Enhanced D2 Agonism Induces Conditioned Appetitive Sexual Responses Toward Non-reproductive Conspecifics.

Brain mechanisms of sexual attraction toward reproductive partners develop from a systematic interrelationship between biology (nature) and learning (nurture). However, the causes of attraction toward non-reproductive partners are poorly understood. Here, we explored the role of Pavlovian learning under dopaminergic agonism on the development of sexual preference and brain activation for young male rats. During conditioning, adult sexually naïve males received either Saline (Saline-Paired) or the D2-receptor agonist quinpirole (QNP-Paired) and cohabited in contingency, or out of contingency (QNP-Unpaired) during 24 h with an almond-scented prepubertal juvenile male (PD25). Conditioning occurred every 4 days for three trials. Social and sexual responses were assessed four days after the last conditioning trial in a drug-free test, and males chose freely between a scented young male (PD37) and a novel receptive female. Four days later, males were exposed to the conditioned odor only and brain Fos-IR and serum testosterone were analyzed. Saline-Paired and QNP-Unpaired males displayed more non-contact erections (NCEs) and genital investigations for females, whereas QNP-Paired males expressed more NCEs and genital investigations for young males. In the QNP-Paired group, exposure to the young male-paired odor evoked more Fos-IR in limbic, hypothalamic and cortical areas, but no differences in serum testosterone were observed. Cohabitation with juvenile males during enhanced D2 agonism results in atypical appetitive sexual responses and a higher pattern of brain response for the young male-paired odor, with no changes in serum testosterone. We discuss the potential implications for the development of pedophilic disorder and perhaps other paraphilias.

Brain activation associated to olfactory conditioned same-sex partner preference in male rats.

Sexual preferences can be strongly modified by Pavlovian learning. For instance, olfactory conditioned same-sex partner preference can occur when a sexually naïve male cohabits with an scented male during repeated periods under the effects of enhanced D2-type activity. Preference is observed days later via social and sexual behaviors. Herein we explored brain activity related to learned same-sex preference (Fos-Immunoreactivity, IR) following exposure to a conditioned odor paired with same-sex preference. During conditioning trials males received either saline or the D2-type receptor agonist quinpirole (QNP) and cohabitated during 24 h with a stimulus male that bore almond scent on the back as conditioned stimulus. This was repeated every 4 days, for a total of three trials. In a drug-free final test we assessed socio/sexual partner preference between the scented male and a receptive female. The results indicated that QNP-conditioned males developed a same-sex preference observed via contact, time spent, olfactory investigations, and non-contact erections. By contrast, saline-conditioned and intact (non-exposed to conditioning) males expressed an unconditioned preference for the female. Four days later the males were exposed to almond scent and their brains were processed for Fos-IR. Results indicated that the QNP-conditioned group expressed more Fos-IR in the nucleus accumbens (AcbSh), medial preoptic area (MPA), piriform cortex (Pir) and ventromedial nucleus of the hypothalamus (VMH) as compared to saline-conditioned. Intact males expressed the lowest Fos-IR in AcbSh and VMH, but the highest in MPA and Pir. We discuss the role of these areas in the learning process of same-sex partner preferences and olfactory discrimination.

Olfactory conditioned same-sex partner preference in female rats: Role of ovarian hormones.

The dopamine D2-type receptor agonist quinpirole (QNP) facilitates the development of conditioned same-sex partner preference in males during cohabitation, but not in ovariectomized (OVX) females, primed with estradiol benzoate (EB) and progesterone (P). Herein we tested the effects of QNP on OVX, EB-only primed females. Females received a systemic injection (every four days) of either saline (Saline-conditioned) or QNP (QNP-conditioned) and then cohabited for 24h with lemon-scented stimulus females (CS+), during three trials. In test 1 (female-female) preference was QNP-free, and females chose between the CS+ female and a novel female. In test 2 (male-female) they chose between the CS+ female and a sexually experienced male. In test 1 Saline-conditioned females displayed more hops & darts towards the novel female, but QNP-conditioned females displayed more sexual solicitations towards the CS+ female. In test 2 Saline-conditioned females displayed a clear preference for the male, whereas QNP-conditioned females displayed what we considered a bisexual preference. We discuss the effect of dopamine and ovarian hormones on the development of olfactory conditioned same-sex preference in females.

Involving the cerebellum in cocaine-induced memory: pattern of cFos expression in mice trained to acquire conditioned preference for cocaine.

Because of its primary role in drug-seeking, consumption and addictive behaviour, there is a growing interest in identifying the neural circuits and molecular mechanisms underlying the formation, maintenance and retrieval of drug-related memories. Human studies, which focused on neuronal systems that store and control drug-conditioned memories, have found cerebellar activations during the retrieval of drug-associated cue memory. However, at the pre-clinical level, almost no attention has been paid to a possible role of the cerebellum in drug-related memories. In the present study, we ought to fill this gap by aiming to investigate the pattern of neuronal activation (as revealed by cFos expression) in different regions of the prefrontal cortex and cerebellum of mice trained to develop conditioned preference for an olfactory stimulus (CS+) paired with cocaine. Our results indicate that CS+ preference was directly associated with cFos expression in cells at the apical region of the granule cell layer of the cerebellar vermis; this relationship being more prominent in some specific lobules. Conversely, cFos+ immunostaining in other cerebellar regions seems to be unrelated to CS+ preference but to other aspects of the conditioning procedure. At the prefrontal cortex, cFos expression seemed to be related to cocaine administration rather than to its ability to establish conditioned preference. The present results suggest that as it has been observed in some clinical studies, the cerebellum might be an important and largely overlooked part of the neural circuits involved in generating, maintaining and/or retrieving drug memories.

Cohabitation with receptive females under D2-type agonism in adulthood restores partner preference and brain dimorphism in the SDN-POA following neonatal gonadectomy in male rats.

Perinatal testosterone, or its metabolite estradiol, organize the brain toward a male phenotype. Male rodents with insufficient testosterone during this period fail to display sexual behavior and partner preference for receptive females in adulthood. However, cohabitation with non-reproductive conspecifics under the influence of a D2 agonist facilitates the expression of conditioned partner preference via Pavlovian learning in gonadally intact male rats. In the present experiment, three groups of neonatal PD1 males (N = 12/group) were either gonadectomized (GDX), sham-GDX, or left intact and evaluated for social preferences and sexual behaviors as adults. We then examined whether the effects of GDX could be reversed by conditioning the males via cohabitation with receptive females under the effects of the D2 agonist quinpirole (QNP) or saline, along with the size of some brain regions, such as the sexually dimorphic nucleus of the preoptic area (SDN-POA), suprachiasmatic nucleus (SCN), posterior dorsal medial amygdala (MeApd) and ventromedial hypothalamus (VMH). Results indicated that neonatal GDX resulted in the elimination of male-typical sexual behavior, an increase in same-sex social preference, and a reduction of the area of the SDN-POA. However, GDX-QNP males that underwent exposure to receptive females in adulthood increased their social preference for females and recovered the size in the SDN-POA. Although neonatal GDX impairs sexual behavior and disrupts partner preference and brain dimorphism in adult male rats, Pavlovian conditioning under enhanced D2 agonism ameliorates the effects on social preference and restores brain dimorphism in the SDN-POA without testosterone.

Pubertal stress in male rats: Effects on juvenile play behavior and adult sexual partner preference.

Puberty is a period of brain organization impacting the expression of social and sexual behaviors. Here, we assessed the effects of an acute pubertal stressor (immune challenge) on the expression of juvenile play (short-term) and sexual partner preference (long-term) in male rats. Juvenile play was assessed over ten trials at postnatal days (PND) (31-40) with age- and sex-matched conspecifics, and at PND35 males received a single injection of lipopolysaccharide (LPS, 1.5 mg/kg i.p.) or saline. Then, sexual partner preference was assessed at PND 60, 64, and 68, in a three-compartment chamber with a sexually receptive female and a male as potential partners simultaneously. The results confirmed that a single injection of LPS during puberty induced sickness signs indicative of an immune challenge. However, juvenile play was not affected by LPS treatment during the following days (PND36-40), nor was sexual behavior and partner preference for females in adulthood. These findings highlight that, while other studies have shown that LPS-induced immunological stress during puberty affects behavior and neuroendocrine responses, it does not affect juvenile play and sexual behavior in male rats. This suggests a remarkable resilience of these behavioral systems for adaptation to stressful experiences mediated by immune challenges during critical periods of development. These behaviors, however, might be affected by other types of stress.

The impact of subchronic hypercaloric and restriction diets on sexual behavior, serum testosterone, and prostate histology in rats.

The role of diet in health is crucial, with calorie intake playing a significant role. Hypercaloric diets (HD) often lead to adipose tissue accumulation and increased risk of chronic diseases, including reproductive impairments. By contrast, restriction diets (RD) help with weight loss, improve cardiovascular function, and ameliorate reproduction. Herein we sought to investigate the impact of subchronic HD and RD on body weight, sexual behavior, serum testosterone and prostate histology in rats. Hence, 10-week old male rats gained sexual experience during five trials with ovariectomized, hormone-primed females. Then at postnatal week PW15 the males were organized in three groups, depending on the feeding they received until PW18: HD, RD and standard diet (SD). During PW19-22 they were tested for sexual behavior, and at PW23 were euthanized for prostate histology (hematoxylin & eosin stain) and hormone analysis. Results indicated that HD males increased their body weight (16-23%) compared to SD and RD. Furthermore, HD males showed 65% less testosterone than RD males. The prostate of HD males revealed histological alterations, including a notable increase in epithelium height and other abnormal features, while no changes were observed in the performance of sexual behavior between HD and RD, although HD appeared to facilitate ejaculation when compared to SD. The histological features of RD males were comparable to SD males. Accordingly, we argue that subchronic modifications in calorie intake can alter body weight (in HD), serum testosterone levels (HD and RD in opposite directions), and prostate histology (in HD), while having no immediate effect on male sexual behavior.

Preterm rat survival is enhanced by gestational environmental enrichment.

Throughout the last decade, the estimated global human preterm birth rate was 10.6 %, with higher rates in Asia, South America, and Africa. Preterm individuals, even in adulthood, are more likely to develop cardiorespiratory, renal, and metabolic disorders. On the other hand, when experimental animals are housed in an enriched environment during gestation, the development of the progeny in utero is accelerated, compared to standard housing conditions. By terminating gestation one and a half days before parturition, we investigated whether environmental enrichment restricted to gestation may have an impact on progeny survival. Our results demonstrate that the gestational enriched environment tripled the rat´s offspring survival, which was associated with decreased expression of anxiety-like behaviors in the pregnant mother. Sex of the offspring was not a factor in determining survival. We discuss the effect of increased secretion of various trophic factors and hormones induced by the enriched environment on progeny survival.

Electrophysiological Characterization of Cerebellar Responses during Exploration and Grooming Behaviors in a Rat Model of Parkinsonism.

Parkinson's disease is currently a global public health challenge due to the rapid growth of aging populations. To understand its pathophysiology is necessary to study the functional correlation between the basal ganglia (BG) and the cerebellum, which are involved in motor control. Herein, we explored multiunit electrical activity (MUA) in the cerebellum of rats with induced Parkinsonism as a result of lesions following bilateral placement of electrodes and passing of current in the ventrolateral striatum (VLS). In one control group, the electrodes descended without electrical current, and another group was left intact in VLS. MUA was recorded in Sim B and Crus II lobes, and in the dentate nucleus (DN) during the execution of exploration behaviors (horizontal and vertical) and grooming. The lesioned and sham groups showed a decrease in MUA amplitude in the Crus II lobe compared to the intact group in all recorded behaviors. However, Sim B and DN did not express differences. Both electrical and physical insults to the VLS induced Parkinsonism, which results in less MUA in Crus II during the execution of motor behaviors. Thus, this type of Parkinsonism is associated with a decrease in the amplitude of Crus II.

Who, what, where, when (and maybe even why)? How the experience of sexual reward connects sexual desire, preference, and performance.

Although sexual behavior is controlled by hormonal and neurochemical actions in the brain, sexual experience induces a degree of plasticity that allows animals to form instrumental and Pavlovian associations that predict sexual outcomes, thereby directing the strength of sexual responding. This review describes how experience with sexual reward strengthens the development of sexual behavior and induces sexually-conditioned place and partner preferences in rats. In both male and female rats, early sexual experience with partners scented with a neutral or even noxious odor induces a preference for scented partners in subsequent choice tests. Those preferences can also be induced by injections of morphine or oxytocin paired with a male rat's first exposure to scented females, indicating that pharmacological activation of opioid or oxytocin receptors can "stand in" for the sexual reward-related neurochemical processes normally activated by sexual stimulation. Conversely, conditioned place or partner preferences can be blocked by the opioid receptor antagonist naloxone. A somatosensory cue (a rodent jacket) paired with sexual reward comes to elicit sexual arousal in male rats, such that paired rats with the jacket off show dramatic copulatory deficits. We propose that endogenous opioid activation forms the basis of sexual reward, which also sensitizes hypothalamic and mesolimbic dopamine systems in the presence of cues that predict sexual reward. Those systems act to focus attention on, and activate goal-directed behavior toward, reward-related stimuli. Thus, a critical period exists during an individual's early sexual experience that creates a "love map" or Gestalt of features, movements, feelings, and interpersonal interactions associated with sexual reward.

Sexual experience increases oxytocin, but not vasopressin, receptor densities in the medial preoptic area, ventromedial hypothalamus, and central amygdala of male rats.

Oxytocin (OT) and vasopressin (VP) are considered to be principal neurochemical substrates of bonding in monogamous species. We have reported previously that conditioning of a sexual partner preference in male rats resulted in conditioned activation of OT and VP neurons in hypothalamic paraventricular and supraoptc nuclei. Here we asked whether such conditioning would also alter OT or VP receptor densities. Sexually naïve male rats were assigned to one of three groups (n = 15/group). The Paired group received 9 copulatory training trials with sexually receptive females scented with a neutral almond odor. The Unpaired group received 9 copulatory training trials with unscented sexually receptive females. The Naïve group were not given sexual experience. Paired and Unpaired males were given a final test in an open field with two receptive females, one scented and the other unscented, to assess the development of conditioned ejaculatory preference (CEP), which was expressed significantly in the Paired group. Brains from rats in the three groups were then assessed for OT receptor (OTR) or VP1a receptor (VPR) densities within cortical, limbic and hypothalamic structures using autoradiography with selective 125I-labeled receptor ligands. Sexual experience alone increased OTR significantly in the medial preoptic area (mPOA), ventromedial hypothalamus (VMH), and central nucleus of the amygdala (CeA) in both Paired- and Unpaired-trained males compared to sexually Naïve males. No differences were found for experience on VPR densities in any region. These data add to a growing body of evidence that sexual experience alters brain function and processing of sex-related cues, and suggest that enhanced activation of OTRs in the mPOA, VMH, and CeA by conditioned OT release in those regions may underlie CEP in the male rat.

The Neurobiology of Behavior and Its Applicability for Animal Welfare: A Review.

Understanding the foundations of the neurobiology of behavior and well-being can help us better achieve animal welfare. Behavior is the expression of several physiological, endocrine, motor and emotional responses that are coordinated by the central nervous system from the processing of internal and external stimuli. In mammals, seven basic emotional systems have been described that when activated by the right stimuli evoke positive or negative innate responses that evolved to facilitate biological fitness. This review describes the process of how those neurobiological systems can directly influence animal welfare. We also describe examples of the interaction between primary (innate) and secondary (learned) processes that influence behavior.

Neurobiology of Maternal Behavior in Nonhuman Mammals: Acceptance, Recognition, Motivation, and Rejection.

Among the different species of mammals, the expression of maternal behavior varies considerably, although the end points of nurturance and protection are the same. Females may display passive or active responses of acceptance, recognition, rejection/fear, or motivation to care for the offspring. Each type of response may indicate different levels of neural activation. Different natural stimuli can trigger the expression of maternal and paternal behavior in both pregnant or virgin females and males, such as hormone priming during pregnancy, vagino-cervical stimulation during parturition, mating, exposure to pups, previous experience, or environmental enrichment. Herein, we discuss how the olfactory pathways and the interconnections of the medial preoptic area (mPOA) with structures such as nucleus accumbens, ventral tegmental area, amygdala, and bed nucleus of stria terminalis mediate maternal behavior. We also discuss how the triggering stimuli activate oxytocin, vasopressin, dopamine, galanin, and opioids in neurocircuitries that mediate acceptance, recognition, maternal motivation, and rejection/fear.

Maternal behavior, novelty confrontation, and subcortical c-Fos expression during lactation period are shaped by gestational environment.

The environmental context during gestation may modulate the postpartum variations in maternal behaviors observed within different animal species. Most of our experimental knowledge on this phenomenon and its physiological effects have been gained by confronting the pregnant mother with stressful situations, with the consensual results indicating a reduced maternal behavior and a hyper reactivity of stress-related neural paths. Here, in contrast, by exposing nulliparous rats strictly during pregnancy to a standard laboratory environment (STD) or a highly stimulating sensory and social environment (EE), we investigated the hypothesis that subjects frequently exposed to social stimuli and novel situations during pregnancy will show postpartum changes in subcortical brain areas' activity related to the processing of social stimuli and novelty, such that there will be modifications in maternal behavior. We found that EE mothers doubled the levels of licking and grooming, and active hovering over pups during the first postpartum week than STD dams, without a difference in the time of contact with the pups. Associated with these behaviors, EE dams showed increased c-Fos immunoreaction in hypothalamic nuclei and distinct responses in amygdalar nuclei, than STD dams. In the maternal defensive test, EE dams tripled the levels of aggressive behaviors of the STD rats. Additionally, in two different tests, EE mothers showed lower levels of postpartum anxiety-like behaviors when confronted with novel situations. Our results demonstrate that the activity of brain areas related to social behavior is adaptable by environmental circumstances experienced during gestation, presumably to prepare the progeny for these particular conditions.

Multiunit recording of the cerebellar cortex, inferior olive, and fastigial nucleus during copulation in naive and sexually experienced male rats.

The sexual behavior of male rats constitutes a natural model to study learning of motor skills at the level of the central nervous system. We previously showed that sexual behavior increases Fos expression in granule cells at lobules 6 to 9 of the vermis cerebellum. Herein, we obtained multiunit recordings of lobules 6a and 7 during the training period of naive subjects, and during consecutive ejaculations of expert males. Recordings from both lobules and the inferior olive showed that the maximum amplitude of mount, intromission, and ejaculation signals were similar, but sexual behavior during training tests produced a decrease in the amplitude for mount and intromission signals. The fastigial nucleus showed an inverse mirror-like response. Thus, the cerebellum is involved in the neural basis of sexual behavior and the learning of appropriate behavioral displays during copulation, with a wiring that involves the cerebellar cortex, inferior olive, and fastigial nucleus.

Appetitive olfactory conditioning in the neonatal male rat facilitates subsequent sexual partner preference.

Pairing a neutral odor with a male rat's initial sexual experiences to ejaculation produces a subsequent conditioned ejaculatory preference (CEP) in which males ejaculate preferentially with receptive females that bear the odor relative to unscented receptive females. In 1986, Fillion and Blass reported that neonatal male rats exposed to a neutral lemon odor (citral) painted on their mother's ventrum while nursing ejaculated faster as adults with sexually receptive, citral-scented females compared to unscented receptive females. The present study examined whether the same odor paired with tactile reward in neonatal male rats would alter the subsequent expression of a CEP. Newborn Long-Evans male rats were separated from their mothers each day beginning on Postnatal Day 1 and placed into a Plexiglas cage that contained either unscented or citral-scented bedding (N = 8/group). During each trial, rats were stroked from head to toe with a soft, narrow paintbrush, after which they were returned to their mothers. Males were weaned at 21 days of age and housed in same-treatment pairs for an intervening 50 days. Following habituation to a large open field, males were presented with two sexually receptive Long-Evans females, one scented with citral, and the other unscented, for a 30-min test of copulation. Males in the Paired group copulated and ejaculated preferentially with the scented female whereas males in the Unpaired group showed no preference. Pairing a neutral odor with a reward state in infancy generates a preference in male rats to ejaculate with sexually receptive females bearing the same odor in adulthood.

Clicker Training Accelerates Learning of Complex Behaviors but Reduces Discriminative Abilities of Yucatan Miniature Pigs.

Animal training is meant to teach specific behavioral responses to specific cues. Clicker training (CT) is a popular training method based on the use of a device that emits a sound of double-click to be associated as a first-order conditioned stimulus in contingency with positive reinforcements. After some repetitions, the clicker sound gains some incentive value and can be paired with the desired behavior. Animal trainers believed that CT can decrease training time compared to other types of training. Herein, we used two-month old miniature piglets to evaluate whether CT decreased the number of repetitions required to learn complex behaviors as compared with animals trained with voice instead of the clicker. In addition, we compared the number of correct choices of animals from both groups when exposed to object discriminative tests. Results indicated that CT decreased the number of repetitions required for pigs to learn to fetch an object but reduced the ability of animals to make correct choices during the discriminate trials. This suggests that CT is more efficient than voice to teach complex behaviors but reduces the ability of animals to use cognitive processes required to discriminate and select objects associated with reward.

The Sleep of Shelter Dogs Was Not Disrupted by Overnight Light Rather than Darkness in a Crossover Trial.

Dogs in shelters may be unattended at night. The purpose of this study is to describe the night-time behavior of dogs in a shelter and to determine if artificial light affected their sleeping patterns. Ten dogs were video-recorded under both light and dark conditions and their behavior recorded using focal animal sampling. The dogs were lying down 649 ± 40 min (mean ± SD) in the light condition and 629 ± 58 min in the dark condition each night. They awoke, stood up, turned around and then lay down again every 48 to 50 min. There was no significant difference in time spent lying between the two conditions (p > 0.05). Light did not seem to affect their behavior. The conclusion is that dogs in shelters may sleep in the absence of people and that light does not disrupt their sleep patterns.

Disruptive effects of neonatal gonadectomy on adult sexual partner preference and brain dimorphism in male rats: partial restoration with pubertal testosterone.

According to the organizational-activational hypothesis, testosterone or its metabolite estradiol, can organize the brain in a male direction (permanently or for long periods) if exposure occurs during a critical (sensitive) time of brain development like the prenatal period. Male rodents with insufficient levels of testosterone during such critical period would irreversibly fail to display sexual partner preference for receptive females in adulthood. However, exposure to testosterone during puberty is believed to function as a second critical period for organization of brain and behavior. Thus, in the present study we explored the effects of neonatal gonadectomy at postnatal day 1 (GNX) on the partner preference of adult males and the size of some sexually dimorphic regions in the brain like the SDN-MPOA, SCN, MeApd and VMH; and challenged its irreversibility by providing exogenous testosterone during puberty. Our results indicated that neonatal GNX impaired partner preference for females and reduced the size of SDN-MPOA, MeApd and VMH, but not SCN. GNX males restored with testosterone in PD30-PD59 (GNX + T) expressed partner preference for sexually receptive females and increased the size of SDN-MPOA and VMH, but not MeApd in adulthood. We conclude that neonatal castration and the lack of testosterone during the first month of life alters sexual behavior and brain dimorphism in adult male rats, but pubertal testosterone reverses the effects on behavior and brain dimorphism to some extent.