Operant conditioning with a stimulus discrimination: An alternative method for evaluating alcohol reinforcement in preweaning rats.

BACKGROUND: Ethanol exposure at early ontogeny promotes further predisposition to consume the drug. Operant conditioning allows motivational alcohol properties to be assessed. To date, the operant conditioning approach used during infancy consisted in paired subjects being trained to learn an operant response, using simultaneously a yoked partner, which received reinforcer solution as a result of a paired animal instrumental response (OYS). NEW METHOD: In our study, we attempted to evaluate ethanol reinforcing effects during PDs 15-18 in an operant conditioning schedule with a stimulus discrimination procedure (OSD), as an alternative control learning. This new proposal includes a single subject, who has to choose between an S+ nose-poke hole, which delivers the reinforcer into the mouth, or an S- nose-poke hole with no reinforcement effect. RESULTS: The OSD results seemed to be more reliable than those obtained using the OYS procedure, since some data appeared to be more robust when using a yoked nose-poke hole than when employing a yoked subject, such as in control learning. Consequently, OSD has the following advantages compared to the OYS procedure: a) the operant response learned is controlled by the overall behavior of the same subject, resulting in a relatively clearer data; b) a yoked animal is not necessary, thereby reducing the number of rats used in the operant conditioning procedure. COMPARISON WITH EXISTING METHODS AND CONCLUSIONS: A novel technique of operant conditioning adapted to infancy was developed by training animals to emit a particular response to gain access to alcohol solution as a reinforcer.

Fetal Alcohol Programming of Subsequent Alcohol Affinity: A Review Based on Preclinical, Clinical and Epidemiological Studies.

The anatomo-physiological disruptions inherent to different categories of the Fetal Alcohol Spectrum Disorder do not encompass all the negative consequences derived from intrauterine ethanol (EtOH) exposure. Preclinical, clinical and epidemiological studies show that prenatal EtOH exposure also results in early programming of alcohol affinity. This affinity has been addressed through the examination of how EtOH prenatally exposed organisms recognize and prefer the drug's chemosensory cues and their predisposition to exhibit heightened voluntary EtOH intake during infancy and adolescence. In altricial species these processes are determined by the interaction of at least three factors during stages equivalent to the 2nd and 3rd human gestational trimester: (i) fetal processing of the drug's olfactory and gustatory attributes present in the prenatal milieu; (ii) EtOH's recruitment of central reinforcing effects that also imply progressive sensitization to the drug's motivational properties; and (iii) an associative learning process involving the prior two factors. This Pavlovian learning phenomenon is dependent upon the recruitment of the opioid system and studies also indicate a significant role of EtOH's principal metabolite (acetaldehyde, ACD) which is rapidly generated in the brain via the catalase system. The central and rapid accumulation of this metabolite represents a major factor involved in the process of fetal alcohol programming. According to recent investigations, it appears that ACD exerts early positive reinforcing consequences and antianxiety effects (negative reinforcement). Finally, this review also acknowledges human clinical and epidemiological studies indicating that moderate and binge-like drinking episodes during gestation result in neonatal recognition of EtOH's chemosensory properties coupled with a preference towards these cues. As a whole, the studies under discussion emphasize the notion that even subteratogenic EtOH exposure during fetal life seizes early functional sensory and learning capabilities that pathologically shape subsequent physiological and behavioral reactivity towards the drug.

Prenatal ethanol exposure and enkephalinergic neurotransmission.

Endogenous opioids (enkephalins, endorphins and dynorphins) are small peptides that play a main role in pain perception and analgesia, as well as in alcohol (ethanol) reinforcement and reward. Alcohol reinforcement involves the ethanol-induced activation of the endogenous opioid system, a process that may augment the hedonic value and the reinforcing properties of the drug, which in turn increases substance consumption. Changes in opioidergic transmission may contribute to alcohol intoxication and to the neuroadaptive responses produced by the long-lasting exposure to ethanol. Opioidergic transmission may be altered by ethanol at distinct levels, including the expression of precursor mRNAs, biosynthesis and release of opioid peptides, as well as ligand binding to opioid receptors. In adult rats, ß-endorphinergic and enkephalinergic transmission, through activation of mu and delta opioid receptors, mediate ethanol reinforcement and high alcohol drinking behavior. Prenatal ethanol exposure (PEE) selectively modifies Methionine-enkephalin (Met-enk) content in several brain regions of infant and adolescent rats, particularly those of the reward circuits. In preweanling rats, Met-enk content is decreased in the ventral tegmental area but is increased in the prefrontal cortex and the nucleus accumbens and other brain areas, as a consequence of a short and moderate ethanol exposure during late gestation. PEE also increases Met-enk levels in the prefrontal cortex and other brain regions of 30-day-old adolescent rats. These findings suggest that mesocorticolimbic enkephalins are essential in ethanol reinforcement in offspring, as previously reported in adult rats.

Hyperlocomotion and anxiety- like behavior induced by binge ethanol exposure in rat neonates. Possible ameliorative effects of Omega 3.

Maternal alcohol consumption during pregnancy may cause neurocognitive and behavioral disorders that can persist until adulthood. Epidemiological data has revealed an alarming increase in the frequency of alcohol intake in pregnant women. Nutritional variables may also have an impact on the behavioral alterations occasioned by alcohol during development. Moreover, omega-3, a polyunsaturated fatty acid necessary for normal brain development, is deficient in ethanol-treated animals. Although studies have shown that omega-3 supplementation after prenatal ethanol (EtOH) treatment improves some disorders, there are no reports about acute treatment with omega-3 in binge alcohol neurotoxic models during postnatal development. The goal of this study was to determine whether an administration of omega-3, after an acute ethanol dose in neonates, would be able to attenuate alcohol effects in offspring. Male/ female rats were administered ethanol (2.5 g/kg s.c. at 0 and 2 h) or saline on postnatal day (PND) 7, with a single dose of omega-3 (720 mg/kg) 15 min after the last alcohol injection. It was have found that EtOH-treated animals showed hyperlocomotion on PND 14 (pre-juvenile), and anxiety-like behavior was observed at all the three ages studied. Administration of omega-3 after EtOH treatment reduced hyperlocomotion and the anxiety-like behaviors on PND 14, but did not diminish the anxiety on either PND 20 or 30 (juvenile). In conclusion, acute ethanol exposure produced neurobehavioral alterations that persisted in the offspring, with omega-3 able to ameliorate these effects on PND 14. These data are relevant considering that omega-3 administration may have therapeutic effects through mitigating some of ethanol´s damaging consequences.

Maternal manipulation during late gestation (GDs 17-20) enhances ethanol consumption and promotes changes and opioid mRNA expression in infant rats.

Fetal ethanol experience generates learning and memories capable to increase ethanol consummatory behaviors during infancy. Opioid system seems to be involved in mediating those alcohol-related behaviors. In this work, we proposed to study the impact of prenatal exposure to a moderate ethanol dose, upon ingestion of the drug and possible ethanol-induced molecular changes on opioid precursor peptides (POMC, Pro-enk and Pro-DYN) and receptors (MOR, DOR and KOR) mRNA expression, in hypothalamus. Pregnant rats received during gestational days (GDs) 17-20, a daily intragastric (i.g.) administration with 2g/kg ethanol or water. A third group of dams was left undisturbed during pregnancy (Unmanipulated group). Intake test was conducted at postnatal days (PDs) 14-15. Three groups of pups were performed: control (no intake test), water (vehicle) and 5% ethanol. At the end of intake test blood samples were taken to quantify blood ethanol concentrations (BECs) and hypothalamus sections were obtained to perform qRT-PRC assessment of opioid precursor peptides and receptors. The analysis of the consummatory responses (% of consumption) and pharmacokinetic profiles (BECs) suggested that maternal manipulation induced by i.g. intubations, during the last four days of gestation (whenever ethanol or water), are sufficient to induce infantile ethanol intake during infancy. Gene expression from the hypothalamus of unmanipulated group revealed that infantile ingestive experiences with ethanol can down-regulate expression of mRNA Pro-Dyn and up-regulate mRNA expression of MOR and KOR. Finally, MOR mRNA expression was attenuated by prenatal i.g. manipulation in pups exposed to 5% ethanol.

Context-dependent reinstatement of an extinguished operant response in preweanling rats.

It is frequently assumed that infants are impaired in contextual memory and consequently, in recovery from extinction, a phenomenon considered to be context dependent. However, the evidence in the field is far from consistent with this interpretation, since several studies have shown context-dependent extinction in infant rats using a variety of procedures and behavioral measures. This discussion has primarily been based on studies using Pavlovian conditioning tasks. Three experiments were conducted to study reinstatement of an extinguished operant response and additionally to evaluate the context dependence of such an effect. Preweanling rats were trained on postnatal days (PDs) 16 and 17 to acquire an appetitive operant response using chocolate milk as reinforcer. At PD18 the operant response was extinguished and at PD19 animals received a reminder in the same (Experiment 1 and 2) or in a different context than the one used during the extinction phase (Experiment 3). Infant rats showed recovery from extinction after a reminder treatment (Experiments 2 and 3). This effect was dependent on the context in which the reminder was delivered (Experiment 3). Our results indicate that infant rats can recover an extinguished operant response by means of a reminder treatment in a context-dependent way, highlighting the importance of the context for modulating learning processes during infancy. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Context-dependent extinction of an appetitive operant conditioned response in infant rats.

The present study evaluated context-dependent learning under an operant conditioning procedure in infant rats. Preweanling rats were trained in context A during postnatal days (PDs) 16 and 17 to learn an appetitive operant conditioning task, employing milk chocolate as appetitive reinforcer. On PD18 the operant response was extinguished in context A, or in an alternative context B. The change from context A to B between acquisition and extinction did not affect the number of responses during extinction, but slightly modified the shape of the extinction curve. On PD19, a renewal test conducted in context A clearly showed ABA-renewal of the extinguished operant response. These results add to the body of evidence indicating that infants are able to acquire and retain contextual information, and support the notion that extinction during this ontogenetic period involves new learning.

Maternal isolation during the first two postnatal weeks affects novelty-induced responses and sensitivity to ethanol-induced locomotor activity during infancy.

Animals exposed to chronic maternal separation (MS) exhibit enhanced ethanol self-administration and greater hormonal and behavioral responsiveness to stress in adulthood. Whether the effects of MS are immediately evident in infancy or whether they appear only later on development is still an unanswered question This study tested sensitivity to ethanol's behavioral stimulating effects in infant rats that experienced MS from postnatal Day 1-14. MS infants exhibited significantly greater reactivity to the motor stimulating effects of 1.25 g/kg ethanol than control animals, yet greater motor suppression after 2.5 g/kg ethanol. Baseline level of response to novelty was altered in MS infants, in a nor-binaltorphimine insensitive manner, that is, despite modified activity of the kappa-opioid system. These results indicate that the consequences of chronic maternal isolation emerge early in ontogeny, affecting ethanol sensitivity in infancy.

Acetaldehyde reinforcement and motor reactivity in newborns with or without a prenatal history of alcohol exposure.

Animal models have shown that early ontogeny seems to be a period of enhanced affinity to ethanol. Interestingly, the catalase system that transforms ethanol (EtOH) into acetaldehyde (ACD) in the brain, is more active in the perinatal rat compared to adults. ACD has been found to share EtOH's behavioral effects. The general purpose of the present study was to assess ACD motivational and motor effects in newborn rats as a function of prenatal exposure to EtOH. Experiment 1 evaluated if ACD (0.35 µmol) or EtOH (0.02 µmol) supported appetitive conditioning in newborn pups prenatally exposed to EtOH. Experiment 2 tested if prenatal alcohol exposure modulated neonatal susceptibility to ACD's motor effects (ACD dose: 0, 0.35 and 0.52 µmol). Experiment 1 showed that EtOH and ACD supported appetitive conditioning independently of prenatal treatments. In Experiment 2, latency to display motor activity was altered only in neonates prenatally treated with water and challenged with the highest ACD dose. Prenatal EtOH experience results in tolerance to ACD's motor activity effects. These results show early susceptibility to ACD's appetitive effects and attenuation of motor effects as a function of prenatal history with EtOH, within a stage in development where brain ACD production seems higher than later in life.

The role of acetaldehyde in ethanol reinforcement assessed by Pavlovian conditioning in newborn rats.

RATIONALE: Animal studies indicate that central acetaldehyde, dependent on catalase metabolism of ethanol (EtOH), modulates ethanol reinforcement. Brain catalase activity and acetaldehyde (ACD) production are significantly higher in rat pups compare d with adults. Interestingly, infant rats show high EtOH affinity for alcohol consumption and are particularly sensitive to the drug's reinforcing effects. OBJECTIVES: We tested whether central ACD is necessary and sufficient to induce appetitive conditioning in newborn rats through the artificial nipple technique. METHODS: Vehicle, EtOH (100 mg%), and acetaldehyde (0.35 µmol) were administered into the cisterna magna (1 µl). Half of the animals also received a central administration of 75 µg (experiment 1) or 40 µg of D-penicillamine (experiment 2). Afterwards, pups were exposed to an olfactory cue (conditioned stimulus). One hour later, neonates were tested with an artificial nipple in the presence of the conditioned cue. Nipple attachment duration, mean grasp duration, and number of nipple disengagements served as dependent variables. RESULTS: Positive responses to the scented nipple occurred in neonates conditioned with EtOH or ACD (experiments 1 and 2). In experiment 1, there were indications that D-penicillamine weakened the reinforcing effects of EtOH and ACD. In experiment 2, D-penicillamine (40 µg) significantly inhibited appetitive conditioned responses dependent upon EtOH or ACD. CONCLUSIONS: Appetitive conditioning was observed when employing either central EtOH or ACD as unconditioned stimuli. Central abduction of ACD inhibited conditioned appetitive responsiveness to the surrogate nipple. Central ACD is involved in the determination or modulation of EtOH's motivational properties during early stages in development.

Role of mu, delta and kappa opioid receptors in ethanol-reinforced operant responding in infant rats.

We recently observed that naloxone, a non-specific opioid antagonist, attenuated operant responding to ethanol in infant rats. Through the use of an operant conditioning technique, we aimed to analyze the specific participation of mu, delta, and kappa opioid receptors on ethanol reinforcement during the second postnatal week. In Experiment 1, infant rats (PDs 14-17) were trained to obtain 5, 7.5, 10, or 15% ethanol, by operant nose-poking. Experiment 2 tested blood ethanol levels (BELs) attained by operant behavior. In Experiment 3, at PDs 16-18, rats received CTOP (mu antagonist: 0.1 or 1.0 mg/kg), naltrindole (delta antagonist: 1.0 or 5.0 mg/kg) or saline before training. In Experiment 4, rats received nor-binaltorphimine (kappa antagonist: 10.0 or 30.0 mg/kg, a single injection after completion of PD 15 operant training), spiradoline mesylate (kappa agonist: 1.0 or 5.0 mg/kg; at PDs 16-18) or saline (PDs 16-18), before the conditioning. Experiments 5 and 6 assessed possible side effects of opioid drugs in locomotor activity (LA) and conditioned taste aversion (CTA). Ethanol at 7.5 and 10% promoted the highest levels of operant responding. BELs were 12-15 mg/dl. In Experiment 3 naltrindole (dose-response effect) and CTOP (the lowest dose) were effective in decreasing operant responding. Nor-binaltorphimine at 10.0 mg/kg and spiradoline at 5.0 mg/kg also blocked ethanol responding. The effects of opioid drugs on ethanol reinforcement cannot be explained by effects on LA or CTA. Even though particular aspects of each opioid receptor require further testing, a fully functional opioid system seems to be necessary for ethanol reinforcement, during early ontogeny.

Naloxone attenuation of ethanol-reinforced operant responding in infant rats in a re-exposure paradigm.

RATIONALE: Early ethanol exposure promotes ethanol reinforcement, mediated perhaps by ethanol's motivational effects. The opioid system mediates ethanol reinforcement, at least in part. OBJECTIVES: Modulation of consummatory and seeking behaviors by the opioid system was tested in terms of ethanol or sucrose operant self-administration. METHODS: Wistar-derived infant rats were tested in an operant conditioning task. (1) Infants were trained on postnatal days (PDs) 14-17 to obtain 5% sucrose and 3.75% ethanol or water, and evaluated in an extinction session at PD 18. (2) Ethanol (3.75%) was used as reinforcer. At PDs 16-17, 6 h before operant task, pups were re-exposed to ethanol after naloxone injection (0 or 1 mg/kg). (3) Sucrose (5%) acted as reinforcer. Pups were re-exposed to sucrose after naloxone injection. (4) A PD 18 re-exposure trial in which pups were injected with naloxone and re-exposed to ethanol was added. RESULTS: Sucrose and ethanol promoted higher levels of operant responding than water during training and extinction. Re-exposure to ethanol preceded by naloxone decreased nose-poking. A similar profile was observed towards sucrose. No seeking behavior was observed in pups re-exposed to ethanol following naloxone injection during PDs 16-18. CONCLUSIONS: Self-administration of ethanol was established in terms of operant responding in preweanling rats with no previous exposure to the drug. Pairing of naloxone with ethanol, at a point separate in time from operant responding, reduced ethanol reinforcement. This indicated participation of the opioid system in ethanol reinforcement. This effect seems not to be unique to ethanol but also is observable when sucrose acts as reinforcer.

Prenatal and postnatal ethanol experiences modulate consumption of the drug in rat pups, without impairment in the granular cell layer of the main olfactory bulb.

The effect of moderate exposure to ethanol during late gestation was studied in terms of its interaction with moderate exposure during nursing from an intoxicated dam. A further issue was whether behavioral effects of ethanol, especially the enhanced ethanol intake known to occur after moderate ethanol prenatally or during nursing, depend upon teratological effects that may include death of neurons in the main olfactory bulb (MOB). During gestational days 17-20 rats were given 0, 1 or 2g/kg ethanol doses intragastrically (i.g.). After parturition these dams were given a dose of 2.5g/kg ethanol i.g. each day and allowed to perform regular nursing activities. During postnatal days (PDs) 15 and 16, ethanol intake of pups was assessed along with aspects of their general activity. In a second experiment pups given the same prenatal treatment as above were tested for blood ethanol concentration (BEC) in response to an ethanol challenge on PD6. A third experiment (Experiment 2b) assessed stereologically the number of cells in the granular cell layer of the MOB on PD7, as a function of analogous pre- and postnatal ethanol exposures. Results revealed that ethanol intake during the third postnatal week was increased by prenatal as well as postnatal ethanol exposure, with a few interesting qualifications. For instance, pups given 1g/kg prenatally did not have increased ethanol intake unless they also had experienced ethanol during nursing. There were no effects of ethanol on either BECs or conventional teratology (cell number). This increases the viability of an explanation of the effects of prenatal and early postnatal ethanol on later ethanol intake in terms of learning and memory.

Participation of the endogenous opioid system in the acquisition of a prenatal ethanol-related memory: effects on neonatal and preweanling responsiveness to ethanol.

The present study tested the involvement of the opioid system in the acquisition and expression of prenatal ethanol-related memories. We evaluated how this prenatal experience modulates ethanol self-administration in newborn rats, and preweanling's ingestion of the drug. During Gestational Days (GDs) 17-20, four groups of dams were treated with ethanol (2 g/kg) or water, followed immediately by naloxone (10 mg/kg) or saline administration. A fifth group received a similar dose of naloxone 20min before ethanol administration. On PD 1, pups were tested on an operant learning procedure to obtain milk or 3% ethanol. One hour later, an extinction session was performed. At Postnatal Days (PDs) 14 and 15, preweanlings representing each prenatal treatment were evaluated in an intake test with infusions of 5% ethanol or water. Prior to the intake test on PD14, preweanlings were administered naloxone (1 mg/kg), saline or remained untreated. In both tests, animals representative of both genders were utilized. One-day-old pups rapidly learned the operant behavior to gain access to milk. In contrast, only pups prenatally treated with ethanol (administered immediately before naloxone or saline injection) increased operant responding to gain access to ethanol. On an intake test at PDs 14 and 15, those animals prenatally exposed to naloxone 20 min before ethanol administration consumed significantly lower ethanol levels than the remaining prenatal ethanol groups. Postnatal treatment with naloxone diminished intake of all solutions at PD14. These results suggest that prenatal ethanol exposure facilitates neonatal operant learning reinforced by intraoral administration of ethanol and increases ethanol consumption during PDs 14-15. The endogenous opioid system apparently is involved in the acquisition of prenatal ethanol memories, which can modulate the reinforcing attributes of the drug in neonatal and preweanling rats.

Fetal exposure to moderate ethanol doses: heightened operant responsiveness elicited by ethanol-related reinforcers.

BACKGROUND: Prenatal exposure to moderate ethanol doses during late gestation modifies postnatal ethanol palatability and ingestion. The use of Pavlovian associative procedures has indicated that these prenatal experiences broaden the range of ethanol doses capable of supporting appetitive conditioning. Recently, a novel operant technique aimed at analyzing neonatal predisposition to gain access to ethanol has been developed. Experiment 1 tested the operant conditioning technique for developing rats described by Arias and colleagues (2007) and Bordner and colleagues (2008). In Experiment 2, we analyzed changes in the disposition to gain access to ethanol as a result of moderate prenatal exposure to the drug. METHODS: In Experiment 1, newborn pups were intraorally cannulated and placed in a supine position that allowed access to a touch-sensitive sensor. Paired pups received an intraoral administration of a given reinforcer (milk or quinine) contingent upon physical contact with the sensor. Yoked controls received similar reinforcers only when Paired pups activated the circuit. In Experiment 2, natural reinforcers (water or milk) as well as ethanol (3% or 6% v/v) or an ethanol-related reinforcer (sucrose compounded with quinine) were tested. In this experiment, pups had been exposed to water or ethanol (1 or 2 g/kg) during gestational days 17 to 20. RESULTS: Experiment 1 confirmed previous results showing that 1-day-old pups rapidly learn an operant task to gain access to milk, but not to gain access to a bitter tastant. Experiment 2 showed that water and milk were highly reinforcing across prenatal treatments. Furthermore, general activity during training was not affected by prenatal exposure to ethanol. Most importantly, prenatal ethanol exposure facilitated conditioning when the reinforcer was 3% v/v ethanol or a psychophysical equivalent of ethanol's gustatory properties (sucrose-quinine). CONCLUSIONS: The present results suggest that late prenatal experience with ethanol changes the predisposition of the newborn to gain access to ethanol-related stimuli. In conjunction with prior literature, this study emphasizes the fact that intrauterine experience with ethanol not only augments ethanol's palatability and ingestion, but also facilitates the acquisition of response-stimulus associations where the drug acts as an intraoral reinforcer.

Ethanol exposure during late gestation and nursing in the rat: effects upon maternal care, ethanol metabolism and infantile milk intake.

Ethanol experiences, during late gestation as well as during nursing, modify the behavioral dynamics of the dam/pup dyad, and leads to heightened ethanol intake in the offspring. This study focuses on: a) behavioral and metabolic changes in intoxicated dams with previous exposure to ethanol during pregnancy and b) infantile consumption of milk when the dam is either under the effects of ethanol or sober. Pregnant rats received water, 1.0 or 2.0 g/kg ethanol, and were administered with water or ethanol during the postpartum period. Intoxication during nursing disrupted the capability of the dam to retrieve the pups and to adopt a crouching posture. These disruptions were attenuated when dams had exposure to ethanol during pregnancy. Ethanol experiences during gestation did not affect pharmacokinetic processes during nursing, whereas progressive postpartum ethanol experience resulted in metabolic tolerance. Pups suckling from intoxicated dams, with previous ethanol experiences, ingested more milk than did infants suckling from ethanol-intoxicated dams without such experience. Ethanol gestational experience results in subsequent resistance to the drug's disruptions in maternal care. Consequently, better maternal care by an intoxicated dam with ethanol experience during gestation facilitates access of pups to milk which could be contaminated with ethanol.

Fetal learning about ethanol and later ethanol responsiveness: evidence against "safe" amounts of prenatal exposure.

Near-term fetuses of different mammalian species, including humans, exhibit functional sensory and learning capabilities. The neurobiological literature indicates that the unborn organism processes sensory stimuli present in the amniotic fluid, retains this information for considerable amounts of time, and is also capable of associating such stimuli with biologically relevant events. This research has stimulated studies aimed at the analysis of fetal and neonatal learning about ethanol, a topic that constitutes the core of the present review. Ethanol has characteristic sensory (olfactory, taste, and trigeminal) attributes and can exert pharmacologic reinforcing effects. The studies under examination support the hypothesis that low to moderate levels of maternal ethanol intoxication during late pregnancy set the opportunity for fetal learning about ethanol. These levels of prenatal ethanol exposure do not generate evident morphologic or neurobehavioral alterations in the offspring, but they exert a significant impact upon later ethanol-seeking and intake behaviors. Supported by preclinical and clinical findings, this review contributes to strengthening the case for the ability of prenatal ethanol exposure to have effects on the postnatal organism.

Heightened ethanol intake in infant and adolescent rats after nursing experiences with an ethanol-intoxicated dam.

BACKGROUND: Preweanling rats detect ethanol (175 mg/100 ml) in maternal milk when the dam is moderately intoxicated. Repeated experiences with the intoxicated dam facilitate subsequent recognition of ethanol's chemosensory attributes and promote ethanol-related memories with a negative hedonic content. This memory has been attributed to the infant's acquired association between ethanol's chemosensory attributes and its disruptive effects on maternal care. In this study, infant and adolescent ethanol intake patterns were analyzed as a function of prior interactions, during early infancy, with their intoxicated dams. METHODS: During postpartum days 3, 5, 7, 9, 11, and 13, breast-feeding dams received an intragastric administration of either 2.5 g/kg of ethanol or water. Pups whose dams had been given one of these two maternal treatments were tested on postnatal day 15 for ingestion of 0% (water), 2.5, 5.0, or 10% v/v ethanol solution. During adolescence, remaining animals from these litters were first adapted to ingest water from drinking tubes and then were given simultaneous access to tap water and a given ethanol solution. The first day, a 3% v/v ethanol solution was used. This solution was increased by 1% ethanol each following day until the solution was 6% v/v ethanol. RESULTS: Maternal drug treatment did not affect the body weights of dams, infants, or adolescents. Water intake during infancy and adolescence also was unaffected by prior maternal treatment. However, infants that had previously interacted with ethanol-intoxicated dams exhibited heightened ethanol intake scores (grams per kilogram and percentage body weight gains), especially when tested with 5 or 10% v/v ethanol solutions. Similarly, adolescent males (but not females) that had interacted with an intoxicated dam during infancy also had higher ethanol consumption levels than those that had interacted with a nonintoxicated dam. CONCLUSIONS: Contrary to what might be expected in animals that acquire an aversive memory for ethanol's chemosensory cues as a function of prior interactions with an intoxicated mother, these results indicate that such interactions promote a long-lasting increase in ethanol intake. These results suggest that rats reared by intoxicated dams become sensitive to the negative reinforcing properties of ethanol.

Fetal learning with ethanol: correlations between maternal hypothermia during pregnancy and neonatal responsiveness to chemosensory cues of the drug.

BACKGROUND: Fetuses learn about ethanol odor when the drug is present in the amniotic fluid. Prenatal learning comprising ethanol's chemosensory cues also suggests an acquired association between ethanol's chemosensory and postabsorptive properties. Ethanol-related thermal disruptions have been implicated as a significant component of the drug's unconditioned properties. In the present study, ethanol-induced thermal changes were analyzed in pregnant rats subjected to a moderate ethanol dose. This thermal response was later tested for its correlation with the responsiveness of the progeny to ethanol and nonethanol chemosensory stimuli. METHODS: During gestational day (GD) 14, pregnant rats were subjected to a minor surgical procedure to place a subcutaneous telemetric thermal sensor in the nape of the neck. During GDs 17 to 20, females received a daily intragastric administration of ethanol (2 g/kg) or water, using solutions kept at room temperature. Maternal body temperatures were recorded before and after (4 consecutive hours) the administration of water or ethanol. Newborns representative of both prenatal treatments were tested in terms of behavioral activity elicited by the smell of ethanol or of a novel odorant (cineole). A third group of pups were tested in response to unscented air stimulation. RESULTS: Ethanol administration during late gestation induced reliable maternal hypothermia, a thermal disruption greater than that observed in water-treated females. It was systematically observed that maternal ethanol-induced hypothermia negatively correlated with neonatal motor reactivity elicited by ethanol olfactory stimulation. No other significant correlations were observed in terms of responsiveness to cineole or to unscented air in animals prenatally exposed to ethanol or water. CONCLUSIONS: In conjunction with prior research, the present results indicate that fetal ethanol exposure may yield learning of an association between ethanol's sensory and unconditioned properties. Ethanol-induced hypothermia during late gestation seems to represent a significant component of ethanol's unconditioned consequences. Specifically, ethanol-related thermal disruptions in the womb are highly predictive of neonatal responsiveness to ethanol's chemosensory cues that are known to be processed by the near-term fetus.

Neonatal activation of alcohol-related prenatal memories: impact on the first suckling response.

BACKGROUND: Rat fetuses seem to be capable of associative learning mediated through alcohol's unconditioned properties. The newborn's suckling response immediately after birth is dependent on olfactory stimuli present in the fetal milieu and/or odorants perceived immediately after birth. The present study analyzed the impact of olfactory fetal learning supported by maternal-fetal alcohol intoxication on the newborn's first postnatal suckling response, in the presence or absence of chemosensory cues originally associated with alcohol during prenatal life. METHODS: During gestational days 17 to 20, fetuses experienced a salient novel cue (cineole) explicitly paired or not with the induction of alcohol intoxication resulting from maternal intragastric intubation of a 2 g/kg ethanol dose. A separate set of dams were intubated with only water. In experiment 1, cesarean-delivered pups were tested shortly after birth for their response to a surrogate nutritive nipple scented with cineole or with no explicit odor. In experiment 2, pups were similarly evaluated after the prenatal treatment when cineole, a novel odorant (lemon), or no explicit odor was presented either in conjunction with the nipple or before being tested with the nipple. RESULTS: In both experiments, fetal olfactory conditioning supported by alcohol intoxication had a significant effect on the newborn's first suckling episode, depending on the olfactory cues presented before or during suckling. Presence of cineole but not of novel odorants seemed to activate the associative memory acquired in utero. Once the memory was activated, pups that were subjected to fetal odor-alcohol pairings were found to attach significantly more to surrogate nutritive nipples than did fetal control animals. CONCLUSIONS: Prenatal experience with alcohol intoxication can result in chemosensory associative learning mediated by the drug's postabsorptive effects. This learning determines attachment patterns of newborns when they reexperience olfactory cues that had signaled onset of the state of acute alcohol intoxication when they were fetuses. Considered in view of previous experiments, the present results generate two alternative hypotheses relative to the affective component of the memory established in utero: (1) alcohol intoxication is an appetitive reinforcer during fetal life, or (2) the calming effects of postnatal suckling behavior counteract negative hedonic components of the memory accrued in utero.