Adolescent development of anxiety-related behavior and shifts in behavioral responsiveness to estradiol in female mice.

Early pubertal onset during adolescence is consistently linked with increased risk of anxiety and depression in girls. Although estradiol tends to have anxiolytic effects in adulthood, whether sensitivity to estradiol's anxiolytic actions increases during adolescence is not clear. Using a rodent model, the current study tested the hypothesis that a shift in sensitivity to the anxiolytic effects of estradiol occurs during adolescence. To test this hypothesis, prepubertal and adult C57BL/6 female mice were ovariectomized, implanted with vehicle- or estradiol-filled silastic capsules, and behavioral tested one week later in the open field and elevated zero maze. Our hypothesis predicted that estradiol would decrease anxiety-related behavior to a greater extent in adults than in adolescent females, however, our results did not support this hypothesis. In the open field, estradiol implants significantly decreased anxiety-like behavior in adolescent females (relative to vehicle) and had little to no effect on the behavior of adults. These data suggest that adolescence is associated with a downward shift in sensitivity to the anxiolytic effects of estradiol on behavior in the open field. In contrast, although estradiol treatment did not influence anxiety-like responses in the elevated zero maze in early adolescent or adult females, adolescent females displayed significantly higher levels of anxiety-like behavior than adults. These findings demonstrate that substantial changes in anxiety-related behavior occur during adolescence, including a context-dependent shift in behavioral responsiveness to estradiol.

Developmental stress has sex-specific effects on contextual and cued fear conditioning in adulthood.

Stress-induced deviations in central nervous system development has long-term effects on adult mental health. Previous research in humans demonstrates that prenatal or adolescent stress increases the risk for psychiatric disorders. Animal models investigating the effects of stress during prenatal or adolescent development produces behavioral outcomes analogous to those observed in humans. However, whether adolescent stress exposure potentiates the effects of prenatal stress is currently unknown. Thus, the current study tested whether adolescent stress increases the impact of prenatal stress on contextual and cued fear memory in adulthood. Male and female Sprague Dawley rats were exposed to a chronic variable stress schedule during the last week of gestation, during adolescence, or during both developmental periods before undergoing fear conditioning training in adulthood. Our hypothesis predicted that the combined effects of prenatal and adolescent stress on contextual and cued fear memory would be greater than the effects of stress during either time period alone. In contrast to our hypothesis, however, we found independent effects of prenatal and adolescent stress on contextual and cued fear memory in both sexes, with no additional combined impact of stress exposure during both developmental phases. In males, developmental stress increased freezing behavior during contextual and cued testing regardless of whether stress exposure was prenatal, adolescent, or combined prenatal and adolescent stress exposure. In contrast, the effects of developmental stress in females were both test- and ovarian hormone status-dependent. During cued testing, nonstressed female freezing behavior depended on estrous cycle phase, whereas freezing behavior in stressed females did not, suggesting that developmental stress interferes with hormone-dependent cued fear memory. No effects of developmental stress or estrous cycle phase were observed for contextual fear memory in females. The results of the current study suggest that the effects of prenatal and adolescent stress on contextual and cued fear memory are not cumulative, but the effects of developmental stress on associative memory differ between males and females.

Back to the future: The organizational-activational hypothesis adapted to puberty and adolescence.

Phoenix, Goy, Gerall, and Young first proposed in 1959 the organizational-activational hypothesis of hormone-driven sex differences in brain and behavior. The original hypothesis posited that exposure to steroid hormones early in development masculinizes and defeminizes neural circuits, programming behavioral responses to hormones in adulthood. This hypothesis has inspired a multitude of experiments demonstrating that the perinatal period is a time of maximal sensitivity to gonadal steroid hormones. However, recent work from our laboratory and others demonstrates that steroid-dependent organization of behavior also occurs during adolescence, prompting a reassessment of the developmental time-frame within which organizational effects are possible. In addition, we present evidence that adolescence is part of a single protracted postnatal sensitive period for steroid-dependent organization of male mating behavior that begins perinatally and ends in late adolescence. These findings are consistent with the original formulation of the organizational/activational hypothesis, but extend our notions of what constitutes "early" development considerably. Finally, we present evidence that female behaviors also undergo steroid-dependent organization during adolescence, and that social experience modulates steroid-dependent adolescent brain and behavioral development. The implications for human adolescent development are also discussed, especially with respect to how animal models can help to elucidate the factors underlying the association between pubertal timing and adult psychopathology in humans.

Pubertal hormones, the adolescent brain, and the maturation of social behaviors: Lessons from the Syrian hamster.

Conventional wisdom holds that gonadal steroid hormones organize and sexually differentiate neural circuits perinatally, and at puberty they activate these circuits to facilitate expression of social behaviors. Using the Syrian hamster to study the role of pubertal hormones in behavioral maturation, we have found that pubertal hormones also organize the adolescent brain. Initial studies revealed that male reproductive behavior cannot be activated by gonadal steroids prepubertally, indicating that the brain acquires behavioral responsiveness during adolescence. Subsequent experiments demonstrated that the presence of gonadal hormones during adolescence masculinize and defeminize behavioral responses of males to hormones in adulthood. Preliminary data also suggest that ovarian hormones defeminize but do not masculinize behavioral responses of females to hormones in adulthood. Furthermore, pubertal hormones program the adult expression of agonistic behaviors that are both steroid-dependent and steroid-independent in adulthood. Thus, the interaction between pubertal hormones and the adolescent brain is key for the maturation of adult social behaviors, and perturbations in the timing of this interaction have long-lasting consequences on adult behavior.

Effects of prenatal stress on sexual partner preference in mice.

Three-month old, male Swiss Webster mice were born to either control dams or dams who had been prenatally stressed with light, heat, noise and handling during the last week of gestation. As adults, male offspring were tested on sexual partner preference and sexual behavior (mounting, intromissions and lordosis) with a sexually experienced male stimulus animal and a stimulus estrous female. In comparison to males born to control dams, prenatally stressed males showed a sexual partner preference for the sexually active male as demonstrated by a negative partner preference score, more and longer visits to the male's compartment, fewer and shorter visits to the female's compartment and longer latencies to and lower frequencies of mounts and intromissions of females. In addition, stressed males showed a greater frequency of lordosis and a higher lordosis quotient than did control males. This study is the first to investigate the effects of prenatal stress alone, without hormonal manipulation, on sexual partner preference using both a partner preference paradigm and measures of sexual behavior such as mounting, intromissions and lordosis. These findings support the suggestion that prenatal stress alone is enough to significantly affect sexual partner preference in male mice.

The organizing actions of adolescent gonadal steroid hormones on brain and behavioral development.

Adolescence is a developmental period characterized by dramatic changes in cognition, risk-taking and social behavior. Although gonadal steroid hormones are well-known mediators of these behaviors in adulthood, the role gonadal steroid hormones play in shaping the adolescent brain and behavioral development has only come to light in recent years. Here we discuss the sex-specific impact of gonadal steroid hormones on the developing adolescent brain. Indeed, the effects of gonadal steroid hormones during adolescence on brain structure and behavioral outcomes differs markedly between the sexes. Research findings suggest that adolescence, like the perinatal period, is a sensitive period for the sex-specific effects of gonadal steroid hormones on brain and behavioral development. Furthermore, evidence from studies on male sexual behavior suggests that adolescence is part of a protracted postnatal sensitive period that begins perinatally and ends following adolescence. As such, the perinatal and peripubertal periods of brain and behavioral organization likely do not represent two discrete sensitive periods, but instead are the consequence of normative developmental timing of gonadal hormone secretions in males and females.

Puberty: a finishing school for male social behavior.

The classical view of steroid-dependent organization of brain and behavior holds that gonadal steroid hormones, acting during an early critical period of development, cause permanent structural changes in neural circuits that determine behavioral responses to hormones in adulthood. This classical view has been modified to incorporate evidence that organizational effects of steroids can occur outside of the established perinatal critical period and that multiple critical periods may exist during development. Experiments in this laboratory indicate that steroid-dependent organization of neural circuits underlying male social behaviors occurs during puberty. This work shows that adult-typical reproductive and flank marking behaviors cannot be activated by gonadal steroids in male Syrian hamsters prior to puberty, suggesting that developmentally timed processes during puberty render the nervous system responsive to activating effects of gonadal steroids in adulthood. Additional experiments demonstrate that the presence or absence of gonadal hormones during puberty is a major factor in the ability of steroids to activate reproductive and flank marking behavior in adult male hamsters and in androgen receptor expression within the neural circuit underlying these behaviors. Thus, gonadal hormones during puberty appear to exert long-lasting changes in neural circuits that are responsible for the programming of activational responses to steroids later in adulthood. A two-stage model for maturation of male social behaviors is proposed: a perinatal critical period for sexual differentiation of neural circuits, followed by the pubertal period, during which gonadal steroids further organize the circuits to enhance behavioral responsiveness to hormones in adulthood. Whether puberty is a critical period for the proposed second wave of steroid-dependent organization of behavioral circuits remains to be determined.

Medial preoptic area dopaminergic responses to female pheromones develop during puberty in the male Syrian hamster.

Chemosensory cues from receptive females do not elicit similar reactions before and after puberty in male hamsters. While pheromones facilitate a complex display of reproductive behavior in adults, prepubertal males do not engage in these same behaviors. Dopamine (DA) released from the medial preoptic area (MPOA) in response to a receptive female or her odors is an important component of the neural events underlying adult male rat sexual behavior. The current experiment investigated whether increased dopaminergic activity occurs in the adult male hamster MPOA in response to female pheromones, and if so, whether this response is absent in prepubertal males, which do not mate. Sexually nai;ve prepubertal and adult male hamsters were exposed to cotton swabs with or without pheromone from an estrous female for 0, 5, 15, or 25 min, after which brains were collected and frozen on dry ice. The MPOA was micropunched from frozen coronal sections (500 microm), and concentrations of DA and its primary metabolite DOPAC were determined by high-performance liquid chromatography-electrochemical detection. DOPAC was used as an index of dopaminergic activity. DOPAC levels significantly increased in adults after 15 min exposure to pheromone. In contrast, MPOA DOPAC concentrations did not increase in prepubertal males exposed to pheromone. These data demonstrate that the neural processing of sexually relevant chemosensory stimuli matures during puberty. The absence of a DA response to female pheromones prior to puberty may contribute to the inability of prepubertal males to display reproductive behavior.

Specific alterations in the performance of learning and memory tasks in models of chemoconvulsant-induced status epilepticus.

Cognitive impairment is a common comorbidity in patients with Temporal Lobe Epilepsy (TLE). These impairments, particularly deficits in learning and memory, can be recapitulated in chemoconvulsant models of TLE. Here, we used two relatively low-stress behavioral paradigms, the novel object recognition task (NOR) and a spatial variation, the novel placement recognition task (NPR) to reveal deficits in short and long term memory, in both kainic acid (KA) and pilocarpine (Pilo) treated animals. We found that both KA- and Pilo-induced significant deficits in long term recognition memory but not short term recognition memory. Additionally, KA impaired spatial memory as detected by both NPR and Morris water maze. These deficits were present 1 week after SE. The characterization of memory performance of two chemoconvulsant-models, one of which is considered a surrogate organophosphate, provides an avenue for which targeted cognitive therapeutics can be tested.

Dietary choline supplementation to dams during pregnancy and lactation mitigates the effects of in utero stress exposure on adult anxiety-related behaviors.

Brain cholinergic dysfunction is associated with neuropsychiatric illnesses such as depression, anxiety, and schizophrenia. Maternal stress exposure is associated with these same illnesses in adult offspring, yet the relationship between prenatal stress and brain cholinergic function is largely unexplored. Thus, using a rodent model, the current study implemented an intervention aimed at buffering the potential effects of prenatal stress on the developing brain cholinergic system. Specifically, control and stressed dams were fed choline-supplemented or control chow during pregnancy and lactation, and the anxiety-related behaviors of adult offspring were assessed in the open field, elevated zero maze and social interaction tests. In the open field test, choline supplementation significantly increased center investigation in both stressed and nonstressed female offspring, suggesting that choline-supplementation decreases female anxiety-related behavior irrespective of prenatal stress exposure. In the elevated zero maze, prenatal stress increased anxiety-related behaviors of female offspring fed a control diet (normal choline levels). However, prenatal stress failed to increase anxiety-related behaviors in female offspring receiving supplemental choline during gestation and lactation, suggesting that dietary choline supplementation ameliorated the effects of prenatal stress on anxiety-related behaviors. For male rats, neither prenatal stress nor diet impacted anxiety-related behaviors in the open field or elevated zero maze. In contrast, perinatal choline supplementation mitigated prenatal stress-induced social behavioral deficits in males, whereas neither prenatal stress nor choline supplementation influenced female social behaviors. Taken together, these data suggest that perinatal choline supplementation ameliorates the sex-specific effects of prenatal stress.

The effects of prenatal stress on alpha4 beta2 and alpha7 hippocampal nicotinic acetylcholine receptor levels in adult offspring.

Prenatal stress in humans is associated with psychiatric problems in offspring such as anxiety, depression, and schizophrenia. These same illnesses are also associated with neuronal nicotinic acetylcholine receptor (nAChR) dysfunction. Despite the known associations between prenatal stress exposure and offspring mental illness, and between mental illness and nAChR dysfunction, it is not known whether prenatal stress exposure impacts neuronal nAChRs. Thus, we tested the hypothesis that maternal stress alters the development of hippocampal alpha4 beta2 (α4ß2∗) and alpha7 (α7∗) nicotinic receptor levels in adult offspring. Female Sprague-Dawley rats experienced unpredictable variable stressors two to three times daily during the last week of gestation. At weaning (21 days) the offspring of prenatally stressed (PS) and nonstressed (NS) dams were assigned to same-sex PS or NS groups. In young adulthood (56 days), the brains of offspring were collected and adjacent sections processed for quantitative autoradiography using [125I]-epibatidine (α4ß2* nicotinic receptor-selective) and [125I]-α-bungarotoxin (α-BTX; α7* nicotinic receptor-selective) ligands. We found that PS significantly increased hippocampal α4ß2* nAChRs of males and females in all subfields analyzed. In contrast, only females showed a trend toward PS-induced increases in α7* nAChRs in the dentate gyrus. Interestingly, NS females displayed a significant left-biased lateralization of α7* nAChRs in the laconosum moleculare of area CA1, whereas PS females did not, suggesting that PS interfered with normal lateralization patterns of α7* nAChRs during development. Taken together, our results suggest that PS impacts the development of hippocampal nAChRs, which may be an important link between PS exposure and risk for neuropsychiatric illness.

Advancing addiction treatment: what can we learn from animal studies?

Substance addiction is a maladaptive behavior characterized by compulsive and uncontrolled self-administration of a substance (drug). Years of research indicate that addictive behavior is the result of complex interactions between the drug, the user, and the environment in which the drug is used; therefore, addiction cannot simply be attributed to the neurobiological actions of a drug. However, despite the obvious complexity of addictive behavior, animal models have both advanced understanding of addiction and contributed importantly to the development of medications to treat this disease. We briefly review recent animal models used to study drug addiction and the contribution of data generated by these animal models for the clinical treatment of addictive disorders.

Pubertal testosterone organizes regional volume and neuronal number within the medial amygdala of adult male Syrian hamsters.

The medial amygdala plays a key role in regulating adult social behavior and undergoes structural changes during puberty that may be driven by gonadal hormone secretion during this developmental period. The current study sought to investigate potential organizational effects of testosterone during puberty, activational effects of testosterone in adulthood, and any interactions on regional volume and neuronal number of the medial amygdala. Male Syrian hamsters either did or did not experience endogenous testosterone during pubertal brain development, and then received either testosterone-filled or blank capsules during adulthood 2 weeks before tissue collection. The results show that pubertal testosterone has long-term organizational effects on volume of specific subregions of the medial amygdala such that the presence of pubertal testosterone resulted in 1) decreased volume of the anterior ventral amygdala and, to a lesser extent, the anterior dorsal medial amygdala; and 2) increased volume of the posterior dorsal medial amygdala. Both effects were independent of the presence of testosterone during adulthood. Pubertal testosterone also decreased neuronal number in the anterior dorsal medial amygdala, suggesting a possible mechanism by which pubertal testosterone decreases volume in this subregion. In addition, there was a significant interaction between pubertal and adult testosterone, such that testosterone in adulthood increased the number of neurons in the posterior ventral medial amygdala only in males that did not experience endogenous pubertal testosterone. In conclusion, pubertal testosterone organizes the medial amygdala in a subregion-specific manner, which may contribute to the maturation of adult-typical social behavior.

Maternal stress during pregnancy causes sex-specific alterations in offspring memory performance, social interactions, indices of anxiety, and body mass.

Prenatal stress (PS) impairs memory function; however, it is not clear whether PS-induced memory deficits are specific to spatial memory, or whether memory is more generally compromised by PS. Here we sought to distinguish between these possibilities by assessing spatial, recognition and contextual memory functions in PS and nonstressed (NS) rodents. We also measured anxiety-related and social behaviors to determine whether our unpredictable PS paradigm generates a behavioral phenotype comparable to previous studies. Female Sprague-Dawley rats were exposed to daily random stress during the last gestational week and behavior tested in adulthood. In males but not females, PS decreased memory for novel objects and novel spatial locations, and facilitated memory for novel object/context pairings. In the elevated zero maze, PS increased anxiety-related behavior only in females. Social behaviors also varied with sex and PS condition. Females showed more anogenital sniffing regardless of stress condition. In contrast, prenatal stress eliminated a male-biased sex difference in nonspecific bodily sniffing by decreasing sniffing in males, and increasing sniffing in females. Finally, PS males but not females gained significantly more weight across adulthood than did NS controls. In summary, these data indicate that PS differentially impacts males and females resulting in sex-specific adult behavioral and bodily phenotypes.

Testosterone programs adult social behavior before and during, but not after, adolescence.

Whereas the adolescent brain is a major target for gonadal hormones, our understanding of hormonal influences on adolescent neural and behavioral development remains limited. These experiments investigated how variations in the timing of testosterone (T) exposure, relative to adolescence, alters the strength of steroid-sensitive neural circuits underlying social behavior in male Syrian hamsters. Experiment 1 simulated early, on-time, and late pubertal development by gonadectomizing males on postnatal d 10 and treating with SILASTIC brand T implants for 19 d before, during, or after adolescence. T treatment before or during, but not after, adolescence facilitated mating behavior in adulthood. In addition, preadolescent T treatments most effectively increased mating behavior overall, indicating that the timing of exposure to pubertal hormones contributes to individual differences in adult behavior. Experiment 2 examined the effects of preadolescent T treatment on behavior and brain regional volumes within the mating neural circuit of juvenile males (i.e. still preadolescent). Although preadolescent T treatment did not induce reproductive behavior in juvenile males, it did increase volumes of the bed nucleus of the stria terminalis, sexually dimorphic nucleus, posterodorsal medial amygdala, and posteroventral medial amygdala to adult-typical size. In contrast, juvenile anterodorsal medial amygdala and ventromedial hypothalamus volumes were not changed by preadolescent T treatment yet differed significantly in volume from adult controls, suggesting that further maturation of these brain regions during adolescence is required for the expression of male reproductive behavior. Thus, adolescent maturation of social behavior may involve both steroid-independent and -dependent processes, and adolescence marks the end of a postnatal period of sensitivity to steroid-dependent organization of the brain.

Adolescents and androgens, receptors and rewards.

Adolescence is associated with increases in pleasure-seeking behaviors, which, in turn, are shaped by the pubertal activation of the hypothalamo-pituitary-gonadal axis. In animal models of naturally rewarding behaviors, such as sex, testicular androgens contribute to the development and expression of the behavior in males. To effect behavioral maturation, the brain undergoes significant remodeling during adolescence, and many of the changes are likewise sensitive to androgens, presumably acting through androgen receptors (AR). Given the delicate interaction of gonadal hormones and brain development, it is no surprise that disruption of hormone levels during this sensitive period significantly alters adolescent and adult behaviors. In male hamsters, exposure to testosterone during adolescence is required for normal expression of adult sexual behavior. Males deprived of androgens during puberty display sustained deficits in mating. Conversely, androgens alone are not sufficient to induce mating in prepubertal males, even though brain AR are present before puberty. In this context, wide-spread use of anabolic-androgenic steroids (AAS) during adolescence is a significant concern. AAS abuse has the potential to alter both the timing and the levels of androgens in adolescent males. In hamsters, adolescent AAS exposure increases aggression, and causes lasting changes in neurotransmitter systems. In addition, AAS are themselves reinforcing, as demonstrated by self-administration of testosterone and other AAS. However, recent evidence suggests that the reinforcing effects of androgens may not require classical AR. Therefore, further examination of interactions between androgens and rewarding behaviors in the adolescent brain is required for a better understanding of AAS abuse.

Pubertal hormones modulate the addition of new cells to sexually dimorphic brain regions.

New cells, including neurons, arise in several brain regions during puberty in rats. Sex differences in pubertal addition of cells coincide with adult sexual dimorphisms: for each region, the sex that gains more cells during puberty has a larger volume in adulthood. Removing gonadal hormones before puberty eliminates these sex differences, indicating that gonadal steroids direct the addition of new cells during puberty to maintain and accentuate sexual dimorphisms in the adult brain.

Adolescent development of neuron structure in dentate gyrus granule cells of male Syrian hamsters.

Hippocampal function, including spatial cognition and stress responses, matures during adolescence. In addition, hippocampal neuron structure is modified by gonadal steroid hormones, which increase dramatically at this time. This study investigated pubertal changes in dendritic complexity of dentate gyrus neurons. Dendrites, spines, and cell bodies of Golgi-impregnated neurons from the granule cell layer were traced in pre-, mid-, and late-pubertal male Syrian hamsters (21, 35, and 49 days of age). Sholl analysis determined the number of intersections and total dendritic length contained in concentric spheres set at 25-microm increments from the soma. Spine densities were quantified separately in proximal and distal segments of a subset of neurons used for the Sholl analysis. We found that the structure of neurons in the lower, but not upper, blade of the dentate gyrus changed during adolescence. The lower, infrapyramidal blade showed pruning of dendrites close to the cell body and increases in distal dendritic spine densities across adolescence. These data demonstrate that dentate gyrus neurons undergo substantial structural remodeling during adolescence and that patterns of maturation are region specific. Furthermore, these changes in dendrite structure, which alter the electrophysiological properties of granule cells, are likely related to the adolescent development of hippocampal-dependent cognitive functions such as learning and memory, as well as hippocampus-mediated stress responsivity.

Testicular hormone exposure during adolescence organizes flank-marking behavior and vasopressin receptor binding in the lateral septum.

Adolescence is a period during which many social behaviors emerge. One such behavior, flank marking, is a testosterone-modulated scent marking behavior that communicates dominance status between adult male Syrian hamsters. Testosterone modulates flank-marking behavior by altering neural transmission of vasopressin within a forebrain circuit. This study tested whether testicular hormones secreted during adolescence play purely a transient activational role in the display of flank-marking behavior, or whether adolescent steroid hormone secretions also cause long-term organizational changes in vasopressin binding within brain regions underlying flank-marking behavior. We tested this hypothesis by manipulating whether testicular secretions were present during adolescent development and then tested for flank-marking behavior and vasopressin receptor binding within the flank-marking neural circuit in young adulthood. Specifically, males were gonadectomized immediately before or after adolescence, replaced with testosterone 6 weeks following gonadectomy in young adulthood, and behavior tested 1 week later. Adult testosterone treatment activated flank-marking behavior only in males that were exposed to testicular hormones during adolescence. In addition, males exposed to testicular hormones during adolescence exhibited significantly less vasopressin receptor binding within the lateral septum than males deprived of adolescent hormones, suggesting that hormone-dependent remodeling of synapses normally occurs in the lateral septum during adolescence. These data highlight the importance of gonadal steroid hormone exposure during adolescence for the organization of neural circuits and social behavior.

Dendritic pruning of the medial amygdala during pubertal development of the male Syrian hamster.

The medial amygdala (Me), a brain region essential for mating behavior, changes in size during puberty. In pre-, mid-, and late pubertal (21, 35, and 49 days of age) male Syrian hamsters, we examined neuronal structure in Me and protein levels of spinophilin and synaptophysin in the amygdaloid complex for evidence of synaptic plasticity coincident with behavioral and physiological development. Body weight, testes weight, and testosterone levels increased during puberty. Mounting behavior, including ectopic, nonintromittive, and intromittive mounts, also increased. Neuronal structure in the posterodorsal medial amygdala (MePD) was assessed in Golgi-impregnated neurons. Pruning occurred during puberty in the number of dendrites emanating from the cell body and in terminal dendritic spine densities. Approximately half of all MePD neurons analyzed had an axon emanating from a dendrite rather than the cell body. However, prepubertal males were more likely to have the axon emanating from a higher order dendritic segment (secondary or tertiary) than were mid- and late pubertal males. Finally, protein levels in the amygdaloid complex varied with pubertal age. Spinophilin decreased, while synaptophysin and GAPDH protein levels increased. These results suggest that puberty is a period of dramatic synaptic plasticity in Me. Specifically, pruning of dendrites and spines, in combination with axonal changes, is likely to modify the afferent influences and electrophysiological properties of Me neurons. Because the Me is an integral component of a social behavior neural network, these changes may be related not only to sexual behavior, but also to other behaviors that mature during puberty, including aggressive, risk-taking, fear-related, and parental behaviors.