Placentophagia and the Tao of POEF.

Placentophagia, ingestion of placenta and amniotic fluid, usually during parturition, is a behavioral feature of nearly all nonaquatic, placental mammals, and is a nexus for several interlocking behavioral phenomena. Placentophagia has not been typical of human cultures, but in recent years, some women in affluent societies have engaged in it, thereby bringing publicity to the behavior. First, we summarized benefits of placentophagia for nonhuman mammals, which include increased attractiveness of neonates, enhanced onset of maternal behavior, suppression of pseudopregnancy, and enhancement of opioid hypoalgesia by Placental Opioid-Enhancing Factor (POEF), a benefit that may extend well outside the context of parturition. The research on POEF in animals was discussed in detail. Then we discussed placentophagia (placentophagy) in humans, and whether there is validity to the claims of various benefits reported primarily in the pro-placentophagy literature, and, although human afterbirth shows POEF activity, the POEF effect has not yet been tested in humans. Finally, we discussed the general possible implications, for the management of pain and addiction, of isolating and characterizing POEF.

Amniotic-fluid ingestion enhances central δ-opioid-induced hypoalgesia in rats in the cold-water tail-flick assay in a repeated-measures design.

Placental Opioid Enhancing Factor (POEF) is found in amniotic fluid (AF) and placenta. When ingested, it enhances opioid-mediated pain relief. Our laboratory has shown that ingestion of AF specifically enhances the hypoalgesia associated with δ-opioid receptor activation in the brain. The specific biochemical compound in AF responsible for the enhancement of δ-opioid activity is of great interest as an analgesic adjunct for pain but is unknown at this time. Research efforts to isolate and characterize this biochemical compound are hampered by the lack of an algesiometric assay that allows repeated measurement of pain threshold and repeated exposure to δ-opioid receptor activation. The cold water tail-flick assay (CWTF) may be a sensitive and reliable pain threshold test of (a) all species of opioids that is (b) not subject to repeated-testing effects. Therefore the CWTF test is potentially ideal for the study of δ opioid systems in a repeated measures design. Here, we confirm these attributes of the CWTF test, and determined that (a) there are no repeated-exposure effects associated with the CWTF assay; (b) there are no repeated-exposure effects associated with repeated central injections of DPDPE ([D-Pen2,D-Pen5]-Enkephalin, a selective δ-opioid agonist) as measured by the CWTF assay; and (c) ingestion of AF in conjunction with a central injection of DPDPE produced the same hypoalgesic enhancement as previously found using another assay.

Placentophagia in humans and nonhuman mammals: causes and consequences.

Afterbirth ingestion by nonhuman mammalian mothers has a number of benefits: (1) increasing the interaction between the mother and infant; (2) potentiating pregnancy-mediated analgesia in the delivering mother; (3) potentiating maternal brain opioid circuits that facilitate the onset of caretaking behavior; and (4) suppressing postpartum pseudopregnancy. Childbirth is fraught with additional problems for which there are no practical nonhuman animal models: postpartum depression, failure to bond, hostility toward infants. Ingested afterbirth may contain components that ameliorate these problems, but the issue has not been tested empirically. The results of such studies, if positive, will be medically relevant. If negative, speculations and recommendations will persist, as it is not possible to prove the negative. A more challenging anthropological question is "why don't humans engage in placentophagia as a biological imperative?" Is it possible that there is more adaptive advantage in not doing so?

Effect of amniotic-fluid ingestion on vaginal-cervical-stimulation-induced Fos expression in female rats during estrus.

Placental Opioid-Enhancing Factor (POEF) is a substance found in amniotic fluid (AF) that, when ingested, potentiates opioid-mediated, but not non-opioid-mediated, hypoalgesia. Vaginal-cervical stimulation (VCS) produces a stimulus-bound, partially opioid-mediated hypoalgesia that previous research has shown to be potentiated by AF ingestion. To understand the mechanism of opioid enhancement by POEF we investigated the pattern of neural activation after a bout of VCS that produced hypoalgesia, with and without co-administration of AF. Specifically, virgin Long-Evans rats showing vaginal estrus were handled briefly (control) or received VCS (75g pressure, 1 min), in a pattern that approximated early parturition rather than copulation, using a spring-loaded glass-rod probe. Rats were given an orogastric infusion (0.25 ml) of either AF or 0.9% saline resulting in four groups (VCS or handling; AF or saline). Rats were perfused 90 min after treatment and tissue was processed by immunohistochemistry for Fos. The number of Fos-immunoreactive cells was counted in structures previously shown to express Fos in response to VCS (the medial preoptic area, MPOA; the ventrolateral portion of the ventromedial hypothalamic nucleus, vlVMH; the arcuate nucleus, ARC). We found that this pattern of VCS did not produce a significant increase in Fos expression in the MPOA and vlVMH unless it was paired with AF. VCS produced a significant increase in Fos in the ARC. The interaction of AF and VCS on Fos expression in the MPOA suggests that POEF may enhance vaginal-cervical sensory input at parturition to facilitate sensitization of the MPOA, and presumably facilitate maternal-behavior onset.

Ingestion of amniotic fluid enhances the facilitative effect of VTA morphine on the onset of maternal behavior in virgin rats.

Previous research has shown that injection of morphine into the ventral tegmental area (VTA) facilitates the onset of maternal behavior in virgin female rats, and injection of the opioid antagonist naltrexone into the VTA disrupts the onset of maternal behavior in parturient rats. Placentophagia -- ingestion of placenta and amniotic fluid, usually at parturition -- modifies central opioid processes. Ingestion of the active substance in placenta and amniotic fluid, Placental Opioid-Enhancing Factor (POEF), enhances the hypoalgesic effect of centrally administered morphine, and more specifically, enhances delta- and kappa-opioid-receptor-mediated hypoalgesia and attenuates mu-opioid-receptor-mediated hypoalgesia. POEF (in placenta or amniotic fluid) ingestion does not, by itself, produce hypoalgesia. In the present study, we tested the hypothesis that ingestion of amniotic fluid enhances the facilitative effect of opioid activity (unilateral morphine injection) in the VTA on the rate of onset of maternal behavior. Virgin female Long-Evans rats were given one intra-VTA injection of morphine sulfate (0.0, 0.01, or 0.03 microg, in saline) and an orogastric infusion of 0.25 ml amniotic fluid or saline once each day of the first three days of the 10-day testing period. Subjects were continuously exposed to foster pups that were replaced every 12 h; replacement of pups was followed by a 15-min observation period. Maternal behavior latency was determined by the first of two consecutive tests wherein the subject displayed pup retrieval, pup licking in the nest, and crouching over all foster pups, during the 15-min observation. We confirmed the previous finding that the VTA injection, alone, of 0.03 microg morphine shortened the latency to show maternal behavior and that 0.0 microg and 0.01 microg morphine did not. Ingestion of amniotic fluid (and therefore POEF) facilitated the onset of maternal behavior in rats receiving an intra-VTA microinjection of an otherwise subthreshold dose of morphine (0.01 microg).

Evaluation of buprenorphine in a postoperative pain model in rats.

We evaluated the commonly prescribed analgesic buprenorphine in a postoperative pain model in rats, assessing acute postoperative pain relief, rebound hyperalgesia, and the long-term effects of postoperative opioid treatment on subsequent opioid exposure. Rats received surgery (paw incision under isoflurane anesthesia), sham surgery (anesthesia only), or neither and were treated postoperatively with 1 of several doses of subcutaneous buprenorphine. Pain sensitivity to noxious and nonnoxious mechanical stimuli at the site of injury (primary pain) was assessed at 1, 4, 24, and 72 h after surgery. Pain sensitivity at a site distal to the injury (secondary pain) was assessed at 24 and 72 h after surgery. Rats were tested for their sensitivity to the analgesic and locomotor effects of morphine 9 to 10 d after surgery. Buprenorphine at 0.05 mg/kg SC was determined to be the most effective; this dose induced isoalgesia during the acute postoperative period and the longest period of pain relief, and it did not induce long-term changes in opioid sensitivity in 2 functional measures of the opioid system. A lower dose of buprenorphine (0.01 mg/kg SC) did not meet the criterion for isoalgesia, and a higher dose (0.1 mg/kg SC) was less effective in pain relief at later recovery periods and induced a long-lasting opioid tolerance, indicating greater neural adaptations. These results support the use of 0.05 mg/kg SC buprenorphine as the upper dose limit for effective treatment of postoperative pain in rats and suggest that higher doses produce long-term effects on opioid sensitivity.

The biopsychology of maternal behavior in nonhuman mammals.

The term "maternal behavior," when applied to nonhuman mammals, includes the behaviors exhibited in preparation for the arrival of newborn, in the care and protection of the newly arrived young, and in the weaning of those young, and represents a complex predictable pattern that is often regarded as a single, comprehensive, species-specific phenomenon. Although the delivering first-time mammalian mother is immediately and appropriately maternal, a "virgin" with no prior exposure to young does not show immediate and appropriate behavior toward foster young. Nevertheless, the virgin female, and indeed the male, possess the neural circuitry that underlies the pattern referred to as maternal behavior, despite not exhibiting the pattern under normal circumstances. At parturition, or after extensive exposure to young, what emerges appears to be a single stereotyped maternal behavior pattern. However, it is actually a smoothly coordinated constellation of simpler actions with proximate causes that, when sequenced properly, have the appearance of a motivated, purposive, adaptive pattern of caretaking. Over the past 50 years, much research has focused on finding the principal external and internal factors that convert the nonmaternal behavior patterns of the nonpregnant nullipara, the virgin, to the almost immediate and intense maternal behavior characteristic of the puerpera, the mother. This review is an attempt to summarize the many comprehensive, even encyclopedic, reviews of these factors, with an emphasis on brain mechanisms, and to highlight the gaps that remain in understanding the processes involved in the almost immediate onset of maternal caretaking behaviors observed in mammals at delivery. Where possible, the reader is directed to some of those excellent reviews.

Lack of analgesic efficacy in female rats of the commonly recommended oral dose of buprenorphine.

Previous work in our laboratory showed that the recommended oral dose of buprenorphine (0.5 mg/kg) was not as effective as the standard therapeutic subcutaneous dose for postoperative analgesia in male Long-Evans (hooded) and Sprague-Dawley (albino) rats. The aim of the current study was to extend this analysis to female rats. We measured the pain threshold in adult female rats in diestrus or proestrus before and 30 and 60 min after oral buprenorphine (0.5 mg/kg,), the standard subcutaneous dose of buprenorphine (0.05 mg/kg), or vehicle only (1 ml/kg each orally and subcutaneously). Female rats showed an increased pain threshold (analgesia) after subcutaneous buprenorphine but no change in pain threshold after either oral buprenorphine or vehicle only. Estrous cycle stage (proestrus versus diestrus) did not affect the analgesic effects of buprenorphine, but rats in proestrus showed significantly lower pain thresholds (less tolerance to pain) than did those in diestrus. These results show that the oral dose of buprenorphine recommended for postoperative analgesic care does not induce significant analgesia in female rats and therefore is not as effective as the standard subcutaneous dose.

Analgesic efficacy of orally administered buprenorphine in rats: methodologic considerations.

Buprenorphine has been widely recommended for treatment of pain in rodents. We have previously documented that the recommended postoperative oral dose of buprenorphine in male Long-Evans rats, 0.5 mg/kg, is not as effective as the recommended parenteral dose of buprenorphine (0.05 mg/kg, s.c.) as an analgesic. In the series of experiments reported here, we compared: the analgesic effect of buprenorphine when prepared in two ways in the laboratory with that of a commercially available injectable solution of buprenorphine; the analgesic effect of buprenorphine in Long-Evans rats with that in Sprague-Dawley rats; and Long-Evans and Sprague-Dawley rats for development of pica, a commonly reported side effect of buprenorphine. We followed the pica experiment with assessment of the effectiveness of buprenorphine in establishing a conditioned flavor aversion. The results indicated that method of preparation did not result in any significant differences in the efficacy of injected buprenorphine. Strain of rat was not associated with a significant difference in the efficacy of buprenorphine. However, a significant strain difference was found in development of pica. Buprenorphine treatment was effective in inducing a conditioned flavor aversion. We concluded that the recommended oral dose of buprenorphine (0.5 mg/kg) is ineffective as an analgesic, and that this was not the result of method of preparation of the buprenorphine or strain of rat used. Furthermore, we concluded that buprenorphine treatment may induce gastrointestinal distress in both strains tested. The results reaffirm our previous conclusion that oral administration of buprenorphine at 0.5 mg/kg, despite the general recommendation, is not a reasonable treatment for postsurgical pain in rats.

Ingested placenta blocks the effect of morphine on gut transit in Long-Evans rats.

Opioids produce antinociception, and ingested placenta or amniotic fluid modifies that antinociception. More specifically, ingested placenta enhances the antinociception produced by selective activation of central kappa-opioid or delta-opioid receptors but attenuates that produced by activation of central mu-opioid receptors. Opioids also slow gut transit by acting on central or peripheral mu-opioid receptors. Therefore, we hypothesized that ingested placenta would reverse the slowing of gut transit that is produced by morphine, a preferential mu-opioid-receptor agonist. Rats were injected with morphine either centrally or systemically and fed placenta, after which gastrointestinal transit was evaluated. We report here that ingested placenta reversed the slowing of gut transit produced by centrally administered morphine but did not affect the slowing of gut transit produced by systemically administered morphine. These results suggest another likely consequence of placentophagia at parturition in mammals--reversal of opioid-mediated, pregnancy-based disruption of gastrointestinal function--as well as an important consideration in opioid-based treatments for pain in humans--enhancement of desirable effects with attenuation of adverse effects.

Placenta ingestion by rats enhances delta- and kappa-opioid antinociception, but suppresses mu-opioid antinociception.

Ingestion of placenta or amniotic fluid produces a dramatic enhancement of centrally mediated opioid antinociception in the rat. The present experiments investigated the role of each opioid receptor type (mu, delta, kappa) in the antinociception-modulating effects of Placental Opioid-Enhancing Factor (POEF-presumably the active substance). Antinociception was measured on a 52 degrees C hotplate in adult, female rats after they ingested placenta or control substance (1.0 g) and after they received an intracerebroventricular injection of a delta-specific ([D-Pen2,D-Pen5]enkephalin (DPDPE); 0, 30, 50, 62, or 70 nmol), mu-specific ([D-Ala2,N-MePhe4,Gly5-ol]enkephalin (DAMGO); 0, 0.21, 0.29, or 0.39 nmol), or kappa-specific (U-62066; spiradoline; 0, 100, 150, or 200 nmol) opioid receptor agonist. The results showed that ingestion of placenta potentiated delta- and kappa-opioid antinociception, but attenuated mu-opioid antinociception. This finding of POEF action as both opioid receptor-specific and complex provides an important basis for understanding the intrinsic pain-suppression mechanisms that are activated during parturition and modified by placentophagia, and important information for the possible use of POEF as an adjunct to opioids in pain management.

Male-induced estrus synchronization in the female Siberian hamster (Phodopus sungorus sungorus).

Olfactory cues play an integral role in the organization of events that mediate reproductive success. In a variety of species, priming pheromones, in particular, are important for ensuring reproductive fitness. To date, very little research has focused on how male-emitted priming pheromones, such as those that regulate the onset of puberty and estrus synchronization in females, affect the reproductive physiology of the female Siberian hamster (Phodopus sungorus sungorus). This lack of research may be due to the physiology of the Phodopus genus; vaginal cytology cannot be used as a reliable indicator of estrus or ovulation. Using a jugular cannulation technique to determine estrous stage by blood analysis of prolactin and luteinizing hormone, we sought to determine if male priming pheromones affect estrous cyclicity in the female Siberian hamster and, if so, whether the production of these priming pheromones is androgen dependent. Our results showed that females exposed to bedding from mature, intact males showed a significantly higher incidence of proestrus 3 days later than did females exposed to the bedding of mature, gonadectomized males. Therefore, we found that not only do male Siberian hamsters emit chemical signals that induce estrus synchronization, but also that this ability is likely to be androgen dependent.