Gonadal steroids modulate the growth-associated protein GAP-43 (neuromodulin) mRNA in postnatal rat brain

Gonadal steroid hormone action during early postnatal life determines the growth and connectivity of certain neuronal populations in the hypothalamus. The results of recent studies indicate that steroid hormones modulate the growth-associated protein GAP-43 mRNA in the adult rodent hypothalamus. Since GAP-43 is concentrated in axonal growth cones and has been implicated in axonal elongation and synaptogenesis, the present study investigated the effect of various gonadal hormonal conditions on GAP-43 mRNA levels in postnatal rat brain. On postnatal day 1, male rats were castrated or sham-operated and injected with sesame oil. Additional intact female rats were also injected with oil, while a group of female pups were injected with testosterone propionate. On postnatal day 6, brains were frozen and 16-microns cryostat sections processed and hybridized with a 35S-labeled antisense riboprobe complimentary to GAP-43 mRNA. Slide-mounted sections were stringently washed, apposed to X-ray film and then dipped in liquid emulsion. Evaluation of slide and film autoradiograms revealed an extensive presence of GAP-43 mRNA in the medial preoptic nucleus, bed nucleus of the stria terminalis and cerebral cortex, while the intensity of hybridization signal in other brain regions including the striatum was low. Quantitative assessment of GAP-43 mRNA in the medial preoptic area revealed that the level of GAP-43 mRNA was highest in the sham-operated male, attenuated after male castration, low in the intact female and markedly augmented in the testosterone-treated female. The pattern of change in the bed nucleus of the stria terminalis and laminae II and III of the frontal cortex was similar to that observed in the preoptic area. The changes in hybridization signal were positively correlated with changes in serum testosterone levels as determined by RIA. The results of these studies indicate that GAP-43 mRNA levels in the medial preoptic area, bed nucleus of the stria terminalis and cerebral cortex are sexually dimorphic and modulated by changes in gonadal steroid hormone levels. The results further suggest that the differential regulation of GAP-43 mRNA by sex steroids in the male and female postnatal brain may influence the phenotype of forebrain neuronal circuitry and thereby determine the phenotype of adult neuronal function.     

Guanine nucleotide regulation and cation sensitivity of agonist binding to rat brain oxytocin receptors

The neuropeptide oxytocin (OT) is synthesized in the hypothalamus and can be released either as a hormone from the neurohypophysis or as a neurotransmitter in various brain regions. The present studies were undertaken to better characterize the pharmacological properties of brain oxytocin receptors (OTRs) using a radioligand selective for OTRs. Based on kinetic analysis, brain membranes obtained from 10-day-old rats display rapid and reversible binding to this ligand. In addition, saturation isotherm studies demonstrated that binding was saturable and of high affinity. Indicative of the selectivity of these receptors, compounds known to be ligands for OTRs in other tissues were able to displace the radioligand with high affinity. Consistent with the divalent cation requirement of OTRs in other tissues, OT binding was greatly reduced in rat brain membranes by the removal of magnesium from the incubation. To examine the possible GTP regulation of these receptors, binding was examined in the presence of a GTP analog. High affinity agonist, but not antagonist, binding was reduced by the GTP analog, indicating that these OTRs are likely to be associated with G proteins.     

Naturally occurring truncated trkB receptors have dominant inhibitory effects on brain-derived neurotrophic factor signaling

trkB encodes a receptor tyrosine kinase activated by three neurotrophins--brain-derived neurotrophic factor (BDNF), neurotrophin-3, and neurotrophin-4/5. In vivo, three isoforms of the receptor are generated by differential splicing--gp145trkB or the full-length trkB receptor, and trkB.T1 and trkB.T2, two cytoplasmically truncated receptors that lack kinases, but contain unique C termini. Although the truncated receptors appear to be precisely regulated during nervous system development and regeneration, their role in neurotrophin signaling has not been directly tested. In this paper, we studied the signaling properties and interactions of gp145trkB, trkB.T1, and trkB.T2 by expressing the receptors in a Xenopus oocyte microinjection assay. We found that oocytes expressing gp145trkB, but not trkB.T1 or trkB.T2, were capable of eliciting 45Ca efflux responses (a phospholipase C-gamma-mediated mechanism) after stimulation by BDNF. When trkB.T1 and trkB.T2 were coexpressed with gp145trkB, they acted as dominant negative receptors, inhibiting the BDNF signal by forming nonfunctional heterodimers with the full-length receptors. An ATP-binding mutant of gp145trkB had similar dominant inhibitory effects. Our data suggest that naturally occurring truncated trkB receptors function as inhibitory modulators of neurotrophin responsiveness. Furthermore, the homodimerization of gp145trkB appears to be an essential step in activation of the BDNF signaling cascade.     

A PET study of Turner's syndrome: effects of sex steroids and the X chromosome on brain

Women with Turner's syndrome (TS) allow us to study the neurobiological associates of cognitive and behavioral abnormalities because they lack one/part of one X chromosome, and endogenous estrogen. We studied 13 healthy controls (mean age +/- SD, 28 +/- 6 years) and 16 TS subjects (mean age +/- SD, 26 +/- 6 years). We measured cognitive abilities using neuropsychological tests, and cerebral metabolic rates for glucose with positron emission tomography. Compared to controls, TS subjects had significant absolute hypermetabolism in most brain areas; however, normalized metabolism was significantly lower in TS subjects than controls in the insula and association neocortices bilaterally, and there were significant differences in functional metabolic associations of brain region pairs originating in occipital cortex bilaterally, and within the right hemisphere. There were significant correlations between right-left cognitive and metabolic asymmetries in the TS group. Also, within TS a preliminary analysis demonstrated "X chromosome dosage" effects in language ability and left temporal metabolism, asymmetry of right-left test scores, and parietal metabolism. We hypothesize that within TS: i) generalized brain hypermetabolism reflects global abnormalities in neuron packing; ii) neuronal abnormalities occur in association neocortex that differ in nature or extent from whole brain and are associated with significant differences in normalized metabolism; iii) cognitive deficits are related to brain metabolic abnormalities; and iv) social-behavioral problems may be related to abnormalities of brain metabolism. Moreover, in human brain the X chromosome involved in development of the association neocortices.     

Vasopressin and oxytocin receptors in the central nervous system

This review concentrates on the pharmacological properties and the regional distribution of the arginine vasopressin (AVP) and oxytocin (OT) receptors that are present in the central nervous system. Of particular interest are the kinetics and the pharmacological profiles of these receptors that resemble those present at the periphery. However, their transduction mechanisms need to be further investigated. This should be rendered easier by molecular biology technology. The current knowledge of the anatomical distribution of AVP and OT receptors in the brain is reviewed. Also of great interest for double labeling studies will be the receptor antibodies, now being developed. The existence of additional receptors (AVP4-9) is examined, and aspects of the regulation of the receptors' expression in relation to the age, the species, and the hormonal status or following injury are also discussed.     

The effects of a paradoxical intervention on therapeutic relationship measures.

Compared paradoxical (PIs) and nondirective interventions (NDIs) along core therapeutic conditions and therapist–client relationship issues (attractiveness, expertness, trustworthiness) in 2 experiments to explore criticisms that PIs are manipulative and not genuine. In Exp I, 4 advanced doctoral students rated 4 tapes (2 NDIs and 2 PIs with high- and low-core conditions) for levels of warmth, empathy, and genuineness. Results show that although both NDI and PI high-core-condition tapes were rated high in empathy, warmth, and genuineness, the NDI high-core-condition tape was rated higher. In Exp II, 133 undergraduates rated the same tapes for therapist–client relationship variables. Results show that the PI was rated higher than the NDI in counselor expertness. It is suggested that findings challenge criticisms that PI is contraindicated for use in therapy because of properties that might interrupt or undermine the therapeutic process. An excerpt from one tape is included. (24 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Cellular receptors for sex steroids in human pituitary adenomas

Cellular receptors for sex steroids (SSRs) were studied in an unselected series of 55 human pituitary tumors. Cytosolic receptors for estrogen (ERcs) and progesterone (PgRcs) were determined in all cases and cytosolic androgen receptors (ARcs) in 47 cases. Nuclear receptors (ERns, PgRns, ARns) were also studied in 33 cases. ERs and PgRs were determined by an ELISA and ARs by [3H]methyltrienolone binding. Where both cytosolic and nuclear receptors were studied (n = 33), ERs, PgRs and ARs were found in at least one subcellular fraction in 66.7, 60.6 and 81.8% of cases respectively, ERs and ARs being mainly recovered from the cytosol and PgRs from the nucleus. No linear correlation was found between pre-operative plasma steroid hormones and their specific cellular receptors. Nonetheless, the differential expression of SSRs according to sex and gonadal status at the time of surgery strongly supports their regulation by the steroid environment in vivo: PgRcs were more frequent in tumors found in women (41.4 vs 15.4%, P < 0.05), whereas a high expression of ERcs and ARcs (> 15 fmol/mg protein) was more common in tumors found in men (34.5 vs 10.3%, P < 0.05 and 54.5 vs 24.0% respectively). PgRs were positively correlated with ERns, indicating the possibility of estrogen priming of their expression, and negatively correlated with ARs in nuclear fractions. SSRs appeared to be widely distributed among pituitary tumors, although, compared with other hormone-secreting groups, prolactinomas displayed a higher ERc expression (34.8 +/- 11.3 vs 4.8 +/- 5.1 fmol/mg protein, P = 0.007) and gonadotroph cell adenomas lower ARc values (1.3 +/- 0.8 vs 38.2 +/- 10.6 fmol/mg protein, P = 0.048). Microadenomas were characterized by a higher PgR expression than macroadenomas, whereas hemorrhagic (macro)adenomas were characterized by a high ER expression (> 90%). The present results indicate that most pituitary tumors are targets for sex steroids, SSR expression being partially triggered by the steroid environment itself. Possible physiopathological and therapeutic implications of these findings are discussed.     

The desensitization of oxytocin receptors in human myometrial cells is accompanied by down-regulation of oxytocin receptor messenger RNA

Clinicians providing care to elderly patients must appreciate the subtle clinical manifestations that herald the onset of life-threatening infectious disease. Aged patients with an infection may have neither fever nor leucocytosis, making diagnosis challenging. Often, the early features of infectious disease are nonspecific and may resemble inflammatory or neoplastic processes, or there may be insufficient time to await definitive laboratory confirmation, and empirical antimicrobial treatment must be initiated. Aging involves inevitable deleterious alterations in biological processes and, in many elderly patients, this is most strongly characterised by diminished renal functional capacity. This has a major influence on antimicrobial prescribing in the elderly, because therapeutic efficacy must be achieved while minimising the risk of drug-related toxicity. Before prescribing an antibiotic to an aged patient with an infection, the clinician must be cognisant of the patient's drug allergy history and the other drugs that the patient is taking. Ignorance of potential drug-drug interactions can result in ineffective treatment or enhanced toxicity. The therapy of elderly patients with infections is being expanded. To reduce costs and enhance the efficiency of care, systems have been developed to provide antimicrobial care in the home and in long term care facilities. Home healthcare has burgeoned, and drugs that are well tolerated, have a broad spectrum of activity and are simple to administer (e.g. ceftriaxone and fluoroquinolones) appear to be eminently suitable for this therapeutic role. Physicians must also be informed of the factors responsible for the emergence of resistant bacteria that are contributing to infections in institutional and community settings. Clinicians should strive to curb inappropriate antibiotic use to stem the tide of infections that are caused by multidrug-resistant bacteria.     

A case of paradoxical brain embolism in a 17-year-old boy

OBJECTIVE: To estimate the proportion of 1-4-year-old New South Wales children immune to measles and compare the documented immunization history with serologically defined immune status. DESIGN: Population based seroprevalence survey piggybacked onto the National Survey of Lead in Children. Immune status was determined by two different enzyme immunoassays on plasma samples from subjects. SETTING: New South Wales, February-March 1995. OUTCOME MEASURES: Documented measles immunization collected by interview survey and serologically defined immunity. RESULTS: Of 689 survey subjects, 430 (62.4%) provided a blood sample. Adequate plasma remained for both assays for 347 children, of whom 279 (80.4%) were immune by both assays. Parents of 330 stated that their children were immunised, of whom 211 (63.9%) were able to produce corroborating records. Of these 211 subjects, 178 (84.4%) were immune compared to 87 (76.3%) of 114 without records (P = 0.07). CONCLUSIONS: We estimate the prevalence of true measles immunity in 1-4-year-old NSW children to be only 80%, a level inadequate to prevent outbreaks of measles in urban populations. Both long term and immediate strategies are required to increase the prevalence of immunity among NSW children; these may include lowering the age of the routine second measles dose and mounting a mass measles immunisation campaign to include preschool aged children.     

Activation of acetylcholine receptors and 5-HT2 receptors have additive effects in the suppression of neocortical high-voltage spindles in aged rats

We investigated if activation of the muscarinic or nicotinic acetylcholine receptors and serotonin (5-hydroxytryptamine; 5-HT) subtype 2 receptors would have additive or synergistic effects on the suppression of thalamocortically generated rhythmic neocortical high-voltage spindles (HVSs) in aged rats. The 5-HT2 receptor antagonist, ketanserin, at a moderate dose (5 mg/kg) prevented the ability of a muscarinic acetylcholine receptor agonist, (oxotremorine 0.1 mg/kg), and a nicotinic acetylcholine receptor agonist (nicotine 0.1 mg/kg), to decrease HVSs. At a higher dose (20 mg/kg), ketanserin completely blocked the decrease in HVSs produced by moderate doses of muscarinic acetylcholine receptor agonists (pilocarpine 1 mg/kg and oxotremorine 0.1 mg/kg), and by a high dose of nicotine (0.3 mg/kg), though not that produced by high doses of pilocarpine (3 mg/kg) and oxotremorine (0.9 mg/kg). The ability of a 5-HT2 receptor agonist, (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.1-1.0 mg/kg), to suppress HVSs was non-significantly modulated by the nicotinic acetylcholine receptor antagonist, mecamylamine (1-15 mg/kg), and the muscarinic acetylcholine receptor antagonist, scopolamine (0.03-0.3 mg/kg). The effects of the drugs on behavioral activity could be separated from their effects on HVSs. The results suggest that activation of the muscarinic or nicotinic acetylcholine receptors plus 5-HT2 receptors has additive effects in the suppression of thalamocortical oscillations in aged rats.     

Demonstration of oxytocin receptors in porcine corpora lutea: effects of the cycle stage and the distribution on small and large luteal cells

Recent investigations have demonstrated an inhibitory effect of oxytocin (OXT) on luteal cell progesterone (P) release under in vitro conditions. This inhibitory effect was counteracted by an OXT antagonist, indicating that it was receptor-mediated. In the present investigation, we demonstrated the presence of OXT binding sites in porcine luteal tissue using a radioiodinated OXT antagonist, [1-(beta mercapto-beta,beta-cyclopentamethylene propionic acid),2-(ortho-methyl)-Tyr2-Thr4-Orn8-Tyr-NH2] vasotocin (OTA), as ligand. For membrane fractions of porcine luteal tissue, Kd values of 0.7-0.8 nM were obtained; these are comparable to those of porcine myometrial fractions, measured under the same experimental conditions. Competition studies with luteal membrane fractions yielded a Ki(OXT) of 10(-9) M. This is a dose of OXT that exerts inhibitory effects on P release under both in vitro and in vivo conditions. To evaluate putative variations of luteal OXT receptor concentrations during the estrous cycle, membrane fractions prepared from corpora lutea (CL) of the early or midluteal (Days 2-6) and late luteal phase (Days 9-11) were used. While no differences in Kd values were observed, OXT binding capacities were significantly (p < 0.05) higher in CL from the early/midluteal phase (Bmax(E/M) = 116 +/- 12 fmol/mg protein) compared to CL from the late luteal phase (Bmax(L) = 65 +/- 10 fmol/mg protein). OXT binding sites were present in both small (SLC) and large luteal cells (LLC). SLC but not LLC responded to hCG with a significant increase of OXT binding sites, whereas E2 augmented OXT receptor binding in SLC as well as in LLC.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gonadal steroids promote glial differentiation and alter neuronal morphology in the developing hypothalamus in a regionally specific manner

One of the more striking sexual dimorphisms in the adult brain is the synaptic patterning in some hypothalamic nuclei. In the arcuate nucleus (ARC) males have twice the number of axosomatic and one-half the number of axodendritic spine synapses as females. The opposite pattern is observed in the immediately adjacent ventromedial nucleus (VMN). In both cases, early exposure to testosterone dictates adult dimorphism, but the exact timing, mechanism, and site of steroid action remain unknown. Astrocytes also exhibit sexual dimorphisms, and their role in mediating neuronal morphology is becoming increasingly evident. Using Golgi-Cox impregnation to examine neuronal morphology and glial fibrillary acidic protein immunoreactivity (GFAP-IR) to characterize astrocytic morphology, we compared structural differences in dendrites and astrocytes from the ARC and VMN in postnatal day 2 rat pups from four hormonally different groups. Consistent with previous observations, testosterone exposure induced a rapid and dramatic stellation response in ARC astrocytes. Coincident with this change in astrocytic morphology was a 37% reduction in the density of dendritic spines on ARC neurons. In contrast, astrocytes in the VMN were poorly differentiated and did not respond to testosterone exposure, nor were there any changes in neuronal dendrite spine density. However, VMN neurons exposed to testosterone had almost double the number of branches compared with that in controls. These data suggest that the degree of maturation and the differentiation of hypothalamic astrocytes in vivo are correlated with the ability of neurons to sprout branches or spines in response to steroid hormones and may underlie regionally specific differences in synaptic patterning.     

Electrophysiological evidence for oxytocin receptors on sympathetic preganglionic neurones--an in vitro study on the neonatal rat

The action of oxytocin (0.01-1 microM) on sympathetic preganglionic neurones was studied by intracellular recording in slices of neonatal rat thoracic spinal cord. In 85% of the cells superfusion induced a slow tetrodotoxin-insensitive depolarization accompanied by the appearance or increase in frequency of repetitive discharges. Oxytocin also caused some cells to switch from silent neurones to spontaneously active ones. These effects were reversibly blocked by a specific oxytocin antagonist.     

Epidermal growth factor and fibroblast growth factor-2 have different effects on neural progenitors in the adult rat brain

Neurons and glia are generated throughout adulthood from proliferating cells in two regions of the rat brain, the subventricular zone (SVZ) and the hippocampus. This study shows that exogenous basic fibroblast growth factor (FGF-2) and epidermal growth factor (EGF) have differential and site-specific effects on progenitor cells in vivo. Both growth factors expanded the SVZ progenitor population after 2 weeks of intracerebroventricular administration, but only FGF-2 induced an increase in the number of newborn cells, most prominently neurons, in the olfactory bulb, the normal destination for neuronal progenitors migrating from the SVZ. EGF, on the other hand, reduced the total number of newborn neurons reaching the olfactory bulb and substantially enhanced the generation of astrocytes in the olfactory bulb. Moreover, EGF increased the number of newborn cells in the striatum either by migration of SVZ cells or by stimulation of local progenitor cells. No evidence of neuronal differentiation of newborn striatal cells was found by three-dimensional confocal analysis, although many of these newborn cells were associated closely with striatal neurons. The proliferation of hippocampal progenitors was not affected by either growth factor. However, EGF increased the number of newborn glia and reduced the number of newborn neurons, similar to the effects seen in the olfactory bulb. These findings may be useful for elucidating the in vivo role of growth factors in neurogenesis in the adult CNS and may aid development of neuronal replacement strategies after brain damage.     

Presence and significance of oxytocin receptors in human neuroblastomas and glial tumors

The potential contribution of cell-cell interactions and extracellular factors to cytoarchitectonic abnormalities in the cerebellum of the reeler mutant mouse was investigated by forming chimeras between the reeler and normal animals. The strain origin of Purkinje cells, granule cells and Golgi epithelial cells was immunohistologically identified with a strain-specific antibody. We analyzed 16 overt coat color chimeras, 10 reeler <--> C3H and 6 reeler <--> Balb/c. Abnormal behavioral traits of reeler were rescued in all chimeras. However, cerebellar histology was more affected in reeler <--> C3H chimeras than in reeler <--> Balb/c. Purkinje cells from the normal genotype occupy ectopic positions, and reeler genotype cells are arranged appropriately in the same chimeric cerebellum. We also obtained histologically normal chimeras with a significantly high contribution of the reeler genotype in Purkinje cells, Golgi epithelial cells and granule cells. These results clearly indicate that the abnormal cell positioning and cytoarchitecture of neurons and glia in the reeler is caused by a deficiency of extracellular environments, but is not determined cell-autonomously. The present data on chimeric mice suggest that Reelin is one of the important extracellular environmental factors that affects indirectly radial glial cells during cerebellar histogenesis.