Altering the sex determination pathway in Drosophila fat body modifies sex-specific stress responses.

The stress response in Drosophila melanogaster reveals sex differences in behavior, similar to what has been observed in mammals. However, unlike mammals, the sex determination pathway in Drosophila is well established, making this an ideal system to identify factors involved in the modulation of sex-specific responses to stress. In this study, we show that the Drosophila fat body, which has been shown to be important for energy homeostasis and sex determination, is a dynamic tissue that is altered in response to stress in a sex and time-dependent manner. We manipulated the sex determination pathway in the fat body via targeted expression of transformer and transformer-2 and analyzed these animals for changes in their response to stress. In the majority of cases, manipulation of transformer or transformer-2 was able to change the physiological output in response to starvation and oxidative stress to that of the opposite sex. Our data also uncover the possibility of additional downstream targets for transformer and transformer-2 that are separate from the sex determination pathway and can influence behavioral and physiological responses.

Comparative approaches to the study of physiology: Drosophila as a physiological tool.

Numerous studies have detailed the extensive conservation of developmental signaling pathways between the model system, Drosophila melanogaster, and mammalian models, but researchers have also profited from the unique and highly tractable genetic tools available in this system to address critical questions in physiology. In this review, we have described contributions that Drosophila researchers have made to mathematical dynamics of pattern formation, cardiac pathologies, the way in which pain circuits are integrated to elicit responses from sensation, as well as the ways in which gene expression can modulate diverse behaviors and shed light on human cognitive disorders. The broad and diverse array of contributions from Drosophila underscore its translational relevance to modeling human disease.

Early manipulation of juvenile hormone has sexually dimorphic effects on mature adult behavior in Drosophila melanogaster.

Hormones are critical for the development, maturation, and maintenance of physiological systems; therefore, understanding their involvement during maturation of the brain is important for the elucidation of mechanisms by which adults become behaviorally competent. Changes in exogenous and endogenous factors encountered during sexual maturation can have long lasting effects in mature adults. In this study, we investigated the role of the gonadotropic hormone, juvenile hormone (JH), in the modulation of adult behaviors in Drosophila. Here we utilized methoprene (a synthetic JH analog) and precocene (a JH synthesis inhibitor) to manipulate levels of JH in sexually immature male and female Drosophila with or without decreased synthesis of neuronal dopamine (DA). Locomotion and courtship behavior were assayed once the animals had grown to sexual maturity. The results demonstrate a sexually dimorphic role for JH in the modulation of these centrally controlled behaviors in mature animals that is dependent on the age of the animals assayed, and present DA as a candidate neuronal factor that differentially interacts with JH depending on the sex of the animal. The data also suggest that JH modulates these behaviors through an indirect mechanism. Since gonadotropic hormones and DA interact in mammals to affect brain development and later function, our results suggest that this mechanism for the development of adult behavioral competence may be evolutionarily conserved.

Social behavior in prepubertal neurexin 1α deficient rats: A model of neurodevelopmental disorders.

Loss-of-function mutations in the synaptic protein neurexin1α (NRXN1α) are associated with several neurodevelopmental disorders, including autism spectrum disorder (ASD), schizophrenia, and attention-deficit hyperactivity disorder (ADHD), and many of these disorders are defined by core deficits in social cognition. Mouse models of Nrxn1α deficiency are not amenable to studying aspects of social cognition because, in general, mice do not engage in complex social interactions such as social play or prosocial helping behaviors. Rats, on the contrary, engage in these complex, well-characterized social behaviors. Using the Nrxn1tm1Sage Sprague Dawley rat, we tested a range of cognitive and social behaviors in juveniles with haplo- or biallelic Nrxn1α mutation. We found a deficit in ultrasonic vocalizations (USVs) of male and female neonatal rats with Nrxn1α deficiency. A male-specific deficit in social play was observed in Nrxn1α-deficient juveniles, although sociability and social discrimination were unaltered. Nurturing behavior induced by exposure to pups was enhanced in male and female juveniles with biallelic Nrxn1α mutation. Performance in tasks of prosocial helping behavior and food retrieval indicated severe deficits in learning and cognition in juveniles with biallelic Nrxn1α mutation, and a less severe deficit in haploinsufficient rats, although Pavlovian learning was altered only in haploinsufficient males. We also observed a male-specific increase in mobility and object investigation in juveniles with complete Nrxn1α deficiency. Together, these observations more fully characterize the Nrxn1tm1Sage Sprague Dawley rat as a model for Nrxn1α-related neurodevelopmental disorders, and support a rationale for the juvenile rat as a more appropriate model for disorders that involve core deficits in complex social behaviors. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Microglial Phagocytosis of Newborn Cells Is Induced by Endocannabinoids and Sculpts Sex Differences in Juvenile Rat Social Play.

Brain sex differences are established developmentally and generate enduring changes in circuitry and behavior. Steroid-mediated masculinization of the rat amygdala during perinatal development produces higher levels of juvenile rough-and-tumble play by males. This sex difference in social play is highly conserved across mammals, yet the mechanisms by which it is established are unknown. Here, we report that androgen-induced increases in endocannabinoid tone promote microglia phagocytosis during a critical period of amygdala development. Phagocytic microglia engulf more viable newborn cells in males; in females, less phagocytosis allows more astrocytes to survive to the juvenile age. Blocking complement-dependent phagocytosis in males increases astrocyte survival and prevents masculinization of play. Moreover, increased astrocyte density in the juvenile amygdala reduces neuronal excitation during play. These findings highlight novel mechanisms of brain development whereby endocannabinoids induce microglia phagocytosis to regulate newborn astrocyte number and shape the sexual differentiation of social circuitry and behavior. VIDEO ABSTRACT.

Activation of Both CB1 and CB2 Endocannabinoid Receptors Is Critical for Masculinization of the Developing Medial Amygdala and Juvenile Social Play Behavior.

Juvenile social play behavior is a shared trait across a wide variety of mammalian species. When play is characterized by the frequency or duration of physical contact, males usually display more play relative to females. The endocannabinoid system contributes to the development of the sex difference in social play behavior in rats. Treating newborn pups with a nonspecific endocannabinoid agonist, WIN55,212-2, masculinizes subsequent juvenile rough-and-tumble play behavior by females. Here we use specific drugs to target signaling through either the CB1 or CB2 endocannabinoid receptor (CB1R or CB2R) to determine which modulates the development of sex differences in play. Our data reveal that signaling through both CB1R and CB2R must be altered neonatally to modify development of neural circuitry regulating sex differences in play. Neonatal co-agonism of CB1R and CB2R masculinized play by females, whereas co-antagonism of these receptors feminized rates of male play. Because of a known role for the medial amygdala in the sexual differentiation of play, we reconstructed Golgi-impregnated neurons in the juvenile medial amygdala and used factor analysis to identify morphological parameters that were sexually differentiated and responsive to dual agonism of CB1R and CB2R during the early postnatal period. Our results suggest that sex differences in the medial amygdala are modulated by the endocannabinoid system during early development. Sex differences in play behavior are loosely correlated with differences in neuronal morphology.

Utilization of same- vs. mixed-sex dyads impacts the observation of sex differences in juvenile social play behavior.

The study of juvenile social play behavior has gained popularity due to the disruption of social behaviors in several psychiatric illnesses. In contrast to many tests currently utilized in animal models of psychiatric illness, juvenile social play behavior is part of the normal behavioral repertoire in the laboratory rat and can be observed in a controlled setting but without evocation by the experimenter. Understanding sources of naturally occurring differences in the juvenile social play behavior of the rat is a fundamental first step to guide future research on identifying factors that disrupt this behavior. One of the most commonly found variations is a sex difference, with male rats displaying higher levels of rough-and-tumble play behavior relative to females. This sex difference is also observed in human play. In our recent paper published in Biology of Sex Differences, we investigated how the sex and familiarity of the play partner can impact different components of rough-and-tumble play behavior (pouncing, pinning, boxing, and chasing) and the observation of sex differences within each of these components. Our findings suggest that juvenile male rough-and-tumble play behavior is impacted by the sex of their play partner, while females are more sensitive to the familiarity of their play partner. In this review, we discuss our recent findings and provide a comprehensive comparison of methodology and the reporting of sex differences in the literature on this topic.

Characterization of juvenile play in rats: importance of sex of self and sex of partner.

BACKGROUND: Juvenile social play is observed in many mammalian species, and its disruption in several neuropsychiatric disorders has greatly increased interest in understanding the origins and sources of variability in this behavior. METHODS: We quantified social play behavior in juvenile rats and investigated the impact of sex and familiarity of the play partner. Sex differences in play behavior were investigated by comparing males and females from either same- or mixed-sex pairs with data pooled over 12 days of analysis. Whether play was altered based on the sex of the play partner was assessed using a paired analysis to compare play with a same- or opposite-sex play partner for both males and females. Additionally, a repeated measures design was utilized to determine whether play changed with increasing age. On postnatal day 33, a novel play partner was introduced. We used a repeated measures analysis to compare postnatal day 33 with the previous day. These approaches were used to assess the effects of age, sex, sex of partner, and familiarity of partner on total social play behavior as well as how play was broken down into components, such as pouncing, pinning, chasing, and boxing. RESULTS: There were sex differences in total frequency of play, and specific parameters of play behavior, such as chasing, pouncing, pinning, and boxing. Additionally, males significantly altered their play behavior in response to the sex of their play partner, whereas females were more sensitive to the familiarity of the play partner. CONCLUSIONS: This study provides critical groundwork for uncovering factors that regulate social play behavior and can be used to guide future mechanistic based work.

Temporally dimorphic recruitment of dopamine neurons into stress response circuitry in Drosophila.

Many studies have pointed to vulnerability to stress and stress-related pathologies at different timepoints during an individual's life span. These sensitive windows are usually during periods of neural development, such as embryogenesis, infancy, and adolescence. It is critical to understand how neural circuitry may change as an individual ages in ways that could affect susceptibility to stress. Here we compare two stages in Drosophila melanogaster: sexual immaturity and sexual maturity. We used the genetic resources available in Drosophila to manipulate pre- and postsynaptic dopamine signaling in sexually immature and mature animals that were then assayed for heart rate and locomotor behavior in response to starvation and oxidative stress. Our results show significant differences in the stress response for sexually immature and mature animals for heart rate, periods of high mobility, mean velocity, and several other parameters of locomotor behavior. Our data show that dopamine neurons are differentially recruited into the stress response circuitry for sexually immature and mature individuals. By observing behaviors that have been previously shown in mammalian models to be affected by stress and altered in models of affective disorders, we provide a genetically tractable model for development and maintenance of the stress response circuitry during sexual maturation.

Sexually dimorphic recruitment of dopamine neurons into the stress response circuitry.

Several previous studies in mammalian systems have shown sexually dimorphic behaviors, neuroendocrine changes, and alterations in neurotransmitter release in response to stress. In addition, men and women are differentially vulnerable to stress-related pathologies, which have led to the hypothesis that the stress response circuitry differs depending on sex. The authors used the genetic tractability of Drosophila to manipulate pre- or postsynaptic dopamine signaling in transgenic animals, which were assayed for several parameters of locomotion and heart rate following exposure to 2 environmental stressors: starvation and oxidative stress. Their results show significant differences in the stress response for males and females by analyzing heart rate, centering time, and high mobility in addition to other locomotor parameters with translational relevance. These data demonstrate that both pre- and postsynaptic neurons are differentially recruited into the dopaminergic stress response circuitry for males and females. The results also show that the response circuits differ depending on the stressor and behavioral output. Furthermore, the authors' studies provide a translatable Drosophila model for further elucidation of factors involved in the sexually dimorphic recruitment of neurons into the stress response circuitry.