which was sustained for 9 h. VP mRNA content in the hypertonic group was 165 +/- 19% of that in control explants (P less than 0.001), but no difference existed in VP content in the posterior pituitary compared to that in time control explants.(ABSTRACT TRUNCATED AT 400 WORDS)

gamma-Aminobutyric acid antagonists stimulate vasopressin release from organ-cultured hypothalamo-neurohypophyseal explants.

gamma-Aminobutyric acid (GABA) has been identified in axon terminals innervating neurons of the supraoptic nucleus and has been shown to inhibit the electrical activity of supraoptic neurons when applied iontophoretically. This study examines the effects of GABA and GABA antagonists on vasopressin (VP) release from organ-cultured explants of the hypothalamo-neurohypophyseal system (HNS). The GABA antagonists bicuculline and picrotoxin stimulated VP release in a concentration-dependent manner. These observations suggest that VP release by HNS explants is tonically inhibited by GABA. Exposure of HNS explants to GABA (10(-8)-10(-3) M) did not consistently alter basal VP release. This was true even when penicillin, which can block GABA-activated chloride channels, was omitted from the medium. Similarly, nipecotic acid, an agent that potentiates GABA activity by inhibiting GABA uptake, did not alter basal VP release; stimulation of VP release by acetylcholine and increases in osmolality was not diminished by the addition of 10(-5) M GABA. The failure of exogenous GABA to modify basal and stimulated VP release suggests that GABAergic inhibition of VP release is maximally activated by endogenously released GABA in cultured HNS explants. This is consistent with evidence for a local source of GABA in the supraoptic nucleus and suggests that one role of GABA in the regulation of VP release is that of a potent local inhibitory neurotransmitter.

Osmotic control of vasopressin release by rat hypothalamo-neurohypophyseal explants in organ culture.

The rat hypothalamo-neurohypophyseal system (HNS) in organ culture has been used as an in vitro system for studying the osmotic control of vasopressin (VP) release. The HNS retains osmotically sensitive components as demonstrated by changes in the rate of VP release following alterations in the osmolality of the culture medium. Increasing the osmolality from 295 to 305 mosmol/kg H2O by the addition of NaCl resulted in a 2.5-fold increase in VP release. VP release was significantly decreased subsequent to reducing the osmolality from 295 to 280 mosmol/kg H2O by the addition of distilled water. Also, VP release was stimulated when the osmolality was increased to 300 mosmol/kg H2O by the addition of mannitol, but not by additions of urea or glucose which resulted in comparable increases in the tonicity of the culture medium. These studies demonstrate that the HNS in organ culture responds appropriately to osmotic challenges within the physiological range, and support Verney's concept of an osmoreceptor inasmuch as both NaCl and mannitol were effective osmotic agents.

Characterization of noradrenergic control of vasopressin release by the organ-cultured rat hypothalamo-neurohypophyseal system.

Evidence is presented indicating that norepinephrine (NE) inhibits vasopressin (VP) release from the rat hypothalamo-neurohypophyseal explant under some, but not all, conditions in vitro. NE at 10(-5) M inhibited basal VP release and inhibited acetylcholine-induced release in a concentration-dependent fashion. However, the induction of VP release caused by the addition of NaCl (sufficient to yield a 10 mosmol/kg H2O increase in culture medium osmolality) was not reduced by NE in concentrations as high as 10(-5) M. An alpha-adrenergic receptor mediation of the inhibition of VP release by NE was suggested by the ability of phentolamine and phenoxybenzamine, but not propranolol, to block this effect. In addition, phentolamine at 10(-4) M, but not equimolar amounts of propranolol, increased VP release when added alone on day 2 of culture, but not on days 3 or 4. Histofluorescence examination of additional explants revealed that endogenous catecholamine was still present on day 2 within varicosities in the supraoptic nucleus, but diminished by days 3 and 4, suggesting that endogenous NE could influence basal VP release. The results indicate that NE can inhibit spontaneous and cholinergically stimulated VP release from the hypothalamo-neurohypophyseal explant, but osmotic stimulation renders the explants insensitive to attenuation of VP release by NE.

Cholinergic stimulation of vasopressin release in spontaneously hypertensive rats.

Plasma vasopressin (VP) concentration is elevated in spontaneously hypertensive rats (SHRs) relative to their normotensive Wistar-Kyoto (WKY) controls. The possibility that this reflects altered responsiveness of the hypothalamo-neurohypophyseal system (HNS) in SHRs was examined by comparing VP release in response to acetylcholine from organ cultured HNS explants obtained from SHR and WKY donors. Explants were prepared from 5-, 8-, and 18-week-old animals. Blood pressure was significantly elevated in the 8- and 18-week-old SHR donors relative to their age-matched WKY donors. VP release was assessed on the 4th day of culture during a control hour and during the subsequent hour in the presence of acetylcholine. Acetylcholine caused a concentration-dependent stimulation of VP release from both types of explants, but the response was significantly greater in the explants from 5- and 8-week-old SHRs than in explants from age-matched WKYs. The explants from 18-week-old SHRs and WKYs demonstrated comparable sensitivity to acetylcholine. Basal VP release was not significantly different in explants from age-matched SHRs and WKYs, but it did increase with donor age in both strains. These studies indicate potential hyperresponsiveness of the HNS to excitatory stimuli in SHRs during the developmental phase of hypertension. The hyperresponsiveness disappears in the chronically hypertensive phase. Thus, increased sensitivity of the HNS during the development of hypertension may contribute to the elevation of plasma VP concentration in SHRs.

Evidence against a pressor role for vasopressin in spontaneous hypertension.

The hypothesis that the vasoconstrictor action of vasopressin may contribute to the development of hypertension in spontaneously hypertensive rats (SHR) was tested by chronic infusion of a specific antagonist of the vascular effects of vasopressin. From 4 to 13 weeks of age, SHR and Wistar-Kyoto rats (WKY) received subcutaneously either isotonic saline or the vasopressin pressor antagonist, d(CH2)5Tyr(Me)arginine vasopressin by osmopump. Systolic blood pressure was measured by tail cuff from 5 to 11 weeks of age. In SHR, the vasopressin analogue did not alter the rate or magnitude of increase in systolic blood pressure. In WKY, systolic blood pressure in the vasopressin analogue group was slightly reduced compared with the saline infusion values until 10 weeks of age (F1, 10 = 10.18, p = 0.008). At 12 to 14 weeks of age, all animals were prepared with indwelling arterial and venous catheters. Resting mean arterial pressure was not altered significantly by the vasopressin analogue infusion in either strain, but the response to an acute vasopressin infusion of 5, 15, or 50 ng/kg body weight was markedly attenuated by the analogue treatment, indicating that plasma levels of the vasopressin analogue were sufficient to block pressor effects of endogenous vasopressin. A bolus injection of the angiotensin II converting enzyme inhibitor teprotide (SQ 20881) resulted in a decrease in mean arterial pressure (p less than 0.05) that was comparable in all groups, and serum renin concentration was not elevated in the vasopressin analogue-treated rat.(ABSTRACT TRUNCATED AT 250 WORDS)

Attenuation of spontaneous hypertension in rats by a vasopressin antagonist.

Although abnormalities in the vasopressin system have been reported in spontaneously hypertensive rats (SHR), neither short-term nor long-term administration of the vasopressin antagonist d(CH2)5-Tyr(Me)arginine vasopressin (AVP), which selectively blocks the action of vasopressin on vascular (V1) receptors, altered the course of hypertension in SHR. In the current study, long-term administration of a different vasopressin antagonist, d(CH2)5-D-Tyr(Me)VAVP, to SHR and Wistar-Kyoto rats (WKY) from 4 to 12 weeks of age significantly attenuated the development of systolic hypertension in SHR (p less than 0.05) without altering blood pressure in normotensive WKY. The antagonist was delivered subcutaneously by osmopump at 0.1 microgram/hr. Systolic blood pressure was monitored twice weekly by tail plethysmography beginning at 5 weeks of age. In a second group of SHR, the drug infusion was continued until 18 weeks of age. In this group, the attenuation of systolic hypertension by the drug was extended and became more prominent (p less than 0.007). Resting mean arterial pressure measured by indwelling catheters in the conscious state at 18 weeks of age was significantly reduced in the antagonist-treated SHR (144 +/- 4 vs 157 +/- 4 mm Hg; p less than 0.05). Heart rate also was significantly reduced by the drug (351 +/- 6 vs 392 +/- 7 beats/min; p less than 0.001). Following measurement of mean arterial pressure in the rats at 18 weeks of age, the osmopumps were removed and systolic blood pressure, mean arterial pressure, and heart rate were observed until 22 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Osmotic regulation of vasopressin and renin in spontaneously hypertensive rats.

Abnormalities in the vasopressin and renin systems have been reported in spontaneously hypertensive rats (SHR). Therefore, studies were performed to evaluate the responsiveness of these systems to changes in plasma osmolality and sodium concentration. These variables were manipulated in vivo by intraperitoneal administration of distilled water, isotonic saline, or hypertonic saline to 8- and 18-week-old SHR and normotensive Wistar-Kyoto rats (WKY). Animals were decapitated 30 minutes later, and trunk blood was collected. The hypertonic saline injections resulted in an increase in plasma osmolality and serum sodium at both ages (p less than 0.001). Serum vasopressin was higher in all groups of animals receiving hypertonic saline (1200 mosm/kg H2O; p less than 0.05), but the magnitude of increase was not significantly different in the SHR and WKY at either age. Serum renin activity was lower in SHR than in WKY following acute decreases in serum sodium at 8 weeks, but it was the same for both strains at 18 weeks. Both kidney renin content and concentration were lower in SHR than in WKY at 18 weeks but not at 8 weeks. Therefore, the suppressed renin response to acute osmotic challenge in 8-week-old SHR is not the consequence of reduced kidney renin content. The vasopressin response to osmotic stimulation also was evaluated in vitro using hypothalamoneurohypophyseal explants obtained from 5-, 8-, and 18-week-old SHR and WKY.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of surgical deafferentation of the hypothalamic supraoptic nucleus on urinary vasopressin levels in twenty-month-old rats.

Indices of water balance (daily water consumption, urine volume, and urine osmolality), and vasopressin (VP) urinary excretion were measured in 3-month-old (m.o.) and 20 m.o. F344 rats after neurosurgical lesion of noradrenergic afferents to the supraoptic nucleus (SON) in the medial forebrain bundle (mfb), in order to assess whether morphological denervation (21) was functionally effective. Lesion of the mfb was accomplished using a mechanical knife cut placed stereotaxically medial and caudal to SON. The experiment was performed three times. Morphological evaluation consistently indicated a marked depletion of noradrenergic afferents in SON after lesion placement. The effect of the lesion on daily excreted VP was assessed by paired t-test, using means of equal numbers of urinary VP measurements representing 13-15 days each pre- and postsurgically for each animal. The postsurgical interval for assessing the functional effect of the lesion was chosen based upon morphological evidence for mfb regeneration at 14 days (21). Three m.o. rats showed comparable urinary VP levels before (mean = 328.4 +/- 97.3 pg/24 hr/100 g b.wt., for 10 rats for 2 weeks) and after (354.7 +/- 63.0 pg/24 hr/100 g b.wt.) lesion placement. In 20 m.o. rats, overall mean postsurgical VP excretion was 199.4 +/- 44.8 pg for 15 rats for 2 weeks, compared with a presurgical mean of 343.2 +/- 86.2 pg/24 hr/100 g b.wt. Mean urine VP was lower in 20 m.o. rats in each of three experiments; VP levels after lesion in 3 m.o. rats was lower in 2, but higher in one experiment; mean differences were not statistically significant for treatment (lesion) effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Vasopressin and renin response to dehydration in aged rats.

Abnormalities in neurohypophyseal function have been postulated to contribute to the alterations in fluid and electrolyte balance observed during aging. In this study, parameters of fluid and electrolyte balance were evaluated during chronic water deprivation in old (30 months) and young (3 months) Fischer 344 rats. The increase in serum vasopressin (VP) and renin concentrations observed in the 3 month animals following chronic water deprivation were absent in the aged rats (p less than 0.05 and p less than 0.02, respectively). This occurred in spite of apparently comparable alterations in fluid volume and osmolality (assessed by changes in body weight, hematocrit and plasma osmolality). Relative to body weight, VP content of the neural lobe was significantly reduced and was more severely depleted by dehydration in aged rats than in young rats. Thus, inadequate neurohypophyseal hormone stores may contribute to the inability of the aged animals to attain elevated serum VP concentrations during chronic stimulation. Several parameters of renal function were examined in the aged rats. Although none of the old rats were in renal failure, they all showed some indication of reduced renal function. In spite of renal abnormalities including reduced concentrating capabilities, the old rats did demonstrate a significant antidiuretic response to dehydration. However, with prolonged fluid deprivation, they were unable to attain serum VP or renin concentrations comparable to that achieved by the young rats.