Prenatal Morphine Research Articles

Disrupted neuregulin 1-ErbB4 signaling: Consequences of prenatal morphine exposure in rat pups and molecular gateway to neurological impairment

ObjectiveMorphine exposure during pregnancy has detrimental effects on both the mother and her offspring, both during and after childbirth. This study aimed to investigate the impact of prenatal morphine exposure on rat pups and dams, specifically focusing on changes in Neuregulin-1 (Nrg-1)/ErbB4 gene expression, inflammation, and brain-derived neurotrophic factor (BDNF) levels. Materials and methodsTwenty female rats were randomized into two experimental groups:1-Morphine Group: Dams received morphine throughout pregnancy. 2-Control Group: Dams received no interventions.At the end of gestation, blood samples were collected from the dams. Subsequently, dams and their pups underwent tissue collection from the cortical area of the brain to evaluate the following parameters: Interleukin-6 (IL-6), Interleukin-10 (IL-10), total antioxidant capacity (TAC), Malondialdehyde (MDA), and Brain-derived neurotrophic factor (BDNF).Additionally, RNA was extracted from the pup's cortical brain tissue for the assessment of gene expression levels of Neuregulin-1 (NRG-1) and ErbB-4 using quantitative real-time polymerase chain reaction (qrt-PCR). ResultsThe molecular investigation revealed a decrease in NRG-1 and ErbB-4 expressions in the brain cortex of offspring exposed to morphine during prenatal development. Additionally, the levels of IL-6 and IL-10 in both the serum and brain of both the mothers and their offspring in the morphine group were significantly higher compared to the control group. The morphine-exposed group also exhibited significantly lower levels of TAC and higher levels of MDA, indicating increased oxidative stress. Furthermore, the levels of BDNF in the morphine group were significantly lower compared to the control group. ConclusionPrenatal morphine exposure in rats has detrimental effects on both the dams and their offspring. This study demonstrates that prenatal morphine exposure disrupts critical molecular pathways involved in neurodevelopment, inflammation, oxidative stress, and neurotrophic signaling. These findings suggest that prenatal morphine exposure can have long-lasting consequences for the offspring, potentially contributing to neurodevelopmental disorders and other health issues later in life.

Enhancement of neuronal excitability in the medial prefrontal cortex following prenatal morphine exposure

The clinical use and abuse of opioids during human pregnancy have been widely reported. Several studies have demonstrated that opioids cross the placenta in rats during late gestation, and prenatal morphine exposure has been shown to have negative outcomes in cognitive function. The medial prefrontal cortex (mPFC) is believed to play a crucial role in cognitive processes, motivation, and emotion, integrating neural information from several brain areas and sending converted information to other structures. Dysfunctions in this area have been observed in numerous psychiatric and neurological disorders, including addiction. This current study aimed to compare the electrophysiological properties of mPFC neurons in rat offspring prenatally exposed to morphine. Pregnant rats were injected with morphine or saline twice a day from gestational days 11–18. Whole-cell patch-clamp recordings were performed in male offspring on postnatal days 14–18. All recordings were obtained in current-clamp configuration from mPFC pyramidal neurons to assess their electrophysiological properties. The results revealed that prenatal exposure to morphine shifted the resting membrane potential (RMP) to less negative voltages and increased input resistance and duration of action potentials. However, the amplitude, rise slope, and afterhyperpolarization (AHP) amplitude of the first elicited action potentials were significantly decreased in rats prenatally exposed to morphine. Moreover, the sag voltage ratio was significantly decreased in the prenatal morphine group. Our results suggest that the changes observed in the electrophysiological properties of mPFC neurons indicate an elevation in neuronal excitability following prenatal exposure to morphine.

Prenatal Morphine Exposure Increases Cardiovascular Disease Risk and Programs Neurogenic Hypertension in the Adult Offspring.

The opioid overdose and opioid use disorder epidemics are concomitant with increased metabolic and CVD risk. Although opioid use disorder causes adverse pregnancy outcomes, the offspring's cardiovascular health is understudied. We hypothesized that offspring exposed to in utero morphine exposure (IUME) would show increased CVD risk factors and endogenous opioid system dysregulation. Sprague Dawley dams were treated with saline (vehicle, n=10) or escalating doses of morphine (5-20 mg/kg per day, SC, n=10) during gestation. Cardiovascular and metabolic parameters were assessed in adult offspring. Litter size and pups' birth weight were not different in response to IUME. Female and male IUME offspring showed reduced body length at birth (P<0.05) and body weight from weeks 1 to 3 of life (P<0.05), followed by a catch-up growth effect. By week 16, female and male IUME rats showed reduced tibia length (P<0.05) and fat mass. IUME increases the mean arterial pressure and the depressor response to mecamylamine (5 mg/kg per day, IP) induced by IUME were abolished by a chronic treatment with an alpha-adrenergic receptor blocker (prazosin; 1 mg/kg per day, IP). Although circulating levels of angiotensin peptides were similar between groups, IUME exacerbated maximal ex vivo Ang (angiotensin) II-induced vasoconstriction (P<0.05) and induced endothelial dysfunction in a sex-specific manner (P<0.05). Proenkephalin, an endogenous opioid peptide that lowers blood pressure and sympathetic-mediated vasoconstriction, showed reduced mRNA expression in the heart, aorta, and kidneys from morphine versus vehicle group (P<0.05). Among the effects of IUME, neurogenic hypertension, vascular dysfunction, and metabolic dysfunction could be associated with the dysregulation of the endogenous opioid system.

Prenatal exposure to morphine enhances excitability in locus coeruleus neurons.

Opioid abuse during pregnancy may have noteworthy effects on the child's behavioral, emotional and cognitive progression. In this study, we assessed the effect of prenatal exposure to morphine on electrophysiological features of locus coeruleus (LC) noradrenergic neurons which is involved in modulating cognitive performance. Pregnant dams were randomly divided into two groups, that is a prenatal saline treated and prenatal morphine-treated group. To this end, ongestational days 11-18, either morphine or saline (twice daily, s.c.) was administered to pregnant dams. Whole-cell patch-clamp recordings were conducted on LC neurons of male offspring. The evoked firing rate, instantaneous frequency and action potentials half-width, and also input resistance of LC neurons significantly increased in the prenatal morphine group compared to the saline group. Moreover, action potentials decay slope, after hyperpolarization amplitude, rheobase current, and first spike latency were diminished in LC neurons following prenatal exposure to morphine. In addition, resting membrane potential, rise slope, and amplitude of action potentials were not changed by prenatal morphine exposure. Together, the current findings show a significant enhancement in excitability of the LC neurons following prenatal morphine exposure, which may affect the release of norepinephrine to other brain regions and/or cognitive performances of the offspring.

Prenatal Morphine Exposure Differentially Alters Addictive and Emotional Behavior in Adolescent and Adult Rats in a Sex-Specific Manner.

The effects of prenatal opioid exposure in adult animals has been widely studied, but little is known about the effects of prenatal opioid on adolescents. Most of the risk behaviors associated with drug abuse are initiated during adolescence. The developmental state of the adolescent brain makes it vulnerable to initiate drug use and susceptible to drug-induced brain changes. In this study, pregnant rats were subcutaneously injected with an increasing dose of morphine (5mg/kg, 7mg/kg, 10mg/kg) for 9 days since the gestation day 11. The effects of prenatal morphine (PNM) on learning and memory, anxiety- and depressive- like behavior, morphine induced conditioned place preference (CPP) as well as locomotor sensitization were tested in both adolescent and adult rats. The results showed that: (1) PNM decreased anxiety-like behavior in both adolescent and adult female rats, but not males; (2) PNM decreased depressive-like behavior in adolescent but increased depressive -like behavior in adult females; (3) PNM increased low dose morphine induced locomotor sensitization in females; (4) PNM decreased tyrosine hydroxylase (TH) expression in the prefrontal cortex but decreased dopamine D1 receptor expression in the nucleus-accumbens (NAc) in female rats. These results suggested that PNM altered the emotional and addictive behavior mainly in female rats, with female rats being less anxiety and depressive during adolescence, but more depressive in adult, and more sensitive to low dose morphine induced locomotor activity sensitization, which might be mediated in part by the differential expression of the TH, dopamine D1 receptors in the female brain.

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

Prenatal morphine exposure increases gamma oscillation and theta coherence in the rat reward system

Previous studies have found that prenatal morphine (PNM) exposure leads to both increased and decreased risk of substance abuse in offspring. Understanding more about the neurobiological changes after the PNM exposure would help to understand more about this issue. Signaling from dopamine neurons of the ventral tegmental area (VTA) in the mesoaccumbal and mesocortical pathways plays a vital role in drug dependency. To provide further knowledge about the effects of PNM on drug seeking behavior and the dopamine system. We recorded local field potentials (LFP) simultaneously in the VTA, NAc (nucleus accumbens), BLA (basolateral amygdala) and mPFC (medial prefrontal cortex) in male adult rats prenatally treated with saline or morphine. Morphine (10 mg/kg) induced conditioned place preference (CPP) establishment, extinction and priming were tested to investigate the effects of PNM on addictive-like behavior. In addition, the expression of nuclear histone deacetylases (HDAC4, HDAC5), which plays essential epigenetic roles in neuroplasticity after drug use were also tested in VTA and NAc. The results showed that PNM did not change the acquisition of morphine CPP in male rats, but impaired CPP extinction and morphine (5 mg/kg) - primed reinstatement of CPP after extinction. PNM increased the low gamma (30–60 Hz) and high (60–90 Hz) gamma LFP powers in NAc and BLA. PNM also leads to increased theta (4–9 Hz) coherence between VTA and NAc, and increased HDAC5 expression in VTA. After chronic morphine administration, coherence between VTA-NAc, mPFC-NAc and mPFC-BLA increased significantly in PNS rats, but no changes were find in PNM rats, indicating impaired plasticity in brain circuits. All these results suggest that PNM exposure increased reward processing in adult male rats, but impaired morphine CPP extinction and reinstatement, which relate to decreases network plasticity and increased HDAC5 expression in the reward system.

Prenatal exposure to morphine impairs attention and impulsivity in adult rats.

An alarming number of neonates born with prenatal exposure to morphine has resulted from the opioid epidemic; however, the long-term effects of prenatal opioid exposure on offspring behavior remain relatively unknown. In this study, we evaluated whether prenatal exposure to the mu opioid receptor agonist, morphine, has enduring effects on cognitive functions in adult life. On embryonic days 11-18 (E11-E18), female pregnant rats were injected subcutaneously with either morphine or saline twice daily. Adult male offspring that was prenatally exposed to saline or morphine was trained in the 5-choice serial reaction time test (5-CSRTT) to test their cognitive abilities under baseline conditions. Next, these rats were treated with saline (1ml/kg), naloxone (1mg/kg), and acute morphine (1, 3, 5mg/kg), subcutaneously, once daily and following drug challenges rats were tested in the 5-CSRTT. Meanwhile, behavioral performance on training days between opioid drug challenges were analyzed to monitor possible drug-induced shifts in baseline performance. As a final experiment in order to investigate subchronic exposure to morphine, rats were injected with 5mg/kg morphine for 5days and then naloxone in the last day of the experiment (day 6). Firstly, during acquisition of a stable baseline in the training phase, rats prenatally exposed to morphine showed delayed learning of the task demands. Furthermore, under baseline responding the rats prenatally exposed to morphine showed declined inhibitory control demonstrated by increased impulsive and compulsive-like responding compared to rats prenatally exposed to saline. Moreover, acute and subchronic morphine challenges in the rats prenatally exposed to morphine caused a deficit in visuospatial attention in comparison with saline treatment as well as the rats prenatally exposed to saline. These effects were abolished by naloxone. The current findings indicate a direct causal effect of prenatal morphine exposure on inhibitory control and task learning later in life, as well as deficits in attention following morphine exposure in adulthood.

Enhanced H3K4 Trimethylation in TNF-α Promoter Gene Locus with Cell Apoptosis in the Ventral-Medial Striatum following Opioid Withdrawal of Neonatal Rat Offspring from Morphine-Addicted Mothers.

Prenatal opioid exposure might disturb epigenetic programming in the brain of neonatal offspring with various consequences for gene expressions and behaviors. This study determined whether altered trimethylation of histone 3 at lysine 4 (H3K4me3) in the promoter of the tumor necrosis factor-α (tnf-α) gene with neural cell apoptosis was involved in the ventral-medial striatum, an important brain region for withdrawal symptoms, of neonatal rat offspring from morphine-addicted mothers. Female adult rats were injected with morphine before gestation and until 14 days after giving birth. On postnatal day 14 (P14), rat offspring from morphine-addicted mothers were subjected to an opioid-withdrawal protocol and were analyzed 2 or 8 h after administration of that protocol. Expressions of the TNF-α protein, H3K4me3 in the tnf-α promoter gene, and neural cell apoptosis within the ventral-medial striatum of neonatal rat offspring were evaluated. In the absence of significant opioid withdrawal (2 h after initiation of the opioid-withdrawal protocol on P14), prenatal morphine exposure led to increased levels of H3K4me3 in the tnf-α promoter gene, of the TNF-α protein, and of neural cell apoptosis within the ventral-medial striatum of neonatal rat offspring. Following opioid withdrawal (8 h after initiation of the opioid-withdrawal protocol on P14), differential expression of H3K4me3 in the tnf-α promoter gene locus and upregulation of the level of TNF-α protein expression were further enhanced in these offspring. In addition, increased levels of caspase-3 and neural cell apoptosis were also observed. Taken together, this study revealed that prenatal opioid exposure can activate an epigenetic histone mechanism which regulates proinflammatory factor generation, which hence, led to cell apoptotic damage within the ventral-medial striatum of neonatal rat offspring from morphine-addicted mothers. More importantly, the opioid-withdrawal episode may provide augmented effects for the abovementioned alterations and could lead to deleterious effects in the neonatal brain of such offspring.

Opioid withdrawal behavior in spiny mice: A novel preclinical model of neonatal opioid withdrawal syndrome (NOWS)

As the opioid epidemic continues to grow, opioid use among pregnant women is increasing significantly. This has led to a steady rise in the number of infants born with neonatal opioid withdrawal syndrome (NOWS). Although short-term withdrawal symptoms associated with NOWS are well characterized, there are many gaps in our understanding of the short and long-term effects of prenatal opioid exposure. Current animal models of NOWS are limited by shortened gestational periods, large litter sizes, and primary organogenesis occurring after birth. This often leads to postnatal treatment to mimic drug exposure during third-trimester development. Using the unique rodent species Acomys cahirinus, more commonly known as spiny mice, which have an extended 40-day gestation period, small litter sizes, and increased in utero organogenesis we aim to study the short-term effects of prenatal morphine exposure by assessing withdrawal behavior. To model maternal opioid use, dams were treated daily with morphine (10 and 30 mg/kg S.C.) beginning on gestation day 19 until the day of birth; this resulted in a cumulative exposure of 19–21 days. Withdrawal behaviors for each pup were recorded daily between postnatal days 0–7 (PND 0–7). Our study found that prenatal morphine exposure in spiny mice led to an increase in withdrawal behavior throughout the early postnatal period and validated the use of this species as a novel pre-clinical model of NOWS. We are hopeful this rodent model will further our understanding of the short and long-term consequences of prenatal opioid exposure on neurodevelopment and behavior.

Long‐term Cardiorespiratory Anomalies in Rat Offspring Following Prenatal Morphine Exposure

The incidence of opioid‐related pregnancies is a growing public health concern. Although not all infants born from opioid using mothers require hospitalization, many exhibit a variety of withdrawal symptoms (Neonatal Opioid Withdrawal Syndrome, NOWS) often requiring weeks of neonatal intensive care. There are longer‐term effects of prenatal opioid use including altered CNS function comprising cognitive impairments, neurobehavioral abnormalities and increased risk of Sudden Infant Death Syndrome (SIDS). In fact, abnormal cardiorespiratory regulation in infants born following prenatal opioid exposure includes reduced vagal tone, tachypnea, delayed arousal, and altered ventilatory responses to hypoxia and hypercapnia. In the present study, we used a rat model to characterize long‐term (postnatal (P) 10 days of age) cardiorespiratory responses of the offspring following in utero morphine exposure. Pregnant dams received twice daily (morning and evening) subcutaneous injections of various doses of morphine (1, 5 or 10mg/kg) during the 3rd week of pregnancy to mimic 3rd trimester maternal opioid use. Following birth, rat pups (and the nursing dam) were left untreated and whole‐body plethysmography was used to assess the acute hypoxic (HVR) and hypercapnic (HCVR) ventilatory responses at P10 days of age. Heart rate was also assessed in rats fitted with a custom‐made jacket with built‐in ECG electrodes. Generally, the postnatal effects of morphine were dose‐dependent: 1) increasing doses of in utero morphine exposure caused a progressive attenuation of the HCVR in 10 day old rats; 2) there was a high rate of neonatal (within 24hrs of birth) mortality in the highest (10mg/kg) prenatal dose; 3) however, the rats who survived exhibited both an attenuated HVR and HCVR; and 4) rats also exhibited resting tachycardia and tachypnea (at 5–10 mg/kg) compared to in utero saline treated rats. These data demonstrate a long‐term (10 days) effect of prenatal morphine exposure on resting cardiorespiratory function, including impairments in ventilatory defense responses to acute hypoxia and hypercapnia. These cardiorespiratory anomalies could offer a window of insight into the pathophysiology of the increased risk of SIDS seen in infants born from mothers of substance use.

Long-time effects of prenatal morphine, tramadol, methadone, and buprenorphine exposure on seizure and anxiety in immature rats

Purpose: This study aimed to investigate seizures and anxiety-like behaviors in immature rats prenatally exposed to opiate drugs.Materials and methods: Pregnant rats were randomly divided into five groups: saline, morphine, tramadol, methadone, and buprenorphine. Administrations were performed intraperitoneally once a day for the last seven days of pregnancy. Neonatal rats were subdivided into ten groups, split according to sex. Anxiety-like behavior was tested on postnatal day (PD) 19. On PD 20, seizure was induced by PTZ injection.Results: Morphine in male rats had an increased time to onset (p < 0.005), whereas there was a decreased number of tonic-clonic seizures in females (p < 0.05). Tramadol had an increased duration of tonic-clonic seizures compared to morphine and methadone in males (p < 0.005). Moreover, tramadol decreased open arm time and locomotor activity in males more than in females (p < 0.05). Methadone decreased open arm time in males (p < 0.05). Furthermore, buprenorphine and tramadol decreased open arm entrance in male rats (p < 0.05).Conclusions: It was demonstrated that prenatal tramadol significantly increases both the duration of seizures and anxiety-like behaviors in immature male rats, whereas morphine decreases both of them. The effects of tramadol on seizure and anxiety-like behavior may be due to the comorbid occurrence of the symptoms of these two disorders.

Long-Lasting Alterations in Gene Expression of Postsynaptic Density 95 and Inotropic Glutamatergic Receptor Subunit in the Mesocorticolimbic System of Rat Offspring Born to Morphine-Addicted Mothers.

Prenatal exposure to morphine causes altered glutamatergic neurotransmission, which plays an important pathophysiological role for neurobiological basis of opiate-mediated behaviors in such offspring. However, it is still not clear whether such alteration involves gene expression of ionotropic glutamate receptor subunits. In this study, we further studied whether prenatal morphine exposure resulted in long-term changes in the gene expression of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor, N-methyl-d-aspartate (NMDA) receptor, and postsynaptic density 95 in the mesocorticolimbic area (an essential integration circuitry for drug craving behavior), nucleus accumbens (NAc), ventral tegmental area (VTA), and prefrontal cortex (PFC), of rat offspring from morphine-addicted mothers. Experimental results showed that prenatal morphine exposure led to a persistent downregulation of gene expression in the AMPA and NMDA receptor subunit, with a differential manner of decreased magnitudes, at the age of postnatal days 14 (P14) and P30. However, in PFC, the gene expression of the AMPA receptor subunit was not synchronized in observed rat offspring subjected to prenatal morphine exposure. An upregulation of gene expression in the AMPA receptor subunit 3 (GluR3) was persistently observed at P14 and P30. Furthermore, the gene expressions of PSD-95 in NAc, VTA, and PFC were all decreased concurrently. Collectively, the results suggest that prenatal exposure to morphine may initiate molecular mechanisms leading to a long-lasting, differential alteration in gene expression of the inotropic glutamate receptor subunit and PSD-95 in the mesocorticolimbic circuitry in rat offspring. This study raises a possibility in which differential changes in gene expression with a long-lasting manner may play a role for the development of nearly permanent changes in opiate-mediated behaviors, at least in part for the neurobiological pathogenesis in offspring.

Deleterious effects of prenatal exposure to morphine on the spatial learning and hippocampal BDNF and long-term potentiation in juvenile rats: Beneficial influences of postnatal treadmill exercise and enriched environment

Prenatal morphine exposure causes a variety of neurobehavioral alterations observed in later life. The present study investigated the effects of postnatal exercise and enriched environment (EE) on alterations in water maze learning and hippocampal long-term potentiation (LTP) and brain derived neurotrophic factor (BDNF) levels induced by exposure to morphine during prenatal period in rats. On gestation days 11–18, pregnant rats were injected twice daily with saline or morphine. Offspring were subjected to postnatal exercise and EE for 30 days and afterward, spatial learning and hippocampal LTP and BDNF levels were investigated. Prenatal morphine-exposure impaired the spatial learning and hippocampal LTP in both male and female offspring. Interestingly, postnatal exercise and EE increased performance in the water maze and improved LTP in both prenatally saline and morphine-exposed male and female rats. Prenatal morphine exposure also caused a reduction in the hippocampal BDNF levels in the female, but not male rats, and postnatal exercise and EE alleviated this deficit. Our results demonstrate that postnatal exercise and EE can improve deficits in water maze learning and hippocampal LTP and BDNF levels caused by prenatal morphine exposure.

Prenatal morphine exposure during late embryonic stage enhances the rewarding effects of morphine and induces the loss of membrane-bound protein kinase C-α in intermediate medial mesopallium in the chick

The susceptibility to drug abuse may be associated with the structural and/or functional changes in the reward-related brain regions induced by drug exposure during sensitive periods of embryonic development. Previously, we have found that prenatal morphine exposure during embryonic days 17–20 may be crucial for developing the susceptibility to morphine reward after hatching. However, the underlying structure and cellular mechanisms need further investigation. In the present study, the chicks of a few days old, which were prenatally exposed to morphine during E17-20, obviously showed higher preference for the morphine-paired chamber and hyperactivity during the expression of morphine conditioned place preference (CPP), and the reduction in membrane-bound of PKCα of the bilateral intermediate medial mesopallium (IMM) assayed immunologically. These results indicate that the decreased expression of PKCα in IMM may participate in the development of the susceptibility to the rewarding effects of morphine in chicks prenatally exposed to morphine, and provide further support for the cross-species evolutionary concordance among amniotes.

Implication of protein kinase C of the left intermediate medial mesopallium in memory impairments induced by early prenatal morphine exposure in one-day old chicks

Previously we reported that prenatal morphine exposure during embryonic days 5–8 can cause cognitive deficits of one-trial passive avoidance learning (PAL) in one-day old chicks. Because protein kinase C (PKC) has been associated with memory capacity, we investigated the effects of prenatal morphine exposure on PKC isoforms expression in the left intermediate medial mesopallium (IMM) of chick brain at a time when memory tests were performed at 30, 120 and 360min respectively following training in PAL paradigm. We found that the level of PKCα in the membrane fractions in left IMM was decreased but that in the cytosol fractions showed a increased trend in prenatally morphine-exposed chicks with impaired long-term memory (120 and 360min). Moreover, the translocation of PKC δ from cytosol to membrane in left IMM was shown in prenatal morphine group which had significantly impaired long-term memory at 360min after training. Furthermore, there were no statistical differences between the two groups regarding the expressions of PKCα and PKC δ in the membrane fraction, although their levels in the cytosol fraction of prenatal morphine group which showed impaired intermediate-term memory at 30min after training, were quite different from that of prenatal saline group. Taken together, these results indicate that PKCα and PKC δ in the left IMM are differentially involved in the impairments of long-term memory induced by prenatal morphine exposure. Neither PKCα nor PKC δ in left IMM may be associated with the disruption of intermediate-term memory of chicks prenatally exposed to morphine.

Delayed extinction and stronger drug-primed reinstatement of methamphetamine seeking in rats prenatally exposed to morphine

Prenatal morphine (PM) affects the development of brain reward system and cognitive function. The present study aimed to determine whether PM exposure increases the vulnerability to MA addiction. Pregnant Sprague-Dawley rats were administered saline or morphine during embryonic days 3–20. The acquisition, extinction and reinstatement of methamphetamine (MA) conditioned place preference (CPP) and intravenous self-administration (SA) paradigms were assessed in the male adult offspring. There was no difference in the acquisition and expression of MA CPP between saline- and PM-exposed rats, whereas PM-exposed rats exhibited slower extinction and greater MA priming-induced reinstatement of drug-seeking behavior than controls. Similarly, MA SA under progressive ratio and fixed ratio schedules was not affected by PM exposure, but PM-exposed rats required more extinction sessions to reach the extinction criteria and displayed more severe MA priming-, but not cue-induced, reinstatement. Such alterations in extinction and reinstatement were not present when PM-exposed rats were tested in an equivalent paradigm assessing operant responding for food pellets. Our results demonstrate that PM exposure did not affect the association memory formation during acquisition of MA CPP or SA, but impaired extinction learning and increased MA-primed reinstatement in both tasks. These findings suggest that the offspring of women using morphine or heroin during pregnancy might predict persistent MA seeking during extinction and enhanced propensity to MA relapse although they might not be more susceptible to the reinforcing effect of MA during initiation of drug use.

Effects of environmental enrichment on behavioral deficits and alterations in hippocampal BDNF induced by prenatal exposure to morphine in juvenile rats

Prenatal morphine exposure throughout pregnancy can induce a series of neurobehavioral and neurochemical disturbances by affecting central nervous system development. This study was designed to investigate the effects of an enriched environment on behavioral deficits and changes in hippocampal brain-derived neurotrophic factor (BDNF) levels induced by prenatal morphine in rats. On pregnancy days 11–18, female Wistar rats were randomly injected twice daily with saline or morphine. Offspring were weaned on postnatal day (PND) 21. They were subjected to a standard rearing environment or an enriched environment on PNDs 22–50. On PNDs 51–57, the behavioral responses including anxiety and depression-like behaviors, and passive avoidance memory as well as hippocampal BDNF levels were investigated. The light/dark (L/D) box and elevated plus maze (EPM) were used for the study of anxiety, forced swimming test (FST) was used to assess depression-like behavior and passive avoidance task was used to evaluate learning and memory. Prenatal morphine exposure caused a reduction in time spent in the EPM open arms and a reduction in time spent in the lit side of the L/D box. It also decreased step-through latency and increased time spent in the dark side of passive avoidance task. Prenatal morphine exposure also reduced immobility time and increased swimming time in FST. Postnatal rearing in an enriched environment counteracted with behavioral deficits in the EPM and passive avoidance task, but not in the L/D box. This suggests that exposure to an enriched environment during adolescence period alters anxiety profile in a task-specific manner. Prenatal morphine exposure reduced hippocampal BDNF levels, but enriched environment significantly increased BDNF levels in both saline- and morphine-exposed groups. Our results demonstrate that exposure to an enriched environment alleviates behavioral deficits induced by prenatal morphine exposure and up-regulates the decreased levels of BDNF. BDNF may contribute to the beneficial effects of an enriched environment on prenatal morphine-exposed to rats.

Effects of treadmill running exercise during the adolescent period of life on behavioral deficits in juvenile rats induced by prenatal morphine exposure

Prenatal exposure to morphine throughout pregnancy results in an array of prolonged or permanent neurochemical and behavioral deficits, including deficits in learning and memory in children of addicted mothers. This study investigated the effects of forced exercise on behavioral deficits of pups born to mothers addicted to morphine in rats. After mating and ensuring of pregnancy of female Wistar rats, they were divided into morphine or saline groups and in the second half of pregnancy (on days 11–18 of gestation) were injected subcutaneously with morphine or saline, respectively. Pups were weaned at postnatal day (PND) 21 and trained at mild intensity on a treadmill 20days. On PND 41–47, the behavioral responses were studied. Light/dark (L/D) box and elevated plus maze (EPM) apparatus were used for investigation of anxiety, shuttle box and forced swimming tests were used to assess passive avoidance learning and memory and depression behavior, respectively. The results showed that prenatal morphine exposure caused reductions in time spent in light compartment of L/D box and EPM open arm, while postnatal exercise reversed these effects. We also found that prenatal morphine exposure caused a reduction in step through latency in passive avoidance memory test and exercise counteracted with this effect. Performance in the forced swimming test did not affected by prenatal morphine exposure or postnatal exercise. Exercise seems to be one of the strategies in reduction of behavioral deficits of children born to addicted mothers to morphine.

Impaired contextual fear extinction and hippocampal synaptic plasticity in adult rats induced by prenatal morphine exposure.

Prenatal opiate exposure causes a series of neurobehavioral disturbances by affecting brain development. However, the question of whether prenatal opiate exposure increases vulnerability to memory-related neuropsychiatric disorders in adult offspring remains largely unknown. Here, we found that rats prenatally exposed to morphine (PM) showed impaired acquisition but enhanced maintenance of contextual fear memory compared with control animals that were prenatally exposed to saline (PS). The impairment of acquisition was rescued by increasing the intensity of footshocks (1.2 mA rather than 0.8 mA). Meanwhile, we also found that PM rats exhibited impaired extinction of contextual fear, which is associated with enhanced maintenance of fear memory. The impaired extinction lasted for 1 week following extinction training. Furthermore, PM rats exhibited reduced anxiety-like behavior in the elevated plus-maze and light/dark box test without differences in locomotor activity. These alterations in PM rats were mirrored by abnormalities in synaptic plasticity in the Schaffer collateral-CA1 synapses of the hippocampus in vivo. PS rats showed blocked long-term potentiation and enabled long-term depression in CA1 synapses following contextual fear conditioning, while prenatal morphine exposure restricted synaptic plasticity in CA1 synapses. The smaller long-term potentiation in PM rats was not further blocked by contextual fear conditioning, and the long-term depression enabled by contextual fear conditioning was abolished. Taken together, our results provide the first evidence suggesting that prenatal morphine exposure may increase vulnerability to fear memory-related neuropsychiatric disorders in adulthood.