Biomedical subjects

Donald W Pfaff

Publications and source records attributed to Donald W Pfaff.

At least 19 recordsLinked to original sources

PubMed2006

Mathematical analysis of locomotor behavior by mice in a radial maze.

We investigated the effects of beta-estradiol on the locomotor behavior of female mice in a radial maze. Data comprising the total distance traveled during each arm entry were obtained from video records of six consecutive daily recording sessions. Distributions of these data were bimodal for both ovariectomized control and beta-estradiol-treated ovariectomized subjects. Data were fit with the sum of two gamma probability distributions. Three parameters of the analytic fits were useful for quantifying the effect of beta-estradiol on locomotor behavior: (i) the sampling distance (median of the total distance traveled during each arm entry in the short-distance peak of a bimodal distribution), (ii) the committed distance (median of the total per-arm-entry distance traveled in the long-distance peak), and (iii) the partition distance (distance represented by the minimum between the two peaks). Analysis showed that for sampling-distance arm entries beta-estradiol typically had little if any significant effect on female locomotor behavior, whereas it significantly increased the total distance traveled during committed-distance arm entries on the first 2 days of exposure to the empty maze. beta-Estradiol also increased the ability of females to discriminate between empty maze arms and arms that contained intact or castrated male mice and partially prevented loss of this capacity after removal of the males.

Animals↗

PubMed2006

Membrane-initiated actions of estrogens in neuroendocrinology: emerging principles.

Hormonal ligands for the nuclear receptor superfamily have at least two interacting mechanisms of action: 1) classical transcriptional regulation of target genes (genomic mechanisms); and 2) nongenomic actions that are initiated at the cell membrane, which could impact transcription. Although transcriptional mechanisms are increasingly well understood, membrane-initiated actions of these ligands are incompletely understood. Historically, this has led to a considerable divergence of thought in the molecular endocrine field. We have attempted to uncover principles of hormone action that are relevant to membrane-initiated actions of estrogens. There is evidence that the membrane-limited actions of hormones, particularly estrogens, involve the rapid activation of kinases and the release of calcium. Membrane actions of estrogens, which activate these rapid signaling cascades, can also potentiate nuclear transcription. These signaling cascades may occur in parallel or in series but subsequently converge at the level of modification of transcriptionally relevant molecules such as nuclear receptors and/or coactivators. In addition, other hormones or neurotransmitters may also activate cascades to crosstalk with estrogen receptor-mediated transcription. The idea of synergistic coupling between membrane-initiated and genomic actions of hormones fundamentally revises the paradigms of cell signaling in neuroendocrinology.

Animals↗

PubMed2006

Abolition of sex-dependent effects of prenatal exposure to diethylstilbestrol on emotional behavior in estrogen receptor-alpha knockout mice.

To investigate the contribution of estrogen receptor-alpha in the effects of prenatal exposure to diethylstilbestrol on emotionality, estrogen receptor-alpha knockout heterozygous pregnant mice were orally given 0.1 microg/animal of diethylstilbestrol from gestational day 11 to 17. Emotional behavior of the offspring was assessed at 5 weeks in light-dark transition tests. Time spent in the light area was significantly decreased (i.e. decrease of emotionality) by diethylstilbestrol exposure in wild-type female mice, whereas in wild-type male mice this measurement tended to be increased (i.e. increase of emotionality) by diethylstilbestrol treatment. These sex-dependent effects of diethylstilbestrol were completely abolished in estrogen receptor-alpha knockout mice. These results suggest that the sex-dependent effects of diethylstilbestrol on emotionality are mainly produced by its action on estrogen receptor-alpha.

Animals↗

PubMed2006

CNS arousal mechanisms bearing on sex and other biologically regulated behaviors.

It now seems possible to move beyond analyzing only the mechanisms for specific sexual behaviors to the analysis of 'generalized arousal' that underlies all motivated behaviors. Our science has advanced sufficiently to attack mechanisms linking specific motivations to these general arousal mechanisms that intrinsically activate all biologically-regulated behaviors including ingestive behaviors. Learning from the well-developed reproductive behavior paradigm, we know that sex hormone effects on hypothalamic neurons have been studied to a point where receptor mechanisms are relatively well understood, a neural circuit for a sex steroid-dependent behavior has been worked out, and several functional genomic regulations have been discovered. Here we focus for the first time on three chemical systems that signal 'generalized arousal' and which impact hormone-dependent hypothalamic neurons of importance to sexual arousal: histamine, norepinephrine and enkephalin. Progress in linking generalized arousal to specific motivational mechanisms is reviewed.

Animals↗

PubMed2006

RNAi-mediated silencing of estrogen receptor {alpha} in the ventromedial nucleus of hypothalamus abolishes female sexual behaviors.

Estrogen receptor alpha (ERalpha) plays a major role in the regulation of neuroendocrine functions and behaviors by estrogens. Although the generation of ERalpha knockout mice advanced our knowledge of ERalpha functions, gene deletion using this method is global and potentially confounded by developmental consequences. To achieve a site-specific knockdown of ERalpha in the normally developed adult brain, we have generated an adeno-associated virus vector expressing a small hairpin RNA targeting ERalpha. After bilateral injection of this vector into the hypothalamic ventromedial nucleus in ovariectomized female mice, expression levels of ERalpha as well as the estrogen-inducible progesterone receptor were profoundly reduced despite the continued presence of this receptor elsewhere in the brain. Functionally, silencing of ERalpha in the ventromedial nucleus abolished female proceptive and receptive sexual behaviors while enhancing rejection behavior. These results provide evidence that adeno-associated virus-mediated long-term knockdown of genes can be used to delineate their effects on complex behaviors in discrete brain regions.

Adenocarcinoma↗

PubMed2006

Inadvertent social information and the avoidance of parasitized male mice: a role for oxytocin.

Social information can be acquired either directly or indirectly from cues inadvertently produced by individuals with similar interests and requirements ("inadvertent social information," ISI). These inadvertent cues provide "public information" that other individuals can use to guide their behavior. We show here that female mice use olfactory ISI to determine their choice of, and responses to, males and that the use of this ISI involves the gene for oxytocin (OT). Female mice (OT wild type and CF-1 strain) displayed a significant interest in, and choice of, the odors of uninfected males of varying sexual status that were associated with the odors of an another estrous female. This recognition of, and choices for, specific, individual male odors was evident 24 h later. Female mice also distinguished between males subclinically infected with the gastrointestinal nematode parasite, Heligimosomoides polygyrus, and nonparasitized males, displaying aversive responses (analgesia, increased corticosterone) to, and avoidance of, the odors of infected males. The presence of the odors of another estrous female with that of the infected male, which are indicative of potential mate interests, attenuated these aversive responses and resulted in a choice for the odors of infected male. OT gene-deficient (knockout) females were impaired in their use of this ISI to modulate their responses to either uninfected males of differing sexual states or infected males. These findings suggest that OT genes are necessary for the processing of inadvertent social information and likely the integration of both direct and indirect social information.

Animals↗

PubMed2006

Functional genomics of sex hormone-dependent neuroendocrine systems: specific and generalized actions in the CNS.

Sex hormone effects on hypothalamic neurons have been worked out to a point where receptor mechanisms are relatively well understood, a neural circuit for a sex steroid-dependent behavior has been determined, and several functional genomic regulations have been discovered and conceptualized. With that knowledge in hand, we approach deeper problems of explaining sexual arousal and generalized CNS arousal. After a brief summary of arousal mechanisms, we focus on three chemical systems which signal generalized arousal and impact hormone-dependent hypothalamic neurons of behavioral importance: histamine, norepinephrine and enkephalin.

Animals↗

PubMed2006

Estrogen receptor-beta gene disruption potentiates estrogen-inducible aggression but not sexual behaviour in male mice.

Aggressive behaviour of gonadally intact male mice is increased by estrogen receptor (ER)-beta gene disruption, whereas sexual behaviour remains unchanged. The elevated aggression levels following ER-beta gene disruption is pronounced during repeated aggression tests in young animals and the first aggression test in adults. In the present study, the roles of ER-beta activation in the regulation of aggressive and sexual behaviour were investigated in gonadectomized ER-beta knockout (betaERKO) and wild-type (WT) male mice treated with various doses of estrogen. Overall, estradiol benzoate (EB) treatment induced higher levels of aggression in betaERKO mice than in WT mice. In WT mice, the levels of aggression induced by EB were highest in the lowest-dose (2.5 microg/day) group and gradually decreased in higher-dosage groups. On the other hand, equally high levels of aggressive behaviour were induced by all three doses of EB in betaERKO mice. A marked genotype difference in dose responses is inferred, such that the ER-alpha-mediated facilitatory action of estrogen is more pronounced at lower and physiological doses and the ER-beta-mediated inhibitory action is only unveiled at higher doses of estrogen. In contrast to aggression, the levels of sexual behaviour induced by EB were not different between betaERKO and WT at either dose of EB (2.5 and 12.5 microg/day) examined. These findings support the notion that ER-beta activation may exert an attenuating action on male aggression induced by estrogen through ER-alpha-mediated brain mechanisms, whereas its effect on male sexual behaviour is relatively small.

Aggression↗

PubMed2006

Cybernetic principles in the systematic concept of hypothalamic feeding control.

Research on biological mechanisms of eating behavior and related disorders, such as obesity and anorexia nervosa, has become a large field of research in the last 15 years. With the discovery of peptides related to hypothalamic control of food intake (e.g. leptin and ghrelin) the search for the biological 'master key' of feeding control was renewed. As a result, mid-20th century biological concepts based on systematic and cybernetic thoughts fell into oblivion. This review highlights discoveries of hypothalamic-controlled feeding and eating behavior with a cybernetic and systematic perspective. Interestingly, older ideas of hypothalamic function offer possibilities for the incorporation of new molecular discoveries into systematic concepts of feeding behavior.

Animals↗

PubMed2005

Sex differences in estrogenic regulation of neuronal activity in neonatal cultures of ventromedial nucleus of the hypothalamus.

Estrogenic effects have been implicated in sexual differentiation of brain and behavior, in part by affecting neuronal activity in the ventromedial nucleus of the hypothalamus (VMN). We report here a remarkable sex difference in estrogenic regulation of neuronal activity in male vs. female neural networks. Spontaneous synaptic currents originating from a population of neurons were recorded in primary VMN cultures using the whole-cell patch-clamp technique. Treatment with 17beta-estradiol (E2, 10 nM) for 24 h induced opposite effects in the two sexes: the frequency of spontaneous synaptic events decreased significantly in neurons derived from males but increased in those from females. Interestingly, the 24-hour E2 effect was partially reversed by an acute application (5 min) of a second dose of E2 (10 nM), suggesting an interaction between extended (24-hr) and acute (5-min) effects of E2 in VMN neurons. To understand the underlying mechanism of this sexually dimorphic action of E2, we analyzed the E2 effect on GABAergic neurotransmission by recording miniature inhibitory postsynaptic currents. After 24-hour E2 treatment, both the amplitude and frequency of miniature inhibitory postsynaptic currents increased in neurons derived from males but decreased in those from females. These results suggest that E2-induced changes in GABAergic inhibition could at least partially explain E2 effects on neuronal activity. We conclude that E2 may have sexually dimorphic effects on the synaptic output of VMN neurons by modulating GABAergic neurotransmission.

Animals↗

PubMed2005

Estrogen facilitates fear conditioning and increases corticotropin-releasing hormone mRNA expression in the central amygdala in female mice.

Estrogens exert important actions on fear and anxiety-like behavior both in humans and non-human animals. Currently, the mechanisms underlying estrogenic modulation of fear are not known. However, evidence suggests that estrogens may exert their influence on fear within the amygdala. The purpose of the present study was to examine effects of estrogen on fear conditioning. Specifically, the present study examined whether long-term estrogen treatment in ovariectomized female mice via Silastic capsule implantation would facilitate both contextual and cued fear conditioning. In a separate set of experiments, we then examined whether estrogen treatment in ovariectomized female mice would modulate corticotropin-releasing hormone (CRH) gene expression within the amygdala. Long-term estrogen treatment facilitated both contextual and cued fear. Ovariectomized mice treated with estrogen froze significantly more to a context as well as to a discrete auditory cue. In addition, estrogen treatment significantly increased CRH mRNA expression within the central nucleus of the amygdala as measured by in situ hybridization and quantitative PCR. These data raise the possibility that estrogens could influence fear responses in females through their actions in the amygdala.

Amygdala↗

PubMed2005

Recognition and avoidance of the odors of parasitized conspecifics and predators: differential genomic correlates.

In many species of animals chemical stimuli are an important source of information about the threats and dangers present in the social and non-social world. Olfactory cues play a fundamental role in modulating social recognition and interactions in a wide variety of mammals. Rodents, in particular, utilize chemical signals, to recognize and avoid conspecifics infected with parasites and other pathogens. Animals also respond to, and utilize, predator odor related information to assess and minimize their risk of predation. In this review, we briefly focus on the relations between odors, parasite recognition and avoidance and consider some of the associated hormonal, neural and genomic mechanisms. We describe how both male and female rodents distinguish between infected and uninfected males on the basis of odors, displaying aversive response to, and avoidance of, the urine odors of infected individuals. We further describe how the recognition and avoidance of the odors of infected individuals involves genes for the neuropeptide, oxytocin, (OT), and estrogenic mechanisms. We show that mice with deletions of the oxytocin gene (OT knockout mice (OTKO)) and mice whose genes for estrogen receptor (ER)-alpha or ER-beta [ER knockout mice (ERKO), alphaERKO and betaERKO] have been disrupted are specifically impaired in their recognition, avoidance, and memory of the odors of infected individuals. We contrast this with the recognition and display of aversive responses to predator (cat) odor that are insensitive to these genetic manipulations. These findings reveal some of the mechanisms associated with the olfactory mediated recognition of parasitized individuals and predators.

Animals↗

PubMed2005

Calcium flux in neuroblastoma cells is a coupling mechanism between non-genomic and genomic modes of estrogens.

Estrogens have been demonstrated to rapidly modulate calcium levels in a variety of cell types. However, the significance of estrogen-mediated calcium flux in neuronal cells is largely unknown. The relative importance of intra- and extracellular sources of calcium in estrogenic effects on neurons is also not well understood. Previously, we have demonstrated that membrane-limited estrogens, such as E-BSA given before an administration of a 2-hour pulse of 17beta-estradiol (E2), can potentiate the transcription mediated by E2 from a consensus estrogen response element (ERE)-driven reporter gene. Inhibitors to signal transduction cascades given along with E-BSA or E2 demonstrated that calcium flux is important for E-BSA-mediated potentiation of transcription in a transiently transfected neuroblastoma cell line. In this report, we have used inhibitors to different voltage-gated calcium channels (VGCCs) and to intracellular store receptors along with E-BSA in the first pulse or with E2 in the second pulse to investigate the relative importance of these channels to estrogen-mediated transcription. Neither L- nor P-type VGCCs seem to play a role in estrogen action in these cells; while N-type VGCCs are important in both the non-genomic and genomic modes of estrogen action. Specific inhibitors also showed that the ryanodine receptor and the inositol trisphosphate receptor are important to E-BSA-mediated transcriptional potentiation. This report provides evidence that while intracellular stores of calcium are required to couple non-genomic actions of estrogen initiated at the membrane to transcription in the nucleus, extracellular sources of calcium are also important in both non-genomic and genomic actions of estrogens.

Calcium Channel Blockers↗

PubMed2005

Nonmammalian gonadotropin-releasing hormone molecules in the brain of promoter transgenic rats.

Mammalian gonadotropin-releasing hormone (GnRH1) and nonmammalian immunoreactive GnRH subtypes were examined in transgenic rats carrying an enhanced GFP (EGFP) reporter gene driven by a rat GnRH1 promoter. Double-label immunocytochemistry was performed on EGFP(+)/GnRH1 brain sections by using antisera against GnRH1, GnRH2 (chicken II), GnRH3 (salmon), or seabream GnRH. EGFP(+)/GnRH1 neurons were in the septal-preoptic hypothalamus but not in the midbrain, consistent with GnRH1-immunopositive neurons in WT rats. Apparent coexpression of EGFP(+)/GnRH1 with other GnRH subtypes was observed. All EGFP(+) neurons in the septal-preoptic hypothalamus were GnRH1-immunopositive. However, only approximately 80% of GnRH1-immunopositive neurons were EGFP(+), which awaits further elucidation. GnRH subtypes-immunopositive fibers and EGFP(+)/GnRH1 fibers were conspicuous in the organum vasculosum of the lamina terminalis, median eminence, and surrounding the ependymal walls of the third ventricle and the aqueduct in the midbrain. These results demonstrate that the expression of the EGFP-GnRH1 transgene is restricted to the bona fide GnRH1 population and provide clear morphological evidence supporting the existence of GnRH1 neuronal subpopulations in the septal-preoptic hypothalamus, which might be driven by different segments of the GnRH promoter. This genetic construct permits analyses of promoter usage in GnRH neurons, and our histochemical approaches open questions about functional relations among isoforms of this peptide, which regulates reproductive physiology in its behavioral and endocrine aspects.

Animals↗

PubMed2005

Genes, odours and the recognition of parasitized individuals by rodents.

Social recognition, whereby animals identify and recognize other individual conspecifics, is a crucial prerequisite for a wide range of social behaviours. There are relationships among social odours (chemical signals), parasite recognition and avoidance that are associated with hormonal, neural and genomic mechanisms in rodents. Rodents use social odours to: (i) distinguish between infected and uninfected individuals; (ii) recognize specific infected individuals; and (iii) avoid and display aversive responses to infected individuals. There are genomic correlates of this parasite recognition and avoidance in which genes expressing the neuropeptide oxytocin have roles. In this article, we provide a framework ("micronet") by which the genetic, hormonal and neural interactions associated with social behaviours and recognition and avoidance of parasitized individuals can be explored.

Animals↗

PubMed2005

Social and sexual motivation in the mouse.

Does a sexual encounter have reward value for a learned operant response? Ovariectomized female mice with or without estradiol replacement were trained to perform a bar-contact operant response for either male or female targets. Response rates of females with estradiol replacement did not differ from those of females without estradiol replacement or females responding for access to females. Reflexive receptive sexual behavior remained responsive to estradiol replacement. Experiment 2 demonstrated that socially isolated females would respond faster for access to a female target than when group housed. Finally, the oxytocin blocker, atosiban, reduced both operant and reflexive social behavior. These results converge on the conclusion that the operant reward value of social and sexual contact is primarily social.

Animals↗

PubMed2005

Potentiation of genomic actions of estrogen by membrane actions in mcf-7 cells and the involvement of protein kinase C activation.

It is now well established that estrogens (E) have at least two kinds of actions: genomic and nongenomic. But the relationship between these actions has hardly been explored. In this study we investigated this relationship in MCF-7 cells, a human breast cancer cell line, and explored the possible involvement of protein kinase C (PKC) signaling pathways. For this purpose a two-pulse paradigm was used: cells were treated with 17beta-estradiol (E), E conjugated with bovine serum albumin (E-BSA or fE'), or other test agents in the first pulse and with E in the second pulse following a 4-h interval. An E-BSA+E paradigm was used to show that replacement of E with the membrane-impermeable E-BSA in the first pulse could potentiate genomic actions of E, in the second pulse. To investigate involvement of signaling pathways, two PKC activators, phorbol 12,13-diacetate (PDAc) or phorbol 12-myristate 13-acetate (PMA), and inhibitors (chelerythrine chloride and H7-dihydrochloride) were used to replace E or E-BSA in the first pulse. PDAc was as effective as E or E-BSA in potentiating the genomic action of E in the second pulse, while PMA was almost without an effect. Conversely, the potentiating effects of E-BSA and PDAc were blocked by chelerythrine chloride but, interestingly, not by H7. The exact reason underlying these differences is not known. In summary, in MCF-7 cells a membrane action of E can potentiate a later genomic action and involves PKC signaling.

Breast Neoplasms↗

PubMed2004

Vasopressin stimulates ventromedial hypothalamic neurons via oxytocin receptors in oxytocin gene knockout male and female mice.

A wealth of neuropharmacological data demonstrates that oxytocin (OT) actions in the mammalian forebrain support a wide variety of affiliative behaviors and repress aggressive behaviors. Based on that literature, it was expected that reproductive and affiliative behaviors would be vastly decreased and aggression markedly increased in OT gene knockout (OTKO) mice. The initial publications reporting the behaviors of these mice did not include such phenotypes. Here, we compared single-unit activities recorded from the ventromedial hypothalamus in tissue slices of male and female OTKO mice and their wild-type littermate to test two hypotheses about OT functional genomics. First, we proposed that in OTKO mice, a very similar 9-amino-acid neuropeptide, arginine vasopressin (a likely gene duplication product), can 'cross over' and compensate for the lack of OT. This hypothesis was confirmed in both males and females. Further, we proposed that because of the lifelong absence of OT in OTKO, OT receptors would be more sensitive to OT in the knockout animals. We tested this idea in males and found that it was correct. Thus, an answer to the 'OTKO paradox' is put forth, with implications for OT-sensitive behaviors in a variety of species.

Animals↗