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• AgRP and POMC terminals project to an identical set of brain nuclei. • Several brain regions exhibit AgRP and α-MSH terminals, but do not express MC4R. • Other areas contain MC4R cells and receive no innervation of AgRP and POMC neurons. • The amount of AgRP and POMC presumptive appositions on MC4R soma is area-specific. • Several brain regions receive more AgRP appositions than α-MSH appositions on MC4R soma. The central melanocortin system is composed of neurons that express either the proopiomelanocortin (POMC) or the agouti-related protein (AgRP). POMC is cleaved in bioactive peptides, including the α-melanocyte-stimulating hormone (α-MSH). α-MSH activates the melanocortin-4 receptor (MC4R) inducing satiety, whereas AgRP acts as an inverse agonist of MC4R. However, only limited information is available regarding possible area-specific differences in the interaction between α-MSH and AgRP terminals on MC4R-expressing cells. Therefore, the objective of the present study was to compare the distribution pattern of α-MSH and AgRP terminals on the perikarya of MC4R-expressing neurons. We performed a triple-label immunofluorescence reaction in brain series of MC4R-reporter mice to visualize MC4R-expressing neurons together with AgRP and α-MSH terminals. POMC and AgRP neurons project to areas that contain MC4R-expressing cells, although several brain nuclei exhibit AgRP and α-MSH terminals, but they do no express MC4R, while other brain areas contain MC4R-expressing cells and receive no apparent innervation of AgRP and POMC neurons. AgRP terminals make more presumptive appositions than α-MSH on the soma of MC4R-expressing neurons of the medial preoptic area and paraventricular nucleus of the hypothalamus (Pa). Additionally, a higher percentage of MC4R cells receive at least one presumptive apposition from AgRP terminals in the median preoptic nucleus and Pa, compared to α-MSH appositions. Thus, our study revealed area-specific differences in the interaction between α-MSH and AgRP terminals and the soma of MC4R-expressing neurons. These findings provide new insights about the relationship between first- and second-order neurons of the central melanocortin system. [ABSTRACT FROM AUTHOR]

Highlights • Neonatal PR and ERβ expression in the POA is decreased in SF-1 KO males. • POA kisspeptin expression at P14 and P21 is decreased in SF-1 KO females. • ARC kisspeptin expression at P7–P21 is decreased in WT but not SF-1 KO males. • These alterations in SF-1 KO mice eliminate sexually dimorphic expression. • ERα levels in the POA and ARC are gonad-independent and similar in all groups. Abstract Gonadal hormones contribute to brain sexual differentiation. We analyzed expression of progesterone receptor (PR), estrogen receptor-α (ERα), ERβ, and kisspeptin, in the preoptic area (POA) and/or the arcuate nucleus (ARC), in gonad-lacking steroidogenic factor-1 knockout (KO) mice during perinatal development. At postnatal-day (P) 0-P7, POA PR levels were higher in wild-type (WT) males compared with WT females, while those in KO males were lower than in WT males and similar to those in WT and KO females. At P14–P21, PR levels in all groups increased similarly. POA ERα levels were similar in all groups at embryonic-day (E) 15.5-P14. Those in WT but not KO males reduced during postnatal development to be significantly lower compared with females at P21. POA ERβ levels were higher in WT males than in WT females, while those in KO males were lower than in WT males and similar to those in WT and KO females at P0–P21. POA kisspeptin expression was female-biased in WT mice, while levels in KO females were lower compared with WT females and similar to those in WT and KO males. ARC kisspeptin levels were equivalent among groups at E15.5-P0. At P7–P21, ARC levels in WT but not KO males became lower compared with WT females. Diethylstilbestrol exposure during P0–P6 and P7–P13 increased POA PR and ERβ, and decreased POA ERα and ARC kisspeptin levels at P7 and/or P14 in both sexes of KO mice. These data further understanding of gonadal hormone action on neuronal marker expression during brain sexual development. [ABSTRACT FROM AUTHOR]

Highlights • Hypothalamic preoptic area (POA) cooling induces thermogenic responses typically observed during cold exposure in rats. • Short-term POA cooling mimics aspects of the glucoregulatory response to cold exposure. • Long-term POA cooling improves glucose tolerance. Abstract Normal glucose homeostasis depends on the capacity of pancreatic β-cells to adjust insulin secretion in response to a change of tissue insulin sensitivity. In cold environments, for example, the dramatic increase of insulin sensitivity required to ensure a sufficient supply of glucose to thermogenic tissues is offset by a proportionate reduction of insulin secretion, such that overall glucose tolerance is preserved. That these cold-induced changes of insulin secretion and insulin sensitivity are dependent on sympathetic nervous system (SNS) outflow suggests a key role for thermoregulatory neurons in the hypothalamic preoptic area (POA) in this metabolic response. As these POA neurons are themselves sensitive to changes in local hypothalamic temperature, we hypothesized that direct cooling of the POA would elicit the same glucoregulatory responses that we observed during cold exposure. To test this hypothesis, we used a thermode to cool the POA area, and found that as predicted, short-term (8-h) intense POA cooling reduced glucose-stimulated insulin secretion (GSIS), yet glucose tolerance remained unchanged due to an increase of insulin sensitivity. Longer-term (24-h), more moderate POA cooling, however, failed to inhibit GSIS and improved glucose tolerance, an effect associated with hyperthermia and activation of the hypothalamic-pituitary-adrenal axis, indicative of a stress response. Taken together, these findings suggest that POA cooling is sufficient to recapitulate key glucoregulatory responses to cold exposure. [ABSTRACT FROM AUTHOR]

• GABAergic inhibition of shell NAcc evokes a dramatic hypotension and bradycardia. • Glutamatergic stimulation of shell NAcc yields pressor response and tachycardia. • GABAergic and glutamatergic transmission in the shell NAcc affect urinary bladder control. The shell Nucleus Accumbens (NAcc) projects to the lateral preoptic area, which is involved in the central micturition control and receives inputs from medullary areas involved in cardiovascular control. We investigated the role of GABAergic and glutamatergic transmission in the shell NAcc on intravesical pressure (IP) and cardiovascular control. Male Wistar rats with guide cannulas implanted bilaterally in the shell NAcc 7 days prior to the experiments were anesthetized with 2% isoflurane in 100% O 2 and subjected to cannulation of the femoral artery and vein for mean arterial pressure (MAP) and heart rate recordings (HR) and infusion of drugs, respectively. The urinary bladder (UB) was cannulated for IP measurement. A Doppler flow probe was placed around the renal arterial for renal blood flow (RBF) measurement. After the baseline MAP, HR, IP and RBF recordings for 15 min, GABA or bicuculline methiodate (BMI) or L-glutamate or kynurenic acid (KYN) or saline (vehicle) were bilaterally injected into the shell NAcc and the variables were measured for 30 min. Data are as mean ± SEM and submitted to Student́s t test. GABA injections into the shell NAcc evoked a significant fall in MAP and HR and increased IP and RC compared to saline. L-glutamate in the shell NAcc increased MAP, HR and IP and reduced RC. Injections of BMI and KYN elicited no changes in the variables recorded. Therefore, the GABAergic and glutamatergic transmissions in neurons in the shell NAcc are involved in the neural pathways responsible for the central cardiovascular control and UB regulation. [ABSTRACT FROM AUTHOR]

[Display omitted] • A hypothalamic cell line secreted particles consistent with the size of exosomes (sEVs). • Palmitate altered levels of a spectrum of miRNAs associated with exosomes. • Predicted targets of exosome miRNAs included fatty acid metabolism and insulin signalling. • One specific altered secreted miRNAs in hypothalamic neurons was miR-2137. • sEVs from neurons exposed to palmitate mediated differential responses in neuronal co-culture. Exosomes (sEVs) are extracellular vesicles involved in the pathogenesis of obesity. Notably, exosomal microRNAs (miRNAs) have emerged as crucial mediators of communication between cells and are involved in the development of obesity. One region of the brain known to be dysregulated in obesity is the hypothalamus. It coordinates whole-body energy homeostasis through stimulation and inhibition of the orexigenic neuropeptide (NPY)/agouti-related peptide (AgRP) neurons and anorexigenic proopiomelanocortin (POMC) neurons. A role for hypothalamic astrocytic exosomes in communication with POMC neurons was previously elucidated. Yet, it was unknown whether NPY/AgRP neurons secreted exosomes. We previously established that the saturated fat palmitate alters the intracellular levels of miRNAs and we now questioned whether palmitate would also alter the miRNA content of exosomal miRNAs. We found that the mHypoE-46 cell line secreted particles consistent with the size of exosomes and that palmitate altered levels of a spectrum of miRNAs associated with exosomes. The predicted KEGG pathways of the collective miRNA predicted targets included fatty acid metabolism and type II diabetes mellitus. Of note, one of these altered secreted miRNAs was miR-2137, which was also altered within the cells. We also found that while sEVs collected from the mHypoE-46 neurons increased Pomc mRNA in the mHypoA-POMC/GFP-2 cells after 48 h, the effect was absent with sEVs isolated following palmitate treatment, indicating another potential route by which palmitate promotes obesity. Hypothalamic neuronal exosomes may therefore play a role in the control of energy homeostasis that may be disrupted in obese conditions. [ABSTRACT FROM AUTHOR]

Estrogen-related receptor (ERR) is a member of the nuclear receptor superfamily that has strong homology with estrogen receptor (ER) α. Despite the lack of endogenous ligands, ERR serves as transcription factors through their constitutively active structure with or without interaction with ERα. Among the three subtypes of ERR (α, β, and γ), ERRγ is highly expressed in brain, but the distribution of ERRγ is poorly characterized. Therefore, we investigated ERRγ immunoreactivity throughout the rostro-caudal axis in rat brain. Immunohistochemistry revealed localization of ERRγ protein in the cell nucleus, and a ubiquitous distribution of ERRγ in brain regions including the olfactory bulb, cerebrum, brain stem, and cerebellum. Selective intense immunoreactivity was observed in the reticular thalamic nucleus, zona incerta, circular nucleus, interpeduncular nucleus, pontine nucleus, and parasolitary nucleus. Most ERRγ-immunoreactive (ir) regions were also positive for ERα and/or ERβ, which suggests that ERRγ is involved in modulation of estrogen signaling in adult rat brain. Double immunofluorescence demonstrated colocalization of ERRγ with ERα within the anteroventral periventricular nucleus of the preoptic area (AVPV) and medial preoptic nucleus (MPO), which are major target sites for estrogen action. The results of this study suggest that ERRγ function in the brain is affected by estrogens through an interaction with ERα. The findings also provide basic information on brain region-specific ERRγ function. [ABSTRACT FROM AUTHOR]

[Display omitted] • In adult Wistar rats, CVS down-regulate DMH Trh mRNA levels, while moderate exercise enhances them. • CVS suppresses the response of TRHDMH neurons to food restriction but not that to exercise. • There is a strict negative correlation between DMH Trh mRNA levels and serum corticosterone concentration, an effect that is lost in CVS animals. • Trhr cells are homogeneously distributed along the rostro-caudal extent of the medial LH; some are co-expressing Trh. • Fasting reduces DMH Trh mRNA levels and increases LH Trhr and Trhr2 expression, suggesting reduced TRH release from the TRHDMH->LH projection. • Some responses are distinct in male and female rats. The dorsomedial nucleus of the hypothalamus (DMH) is part of the brain circuits that modulate organism responses to the circadian cycle, energy balance, and psychological stress. A large group of thyrotropin-releasing hormone (Trh) neurons is localized in the DMH; they comprise about one third of the DMH neurons that project to the lateral hypothalamus area (LH). We tested their response to various paradigms. In male Wistar rats, food restriction during adulthood, or chronic variable stress (CVS) during adolescence down-regulated adult DMH Trh mRNA levels compared to those in sedentary animals fed ad libitum ; two weeks of voluntary wheel running during adulthood enhanced DMH Trh mRNA levels compared to pair-fed rats. Except for their magnitude, female responses to exercise were like those in male rats; in contrast, in female rats CVS did not change DMH Trh mRNA levels. A very strong negative correlation between DMH Trh mRNA levels and serum corticosterone concentration in rats of either sex was lost in CVS rats. CVS canceled the response to food restriction, but not that to exercise in either sex. TRH receptor 1 (Trhr) cells were numerous along the rostro-caudal extent of the medial LH. In either sex, fasting during adulthood reduced DMH Trh mRNA levels, and increased LH Trhr mRNA levels, suggesting fasting may inhibit the activity of TRHDMH->LH neurons. Thus, in Wistar rats DMH Trh mRNA levels are regulated by negative energy balance, exercise and chronic variable stress through sex-dependent and -independent pathways. [ABSTRACT FROM AUTHOR]

The time course effects of ovarian steroids on kisspeptin and GnRH/LH systems is not totally clarified. We investigated the temporal relationship among kisspeptin and GnRH mRNA and kisspeptin content in the preoptic area (POA), GnRH content and release in the medial basal hypothalamus (MBH) and plasma LH levels under different steroid treatments. Ovariectomized rats treated with oil (OVO O ), oil plus single dose of estradiol (OVO E ), oil plus single dose of progesterone (OVO P ), estradiol for 3 days plus oil (OVE O ) or estradiol for 3 days plus progesterone (OVE P ) were hourly decapitated from 10:00 to 17:00 or had the MBH microdialyzed from 09:00 to 19:00. Estradiol and progesterone acutely increased POA kisspeptin content without altering POA kisspeptin mRNA levels. Short-term exposure to both hormones stimulated MBH GnRH content, although no GnRH/LH surges had occurred. Chronic estradiol-treatment increased both kisspeptin mRNA levels and content in the POA, demonstrating that long exposure to estradiol is required to activate the whole kisspeptin synthesis machinery. This was followed by the peak in the GnRH/LH release. In estradiol-primed rats, progesterone further increased POA kisspeptin content, amplified and advanced GnRH/LH surges, with no additional change on POA kisspeptin mRNA. The data show an estradiol-induced temporal association between kisspeptin increase in the POA and GnRH/LH surges. Interestingly, the classic action of progesterone in amplifying and accelerating the GnRH/LH surges seems to occur by a mechanism which involves POA kisspeptin system. [ABSTRACT FROM AUTHOR]

Mating in rabbits lasts only 3–5 s but profoundly changes the female׳s physiology and behavior (e.g., inhibition of scent-marking and ambulation, changes in EEG, and release of GnRH). The behavioral responsiveness to copulation is reduced in lactating rabbits, relative to estrous does, but is enhanced after suppressing a single nursing bout (“biostimulation”). Little is known about the mechanisms mediating the differential responsiveness to mating among estrous, lactating, and biostimulated rabbits. To begin addressing this issue we quantified the number of c-FOS-immunoreactive (IR) cells in the preoptic area (POA), dorsomedial hypothalamus (DMH), ventromedial hypothalamus (VMH), infundibular nucleus (INF), paraventricular nucleus (PVN), supraoptic nucleus (SON), and lateral septum (LS) in mated and unmated does from the above three reproductive conditions. Mating increased c-FOS-IR cells in the POA and PVN relative to unmated estrous does. Biostimulation increased c-FOS-IR cells in the PVN, relative to lactating does, regardless of mating. Lactation reduced the responsiveness of the LS and INF to copulation but increased it in the DMH. No differences were found in the VMH. Conclusions: a) copulation activates forebrain nuclei that regulate scent-marking (POA), ovulation (INF), and post-coital oxytocin release (PVN); b) lactation and suppression of one nursing bout modulate the magnitude of such changes. [ABSTRACT FROM AUTHOR]

Size matched male green anoles ( Anolis carolinensis ) were paired in a neutral setting and allowed to engage in aggressive displays. Winners and losers were apparent in each pair within 90 min, resulting in stable dominant/subordinate dyads. Androgen receptor (AR) expression was assessed at three time points after the initial pairing, 2 h, 3 days, and 10 days in dominants, subordinates, and two groups of control males housed alone or with a female for an equal period of time. Expression was quantified in three forebrain areas that have been implicated in aggression and reproductive social behavior in this species, the preoptic area (POA), the anterior hypothalamus (AH), septal area (SEP), and ventromedial nucleus of the posterior division of the dorsal ventricular ridge (PDVR VM ). There were significant overall group differences in AR mRNA expression in the POA and AH that appeared to result from higher POA AR expression in dominant males compared to other groups, and generally lower AR expression in subordinate males. Pairwise comparison revealed that dominants′ AR mRNA expression in the POA was significantly higher in the 2 h and 3 day groups compared to that of subordinates, with a similar, but nonsignificant, difference in the 10 day group. Dominants had significantly higher AR mRNA expression in the AH compared to that of subordinates in the 2 h group, but differences were not significant at later times. The results suggest that POA and AH sensitivity to androgens is increased in dominants compared to subordinates, and that the difference can be seen soon after the agonistic interaction establishing winners and losers. [ABSTRACT FROM AUTHOR]

The control of gonadotropin-releasing hormone (GnRH) secretion depends on the action of ovarian steroids and several substances, including nitric oxide (NO). NO in the medial preoptic area (MPOA) stimulates the proestrus surge of luteinizing hormone (LH). We studied the effect of estrogen (Tamoxifen-TMX) and progesterone (RU-486) antagonists on mRNA and protein expression of NO synthase (NOS), the enzyme that produces NO, as well as its activity within MPOA. Female rats received s.c. injections of TMX (3 mg/animal) on first and second days of the estrous cycle (9 am), RU-486 (2 mg/animal) on first, second, (8 am and 5 pm) and third days of the estrous cycle (8 am) or oil (controls) and were killed on the third day (5 pm). Real time-PCR and western blotting were performed to study NOS mRNA and protein expressions. The NOS activity was indirectly assessed by measuring the conversion from [14C]-l-arginine into [14C]-l-citrulline. TMX significantly decreased neuronal NOS (nNOS) mRNA expression (90%), and the activity of NOS, but did not alter nNOS protein expression. Also, TMX significantly decreased LH, FSH, estrogen and progesterone plasma levels. RU-486 nor affected NOS mRNA and protein expressions neither the NOS activity in the MPOA, but reduced FSH levels. The nitrergic system in the MPOA can be stimulated by estrogen whereas TMX decreased NOS activity and mRNA expression. In conclusion, the involvement of the nitrergic system in the MPOA to induce the surge of LH on proestrus depends on the estrogen action to stimulate the mRNA-nNOS expression and the activity of nNOS but it does not seem to depend on progesterone action. [ABSTRACT FROM AUTHOR]

• Chronic perinatal subconvulsive activity leads to sex-related differences in pre-juvenile behavior. • Naïve weanling rat pups exhibit functional alternation asymmetry bias to the left. • Subconvulsive activity removes alternation asymmetry bias. • Convulsive activity induces a compulsive switch in functional asymmetry to the right. • Immobility reduction occurs after subconvulsive or convulsive activity only in males. • Sex differences disappear following single or multiple tonic-clonic convulsion(s). Chronic subconvulsive activity in early life leads to sex-related autistic-like deficits in handling, object recognition, and social performance in pre-pubertal rats. Since autism and epilepsy are common neurodevelopmental disorders with high coincidence, we tested whether early-life chronic subconvulsive activity compared to convulsive activity alters handling, spatial memory, lateralization, coping strategy and the seizure threshold in a sex-dependent manner. A hypothesis is that convulsive seizures may alter sex differences induced by subconvulsive (SC) activity. Serial subconvulsive doses of kainic acid (KA) were administered postnatally (0.25–1 mg/kg) for 15 days to induce the chronic subconvulsive phenotype (SC group). Age-matched controls and a subset of SC pups were exposed to a convulsive dose of KA (KA and SC + KA groups; 7.5 mg/kg) or flurothyl vapors. In our open handling test, controls and the ASD groups escaped to a similar degree whereas after convulsive seizures, the pups exhibited freezing behavior; no escapes occurred. In the spontaneous alternating T-Maze control males and females entered the left arm with higher frequency. The SC males but not SC females entered left and right arms to a similar degree; alternation rates were reduced to chance revealing a sex difference. However, in KA and SC + KA groups, there was a sharp loss of spontaneous alternation rates. The rapid repetitive entries shifted to the right in both sexes possibly be due to hippocampal injury and changes in network activity induced by status epilepticus. In the forced swim test (FST), control and CS females were more active than corresponding males. After convulsions, immobility was reduced and vertical mobility was increased in SC and SC + KA males suggesting an elevated coping strategy compared to females. Onset and severity of KA induced status epilepticus was delayed in SC males and females possibly due to desensitization of KA receptors. Following flurothyl exposure, control males had faster onset of twitches and clonic seizures than control females which disappeared after the sub-convulsive pre-treatment. Data suggest that behavioral manifestations are more readily detectable between males and females when low levels of hyperexcitation are present chronically in early postnatal development but diminished after tonic-clonic convulsions persist. Therefore, therapeutic interventions may benefit patients if initiated upon the initial onset of sex-related autistic pathologies, particularly in males, which may reduce subsequent vulnerability to seizures. [ABSTRACT FROM AUTHOR]

Abstract: Brain structures related to reproduction are thought to depend on the action of gonadal steroids acting either during early life (organizing irreversible effects) or adulthood (activating transient effects). More recently puberty has become a focus of attention and it was demonstrated that action of sex steroid hormones at this time plays a critical role in the final organization of brain and behavior. We studied by BrdU immunohistochemistry the ontogeny from hatching to sexual maturity of a previously identified cell population in the preoptic area labeled by a BrdU injection at the end of embryonic period (E12) of sexual differentiation in male and female Japanese quail. After an initial increase between E12 and hatching, the density of BrdU-immunoreactive cells decreased until the beginning of puberty but then increased again during sexual maturation in the caudal preoptic area specifically. Divisions of these cells took place in the brain parenchyma as indicated by the large numbers of pairs of labeled cells. No sex difference affecting these processes could be detected at any stage of development. Large numbers of new cells thus arise around puberty in the caudal preoptic area and presumably contribute to the reorganization of this structure that precedes the emergence of adult reproductive behaviors. [Copyright &y& Elsevier]

Abstract: Circadian rhythms in behavior and physiology change substantially as female mammals undergo the transition from a non-pregnant to a pregnant state. Here, we examined the possibility that site-specific changes in brain regions known to regulate the sleep/wake cycle and body temperature might reflect altered rhythms in these overt functions. Specifically, we compared daily patterns of immunoreactive FOS in early pregnant and diestrous rats in the medial septum (MS), vertical and horizontal diagonal bands of Broca (VDB and HDB), perifornical lateral hypothalamus (LH), and ventrolateral, medial, and median preoptic areas (VLPO, MPA, and MnPO, respectively). In the pregnant animals, FOS expression was reduced and the daily rhythms of expression were lost or attenuated in the MS, VDB, and LH, areas known to support wakefulness, and in the MPA, a brain region that may coordinate sleep/wake patterns with temperature changes. However, despite the well-documented differences in sleep patterns between diestrous and pregnant rats, reproductive state did not affect FOS expression in the VLPO or MnPO, two brain regions in which FOS expression usually correlates with sleep. These data indicate that plasticity in sleep/wake patterns during early pregnancy may be driven by a reduction in wakefulness-promotion by the brain, rather than by an increase in sleep drive. [Copyright &y& Elsevier]

Abstract: Estrogens are critical for a variety of aspects of brain development and adult processes. These steroids act via receptors within specific tissues. Several estrogen receptors (ER) are thought to exist, including ERα and ERβ, which function via classical, genomic mechanisms. These two ERs are found in a variety of species and are critical to diverse functions, including reproductive behaviors. ERβ was discovered more recently than ERα, and very little work has been done on this receptor in reptiles. Currently no data are available on its distribution in the brain in this vertebrate group. Here, we have cloned ERβ in the green anole lizard, mapped its distribution using in situ hybridization, and quantified expression in three brain areas controlling reproductive behaviors—the preoptic area, ventromedial amygdala (AMY), and ventromedial hypothalamus (VMH). ERβ was detected in discrete areas throughout the anole brain, with high levels in limbic regions and motor nuclei in the brainstem. Females had a greater density of ERβ positive cells in the AMY and VMH than males. While the functional consequences of these differences are not clear, they may result in an increased ability to respond to local levels of estradiol. The present work documents that neural ERβ is distributed similarly in reptilian, rodent and avian species, suggesting that it may perform similar roles. However, more work is necessary to elucidate the function of ERβ in this group. [Copyright &y& Elsevier]

Abstract: Adiponectin can act in the brain to increase energy expenditure and reduce body weight by mechanisms not entirely understood. We found that adiponectin type 1 and type 2 receptors (AdipoR1 and AdipoR2) are expressed in warm sensitive neurons of the hypothalamic preoptic area (POA) which play a critical role in the regulation of core body temperature (CBT) and energy balance. Thus, we tested the ability of adiponectin to influence CBT in wild-type mice and in mice deficient for AdipoR1 or AdipoR2. Local injection of adiponectin into the POA induced prolonged elevation of core body temperature and decreased respiratory exchange ratio (RER) indicating that increased energy expenditure is associated with increased oxidation of fat over carbohydrates. In AdipoR1 deficient mice, the ability of adiponectin to raise CBT was significantly blunted and its ability to decrease RER was completely lost. In AdipoR2 deficient mice, adiponectin had only diminished hyperthermic effects but reduced RER similarly to wild type mice. These results indicate that adiponectin can contribute to energy homeostasis by regulating CBT by direct actions on AdipoR1 and R2 in the POA. [Copyright &y& Elsevier]

Abstract: Intra and interspecific comparisons of arginine vasotocin (AVT) and its mammalian homolog arginine vasopressin (AVP) demonstrate several relationships between these neuropeptides and aggression/dominance behaviors. Prior studies in coral reef butterflyfishes and other fishes indicate that features of AVT neurons in the gigantocellular preoptic area (gPOA) and axon varicosities within the ventral nucleus of the ventral telencephalon should have a positive relationship with aggressive behavior, whereas AVT-ir neuronal features in the parvocellular preoptic area (pPOA) should have a negative relationship. We measured the offensive aggression of wild caught territorial monogamous multiband butterflyfish, Chaetodon multicinctus, in a simple lab paradigm that controlled for social context and variations in social stimuli. Offensive aggression did not follow a clear stereotyped pattern, but rather a complex sequence that includes five action patterns and two approach behaviors. We then used immunohistochemistry to test for associations between AVT immunoreactive features and projections with overall offensive aggression. Our results indicate that gPOA cell number was positively related to aggression while both the size and number of pPOA cells were negatively related to aggression. No association between aggression and the number of axon varicosities in the telencephalic region proposed to be associated with aggression was found. This study provides further support for the relationship between AVT neuronal features and aggression in fishes, and provides preliminary evidence that this relationship may relate to the motivation to produce aggressive behaviors in the immediate future. [Copyright &y& Elsevier]

Abstract: In order to enhance understanding of TRPV1 contributions to thermoregulation, we measured the effects of a TRPV1 receptor antagonist, A-889425, on thermoregulatory neurons in the medial preoptic area of the hypothalamus (mPOA) of rats while simultaneously monitoring rectal temperature (T r). Administration of A-889425 (4 μmol/kg, i.v.) significantly increased T r by 0.42±0.02 °C in anesthetized rats. Warm-sensitive (WS) neurons in the mPOA increase firing in response to body warming, and when active stimulate heat loss and inhibit heat production. WS neurons were initially inhibited by A-889425. Subsequently, WS neuronal activity diverged, differentiating WS neurons into two subgroups. One group of WS neurons continued to be inhibited during the recording period while another group of “biphasic” WS neurons increased firing as T r increased. Cold-sensitive (CS) neurons fire at a higher rate during cooling of the body, and when active, may contribute to heat production. Injection of A-889425 affected CS neurons in a manner opposite to the biphasic WS neurons; activity was initially increased followed by a later decrease. Direct administration of A-889425 into the mPOA (10 and 30 nmol) or spinal cord (30 nmol) did not affect T r. Disruption of abdominal TRPV1 receptor function by injection of the TRPV1 receptor agonist, resiniferatoxin (20 μg/kg, i.p.), 9–15 days prior to experiments, blocked the effects of systemically injected A-889425 on T r and mPOA neuronal activity. These data demonstrate that antagonist block of abdominal TRPV1 receptors indirectly modulates activity of thermoregulatory neurons in the mPOA in a manner that is consistent with producing an acute rise in body temperature. [Copyright &y& Elsevier]

Abstract: The ovine sexually dimorphic nucleus (oSDN) is characterized by high levels of aromatase mRNA expression which can be used to delineate its boundaries. The volume of the oSDN is approximately 2 to 3-fold larger in rams that mate with ewes (female-oriented rams) than in rams that mate with other rams (male-oriented rams) and ewes. The sex difference in oSDN volume is present in late gestation fetuses and can be eliminated before birth by exposing genetic females to exogenous testosterone during midgestation, suggesting that early exposure to androgen masculinizes volume of the oSDN. The present study was performed to determine whether differences in oSDN volume are influenced by the adult hormonal environment. Adult rams, behaviorally characterized as female-oriented or male-oriented, and ewes were gonadectomized and treated with subcutaneous implants of testosterone to achieve physiologic concentrations of serum testosterone. Three weeks after implant placement brain tissue was prepared for histological assessment of oSDN volume using in situ hybridization for detection of aromatase mRNA expression. Quantitative analysis revealed that despite similar serum testosterone levels among the groups, the volume of the oSDN was greater in female-oriented rams than in male-oriented rams and ewes (P <0.05). Differences in oSDN volume were specific and not reflective of differences in preoptic area height or brain size. These results suggest that differences in the size of the oSDN in adult sheep were not influenced by adult exposure to testosterone. [Copyright &y& Elsevier]

Abstract: Although opioids are known to influence sleep–wake regulation, the neuroanatomic substrate(s) mediating these effects remain unresolved. We hypothesized that the influence of opiates on sleep may be mediated, at least in part, by the ventrolateral preoptic nucleus (VLPO), a key cell group for producing behavioral sleep. By combining in situ hybridization for kappa and mu receptor mRNA with immunostaining of Fos expressed by VLPO cells during sleep we show that >85% of sleep-active VLPO neurons contain mRNA for either or both opioid receptors. Microinfusions of a kappa receptor agonist into the VLPO region increased NREM sleep by 51% during the subjective night, whereas a mu receptor agonist increased wakefulness by 60% during the subjective day. The sleep- and wake-promoting effects of the kappa and mu agonists were blocked by prior administration of their respective antagonist. Combining retrograde tracing from the VLPO with immunohistochemistry for dynorphin (Dyn, the endogenous kappa receptor agonist) or endomorphin 1 (EM1, the endogenous mu receptor agonist) we show that the central lateral parabrachial subnucleus (PBcl) provides Dyn inputs to the VLPO, whereas hypothalamic histaminergic neurons provide EM1 inputs to the VLPO. In summary, results from the present study suggest that central opioid inputs to the VLPO may play a role in sleep–wake regulation and that the VLPO likely mediates the hypnotic response to high levels of opioid analgesics. [Copyright &y& Elsevier]

Abstract: Sex and seasonal differences in the brain occur in many species and are often related to behavioral expression. For example, morphology of limbic regions involved in male sex behavior are larger in males than in females, and sometimes are larger in the breeding than non-breeding season. Morphology can often be altered in adulthood by manipulating levels of steroid hormones. In untreated green anole lizards, previous work indicated that neuron soma size and density did not differ between the sexes in the preoptic area (POA) or ventromedial nucleus of the amygdala (AMY), two brain regions involved in the control of male reproductive behaviors [O''Bryant, E.L., Wade, J., 2002. Seasonal and sexual dimorphisms in the green anole forebrain. Horm. Behav. 41, 384–395.]. However, soma size was larger in both areas in breeding than non-breeding animals. The current study examined sex and seasonal differences in estimated brain region volume and total neuron number in the POA, AMY, and the ventromedial hypothalamus (VMH), a region typically involved in female reproductive behaviors. The volume of the POA was larger in males, and the POA and VMH of breeding animals were larger than those of non-breeding individuals. Differences in cell number did not exist in either of these two regions. In contrast, neuron counts in the AMY were greater in non-breeding than breeding animals, but the volume did not differ between the seasons. These data suggest that the structure of limbic brain regions is dynamic in adulthood and that parallels between morphology and the expression of masculine behavior exist for the POA, whereas other relationships are more complicated. [Copyright &y& Elsevier]

Abstract: Estrogens exert a wide variety of actions on reproductive and non-reproductive functions. These effects are mediated by slow and long lasting genomic as well as rapid and transient non-genomic mechanisms. Besides the host of studies demonstrating the role of genomic actions at the physiological and behavioral level, mounting evidence highlights the functional significance of non-genomic effects. However, the source of the rapid changes in estrogen availability that are necessary to sustain their fast actions is rarely questioned. For example, the rise of plasma estrogens at pro-estrus that represents one of the fastest documented changes in plasma estrogen concentration appears too slow to explain these actions. Alternatively, estrogen can be synthesized in the brain by the enzyme aromatase providing a source of locally high concentrations of the steroid. Furthermore, recent studies demonstrate that brain aromatase can be rapidly modulated by afferent inputs, including glutamatergic afferents. A role for rapid changes in estrogen production in the central nervous system is supported by experiments showing that acute aromatase inhibition affects nociception as well as male sexual behavior and that preoptic aromatase activity is rapidly (within min) modulated following mating. Such mechanisms thus fulfill the gap existing between the fast actions of estrogen and their mode of production and open new avenues for the understanding of estrogenic effects on the brain. [Copyright &y& Elsevier]

Abstract: The existence of neural connections between the medial preoptic area (MPOA) and the salivary glands and the increase in salivation by thermal or electrical stimulation of the MPOA have suggested an important role of MPOA in the control of salivary gland function. Although direct cholinergic activation of the salivary glands induces salivation, recent studies have suggested that salivation produced by i.p. pilocarpine may also depend on the activation of central mechanisms. Therefore, in the present study, we investigated the effects of bilateral electrolytic lesions of the MPOA on the salivation induced by i.p. pilocarpine. Adult male Holtzman rats (n = 11–12/group) with bilateral sham or electrolytic lesions of the MPOA were used. One, five, and fifteen days after the brain surgery, under ketamine anesthesia, the salivation was induced by i.p. pilocarpine (1 mg/kg of body weight), and saliva was collected using preweighted small cotton balls inserted into the animal''s mouth. Pilocarpine-induced salivation was reduced 1 and 5 days after MPOA lesion (341 ± 41 and 310 ± 35 mg/7 min, respectively, vs. sham lesions: 428 ± 32 and 495 ± 36 mg/7 min, respectively), but it was fully recovered at the 15th day post-lesion (561 ± 49 vs. sham lesion: 618 ± 27 mg/7 min). Lesions of the MPOA did not affect baseline non-stimulated salivary secretion. The results confirm the importance of MPOA in the control of salivation and suggest that its integrity is necessary for the full sialogogue effect of pilocarpine. However, alternative mechanisms probably involving other central nuclei can replace MPOA function in chronically lesioned rats allowing the complete recovery of the effects of pilocarpine. [Copyright &y& Elsevier]

Abstract: Fever induced by E. coli lipopolysaccharide (LPS) in rats is substantially reduced by blockade of central endothelin ETB receptors. This study explores the role of endothelin-1 as a central mediator of fever in rats, by investigating the effect of a pyrogenic dose of LPS on the levels of big endothelin-1 and endothelin-1 in the cerebrospinal fluid (CSF) and endothelin-1 in the plasma. We further assessed whether the increase in body temperature caused by central injection of endothelin-1 constitutes solely a hyperthermia or a true integrated febrile response. LPS (5 μg kg−1, i.v.) induced fever which peaked at 1.16 ± 0.24 °C within 2 h and remained stable up to 5 h. CSF levels of immunoreactive (ir) big endothelin-1 decreased to undetectable levels at 3 h after LPS, returning only partially at 5 h post-injection. CSF ir-endothelin-1 levels were undetectable in saline-treated animals, but reached 21.9 ± 5.2 fmol ml−1 at 3 h after LPS treatment. Plasma ir-endothelin-1 levels were unchanged after saline or LPS. Central injection of endothelin-1 (1 pmol, i.c.v.) caused long-lasting increases in body temperature (0.81 ± 0.17 °C, 3 h), but simultaneously decreased tail skin temperature (−1.10 ± 0.26 °C), indicating cutaneous vasoconstriction. Moreover, endothelin-1 induced fever (1.0 ± 0.3 °C, 3 h) when injected into the preoptic area of the anterior hypothalamus (100 fmol), but not i.v. (1 or 10 pmol). These data suggest that endothelin-1 is produced in the brain and acts centrally as a mediator of LPS-induced fever. [Copyright &y& Elsevier]

Abstract: In rodents, females but not males, in response to escalating levels of estrogen, express a luteinizing hormone (LH) surge that is prompted by a surge in luteinizing hormone-releasing hormone (LHRH). It cannot take place if estrogen-sensitive afferents located in the anteroventral periventricular nucleus (AVPV) are either absent or disabled. Males appear to lack the ability to exhibit an LH surge, but it is unclear what level of the CNS contributes to this dimorphic response. This study was conducted to determine whether estrogen followed by progesterone treatment (E + P) of gonadectomized males evokes Fos activation in LHRH and AVPV neurons as it does in females. The results indicated that, consistent with the males'' inability to express an LH surge in response to E + P treatment, LHRH and AVPV neurons in males failed to show increased Fos activation. Examination of neuron nuclear antigen (NeuN, a neuron-specific marker), estrogen receptor (ERα) and progesterone receptor (PR) neurons in AVPV neurons indicated that, while essentially all the neurons of the caudal AVPV in males and females are steroid responsive, the male possessed half the number of steroid responsive neurons within the caudal AVPV (where activation of Fos is maximal in females) compared to the female. Together, these data indicate that the male lacks a substantial population of steroid receptive AVPV neurons and is unable to respond to the presence of E and P and activate either AVPV or LHRH neurons. [Copyright &y& Elsevier]

Abstract: Astrocytic gap junctional communication is important in steroid hormone regulation of reproductive processes at the level of the hypothalamus, including estrous cyclicity and sexual behavior. We examined the effects of estradiol and progesterone on the abundance of the gap junctional protein, connexin 43 (CX43), which is highly expressed in astrocytes. Gonadectomized rats received hormone treatments that induce maximal sexual behavior and gonadotropin surges in females (estrogen for 48 h followed by progesterone, estrogen alone or progesterone alone). Control animals received vehicle (oil) injections. In the female rat preoptic area (POA), containing the gonadotropin-releasing hormone (GnRH) cell bodies, treatment with estrogen, progesterone or estrogen + progesterone significantly increased CX43 protein levels in immunoblots. In contrast, estrogen + progesterone significantly decreased CX43 levels in the male rat POA. This sexually dimorphic hormonal regulation of CX43 was not evident in the hypothalamus, which contains primarily GnRH nerve terminals. Treatment with estrogen + progesterone significantly decreased CX43 levels in both the male and female hypothalamus. To examine the role of CX43 in female reproductive function, we studied heterozygous female CX43 (CX43+/−) mice. Most mutant mice did not show normal estrous cycles. In addition, when compared to wild type females, CX43+/− mice had reduced lordosis behavior. These data suggest that hypothalamic CX43 expression is regulated by steroid hormones in a brain-region-specific and sexually dimorphic manner. Therefore, gap junctional communication in the POA and hypothalamus may be a factor regulating the estrous cycle and sexual behavior in female rodents. [Copyright &y& Elsevier]

Abstract: Hypoxia causes a regulated decrease in body temperature (Tb). There is circumstantial evidence that the neurotransmitter serotonin (5-HT) in the anteroventral preoptic region (AVPO) mediates this response. However, which 5-HT receptor(s) is (are) involved in this response has not been assessed. Thus, we investigated the participation of the 5-HT receptors (5-HT1, 5-HT2, and 5-HT7) in the AVPO in hypoxic hypothermia. To this end, Tb of conscious Wistar rats was monitored by biotelemetry before and after intra-AVPO microinjection of methysergide (a 5-HT1 and 5-HT2 receptor antagonist, 0.2 and 2 μg/100 nL), WAY-100635 (a 5-HT1A receptor antagonist, 0.3 and 3 μg/100 nL), and SB-269970 (a 5-HT7 receptor antagonist, 0.4 and 4 μ/100 nL), followed by 60 min of hypoxia exposure (7% O2). During the experiments, the mean chamber temperature was 24.6 ± 0.7 °C (mean ± SE) and the mean room temperature was 23.5 ± 0.8 °C (mean ± SE). Intra-AVPO microinjection of vehicle or 5-HT antagonists did not change Tb during normoxic conditions. Exposure of rats to 7% of inspired oxygen evoked typical hypoxia-induced hypothermia after vehicle microinjection, which was not affected by both doses of methysergide. However, WAY-100635 and SB-269970 treatment attenuated the drop in Tb in response to hypoxia. The effect was more pronounced with the 5-HT7 antagonist since both doses (0.4 and 4 μg/0.1μL) were capable of attenuating the hypothermic response. As to the 5-HT1A antagonist, the attenuation of hypoxia-induced hypothermia was only observed at the higher dose. Therefore, the present results are consistent with the notion that 5-HT acts on both 5-HT1A and 5-HT7 receptors in the AVPO to induce hypothermia, during hypoxia. [Copyright &y& Elsevier]

Abstract: Removal of the vomeronasal organ (VNO) in male mouse lemurs led to an increase in the number of immunoreactive gonadotropin-releasing hormone (GnRH) neurons in the medial preoptic area, compared to control males. No difference was found in the mediobasal hypothalamus. In this primate, which presents a fully functional VNO, the anterior part of the hypothalamus could be the major target for VNO-mediated regulation of GnRH function and the subsequent modulation of chemosensory dependent reproductive behavior. [Copyright &y& Elsevier]

Abstract: The preoptic/anterior hypothalamus (POA) is one of the major brain regions where cytokines and their related mediators (i.e., prostaglandins) exert diverse actions. In the present study, the modulatory effects of prostaglandin E2 (PGE2) on the glutamate-induced membrane currents were examined using perforated-patch clamp method in rat POA neurons that had been mechanically dissociated by vibration without enzyme treatment. Application of glutamate through U-tube induced a slow outward current following fast inward ionotroic current at a holding membrane potential of −30 mV. The slow outward current was also induced by N-methyl-d-aspartate (NMDA), accompanied by an increased membrane conductance, and inhibited by perfusion with Ca2+-free solution, tetraethylammonium chloride (TEA), and apamin, suggesting a Ca2+-dependent K+ current (KCa) activated by Ca2+ entry through NMDA channels. Perfusion with PGE2 at 0.1–10 μM, a principal mediator of fever and neuroendocrine control at the POA, did not produce apparent current by itself, but selectively potentiated the glutamate- or NMDA-induced KCa without affecting inward currents. The KCa induced by activation of NMDA receptors may serve as a feedback mechanism and the modulatory effects of PGE2 on the KCa may have an important physiological significance. [Copyright &y& Elsevier]

Androgen-binding protein (ABP) is known to be expressed in the male and female rat hypothalamus. In the present study, we observed immunocytochemically ABP in neurons of the magnocellular hypothalamic nuclei, in the preoptic region and in the lateral hypothalamus. Dense fiber networks with varicosities, containing ABP immunofluorescence, were visible throughout the hypothalamus, the median eminence and in the posterior pituitary lobe. Double immunostaining revealed a partial coexistence of ABP—and oxytocin immunoreactivity in a portion of the magnocellular perikarya. ABP was isolated by affinity chromatography from hypothalamus homogenates. Western blots resulted in immunoreactive (IR) bands with an approximate molecular weight of 35 and 50 kDa. Mass spectrometry of these preparations confirmed the presence of ABP, which was almost identical to ABP isolated from rat testis. It is likely that ABP, expressed in magnocellular oxytocinergic neurons, is subject to axonal transport and release in the hypothalamo-neurohypophyseal system. [Copyright &y& Elsevier]

We recently showed, using dual tract-tracing, that the suprachiasmatic nucleus (SCN), the site of the principal circadian clock in mammals, may have indirect projections to the sleep-promoting ventrolateral preoptic nucleus (VLPO) via relays in the medial preoptic area (MPA), dorsomedial hypothalamic nucleus (DMH), and, to a lesser extent, the subparaventricular zone (SPVZ). Here, we found that the injection of the rostral MPA, the periventricular nucleus/medial SPVZ, and the caudal DMH with a mixture of anterograde and retrograde tracers resulted in dense anterograde labeling in the median preoptic nucleus (MnPO), another key sleep-promoting nucleus in the preoptic region. The retrograde labeling in the SCN was evident as previously reported. The injections in either the MPA or the DMH produced similar densities of varicose fibers between the MnPO and the VLPO, while the injections in the SPVZ yielded a greater density of varicose fibers in the MnPO than in the VLPO. These results suggest that the MPA and DMH are potential relay nuclei to mediate SCN output to the MnPO, as well as to the VLPO, for the circadian control of sleep–wake states. [Copyright &y& Elsevier]

In many mammals, hormonal fluctuations during pregnancy and parturition produce neurochemical events that are necessary for the transition from a non-maternal state to a maternal state that occurs when infants are born. However, the nature of these events is mostly unknown. We investigated whether changes in dopamine (DA) and serotonin (5-HT) activity within the preoptic area (POA) and striatum, neural sites important for some maternal behaviors, could be part of this process. Female rats were sacrificed as either diestrus virgins, on pregnancy day 10 or 20, on the day of parturition, or on day 7 or 17 of lactation. Bilateral tissue punches from the POA, dorsolateral striatum (STdl), and nucleus accumbens (NA) were obtained and levels of intracellular DA and 5-HT analyzed with high-performance liquid chromatography with electrochemical detection (HPLC–EC). In the POA, DA was high in virgins and during early pregnancy, lowest on the day of parturition, and very high during lactation. Although there were no changes in the DOPAC to DA ratio (i.e., turnover), DOPAC levels also followed this pattern. 5-HT turnover in the POA was lower in virgins compared to other groups. In the STdl, DA turnover was highest during late pregnancy and on the day of parturition, while no changes in 5-HT measures were found. No significant effects were found in the NA. Therefore, decreased DAergic activity in the POA and increased DAergic activity in the STdl occurs around parturition, the time when maternal behavior emerges, and may influence its onset. [Copyright &y& Elsevier]

Prostaglandin E2 (PGE2) is thought to be a principal fever mediator. There are four subtypes of PGE (EP) receptors, EP1–EP4. We investigated which EP receptors mediate PGE2-induced hyperthermia by injecting selective EP receptor agonists into the rat lateral cerebral ventricle under unrestrained condition. ONO-DI-004, an EP1 receptor agonist, increased the core temperature (Tc) in a dose-dependent manner (1.6±0.1 °C at 20 nmol, with the peak 30 min after injection) with a time course similar to PGE2-induced hyperthermia. ONO-AE1-259-01 (20 nmol), an EP2 receptor agonist, did not change the Tc. ONO-AE-248 (20 nmol), an EP3 receptor agonist, also increased the Tc. However, the peak effect was delayed (1.2±0.2 °C, 50 min after injection) compared to PGE2. In contrast, ONO-AE1-329, an EP4 receptor agonist, decreased the Tc. These findings suggest that the EP1, EP3, and EP4 receptors all may contribute to the thermoregulatory response to PGE2, but each may have a different role. [Copyright &y& Elsevier]

Activation of the preoptic area (POA) warm sensitive neurons is known to promote non-REM (NREM) sleep and inhibit neuronal discharge in arousal-related brain structures. The perifornical area of the lateral hypothalamus (PF/LH) was recently recognized to be an additional important arousal promoting region. We studied the behavior of PF/LH neurons in rats during the normal sleep–wake cycle and in response to local POA warming. Most PF/LH neurons were wake-active, and exhibited low discharge throughout NREM. Seventy four percent of these wake-active neurons exhibited moderate or strong activation in REM sleep compared to NREM sleep. A substantial group (26%) exhibited very low discharge in REM as well as NREM sleep. Fifty two percent of units in the PF/LH area were responsive to POA warming; 90% of responsive neurons exhibited a significant reduction (−26.47±2.16% for 1 °C of POA warming) in their discharge rate. The inhibitory effect of POA warming on PF/LH neurons was not associated with EEG slowing. This study supports the hypothesis that sleep induction by POA warm sensitive neurons is mediated through the inhibition of multiple arousal-related structures. [Copyright &y& Elsevier]

The preoptic area (POA) occupies a crucial position among the structures participating in thermoregulation, but we know little about its efferent projections for controlling various effector responses. In this study, we used an immunohistochemical analysis of Fos expression during local warming of the preoptic area. To avoid the effects of anesthesia or stress, which are known to elicit Fos induction in various brain regions, we used a novel thermode specifically designed for chronic warming of discrete brain structures in freely moving rats. At an ambient temperature of 22 °C, local POA warming increased Fos immunoreactivity in the supraoptic nucleus (SON) and the periaqueductal gray matter (PAG). Exposure of animals to an ambient temperature of 5 °C induced Fos immunoreactivity in the magnocellular paraventricular nucleus (mPVN) and the dorsomedial region of the hypothalamus (DMH). Concurrent warming of the POA suppressed Fos expression in these areas. These findings suggest that thermal information from the preoptic area sends excitatory signals to the SON and the PAG, and inhibitory signals to the mPVN and the DMH. [Copyright &y& Elsevier]

• VHb exhibits tonic regular, tonic irregular, rhythmic firing, and silent neurons. • VHb input of auditory, optic, limbic, basal ganglia, and pallial information. • VHb activity pattern can change over time and modulate cell response behavior. • VHb stimulation leads to frequency fluctuation in putative dopaminergic neurons. The habenula – a phylogenetically old brain structure present in all vertebrates – is involved in pain processing, reproductive behaviors, sleep-wake cycles, stress responses, reward, and learning. We performed intra- and extracellular recordings of ventral habenula (VHb) neurons in the isolated brain of anurans and revealed similar cell and response properties to those reported for the lateral habenula of mammals. We identified tonic regular, tonic irregular, rhythmic firing, and silent VHb neurons. Transitions between these firing patterns were observed during spontaneous activity. Electrical stimulation of various brain areas demonstrated VHb input of auditory, optic, limbic, basal ganglia, and pallial information. This resulted in three different response behaviors in VHb neurons: excitation, inhibition, or alternating facilitation and suppression of neuronal activity. Spontaneously changing activity patterns were observed to modulate, reset, or suppress the response behavior of VHb neurons, indicating a gating mechanism. This could be a network status or context dependent selection mechanism for which information are transmitted to task relevant brain areas (i.e., sensory system, limbic system, basal ganglia). Furthermore, alternating facilitation and suppression sequences upon auditory nerve stimulation correlated positively fictive motor activities recorded via the compound potential of the vagal nerve. Stimulation of the auditory nerve or the habenula led to facilitation, suppression, or alternating facilitation and suppression of neuronal activity in putative dopaminergic neurons. Due to complex habenula feedback loops with basal ganglia, limbic, and sensory systems, the habenula involvement in a variety of functions might therefore be explained by a modulatory effect on a task-relevant input stream. [ABSTRACT FROM AUTHOR]

• Adolescence is associated with changes in hormonal stress reactivity. • The neurobiological mechanisms that mediate these changes remain unclear. • Differential activation of the paraventricular nucleus has been implicated in these changes. • Our data indicate similar structural connectivity between the pre- and post-adolescent PVN and its direct afferents. • There were different levels of neuronal activation, however, in two of these direct afferents. The hormonal stress response, mediated by the hypothalamic–pituitary-adrenal (HPA) axis, shows greater responsiveness to various stressors in prepubertal compared to adult animals. Though the implications of this age-related change are unclear, this heightened reactivity might contribute to the increase in stress-related dysfunctions observed during adolescence. Interestingly, prepubertal animals show greater stress-induced neural activation compared to adults in the paraventricular nucleus of the hypothalamus (PVN), the area responsible for initiating the hormonal stress response. Thus, it is possible that direct afferents to the PVN, such as the anterior bed nucleus of the stria terminalis (aBST), nucleus of the solitary tract (NTS), posterior BST (pBST), medial preoptic area (MPOA), and dorsomedial nucleus (DMN), contribute to this age-dependent change in reactivity. To investigate these possibilities, two separate experiments were conducted in prepubertal (30 days old) and adult (70 days old) male rats using the retrograde tracer, Fluoro-Gold (FG), and FOS immunohistochemistry to study neural connectivity and activation, respectively. Though there was no difference in the number or size of FG-positive cells in the PVN afferents we examined, we found a significantly greater number of stress-induced FOS-like-positive cells in the aBST and significantly fewer in the DMN in prepubertal compared to adult animals. Together these data suggest that functional, instead of structural, changes in nuclei that project to the PVN may lead to the greater PVN stress responsiveness observed prior to adolescence. Furthermore, these data indicate that nuclei known to directly modulate HPA stress responsiveness show differential activation patterns before and after adolescent development. [ABSTRACT FROM AUTHOR]

• pSTAT5 can be used as a marker of GH-responsive cells in the brain of GH-injected rats. • Rats and mice exhibit a fairly similar brain distribution of GH-responsive neurons. • However, some species-specific differences were observed. • Cholinergic neurons exhibit a distinct responsiveness to GH comparing rats and mice. A growth hormone (GH) injection is able to induce the phosphorylated form of the signal transducer and activator of transcription 5 (pSTAT5) in a large number of cells throughout the mouse brain. The present study had the objective to map the distribution of GH-responsive cells in the brain of rats that received an intracerebroventricular injection of GH and compare it to the pattern found in mice. We observed that rats and mice exhibited a similar distribution of GH-induced pSTAT5 in the majority of areas of the telencephalon, hypothalamus and brainstem. However, rats exhibited a higher density of GH-responsive cells than mice in the horizontal limb of the diagonal band of Broca (HDB), supraoptic and suprachiasmatic nuclei, whereas mice displayed more GH-responsive cells than rats in the hippocampus, lateral hypothalamic area and dorsal motor nucleus of the vagus (DMX). Since both HDB and DMX contain acetylcholine-producing neurons, pSTAT5 was co-localized with choline acetyltransferase in GH-injected animals. We found that 50.0 ± 4.5% of cholinergic neurons in the rat HDB coexpressed GH-induced pSTAT5, whereas very few co-localizations were observed in the mouse HDB. In contrast, rats displayed fewer cholinergic neurons responsive to GH in the DMX at the level of the area postrema. In summary, pSTAT5 can be used as a marker of GH-responsive cells in the rat brain. Although rats and mice exhibit a relatively similar distribution of GH-responsive neurons, some species-specific differences exist, as exemplified for the responsiveness to GH in distinct populations of cholinergic neurons. [ABSTRACT FROM AUTHOR]

• DBS of the infralimbic cortex attenuated cocaine seeking. • DBS of the prelimbic cortex and anterior cingulate had no effect on cocaine seeking. • DBS of the hippocampus and amygdala blocked cocaine and sucrose seeking. Deep brain stimulation (DBS) is a promising therapeutic modality for the treatment of drug craving and addiction. To date, the nucleus accumbens has received the most attention as a potential target region for examining the impact of DBS on cocaine seeking in preclinical models. The present study investigated the effects of DBS in brain regions that send major glutamatergic projections to the nucleus accumbens including the basolateral amygdala (BLA) and ventral hippocampus (vHipp) as well as subregions of the medial prefrontal cortex (mPFC) including the anterior cingulate, infralimbic and prelimbic cortices. The current results showed that DBS in the infralimbic cortex, but not the prelimbic or anterior cingulate cortices, selectively attenuated cocaine-primed reinstatement of drug seeking in rats. The present data also demonstrated that DBS of the BLA and vHipp attenuated the reinstatement of both cocaine and sucrose seeking. These results indicate that the infralimbic cortex may be a suitable target for DBS to prevent relapse of cocaine taking. [ABSTRACT FROM AUTHOR]