Cellular actions of nesfatin-1 in the subfornical organ.

Nesfatin-1, a centrally acting anorexigenic peptide, is produced in several brain areas involved in metabolic processes and has been implicated in the control of ingestive behaviours and cardiovascular functions. The present study aimed to determine whether the subfornical organ (SFO), a central nervous system (CNS) site that has been extensively implicated in the regulation of appetite and thirst, may represent a potential site for central actions of nesfatin-1. We first used the reverse transcriptase-polymerase chain reaction and were able to confirm the presence of mRNA for the nucleobindin-2 gene in the SFO. We then used whole-cell patch clamp recordings to investigate the influence of nesfatin-1 on the membrane potential of dissociated SFO neurones. A total of 80.3% (49 of 61) of neurones tested showed a response to nesfatin-1 (100 nm, 10 nm and 1 nm). Of these, 47.5% depolarised, with a mean depolarisation of 8.2 ± 0.9 mV (n = 29) and 32.8% hyperpolarised with a mean hyperpolarisation of -8.9 ± 1.2 mV (n = 20). Peak magnitudes were seen at a concentration of 1 nm nesfatin-1, whereas no effect was observed at 100 pm nesftain-1 (n = 3). Furthermore, voltage clamp ramp and step protocols revealed a nesfatin-1 induced activation of the delayed rectifier potassium conductance, IK . Pharmacological blockade of this conductance greatly reduced the magnitude and occurrence of the observed hyperpolarisations. The present study thus demonstrates that nesfatin-1 has the ability to influence the membrane potential of SFO neurones, and thus identifies the SFO as a potential site at which nesfatin-1 may act to regulate ingestive behaviour and cardiovascular control.

α-MSH exerts direct postsynaptic excitatory effects on NTS neurons and enhances GABAergic signaling in the NTS.

The central melanocortin system plays an essential role in the regulation of energy balance. While anorexigenic effects of α-melanocyte-stimulating hormone (α-MSH) acting in the nucleus of the solitary tract (NTS), a critical medullary autonomic control center, have been established, the cellular events underlying these effects are less well characterized. In this study, we used whole-cell patch-clamp electrophysiology to examine firstly whether α-MSH exerts direct postsynaptic effects on the membrane potential of rat NTS neurons in slice preparation, and secondly whether α-MSH influences GABAergic signaling in the NTS. In normal artificial cerebrospinal fluid, perfusion of α-MSH (500 nM) resulted in a depolarization in 39% of cells (n=16, mean 6.14±0.54 mV), and a hyperpolarization in 22% of cells (n=9, -6.79±1.02 mV). Studies using tetrodotoxin to block neuronal communication revealed α-MSH exerts direct depolarizing effects on some NTS neurons, and indirect inhibitory effects on others. A third subset of neurons is simultaneously directly depolarized and indirectly hyperpolarized by α-MSH, resulting in a net lack of effect on membrane potential. The inhibitory inputs influenced by α-MSH were identified as GABAergic, as α-MSH increased the frequency, but not amplitude, of inhibitory postsynaptic currents (IPSCs) in 50% of NTS neurons. α-MSH had no effect on the frequency or amplitude of miniature IPSCs. Furthermore, pharmacological blockade of GABAA and GABAB receptors, and physical removal of all synaptic inputs via cellular dissociation, abolished hyperpolarizations induced by α-MSH. We conclude α-MSH exerts direct, postsynaptic excitatory effects on a subset of NTS neurons. By exciting GABAergic NTS neurons and presynaptically enhancing GABAergic signaling, α-MSH also indirectly inhibits other NTS cells. These findings provide critical insight into the cellular events underlying medullary melanocortin anorexigenic effects, and expand the understanding of the circuitries involved in central melanocortin signaling.

Gender constancy judgments in children with gender identity disorder: evidence for a developmental lag.

Gender constancy judgments in children referred for problems in their gender identity development (N = 206) and controls (N = 95) were compared. On Slaby and Frey's (1975) gender constancy interview, the gender-referred children performed more poorly than the controls at three stage levels: gender identity, gender stability, and gender consistency. On the Boy-Girl Identity Task, a second measure of gender constancy (Emmerich et al., 1977), the gender-referred children also performed more poorly. Gender-referred children who had not attained gender consistency engaged in significantly less same-sex-typed play on a free-play task than the gender-referred children who had, but there were no gender consistency effects for the controls. Two other measures of sex-typed behavior were unrelated to gender consistency. In the gender-referred group alone, children who "failed" the gender identity or gender stability stages were more likely to draw an opposite-sex person first on the Draw-a-Person test and to evince more affective gender confusion on the Gender Identity Interview (Zucker et al., 1993) than children who had "passed." It is concluded that children referred for problems in their gender identity development have a developmental lag in gender constancy acquisition. Possible reasons for the lag are discussed.

A gender identity interview for children.

A 12-item gender identity interview schedule was administered to 85 children referred for concerns regarding their gender identity development and 98 clinical and normal control children. Factor analysis identified two factors, which were labeled Affective Gender Confusion and Cognitive Gender Confusion. The gender-referred group gave significantly more deviant responses than did the controls on both factors. Results were discussed with regard to several diagnostic and assessment issues pertaining to children with gender identity disorder.