Estradiol withdrawal following a hormone simulated pregnancy induces deficits in affective behaviors and increases ∆FosB in D1 and D2 neurons in the nucleus accumbens core in mice.

In placental mammals, estradiol levels are chronically elevated during pregnancy, but quickly drop to prepartum levels following birth. This may produce an "estrogen withdrawal" state that has been linked to changes in affective states in humans and rodents during the postpartum period. The neural mechanisms underlying these affective changes, however, are understudied. We used a hormone-simulated pseudopregnancy (HSP), a model of postpartum estrogen withdrawal, in adult female C57BL/6 mice to test the impact of postpartum estradiol withdrawal on several behavioral measures of anxiety and motivation. We found that estradiol withdrawal following HSP increased anxiety-like behavior in the elevated plus maze, but not in the open field or marble burying tests. Although hormone treatment during HSP consistently increased sucrose consumption, sucrose preference was generally not impacted by hormone treatment or subsequent estradiol withdrawal. In the social motivation test, estradiol withdrawal decreased the amount of time spent in proximity to a social stimulus animal. These behavioral changes were accompanied by changes in the expression of ∆FosB, a transcription factor correlated with stable long-term plasticity, in the nucleus accumbens (NAc). Specifically, estrogen-withdrawn females had higher ∆FosB expression in the nucleus accumbens core, but ∆FosB expression did not vary across hormone conditions in the nucleus accumbens shell. Using transgenic reporter mice, we found that this increase in ∆FosB occurred in both D1- and D2-expressing cells in the NAc core. Together, these results suggest that postpartum estrogen withdrawal impacts anxiety and motivation and increases ∆FosB in the NAc core.

Acute and specific modulation of presynaptic aromatization in the vertebrate brain.

Estrogens affect a diversity of peripheral and central physiological endpoints. Traditionally, estrogens were thought to be peripherally derived transcription regulators (i.e. slow acting). More recently, we have learned that estrogens are also synthesized in neuronal cell bodies and synaptic terminals and have potent membrane effects, which modulate brain function. However, the mechanisms that control local steroid concentrations in a temporal and spatial resolution compatible with their acute actions are poorly understood. Here, using differential centrifugation followed by enzymatic assay, we provide evidence that estrogen synthesis within synaptosomes can be modulated more dramatically by phosphorylating conditions, relative to microsomes. This is the first demonstration of a rapid mechanism that may alter steroid concentrations within the synapse and may represent a potential mechanism for the acute control of neurophysiology and behavior.

Neural distribution of vasotocin receptor mRNA in two species of songbird.

The neurohypophyseal hormones vasopressin and oxytocin are produced and released within the mammalian brain, where they act via multiple receptor subtypes. The neural distributions of these receptors, for example, V1a and oxytocin receptors, have been well described in many mammals. In birds, the distribution of binding sites for the homologous neuropeptides, vasotocin (VT) and mesotocin, has been studied in several species by using synthetic radioligands designed to bind to mammalian receptors. Such binding studies, however, may not reveal the specific distributions of each receptor subtype. To identify and map the receptors likely to bind VT and mesotocin, we generated partial cDNA sequences for four VT receptor subtypes, VT1, VT2 (V1b), VT3 (oxytocin-like), and VT4 (V1a), in white-throated sparrow (Zonotrichia albicollis) and zebra finch (Taeniopygia guttata). These genes shared high sequence identity with the homologous avian and mammalian neurohypophyseal peptide receptors, and we found evidence for VT1, VT3, and VT4 receptor mRNA expression throughout the brains of both species. As has been described in rodents, there was striking interspecific and intraspecific variation in the densities and distribution of these receptors. For example, whereas the VT1 receptor mRNA was more widespread in zebra finch brain, the VT3 (oxytocin-like) receptor mRNA was more prevalent in the sparrow brain. Although VT2 (V1b) receptor mRNA was abundant in the pituitary, it was not found in the brain. Because of their association with brain regions implicated in social behavior, the VT1, VT3, and VT4 receptors are all likely candidates for mediating the behavioral effects of VT.

Assessment of hippocampal adeno-associated viral vector gene delivery via frameless stereotaxis in a nonhuman primate.

BACKGROUND/AIMS: Expression of the neuropeptide galanin in hippocampal neurons reduces seizures in the kainic acid rodent model of epilepsy. In order to translate these findings into a human clinical trial, the safety and feasibility of hippocampal adeno-associated viral (AAV) vector expression must be demonstrated in a nonhuman primate model. METHODS: The Stealth Frameless Stereotactic System and Navigus Biopsy Appliance (Medtronic) were used to inject self-complementary AAV2 carrying the gene for green fluorescent protein (GFP) into monkey hippocampi. Using a single occipital trajectory per side (n = 8 trajectories), multiple injections spaced by 5 mm were delivered to each hippocampus. RESULTS: GFP was expressed in both neuronal and glial cells. Injections led to nonhomogeneous gene expression, suggesting closer spacing of injections may lead to more gene expression. Increasing injection volumes entailed a general increase in volume of expression, but there was no overlap of expression within the 5-mm injection interval. Efforts to avoid the occipital horn failed to prevent leaking of vector into the ventricle, and resulted in deviation of the trajectory at proximal points from the hippocampus. CONCLUSION: Using the occipital approach, adequate cannulation of the monkey hippocampus will require transventricular trajectories.

Comparison of adeno-associated viral vector serotypes for spinal cord and motor neuron gene delivery.

Gene therapy for motor neuron diseases requires efficient gene delivery to motor neurons (MNs) throughout the spinal cord and brainstem. The present study compared adeno-associated viral (AAV) vector serotypes 1, 6, 8, and 9 for spinal cord delivery in adult mice, by the intraparenchymal or intrathecal route of administration. Whereas intraparenchymal injections resulted in local transduction of the lumbar segment of the spinal cord, intrathecal injections led to a broader distribution, transducing cells along the sacral, lumbar, and lower thoracic spinal cord. Overall, AAV6 and AAV9 performed better than the other serotypes. Dramatic differences in cell-specific expression patterns could be observed when constructs bearing the chicken ß-actin (Cba) versus cytomegalovirus (CMV) promoter were compared. In summary, intrathecal delivery of AAV6 or AAV9 vectors containing the CMV promoter yielded the strongest levels of biodistribution and MN transduction in the spinal cord.

Neural distribution of nonapeptide binding sites in two species of songbird.

Vasotocin (VT) and its mammalian homologue, vasopressin (VP), modulate many social behaviors in a variety of vertebrate species. In songbirds, the effects of centrally administered VT vary according to species, which may reflect species-specific distributions of VT binding sites. Different radioligands used to map receptors in previous autoradiographical studies have revealed nonoverlapping distributions of VT binding, suggesting a heterogeneous population of more than one type of VT receptor. For two model songbird species, the white-throated sparrow (Zonotrichia albicollis) and zebra finch (Taeniopygia guttata), we labeled putative VT receptors with two radioligands, [(125)I]ornithine vasotocin analog ([(125)I]OVTA) and [(125)I]linear VP antagonist ([(125)I]HO-LVA). Competitive binding assays in the lateral septum showed that both ligands were effectively displaced by both VT and a related nonapeptide, mesotocin (MT), showing that these radioligands, which were developed to label mammalian nonapeptide receptors, label at least one population of related receptors in songbirds. [(125)I]OVTA labeled receptors throughout the telencephalon, diencephalon, midbrain, and brainstem, with a similar distribution in both species. In contrast, the binding of [(125)I]HO-LVA was restricted to the septal area, dorsal arcopallium, and optic tectum in sparrow and was essentially undetectable in zebra finch. Because the avian brain is likely to express multiple types of VT receptors, we hypothesize that the binding patterns of these radioligands represent a heterogeneous receptor population.

Sex steroids modulate changes in social and sexual preference during juvenile development in zebra finches.

Zebra finches, like many other animals, have close social relationships mainly with the family at young ages but begin to express interest in opposite-sex extra-family animals as they enter the late juvenile period and sexual maturity. This experiment tested a set of hypotheses that sex steroids are involved in this developmental transition. At 25-30 days, subjects were implanted subcutaneously with Silastic tubes that were empty (controls), filled with testosterone propionate, filled with estradiol benzoate, or filled with a combination of ATD (an aromatization inhibitor) and flutamide (an anti-androgen). Once a week between ages 30 and 90 days, they were given three-choice tests where the three stimulus types were the family members, unpaired males, or unpaired females. The preferred category was defined as the one adjacent to the proximity zone in which the subject spent the most time. Control males were more likely to prefer females and less likely to prefer the family as they got older, and control females were increasingly likely to prefer males. Males treated with testosterone or estradiol showed a premature increase in preferences for females. Females treated with ATD plus flutamide failed to show the normal increase in preferences for males shown by controls. These results indicate an involvement of sex steroids in the maturation of sexual preferences in a socially monogamous species that relies on visual and auditory, rather than olfactory, cues for sexual or other social behavior.

Hormonal and social modulation of cloacal muscle activity in female Japanese quail.

Japanese quail are an important model for the discovery of neuroendocrine mechanisms of sexual behavior. In common with most birds and many other vertebrates, there are no genitalia of the mammalian kind (no phallus or vagina). Instead, close contact between the cloacal openings of the male and female is critical for successful mating. Prior research has shown that males produce distinctive rhythmic movements of the foam gland portion of the cloacal sphincter muscle (RCSMs) in response to social stimuli (presentation of a conspecific female or male). These RCSMs enhance the male's fertilization success. Females, unlike males, do not show cloacal sphincter movements to social stimuli (Experiment 1) and female cloacal sphincter muscle activity immediately following copulation does not predict fertilization success (Experiment 2). Females implanted with testosterone propionate (TP), however, show large numbers of male-typical RCSMs in response to social stimuli (Experiment 3). The results of Experiment 3 show that rhythmic movements of the cloacal sphincter muscle shown by TP-treated females are regulated by distal (visual) stimuli and therefore by the brain. They also show that the absence of socially modulated RCSMs in normal females is an activational hormone phenomenon (a sex difference produced by adult hormonal dimorphism), rather than a reflection of prior hormonal organization. These results add to an understanding of the hormonal basis of sex differences in motor mechanisms of sexual behavior in non-mammalian species.