Menstrual cycles in rhesus monkeys (Macaca mulatta) are unaffected by a single dose of the anesthetics ketamine and xylazine administered during the midfollicular phase at laparoscopy

To identify an anesthetic regimen that produces more complete relaxation and analgesia than ketamine hydrochloride (Ketaset®) alone, a combination of ketamine (15 mg/kg body weight) and the hypnotic xylazine (Rompun®, 0.33 mg/kg) was evaluated. Since the desired experimental application required that the anesthetic not interfere with normal hormonal events during the menstrual cycle, this combination administered on day 6 of the cycle was tested to determine whether hormonal surges, incidence of ovulation, or cycle length would be altered relative to the use of ketamine alone. In five of six animals, ketamine plus xylazine had no effect on the occurrence of timely surges of estrogen, luteinizing hormone (LH), or follicle-stimulating hormone (FSH), or on ovulation as determined by the presence of a corpus luteum at laparoscopy and normal serum concentrations of progesterone. There were no significant differences between the cycle during treatment and previous cycles in the same animal for length of the menstrual cycle (26.0 ± 2.3 [5] days; X? ± S.D. [n] or luteal phase (13.4 ± 2.4 [5] days). Likewise, these values did not differ from those of ten control monkeys treated with ketumine only on day 5 or 6 of the cycle (incidence of ovulation, 10/10; cycle length, 27.9 ± 1.8 [10]; luteal phase length, 15.1 ± 1.4 [10], P > 0.05). Patterns of circulating progesterone were not altered by the addition of xylazine anesthesia. These findings indicate that xylazine, given in the midfollicular phase, did not alter ovulatory events or menstrual cycle characteristics in rhesus monkeys. Ketamine plus xylazine apparently provides anesthesia appropriate for laparoscopy.     

An immunoradiometric assay for squirrel monkey prolactin.

Determination of immunoreactive prolactin in squirrel monkeys has been hampered by the lack of specific antibodies. We investigated the adaptability of a commercially available immunoradiometric assay for human prolactin, which employs two separate monoclonal antibodies (MAb I and II) to human prolactin, to determine the presence of squirrel monkey prolactin. We found that immunoreactivity curves for prolactin in squirrel monkey pituitary homogenates and serum were parallel to human prolactin standards, suggesting that the epitopes recognized by these antibodies were common to both human and squirrel monkey prolactin. Both nonglycosylated (23 kD) and glycosylated (26 kD) forms of squirrel monkey prolactin were detected in squirrel monkey pituitary homogenates by Western blot analysis using [125I]-MAb II. Neither sheep nor rat prolactin was recognized by Western blot analysis, indicating that the assay may be specific for primate prolactins. We examined the effect of ketamine HCl, an anesthetic that has been shown to elevate serum prolactin levels in other primates, on prolactin secretion in squirrel monkeys. Serum prolactin levels increased greater than fourfold after the administration of ketamine HCl (30 mg/kg b.w., i.m.) compared with control levels. Serum prolactin levels were unaffected by anesthesia with pentobarbital sodium (15 mg/kg b.w., i.v.). This assay provides a reliable and sensitive method for determining immunoreactive squirrel monkey prolactin.     

Comparison of ketmine with the combination of ketamine and xylazine for effective anesthesia in the rhesus monkey (Macaca mulatta).

The addition of xylazine to ketamine hydrochloride was found to enhance analgesia, anesthesia, and muscle relaxation in rhesus monkeys. At 0.10 ml/kg body weight, this combination provided adequate anesthesia for such procedures as cisternal puncture, lumbar spinal puncture, insertion of urinary catheters, finger amputations, and tattooing. The combination of ketamine and xylazine did depress the heart rate, respiration rate, and body temperature more than the administration of ketamine alone. The period of anesthesia also was prolonged, but the monkeys regained consciousness more rapidly at the end of the anesthetic period.     

Gonadotropin secretion during prolonged hyperprolactinemia: basal secretion and the stimulatory feedback effect of estrogen

Inoculation of cyclic female rats with the prolactin (Prl)/growth hormone-secreting pituitary tumor, MtT.W15, resulted in a cessation of estrous cyclicity within 5--10 days. Associated with this acyclicity was a persistently low serum concentration of estradiol and marked increases in both circulating Prl and progesterone. At Day 26 of acyclicity, basal serum luteinizing hormone (LH) values measured in samples taken every 20 min from 0900--1100 h were significantly reduced when compared to cyclic, nontumor animals on diestrus Day 2. There was no difference in basal follicle-stimulating hormone (FSH) concentrations. In a separate group of acyclic, tumor-bearing females 42--56 days after transplantation, a single s.c. injection of 20 micrograms estradiol benzoate (EB) at 1030 h elicited significant increases in both serum LH and FSH values between 1700 and 1830 h on the next day. The magnitude of the LH surge was reduced and that of FSH was increased in tumor-bearing animals when compared to cyclic, nontumor females given a similar EB injection on diestrus Day 1. These results demonstrate that chronic hyperprolactinemia is associated with inhibition of basal LH secretion and ovarian estrogen production and an increase in circulating progesterone concentrations. Nevertheless, the stimulatory feedback effects of estrogen on LH and FSH release are still present and functioning in acyclic female rats under chronically hyperprolactinemic conditions. These data suggest that the cessation of regular ovulatory cycles associated with hyperprolactinemia may be due to a deficiency of LH and/or estrogen secretion, but not to a lack of central nervous system response to the stimulatory feedback action of estrogen.     

Hematological and serum biochemical values in cynomolgus monkeys anesthetized with ketamine hydrochloride

The effects of ketamine anesthesia on both hematological and serum biochemical variables were investigated in 19 male and 15 female cynomolgus monkeys. Blood samples were obtained from the cephalic vein within 30 minutes of an intramuscular injection of ketamine hydrochloride (10 mg/kg). Ketamine anesthesia caused a reduction in leukocyte counts and a significant reduction in lymphocytes percentages. Ketamine anesthesia also increased the serum activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine phosphokinase (CPK), but reduced the serum concentrations of glucose, inorganic phosphate, sodium and potassium. The alterations of hematological and serum biochemical values will be discussed. These alterations should be considered when designing studies for and interpreting data from cynomolgus monkeys.     

Failure of daily injections of ketamine HCL to adversely alter menstrual cycle length, blood estrogen, and progesterone levels in the rhesus monkey.

Failure of daily injections of ketamine hydrogen chloride (HCL) to adversely alter menstrual cycle length, blood estorgen, and progesterone levels in the rhesus monkey is reported. The study was carried out with 30 adult female monkeys to determine the effects of daily administration of 8-10 mg ketamine HCL/kg. In physically restrained control monkeys there were 14 of 25 ovulatory cycles and inketamine-treated monkeys there were 28 of 32 ovulatory cycles. Menstrual cycle length was the same in both groups. The levels and time course of estrogen and progesterone levels were the same in the ovulatory cycles of both groups. In 30% of the control cycles and in 25% of the ketamine-treated there were luteal phases in which the preovulatory estrogen levels were normal and in which the luteal-phase progesterone levels were low and variable 6-8 days after the preovulatory surge. It is concluded that the daily use of ketamine HCL does not markedly alter menstrual cycle length, or serum estrogen or progesterone levels throughout the menstrual cycle. The incidence of anovulatory cycles and premature menstrual induction was reduced probably by reducing the stress of restraining the monkey for the purpose of taking a blood sample.     

The effects of serotonergic and adrenergic receptor antagonist on prolactin release in the monkey.

The influence of serotonergic and adrenergic antagonists on serum prolactin levels was studied in ketamine anesthetized monkeys. Methysergide, a serotonergic receptor blocker, at 0.035, 0.1 and 1 mg/kg body weight induced a rapid and transient increase in serum prolactin. Cyproheptadine, another serotonergic receptor blocker, at 0.05, 0.5 and 1 mg/kg induced a rapid and sustained increase in serum prolactin. SQ 10631, a third serotonergic receptor blocker, had a minimal effect on increasing basal prolactin levels even at doses as high as 10 mg/kg. Propranolol, a β adrenergic blocker, at a dose of 5 mg/kg induced a small sustained increase in serum prolactin, while a lower dose (1 mg/kg) had a slight but significant effect. Phentolamine, an α adrenergic receptor blocker, at a dose of 5 mg/kg induced a rapid and transient increase in plasma prolactin while a lower dose (1 mg/kg) had no effect. Phenoxybenzamine, a potent α adrenergic receptor blocker, had only a minimal effect on prolactin release even at doses of 3 and 5 mg/kg. It appears that the time course and extent of prolactin release differs among neural antagonists even within the same biogenic amine system.     

Serum prolactin levels of rats under continuous estrogen stimulation and 2 Br-alpha-ergocryptine (CB-154) injection.

The ability of 2 Br-alpha-ergocryptine (CB-154) to suppress serum prolactin levels was examined in animals under the influence of continuous estrogen stimulation. A single injection of polyestradiol phosphate (Estradurin) once every 21 days produced a constant elevation of serum prolactin. The simultaneous administration of 1 mg/day of CB-154 to estrogenized animals suppressed serum prolactin levels below that of Estradurin alone but the levels were significantly greater than those of animals receiving CB-154 alone. It was suggested that, while CB-154 prevents the release of prolactin, estrogen stimulates prolactin synthesis despite the block of its release. Ether anesthesia may be capable of partially overriding the block of CB-154 and released the stored hormone from the gland.     

Novel hyperprolactinemia and hyperprolactinemic anovulation model using the cynomolgus monkey (Macaca fascicularis)

We investigated the relationship between the menstrual cycle and hormone levels in cynomolgus monkeys, and developed a sulpiride-induced hyperprolactinemic anovulation model. On this study, we demonstrated the usefulness of the commercial human prolactin immunoradiometric assay kit for the measurement of cynomolgus monkey serum samples. In the normal menstrual cycle of the cynomolgus monkey, serum prolactin concentrations were not significantly different between luteal and follicular phases. However, the serum prolactin concentration tended to elevate at the ovulation stage. And serum progesterone began to increase after an estradiol surge, and then declined before the ensuing preovulatory rise in estradiol. During the luteal phase, the serum concentration of progesterone was elevated. Moreover, we aimed to develop an anovulation model, using sulpiride-induced hyperprolactinemia in the cynomolgus monkey. The serum prolactin level gradually increased during the twice-daily administration of sulpiride, and the drug produced as big a response at 5 mg/kg. In this study, the length of the menstrual cycle was approximately 29 days in normal cynomolgus monkeys. When treatment with sulpiride had been continued for more than one month, serum progesterone and estradiol levels fell to within the range seen in the follicular phase of the normal cycle, and the absence of ovulation was recognized by laparoscopy. Moreover, in this period we found that amenorrhea or anovulatory menstruation in the experimental animals. We could produce an anovulatory model induced by sulpiride repeatedly administered over a long time period. Our findings suggest that the cynomolgus monkey is useful as a endocrinological model that uses prolactin as a parameter and as an anovulatory model; thus, it could be a useful model for the hyperprolactinemic amenorrhea and/or anovulation seen in humans.     

Measurement of serum prolactin and the effect of ketamine anesthesia on serum prolactin levels in cynomolgus monkeys (Macaca fascicularis)

The effect of an anesthetic, ketamine, on the serum prolactin level was examined in wild-originating female cynomolgus monkeys (Macaca fascicularis) imported from South East Asia. Serum prolactin levels were measured by the homologous radioimmunoassay system which was developed for human prolactin. The validity was confirmed by using an extract of pituitary gland from a female cynomolgus monkey as well as serum and amniotic fluid from a pregnant monkey. Additionally, serum luteinizing hormone (LH) levels were determined by the radioreceptor assay system developed in our laboratory using Leydig cells collected from rat's testes as a receptor fraction. The experiment was repeated three times at one-month interval, using twenty animals that were divided into three groups consisting of 5, 7 and 8 monkeys each. In the first experiment, the first group was injected with physiological saline and the second and third groups were intramuscularly given ketamine at a dose level of 5 mg/kg B.W. and 15 mg/kg B.W., respectively. In the second experiment, the first and second groups were given ketamine at a dose of 5 mg/kg B.W. and of 15 mg/kg B.W., respectively, and the third group was served as control injected with saline. In the third experiment, the first and third groups were administered with 15 mg/kg and 5 mg/kg of ketamine and the second group was injected with saline. In short, all of the twenty monkeys received the three different treatments for two months. The serum prolactin level showed a marked increase after the administration of ketamine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Failure of yohimbine to reverse ketamine anesthesia in rhesus monkeys

Yohimbine hydrochloride has been used experimentally to reverse the anesthetic effects of ketamine and xylazine in dogs, cats, cattle and mule deer, but there are no reports of its use in nonhuman primates. Nine adult female rhesus monkeys were given an intravenous dose of either 0.5 mg/kg yohimbine hydrochloride or saline 10 minutes after intramuscular administration of 10 mg/kg ketamine hydrochloride. There was no difference in the duration of anesthesia between the yohimbine and saline treatments, suggesting yohimbine is not effective in the rhesus monkey.     

Pentobarbital anesthesia: interference in the effects of other pharmacological agents and of electrical stimulation on prolactin secretion

In adult male rats, a pretreatment regimen of serial injections of dexamethasone (1 mg/kg), morphine (20 mg/kg) and pentobarbital (40 mg/kg) was evaluated for use in conjunction with studies on the effects of hypothalamic electrical stimulation on prolactin secretion. Serum prolactin levels were measured before and 15 min after electrical and sham stimulation of the hypothalamic arcuate nucleus in rats subjected to either the pharmacological regimen or to pentobarbital anesthesia alone. Pentobarbital alone caused a transient rise in serum prolactin levels, which obscured any effect of electrical or sham stimulation; this interference was not overcome by the addition of dexamethasone and morphine treatment. Thus, the result indicate that the acute stimulatory affect of pentobarbital anesthesia on prolactin release may interfere with further manipulation of prolactin-controlling mechanisms, by either pharmacological or surgical means. Furthermore, the dexamethasone-morphine-pentobarbital pretreated rat does not provide an adequate preparation for studing the effects of electrical stimulation on prolactin secretion.     

Rapid recovery of estrogen-induced pituitary gonadotropin surges after luteectomy in rhesus monkeys

In the presence of a functional corpus luteum, positive estrogen feedback on the surge modes of gonadotropin secretion is blocked in rhesus monkeys. We investigated the effects of luteectomy (Lx) on the time required for recovery of pituitary responsiveness (LH/FSH surges) to positive estrogen feedback. Estradiol-17 beta-3- benzoate (EB, 50 microgram/kg sc) was given: 1) 24th prior to, 2) the day of, or 3) 24 h after luteal ablation. Daily measurements of serum follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol-17 beta (e2) and progesterone (P) were made on each monkey for 5 days. Serum P fell to undetectable levels within 24 h after Lx, whereas E2 levels in circulation peaked within 24h after injection of EB. Among early follicular phase monkeys, this EB treatment results in typical midcycle type LH/FSH surges within 48h. Lx alone was not soon followed by significant changes in pituitary gonadotropin secretion. When circulating P levels were undetectable the pituitary responded fully to EB; that is, typical midcycle type FSH/LH surges occurred. When serum P was in the midst of declining after Lx, gonadotropin surges were present, but attenuated. However, when P levels remained elevated for more than 24 h after EB injection, the surge modes of FSH/LH secretion remained fully blocked. These results demonstrate that the suppressive influence of luteal secretions (principally progesterone) on positive estrogen feedback regulation of the surge modes of pituitary gonadotropin secretion is quite transient in these primates.     

Rapid elevation of rat serum prolactin concentration by cyclosporine, a novel immunosuppressive drug

Within one hr of the administration of cyclosporine to rats, there was a 4-fold elevation in the serum prolactin concentration. Doses of 0.12, 1.2, and 12 micrograms/100 g body weight cyclosporine significantly elevated the serum prolactin level. Higher doses, 120 or 1200 micrograms/100 g body weight cyclosporine resulted in small but insignificant elevations of the serum prolactin concentration. Bromocriptine, a dopamine agonist which inhibits prolactin release from the anterior pituitary, completely blocked the elevation in serum prolactin in response to cyclosporine alone. These data suggest that the ability of cyclosporine to suppress immune function may involve its ability to rapidly produce hyperprolactinemia.     

Effects of acute and chronic trazodone administration on serum prolactin levels in adult female rats

Trazodone was tested for its ability to elevate serum prolactin levels in mature female rats. When the drug was administered acutely to female rats at doses up to 80 mg/kg ip, it induced a clear rise in serum prolactin levels, with a minimum effective dose of 20 mg/kg; blood trazodone levels at these doses were between 1.6–2.4 μg/ml. However, trazodone could not be considered to be a potent stimulator of prolactin secretion, since the injection of haloperidol at 2 mg/kg elevated serum prolactin to values twice those seen in animals receiving the 80 mg/kg dose of trazodone. When trazodone was administered chronically in the diet for two or four weeks, at an average daily dose of 80 mg/kg, serum trazodone levels were found to be 100–200 ng/ml when measured at each stage of the estrous cycle. Serum prolactin levels in trazodone-treated animals, however, did $\text{not}$ differ from those in control rats. Moreover, drug-treated animals showed normal proestrus surges in serum prolactin. The results of these studies thus indicate that acutely, at very high doses, trazodone probably can stimulate prolactin secretion modestly in female rats. However, when consumed chronically at 80 mg/kg/day, the drug has no effects on serum prolactin levels. Therefore, if trazodone stimulates prolactin secretion by altering neurotransmission across dopamine and/or serotonin synapses in brain, it is probably not potent in these actions, at least as concerns those dopamine and serotonin neurons that influence the secretion of prolactin.     

Effects of physical and chemical restraint on intravenous glucose tolerance test in crested black macaques (Macaca nigra)

The effects of physical and chemical restraint on glucose clearance and insulin secretion were evaluated during intravenous glucose tolerance testing in Macaca nigra. Conscious monkeys placed in plexiglas cylindrical restraining devices (CRD) appeared relaxed, but glucose clearance and insulin secretion were impaired. A combination of midazolam with ketamine, compared to ketamine alone, did not cause detectable changes in the intravenous glucose tolerance tests; midazolam also reduced adverse reactions to ketamine and extended the duration of anesthesia. The cylindrical restraining device can be convenient for examining monkeys, but it is limited by its adverse affects on metabolic and hormonal measurements in intravenous glucose-tolerance tests. Chemical restraint using ketamine with midazolam was more effective than ketamine alone.     

Influence of adrenergic antagonists and apomorphine on prolactin release induced by serotonergic antagonists in the monkey.

The influence of adrenergic receptor blockers on the prolactin releasing effect of methysergide and cyproheptadine was examined in sexually mature female monkeys under ketamine anesthesia. Propranolol, a β-adrenergic blocker, at a dose of 1 mg/kg did not alter the prolactin releasing action of 0.1 mg/kg of methysergide but significantly potentiated (P < 0.025) the prolactin releasing action of 0.5 mg/kg of cyproheptadine. Phentolamine and phenoxybenzamine, both α-adrenergic blockers, at 1 mg/kg blunted the prolactin releasing effect of methysergide and cyproheptadine, but the pattern of prolactin blockade was different between the two putative antiserotonergic drugs. The prior administration of apomorphine, 4 mg/kg, a dopamine receptor stimulator, blocked the prolactin releasing effect of methysergide and cyproheptadine. Evidence presented here and from the literature indicate that the prolactin releasing action of methysergide and cyproheptadine is mediated by an antidopaminergic action directly on the pituitary.     

The effect of ketamine hydrochloride anesthesia on basal and N-methyl-D,L-aspartate induced plasma prolactin secretion in the adult male rhesus monkey

The excitatory amino acids (EAAs), glutamate and aspartate, acting predominantly on N-methyl-D-aspartate (NMDA) receptor, have been shown to be involved in the central regulation of the secretion of several anterior pituitary hormones including prolactin (PRL), whereas ketamine hydrochloride (KH), a widely used anesthetic, has been reported to antagonize a variety of NMDA receptor mediated actions of these EAAs. In the present study, the effect of KH on basal PRL levels as well as on N-methyl-D,L-aspartate (NMA), an agonist of NMDA receptor, induced plasma PRL secretion was investigated in the adult male rhesus monkey. The values were compared to those obtained from the same animals restrained in primate chairs. The plasma PRL concentrations were higher in animals receiving KH administered either intramuscularly (2.5 mg/kg BW at 30 min intervals) or intravenously (10 mg/kg BW) as compared to those observed in the unanesthetized chair-restrained monkeys. NMA induced an unequivocal increase in plasma PRL concentrations in both conscious chair-restrained and KH anesthetized monkeys, but the response was greater in anesthetized animals than the conscious monkeys. The present findings suggest that KH has stimulatory effects on both basal and NMA induced plasma PRL secretion.     

Prolactin modulates steroidogenesis by rat granulosa cells: I. Effects on progesterone

Either testosterone or follicle-stimulating hormone (FSH) stimulates progesterone secretion by granulosa cells from rats but the combination of the two hormones increases progesterone production in a synergistic manner. We have investigated the effects of graded doses of prolactin (0, 0.02, 0.2, 2, or 10 micrograms/ml) alone or in combination with testosterone (0.5 microM), FSH (300 ng/ml), or FSH + testosterone on progesterone secretion by granulosa cells at two stages of differentiation. Relatively undifferentiated granulosa cells from immature, diethylstilbestrol-treated, hypophysectomized (HPX) rats were cultured in defined (serum-free) medium for 3 days. More highly differentiated granulosa cells were obtained on the morning of proestrus from the preovulatory follicles of 30-day-old rats induced to undergo an estrous cycle by injection with 4 IU pregnant mare's serum gonadotropin; these cells were cultured in medium containing 10% fetal bovine serum. Prolactin alone did not enhance the negligible secretion of progesterone by cells from HPX rats, but increased progesterone secretion by cells from proestrous rats. Prolactin significantly enhanced the stimulatory effects of testosterone or FSH alone on cells from both HPX and proestrous rats. When cultures containing both FSH + testosterone were treated with prolactin, progesterone secretion by cells from proestrous rats was significantly enhanced, whereas secretion by cells from HPX rats was significantly depressed. Therefore when cells from HPX rats were cultured with both FSH and testosterone, the direction of the effect of prolactin was reversed from that observed with prolactin + FSH or testosterone alone, and from that observed when cells from proestrous rats were cultured with prolactin + FSH + testosterone.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immobilization with a single dose of ketamine hydrochloride and a combination of xylazine hydrochloride-ketamine hydrochloride and antagonism by yohimbine hydrochloride in the Japanese monkey (Macaca fuscata)

Forty-nine free-ranging Japanese monkeys (Macaca fuscata) were immobilized with 4.3–15.6 mg/kg (mean±S.D.=10.0±2.5 mg/kg) of ketamine hydrochloride (HCl), and 27 Japanese monkeys kept in enclosures were immobilized with a combination of 0.8–1.4 mg/kg (1.0±0.2 mg/kg) of xylazine HCl and 4.0–7.1 mg/kg (5.0±0.6 mg/kg) of ketamine HCl. In the xylazine HCl-ketamine HCl combination, good myorelaxation was induced. The mean induction times for the single dosage of ketamine HCl and the xylazine HCl-ketamine HCl combination were 2.8±1.5 min and 6.9±4.4 min, respectively. The mean immobilization times with the single dosage of ketamine HCl and the xylazine HCl-ketamine HCl combination were 39.3±16.5 min and 58.8±34.2 min, respectively. A half dose of ketamine HCl in combination with xylazine HCl could also immobilize Japanese monkeys successfully. Administrations of 0.5 mg/kg i.v. and 1.0 mg/kg i.m. of yohimbine HCl as an antagonist to xylazine HCl at 30 min after the induction reduced the immobilization time to 31.4±0.5 min and 49.0±22.1 min, respectively. Yohimbine HCl appears to be an effective antagonist to combination anesthesia by xylazine HCl-ketamine HCl in the Japanese monkey.