Maternal behavior deficits in nulliparous oxytocin knockout mice.

The first observations of postpartum oxytocin knockout (OTKO) mice found no maternal behavior deficits. However, it is unclear how detailed those observations were. In this study, we compared maternal behavior exhibited by OTKO and wild-type (WT) nullipara toward six 2-4-day-old foster pups during test sessions conducted on 3 successive days. Each day, subjects were placed in a clean cage 30 min prior to introduction of pups which were deposited in a clump adjacent to the middle of a long wall of each test cage. Behavior was measured for 3.5 h after which pups and test subjects were returned to their home cages. On test days 1 and 3, a significantly smaller proportion of OTKO females retrieved pups to a corner of their cage. Also, significantly fewer pups were retrieved to corners by OTKO females. In contrast to most WTs, most OTKO females mothered pups in the center of the cage where they were initially deposited. Pup-licking frequencies were significantly lower in OTKO females. Their self-grooming frequencies also trended toward being lower. Latencies to retrieve and lick pups, latencies to and frequencies of still crouching over pups and proportion of time in nest did not differ between groups. Our findings suggest that OT stimulates a significant proportion of pup-licking in nulliparous mice, a situation similar to lactating rat mothers. Our results also indicate that OT may play a role in the motivation to retrieve pups to a more secure location.

Clinical and chemical assessment of thyroid function during therapy with amiodarone.

Clinical and chemical variables of thyroid function were studied in 26 patients with symptomatic ventricular tachyarrhythmias before and during long-term oral treatment with amiodarone. The mean (+/-SEM) pretreatment thyroxine (T4) level in the 26 patients was 7.32 +/- 0.33 micrograms/dL, and increased notably to 10.15 +/- 0.47 micrograms/dL by 30 to 120 days after treatment. The free thyroxine index (FT4I) was also notably elevated. Clinical hyperthyroidism or goiter did not develop, but clinical hypothyroidism occurred in four patients during and in one patient after discontinuation of amiodarone treatment. Notable titers of antithyroid antibodies were found in the serum of two of the five and a family history of thyroid disease was present in three of the five hypothyroid patients. An elevation of both the T4 level and the FT4I above the normal range is an expected finding in patients receiving amiodarone and does not by itself indicate hyperthyroidism. Patients with positive antithyroid antibodies or a family history of thyroid disease prior to treatment with amiodarone may be at an increased risk of hypothyroidism developing when treated with this drug.

Diabetic management in patients with renal failure.

The management of diabetes in patients with renal disease requires understanding of the multiple changes in carbohydrate metabolism that accompany renal failure. A decrease in insulin requirements may occur due to changes in insulin clearance or insulin metabolism. With the onset of uremia, a peripheral resistance to insulin action develops that often requires increased insulin administration, and this resistance can be expected to improve with adequate dialysis. Hypoglycemia may occur as a result of decreased gluconeogenesis, excessive insulin action, or decreased caloric intake. Thus, the treatment of diabetes in the settings of renal failures involves anticipation of changes in insulin sensitivity and insulin dosages. An overall management plan that coordinates diet, insulin, and dialysis where appropriate is most likely to yield optimum diabetic care.

Gonadal steroid modulation of oxytocin and vasopressin gene expression in the hypothalamus of the osmotically stimulated rat.

We investigated the modulatory role of gonadal steroids on the expression of oxytocin (OT) and vasopressin (AVP) cytoplasmic mRNAs in the paraventricular nucleus and supraoptic nucleus of the osmotically stimulated rat. We chronically administered an oral salt load (2% sodium chloride solution for 5 days) to intact and gonadectomized female and male Sprague-Dawley rats and measured serum sodium, body weight, pituitary content of OT and AVP immunoreactivities, and size and abundance of hypothalamic cytoplasmic OT and AVP mRNA transcripts. Intact and gonadectomized rats that were administered an osmotic challenge developed comparable degrees of hypernatremia and loss of body weight as well as depletion of posterior pituitary stores of OT and AVP. Hyperosmolality induced elongation of the OT and AVP transcripts in intact and gonadectomized animals, but only intact rats had enhanced hypothalamic cytoplasmic OT and AVP mRNA concentrations to this stimulus. Replacement with gonadal steroids restored the up-regulation in OT and AVP gene expression in gonadectomized animals rendered hyperosmolar. The findings support a modulatory role for gonadal steroids in hypothalamic OT and AVP gene expression during osmotic stimulation.

Effect of progesterone on hypothalamic oxytocin messenger ribonucleic acid levels in the lactating rat.

We recently reported that sequential administration of estrogen and progesterone and subsequent withdrawal of progesterone increased the level of hypothalamic oxytocin (OT) messenger RNA (mRNA) in the female rat. Both estrogen priming and progesterone withdrawal are critical components of this regimen. Rats experience this ovarian steroid pattern during certain lactational events such as on days 10-12 of lactation or with interruption of nursing for 48-72 h. In the present study, we used Northern blot and in situ hybridization to determine the association between the steroid exposure and the level of OT mRNA during these lactational conditions. Week 1 lactating rats that had pups removed for 24 and 48 h, but not 12 h, had significantly higher serum estradiol concentrations than animals continuously nursing their pups on a comparable day of lactation (F4, 15 = 6; P < 0.005). Serum progesterone levels declined significantly during the 48 h after litter removal (F4, 15 = 130.5; P < 0.0001). Significant increases in OT mRNA levels were found at 48 h, but not 12 or 24 h, after litter removal (F4, 15 = 4.3; P < 0.02). Implantation of progesterone-filled capsules to compensate for the spontaneous decline in progesterone that occurs with interruption of nursing attenuated this increase in OT mRNA levels. OT mRNA was significantly higher in the hypothalamic paraventricular nuclei (P < 0.0002) and supraoptic nuclei (P < 0.002) of nursing interrupted animals receiving empty vs. progesterone-filled implants. Implantation of day 12 lactating rats with progesterone capsules for 5 days before being killed blunted the increase in OT mRNA that normally occurs on this day of lactation. The data highlight the pivotal role of estrogen priming and progesterone withdrawal in the increased expression of the OT gene during select lactational events.

Reversal of fasting-induced suppression of luteinizing hormone (LH) secretion in male rhesus monkeys by intragastric nutrient infusion: evidence for rapid stimulation of LH by nutritional signals.

Short periods of fasting (1-2 days) significantly suppress the frequency of pulsatile LH secretion in adult male rhesus monkeys. Resumption of food intake (i.e. refeeding) after fasting rapidly restores pulsatile LH secretion. We hypothesized that the resumption of LH secretion after refeeding results from the metabolic consequences of food intake rather than from relief of the psychological stress associated with fasting. To differentiate between these two possible types of signals, we attempted to provide fasting monkeys with the nutritional signals of a refeed meal without relieving the psychological stress associated with fasting. To do this, we provided monkeys with a refeed meal via indwelling gastric cannulae while continuing to deny them access to monkey chow. Nine adult male rhesus monkeys with indwelling gastric and venous catheters were fasted for 1 day, and on the following day they received a liquid nutrient infusion (equal in caloric content and macronutrient composition to a normal meal) at the normal feeding time (1100 h) via the gastric cannulae. Blood samples were collected from 0600-2400 h at 20-min intervals. Nutrient infusions caused a restoration of pulsatile LH secretion that was not different from the restoration caused by refeeding a normal meal of Purina monkey chow. In contrast, LH secretion remained suppressed on a second day of fasting. Additionally, monkeys receiving nutrient infusions displayed behavioral agitation similar to fasting monkeys, while monkeys refed monkey chow showed very little agitation. These findings support the hypothesis that nutritional/metabolic signals, rather than the relief of psychological stress, lead to the resumption of pulsatile LH secretion after refeeding.

Increased accumulation of oxytocin messenger ribonucleic acid in the hypothalamus of the female rat: induction by long term estradiol and progesterone administration and subsequent progesterone withdrawal.

To examine a possible role for gonadal steroid hormones in the enhanced accumulation of hypothalamic oxytocin (OT) messenger RNA (mRNA) and peptide in late pregnancy, we used an established model (22) in which sequential administration of estradiol (E2) and progesterone (P) SILASTIC capsules to ovariectomized rats is followed by removal of P. Long term and sustained E2 combined with abrupt P withdrawal mimics the gonadal steroid hormone pattern of late gestation in the rat (22). Using this paradigm, we demonstrate that OT mRNA is increased in the rat hypothalamus after long term P treatment, but only in the presence of E2 and only when P capsules are removed 48 h before killing. Furthermore, we show that P replacement in primiparous rats during late pregnancy blunts the increase in OT mRNA normally observed at the end of gestation. Our results support a role for E2 priming and P withdrawal in the enhanced accumulation of OT mRNA in the hypothalamus of the female rat.

Cytoplasmic oxytocin and vasopressin gene transcripts decline postpartum in the hypothalamus of the lactating rat.

Oxytocin (OT) and vasopressin (AVP) gene expression are enhanced in the rat hypothalamus in late gestation and during the second and third weeks of lactation. We report that during the first 3 postpartum days, OT and AVP cytoplasmic mRNAs in the supraoptic and paraventricular nuclei of lactating rats decreased dramatically, reaching less than one fifth of peak gestational levels by day 2 postpartum. Differences in the temporal pattern of OT and AVP expression were observed in the supraoptic and paraventricular nuclei from days 4-10 of lactation. We also compared OT and AVP cytoplasmic mRNAs isolated from the hypothalamus of day 3 lactating rats to cohorts that had litters removed at the time of parturition. Lactating rats had significantly lower OT and AVP cytoplasmic mRNA levels than their nonlactating cohorts. We further compared OT and AVP cytoplasmic mRNAs in the hypothalamus of day 12 lactating rats that had been ovariectomized or sham ovariectomized on day 3 of lactation. Ovariectomized day 12 lactating animals had significantly lower OT and AVP cytoplasmic mRNA levels than their intact cohorts. These data refute the hypothesis that lactation is characterized by persistently elevated hypothalamic cytoplasmic OT and AVP mRNAs produced as a result of continuous stimulation by suckling and suggest that ovarian steroids may exert a modulatory effect on hypothalamic OT and AVP expression during early lactation.

A prospective longitudinal study of the release of oxytocin and prolactin in response to infant suckling in long term lactation.

We studied prospectively eight healthy postpartum breast-feeding women for 6 months during early, middle, and late lactation. Blood for measurement of oxytocin (OT), PRL, and arginine vasopressin was drawn before and every 3 min from each women during 15 min of infant suckling for two consecutive feedings during each stage of lactation. Basal plasma OT was not different in breast-feeding [0.7 +/- 0.1 (+/- SEM) microU/ml] and nonbreast-feeding women (0.8 +/- 0.2 microU/ml). OT increased significantly in response to infant suckling (P less than 0.00001) to 5.9 +/- 0.5 microU/ml and remained elevated throughout a feeding. OT was released during infant suckling in an episodic pattern in some, but not all, women; peak OT varied among women (5.0-23.3 microU/ml). There was no significant difference in the mean stimulated OT or the pattern of release comparing the first and second feedings of the same day. The mean OT (n = 8) released during 15 min of infant suckling was not significantly different in early (3.9 +/- 0.3 microU/ml), middle (4.5 +/- 0.3 microU/ml), and late (5.8 +/- 0.4 microU/ml) lactation. In the four women who breast fed exclusively, the mean stimulated OT was significantly higher (P less than 0.01) during late lactation (8.6 +/- 0.4 microU/ml) vs. early (4.6 +/- 0.4 microU/ml) or middle (6.1 +/- 0.4 microU/ml) lactation. In the other four women who provided formula supplements, OT did not change. Plasma arginine vasopressin did not increase in response to infant suckling. Plasma PRL increased in response to infant suckling, reaching a peak at 15 min. Mean basal PRL decreased progressively from weeks 1-24 postpartum. Mean peak PRL decreased significantly from early (162 +/- 29) to middle (130 +/- 15) to late (77 +/- 10) ng/ml lactation (P less than 0.05). OT release in response to infant suckling continues throughout the first 6 months postpartum in breast-feeding women, and the pattern is reproducible. The maximum release of OT is dependent upon continuous regular nipple stimulation. In contrast to basal and suckling-induced levels of PRL, which decreased with time postpartum, basal and suckling-induced OT release did not decrease from early to late lactation.

Studies of oxytocin in plasma of women during hypocontractile labor.

Oxytocin (OT) was measured by RIA in plasma of women during hypocontractile labor before and during graded doses of iv infused synthetic OT. Based upon in vitro studies of recovery of OT from pregnancy plasma, blood was collected into heparinized tubes which were kept at 4 C. The addition of EDTA and phenanthrolene to an aliquot of each sample resulted in measured levels of OT in plasma that correlated closely with levels measured in the absence of these reagents (r2 = 0.86). Comparison of OT levels in plasma of normal individuals determined in the presence and absence of these reagents also yielded a high degree of linear correlation (r2 = 0.97). The mean level of OT in 11 women during hypocontractile labor before the infusion of OT was 1.01 +/- 0.31 (+/- SEM) microU/ml. There was a linear correlation between the dose of OT infused and the level of OT in plasma with infused doses of OT between 1 and 4 mU/min (r2 = 0.99). The time of onset of adequate uterine contractility was recorded by on-line computer analysis, and the level of OT in plasma obtained simultaneously was variable among the women. The mean OT MCR in these women was 17.4 +/- 9.2 (+/- SEM) ml/kg X min, similar to the MCR in normal men (17.6 +/- 2.1 ml/kg X min). Levels and pharmacokinetics of OT during hypocontractile labor were similar to those in nonpregnant individuals and women in late pregnancy. The variability in OT concentrations at the time of adequate uterine contractility suggests that individual myometrial sensitivity is an important determinant of the response to administered OT in humans.

Neurohypophysial hormones in cerebrospinal fluid of adults: absence of arginine vasotocin and of a diurnal rhythm of arginine vasopressin.

We previously reported a prominent diurnal rhythm of oxytocin (OT) in human cerebrospinal fluid (CSF) similar to the brisk CSF OT rhythm in monkeys. An OT CSF rhythm has not been found in lower species. In contrast, a pronounced arginine vasopressin (AVP) rhythm has been found in lower species, but the AVP CSF rhythm is less marked in subhuman primates. In patients (n = 7) in whom lumbar drains had been temporarily placed for treatment of CSF rhinorrhea, we obtained CSF samples every 6 h. In 6 of these 7 patients, we previously reported (1) finding a prominent CSF OT rhythm, with a peak at 1200 h, by analysis of variance of repeated measures. When the samples of CSF of these same 6 patients (plus 1 additional patient) were assayed for AVP, no AVP rhythm was found. We also measured AVP, OT, and arginine vasotocin (AVT) by RIA in single samples of CSF obtained from 23 other patients. In these single samples of CSF, mean AVP was 0.9 +/- 0.11 (+/- SEM) pg/ml, and OT was 3.7 +/- 0.5 microU/ml. CSF AVT immunoreactivity was 0.6 pg/ml or less in the 23 patients. Two pools of CSF were separated by reverse phase high pressure liquid chromatography. The peak OT and AVP, as determined by RIA, coeluted exactly with synthetic and human posterior pituitary OT and AVP, respectively. No immunoreactive AVT was found in the position of synthetic AVT in the eluates. Thus, OT and AVP are present in human adult CSF, but AVT is not. The lack of a prominent rhythm of AVP in human CSF is in marked contrast to the brisk OT rhythm. The rhythm of neurohypophysial peptides in human and subhuman primates is different from the rhythm in lower species, suggesting different functions for OT and AVP in the central nervous system of various species.

Clearance studies of oxytocin in humans using radioimmunoassay measurements of the hormone in plasma and urine.

Synthetic oxytocin (OT) was infused iv in four men at 3 mU/min, and the rate was doubled every 90 min for a total of three infusion periods. The mean (+/- SEM) OT MCR was 16.4 +/- 1.7 ml/kg X min and was independent of the rate of infusion. A method for measuring OT in urine was developed using an octadecasilyl-silica column for extraction of the hormone. The extracted residue was reconstituted in potassium phosphate buffer, pH 7.4, for RIA. The minimum detectable level of OT in urine was 0.2 microU/ml (defined as a bound to free ratio of approximately 90%). The mean recovery of OT was 77 +/- 2%. The mean (+/- SEM) concentration of endogenous OT in urine was 10.2 +/- 1.4 microU/ml. Endogenous OT in urine eluted from a reverse phase high pressure liquid chromatography column as a single peak of OT immunoreactivity in the position of synthetic OT. Urinary OT excretion during infusion of synthetic OT was linearly correlated with plasma OT concentration whether calculated as microunits of urinary OT per mg creatinine (r = 0.89) or urinary OT per min (r = 0.93). Mean urinary fractional clearance of OT (OT clearance/creatinine clearance) was 3.6% renal clearance of OT (5.5 ml/min or 0.43% of MCR). Thus, OT MCR was constant over a wide range of physiological plasma OT levels and was similar to MCR in pregnant women studied previously in this laboratory. Less than 1% of OT was cleared in urine. This study defines the relationship between urinary and plasma OT during steady state infusion of physiological concentrations of the hormone and indicates that measurements of OT in urine by RIA may prove helpful for pharmacokinetic and physiological studies of OT-related events in humans.

Ampholyte displacement and high pressure liquid chromatographic separation of the estrogen-responsive neurophysin from human plasma.

Estrogen-stimulated neurophysin (ESN) or oxytocin (OT)-neurophysin (Np) was measured in plasma of seven men before and after oral administration of 25 mg diethylstilbestrol (DES). Pre-DES levels of ESN averaged 0.93 +/- 0.3 (+/- SEM) ng/ml and increased to 29.8 +/- 6.5 and 25.4 +/- 5.1 ng/ml 24 and 48 h after DES treatment, respectively. To compare the estrogen-responsive Np in plasma with human OT-Np which is present in the posterior pituitary gland, the Np fraction of post-DES plasma was concentrated by double precipitation with ammonium sulfate and applied to ampholyte displacement and Sephadex G-75 columns. The Np fraction of this plasma extract contained ESN immunoreactivity (IR) but no nicotine-stimulated neurophysin-IR. ESN-IR of plasma and of an extract of human posterior pituitary eluted identically from a Sephadex G-75 column, indicating similar mol wt. The plasma extract containing ESN-IR eluted from the ampholyte displacement column at pH 4.3-4.2. No nicotine-stimulated Np (arginine vasopressin-Np)-IR was found in the plasma samples. ESN-IR in an extract of human posterior pituitary gland eluted from the ampholyte displacement column at the same pH as that of the ESN extracted from plasma. Peak ESN-IR-containing fractions from the ampholyte displacement were pooled, dialyzed, lyophilized, and reconstituted in appropriate carrier buffer for reverse phase high pressure liquid chromatography. The ESN-IR was resolved into two distinct ESN-IR peaks by high pressure liquid chromatography. Plasma and posterior pituitary gave identical pairs of peaks. Thus, the Np that is increased in human plasma in response to estrogen is identical to pituitary OT-Np, providing strong evidence that estrogen stimulates the human neurohypophysis.

A time-dependent peak of oxytocin exists in cerebrospinal fluid but not in plasma of humans.

Oxytocin (OT) and the oxytocin-neurophysin (OT-Np) were measured by RIA in samples of cerebrospinal fluid (CSF) obtained sequentially at 0600, 1200, 1800, and 2400 h from six patients in whom intrathecal catheters were temporarily placed for CSF rhinorrhea. The highest levels of OT in CSF were found at 1200 h. An analysis of variance of sequential measures of the concentration of OT in samples of CSF obtained every 6 h over a 30-h period showed the mean levels (+/- SEM) of OT at 1200 h, 6.41 +/- 1.13 microU/ml and 5.06 +/- 0.58 microU/ml, to be significantly higher (p less than 0.05) than mean levels of OT at 0600 h, 2.50 +/- 0.65 microU/ml; 1800 h, 2.63 +/- 0.61 microU/ml and 2.64 +/- 1.21 microU/ml; and 2400 h, 2.86 +/- 1.13 microU/ml. The levels of OT-Np in CSF did not show a similar peak. In three of the patients simultaneous samples of blood were obtained for measurement of the same peptides, but no corresponding peak of OT or its Np was found in plasma of these three patients. The level of OT in CSF at all times was also significantly higher (p less than 0.05) than the level of OT in plasma of these three patients. Levels of OT and OT-Np were measured by RIA of samples of plasma obtained hourly for a 24-h period from six healthy men and six healthy women. No diurnal variation of OT or its Np in the plasma of men or women was found. This pattern of OT in the CSF of humans is similar to the pattern of OT in the CSF of the Rhesus monkey, but in contrast to the lack of a clearly defined peak of OT in the CSF of the cat or the rat. These observations in humans reinforce the differences among species of the secretion of OT in the CSF.

Oxytocin in human plasma: correlation with neurophysin and stimulation with estrogen.

RIA for the measurement of oxytocin in human plasma is described. Extraction of oxytocin from larger peptides in plasma used acetone precipitation with a 75% +/- 2 SEM recovery of oxytocin. Nonspecific binding of the assay was less than 4%, and the minimum level of detection was 0.2 microunits/tube. No cross-reactivity was noted with neurophysins, arginine, or lysine vasopressin. The mean basal level (+/- SEM) of oxytocin in men was 1.80 +/- 0.07 microunits/ml and was not different in normal women (1.71 +/- 0.07 microunits/ml). Changes in posture had no effect on the levels of oxytocin. Samples obtained every 15 min over 4 h showed no pulsatile secretion of oxytocin. In women chronically receiving estrogen as an oral contraceptive, oxytocin was greater than normal, (4.59 +/- 0.51 microunits/ml; P less than 0.01). Estrogen-stimulated neurophysin was also elevated (8.45 +/- 1.99 ng/ml; P less than 0.005). Acute ingestion of estrogen caused an increase in the level of oxytocin in plasma by 12 h and a concomitant elevation of estrogen-stimulated neurophysin. When the neurophysin was isolated from plasma obtained from a subject after ingestion of estrogen, the neurophysin from plasma comigrated on a polyacrylamide gel with a human pituitary standard of estrogen-stimulated neurophysin. In the studies in which neurophysin was elevated, the correlation between the level of oxytocin and the level of estrogen-stimulated neurophysin in plasma was significant (P less than 0.01). The observation that estrogen administration stimulates the release of oxytocin and estrogen-stimulated neurophysin provides additional evidence that this neurophysin is the oxytocin-neurophysin of man.

PIP: RIA for the measurement of oxytocin in human plasma is described. Extraction of oxytocin from larger peptides in plasma used acetone precipitation with a 75% +or- 2 SEM recovery of oxytocin. Nonspecific binding of the assay was less than 4% and the minimum level of detection was 0.2 mcU/tube. No cross-reactivity was noted with neurophysins, arginine, or lysine vasopressin. The mean basal level (+or- SEM) of oxytocin in men was 1.80 +or- 0.07 mcU/ml and was not different in normal women (1.71 +or- 0.07 mcU/ml). Changes in posture had no effect on the levels of oxytocin. Samples obtained every 15 minutes over 4 hours showed no pulsatile secretion of oxytocin. In women chronically receiving estrogen as an oral contraceptive, oxytocin was greater than normal (4.59 +or- 0.51 mcU/ml; P0.01). Estrogen-stimulated neurophysin was also elevated *8.45 +or- 1.99 ng/ml; P0.005). Acute ingestion of estrogen caused an increase in the level of oxytocin in plasma by 12 hours and a concomitant elevation of estrogen-stimulated neurophysin. When the neurophysin was isolated from plasma obtained from a subject after ingestion of estrogen, the neurophysin from plasma comigrated on a polyacrylamide gel with a human pituitary standard of estrogen-stimulated neurophsin. In the studies in which neurophysin was elevated, the correlation between the level of oxytocin and the level of estrogen-stimulated neurophysin in plasma was significant (P0.01). The observation that estrogen administration stimulates the release of oxytocin and estrogen-stimulated neurophysin provides additional evidence that this neurophysin is the oxytocin-neurophysin of man.

Elevation of oxytocin and the oxytocin-associated neurophysin in the plasma of normal women during midcycle.

A positive correlation was observed between the midcycle elevation of estrogen (E) and the level of oxytocin- and estrogen-stimulated neurophysin (ESN), the protein carrier of oxytocin, in the plasma of five of six women. The time of the maximal level of E was associated with a level of oxytocin significantly greater than that in either the early follicular or late luteal phase (P less than 0.025). Likewise, the level of ESN at midcycle was greater than the level of ESN in the early follicular or late luteal phase (P less than 0.01). Other than states of lactation or pregnancy, this is the only described cyclic secretion of oxytocin in humans. Since oxytocin chronologically correlates with a rise in the level of E at midcycle, a role for oxytocin in ovulation may be considered.

Pattern of oxytocin concentrations in the plasma and cerebrospinal fluid of lactating rhesus monkeys (Macaca mulatta): evidence for functionally independent oxytocinergic pathways in primates.

Two populations of oxytocin-staining neurons have been identified in the paraventricular nucleus: magnocellular neurons that terminate in the posterior pituitary and parvocellular neurons that terminate elsewhere in the central nervous system. Whether these oxytocin neurons are functionally separate was tested by measuring oxytocin concentrations in samples of peripheral blood and cerebrospinal fluid (CSF) obtained simultaneously from lactating rhesus monkeys during suckling. Lactating animals bearing temporary subarachnoid and venous catheters were maintained in a constant photoperiod (0600-1800 h). Samples of CSF were continuously withdrawn by peristaltic pump (0.1 mL/15 min) for 2-4 consecutive days from subarachnoid catheters with the tips placed at the T12-L1 level of the spinal column in four lactating monkeys 4 months postpartum and again after weaning. On 2 of these days, we observed and recorded periods of infant suckling and collected peripheral blood samples (1.2 mL) from the mother at 5-min intervals for 60 min. Oxytocin was measured in blood and CSF by RIA. Oxytocin concentrations increased in the plasma of the lactating monkeys during periods of nursing, with peak concentrations ranging from 4-16.7 microU/mL. No increase in plasma oxytocin was found on the day after the infant was weaned. Variations in the concentrations of oxytocin in CSF were independent of the suckling stimulus and plasma oxytocin concentrations and occurred during observed periods of no nipple contact by the infant and at the time of weaning after the infant had been removed from the mother. Each lactating animal also displayed a normal circadian variation in CSF oxytocin concentrations, with peak and nadir levels during light and dark hours, respectively. We conclude that release of oxytocin into the CSF of lactating monkeys is disassociated from release into the peripheral circulation. The data further support the conclusion that neuronal pathways giving rise to oxytocin in the CSF and the periphery are anatomically and functionally separate in primates.

Dopaminergic stimulation of oxytocin concentrations in the plasma of male and female monkeys by apomorphine and a D2 receptor agonist.

Administration of the dopamine receptor agonist apomorphine causes a dose-dependent increase in plasma oxytocin concentrations and dose-specific behavioral changes in rodents. To investigate whether dopamine receptor agonists will elicit similar neuroendocrine and behavioral effects in primates, we administered graded doses of apomorphine and the respective dopamine D1 and D2 receptor agonists, CY 208-243 and LY 163502, to monkeys and monitored plasma concentrations of oxytocin and behavior. Five female rhesus, two male rhesus, and two male cynomolgus monkeys had chronic indwelling venous catheters implanted and were maintained on standard jacket/tether/swivel systems to allow remote blood sample collection. During experiments, blood samples were collected 10 and 5 min before drug injection and at 2- to 120-min intervals after each injection. Apomorphine (50-400 micrograms/kg) and LY 163502 (10-100 micrograms/kg) elicited dose-dependent stimulations of oxytocin secretion. CY 208-243 (100-400 micrograms/kg) did not significantly affect oxytocin secretion. Low doses of apomorphine (50-100 micrograms/kg) and LY 163502 (10-25 micrograms/kg) elicited yawning, and high doses of apomorphine (200-400 micrograms/kg) and LY 163502 (50-100 micrograms/kg) elicited stereotypic behaviors. No behavioral effects of CY 208-243 (100-400 micrograms/kg) were observed. The magnitude of the oxytocin secretory responses varied among animals, but was similar in male and female monkeys. In summary, apomorphine and LY 163502 both elicited dose-related stimulation of oxytocin secretion coupled with dose-specific behavioral changes in male and female monkeys, while no effects of CY 208-243 on these parameters were observed. We conclude that dopamine receptor agonists, and in particular D2 agonists, may be useful tools for studies exploring the physiological and behavioral actions of oxytocin in primates.

A novel oxytocin-like and vasotocin-like peptide in human plasma after administration of estrogen.

Using three antisera to oxytocin (OT Pitt Ab-1, OT Pitt Ab-2, and TOR OT Ab), we found comparable levels of OT in response to infant suckling and during infusion of synthetic OT, and identical standard curves with biological and synthetic standards of OT. Pitt Ab-1, but not Pitt Ab-2 or TOR OT Ab, measured increased OT in response to estrogen. Using an arginine vasotocin RIA (TOR AVT Ab), we found an increase in AVT immunoreactivity after estrogen treatment. Mean basal OT levels measured with OT Pitt Ab-2 in plasma of men [0.75 +/- 0.06 (+/- SEM) microU/ml] and women (0.8 +/- 0.09 microU/ml) were lower than OT measured with Pitt Ab-1 (1.7 +/- 0.09 microU/ml in men and 1.7 +/- 0.07 microU/al in women; P less than 0.001). Mean OT measured with Pitt Ab-2 in the plasma of women given estrogen chronically (0.8 +/- 0.04 microU/ml) and acutely (0.6 +/- 0.15 microU/ml) were not significantly different from basal levels. OT levels measured with Pitt Ab-1 in the same samples were 4.6 +/- 0.5 and 4.3 +/- 0.5 microU/ml, respectively, both significantly increased from basal levels (P less than 0.001) and significantly higher than OT measured with Pitt Ab-2 (P less than 0.001). Mean OT measured with Pitt Ab-1 in the plasma of pregnant women was 8.6 +/- 1.02 microU/ml, significantly higher than OT measured with Pitt Ab-2 (1.0 +/- 0.3 microU/ml; P less than 0.001). Men given 25 mg diethylstilbestrol had significant increases in OT measured with Pitt Ab-1 and in AVT measured with TOR AVT (P less than 0.01), but not in OT measured with Pitt Ab-2. Plasma from a man given diethylstilbestrol was prepared for high performance liquid chromatography and applied to a C18 muBondapak reverse phase column. The plasma contained two peaks of immunoreactivity detected as OT with Pitt Ab-1 and as AVT using TOR AVT Ab. The material was not detected by Pitt Ab-2 or TOR OT Ab and did not coelute with standards of OT, AVT, or AVP. Pregnancy plasma, thioglycolic acid, chymotrypsin, and trypsin reduced Pitt Ab-1, Pitt Ab-2, and TOR OT immunoreactivity of synthetic OT. The percent recovery of OT immunoreactivity was not significantly different with Pitt Ab-1 vs. Pitt Ab-2. A novel peptide, which is increased in response to administered estrogen, is present in human plasma and is detected by some antisera to OT and AVT. The observation explains the wide variability in OT levels in the estrogen-primed state and provides a new mechanism to study estrogen-related physiology and pathophysiology.

High pressure liquid chromatographic separation of an oxytocin/arginine vasotocin-like peptide from the plasma of patients with chronic renal failure.

Levels of a novel oxytocin (OT)- and arginine vasotocin (AVT)-like peptide detected by one antiserum to OT (Pitt Ab-1) and one antiserum to AVT (Tor AVT) were recently found to rise in human plasma in response to administration of estrogen. The novel peptide rose in parallel with the estrogen-stimulated neurophysin (ESN). The mean level (+/- SEM) of ESN in plasma of 11 individuals with altered renal function (nondialyzed) was significantly higher than the level in individuals with normal renal function (4.2 +/- 0.9 vs. 1.1 +/- 0.04 ng/ml; P less than 0.01). In patients treated with hemo- or peritoneal dialysis, mean (+/- SEM) levels of ESN were 18.1 +/- 3.2 and 16.8 +/- 3.7 ng/ml, respectively. Levels of estradiol and estrone were not elevated and did not correlate with high levels of ESN. Levels of OT Pitt Ab-1, AVT, and ESN immunoreactivity were measured in plasma form nine patients undergoing hemodialysis and eight patients undergoing peritoneal dialysis. Mean (+/- SEM) levels of all three peptides were elevated (12.9 +/- 1.5 microU/ml, 32.1 +/- 6.7 pg/ml, and 13.5 +/- 4.0 ng/ml, respectively). ESN was significantly correlated with OT Pitt Ab-1 and AVT (R2 = 0.80; P less than 0.001). Plasma samples from the same patients were pooled, treated, and separated by reverse phase HPLC. The plasma contained a peak of immunoreactivity detected by Pitt Ab-1 and Tor AVT Ab. The position of the material was distinct from that of synthetic OT, AVT, or AVP and corresponded to the position of the novel OT-like peptide found in plasma of individuals given estrogen. The findings support parallel secretion of the OT-like peptide with ESN and represent the first disease state characterized by high levels of this OT- and AVT-like peptide.