Cell-cell interactions that modulate neuronal development in the leech

Mitotic lineage has been found to determine the cellular identity of leech neurons (reviewed in Stent et al., 1992), Int. Rev. Neurobiol. 33:109-133. However, the details of the adult phenotype of many neurons in the central nervous system of the leech have been shown to be shaped by interactions either with other neurons or with non-neuronal tissues in the environment. Four effects of cell-cell interactions will be considered in this article: stimulation of mitosis that generates new neurons, modulation of cell death or axonal retraction, modification of neurotransmitter metabolism, and modification of other physiological properties. In all cases, the interactions that modify development are thought to occur at a location distant from the soma, requiring that signals be transmitted a significant distance from the site of interaction to the metabolic machinery in the soma.     

Investigating the pharmacological and nonpharmacological factors that modulate drug reinforcement.

Drug use is driven by principles of reinforcement and is sensitive to influences in the environmental context in which it occurs. Although a wide range of factors has been shown to directly influence the reinforcing effects of commonly abused drugs, 2 general types include pharmacological and nonpharmacological factors. Both can assert a powerful impact on a drug's reinforcing effects and, therefore, the degree to which a particular drug comes to be used and abused. This invited review seeks to briefly describe some of the current psychopharmacology research on the interactions between these factors and drug abuse. Several pharmacological influences on drug use will be discussed, including the interactions between psychomotor stimulants and recent advances in the development of pharmacotherapies for opioid abuse. With regard to nonpharmacological factors, there is a large body of research demonstrating that nondrug reinforcers can exert a powerful influence on the reinforcing effects of commonly abused drugs. More specifically, identifying alternative nondrug sources of reinforcement can, if made available contingent on drug abstinence, produce robust decreases in drug self-administration. Presented here is a very brief review of some recent scientific efforts to develop and extend behavioral interventions targeting drug use across a wide range of clinical populations. In summary, understanding the interactions among the variables present in the context of drug use is critical to understanding risk factors for substance use disorders as well as developing efficacious treatments for drug dependence. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Evolution of peptides that modulate the spectral qualities of bound, small-molecule fluorophores

BACKGROUND: Fluorophore dyes are used extensively in biomedical research to sensitively assay cellular constituents and physiology. We have created, as proof of principle, fluorophore dye binding peptides that could have applications in fluorescent dye-based approaches in vitro and in vivo. RESULTS: A panel of Texas red, Rhodamine red, Oregon green 514 and fluorescein binding peptides, termed here 'fluorettes', was selected via biopanning of a combinatorial library of 12-mer peptides fused to a minor coat pIII protein of the filamentous bacteriophage M13. The 'best' fluorette sequences from each of the groups were subjected to further mutagenesis, followed by a second biopanning to select a new generation of improved fluorettes. Phage were selected that had higher avidity for each fluorophore except Rhodamine red. Of these, peptides were characterized that could specifically and with high affinity bind at least one dye, Texas red, in solution. In addition, the binding of certain peptides to Texas red shifted the peak excitation and/or the emission spectra of the bound dye. CONCLUSIONS: Peptides in the context of phage display could readily be selected that could bind to small-molecule fluorophores. The affinities of selected mutant fluorettes could be increased by mutation and further selection. Only a subset of the free peptides could bind free dyes in solution, suggesting that phage context contributed to the selection and ability of certain peptidic regions to independently bind the dyes. Future screens might lead to the creation of other dye-binding peptides with novel characteristics or Texas red derivatives with cross-linking substituents might be designed to increase the utility of the system.     

Injury-induced physiological events that may modulate gene expression in neurons and glia

Damage to the brain triggers a host of reactive responses in neurons and glia which are seen at sites of focal injury as well as at sites that are at a distance from the injury. Although many of these responses have been studied extensively, the signals that initiate the different responses have not been fully characterized, and it is still not understood how focal injury affects neurons and glia in distant sites. The present review summarizes recent findings that suggest that physiological events that occur at the time of the injury or during the early postlesion period can play an important and variable role in modulating neuronal and glial responses to injury. We focus on the events that occur in the hippocampal formation following unilateral lesions of the entorhinal cortex - a model system that has been used extensively for studies of cellular responses following focal brain injury. This lesion destroys the cells of origin of a massive excitatory projection to the dentate gyrus and hippocampus proper. Over time, the denervated neurons in the hippocampal formation are almost completely reinnervated as a result of local sprouting of systems that survive the lesion. Thus, this model system has been useful for studying cellular responses to both denervation and reinnervation. We summarize the information that this injury triggers physiological events that can strongly modulate gene expression in neurons and glia, including episodes of spreading depression that occur at the time of the injury, seizures that occur during the early postlesion period, the loss of afferent drive which leads to decreases in postsynaptic activity, and the restoration of activity that occurs in conjunction with reinnervation. We describe recent studies which suggest that some of these physiological events occur to a variable extent in different animals, especially the episodes of spreading depression and the recurrent seizures. Thus, the spatial pattern and temporal dynamics of altered gene expression following this "model" experimental injury may vary from animal to animal. The fact that physiological events strongly modulate the reactive changes in gene expression that occur following injury has important implications for understanding the sequelae of injury, and offers new opportunities for experimental and therapeutic interventions that may improve cellular repair, regeneration, and recovery of function.     

Discriminative properties of morphine that modulate associations between tastes and lithium chloride.

Wistar rats learned to withhold consumption of a target solution when morphine preceded presentation of the target solution and lithium chloride (LiCl) and to consume the same target solution when saline preceded the presentation of the solution. After this serial feature discrimination training, morphine did not block the formation of a Pavlovian association between saccharin and LiCl but did suppress consumption of familiar tap water. After Pavlovian conditioning, morphine blocked the formation of an association between saccharin and LiCl but did not suppress consumption of a familiar tap water solution. The roles of morphine and saline can be interchanged. It appears that the morphine discriminative stimulus is calling up a representation of neither the conditioned stimulus nor the unconditioned stimulus alone, but rather a modified representation of some aspect of their association. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Structural features that modulate the transmembrane migration of a hydrophobic peptide in lipid vesicles

We have studied the actin-activated ATPase activities of three mutations in the motor domain of the myosin heavy chain that cause familial hypertrophic cardiomyopathy. We placed these mutations in rodent alpha-cardiac myosin to establish the relevance of using rodent systems for studying the biochemical mechanisms of the human disease. We also wished to determine whether the biochemical defects in these mutant alleles correlate with the severity of the clinical phenotype of patients with these alleles. We expressed histidine-tagged rat cardiac myosin motor domains along with rat ventricular light chain 1 in mammalian COS cells. Those myosins studied were wild-type alpha-cardiac and three mutations in the alpha-cardiac myosin heavy chain head (Arg249Gln, Arg403Gln, and Val606Met). These mutations in human beta-cardiac myosin heavy chain have predominantly moderate, severe, and mild clinical phenotypes, respectively. The crystal structure of the skeletal myosin head shows that the Arg249Gln mutation is near the ATP-binding site and the Arg403Gln and Val606Met mutations are in the actin-binding region. Expressed histidine-tagged alpha-motor domains retain physiological ATPase properties similar to those derived from cardiac tissue. All three myosin mutants show defects in the ATPase activity, with the degree of enzymatic impairment of the mutant myosins correlated with the clinical phenotype of patients with the disease caused by the corresponding mutation.     

Musculoskeletal pain syndromes that affect adolescents

Musculoskeletal pain is one of the most common pains of adolescence, along with headache and abdominal pain, and arthralgia is the single most common reason for referral to the pediatric rheumatologist. Not surprisingly, the pediatric rheumatologist is frequently called to distinguish organic from functional symptoms. During the past decade, the pediatric rheumatology community has been evaluating increasing numbers of adolescents and preadolescents who experience musculoskeletal symptoms presumably as a defense against emotional stress from achievement either in academic work or in sports. To complicate the challenge further, coexistent organic and psychologic disturbance is not rare. Clearly, organic illness does not protect a patient from emotional plan, and it may be most difficult to differentiate nonorganic pain in a patient with a known organic illness. Conversely, adolescents with organic illness may use their disease for secondary gain. Fear of misdiagnosis of physical illness as psychiatric and the notion that all of the patient's complaints should be explained by a unifying diagnosis cause diagnostic error in both psychogenic illness with physical manifestations and physical illness with psychogenic symptoms.     

Signal transduction pathways that regulate cell survival and cell death

Apoptosis or programmed cell death (PCD) is a physiological process critical for organ development, tissue homeostasis and elimination of defective or potentially dangerous cells in complex organisms. Apoptosis permits cell death without a concomitant inflammatory response in the surrounding tissues. The process of apoptosis depends on the reception of multiple extracellular and intracellular signals, integration and amplification of these signals by second messengers and finally, activation of the death effector proteases. Defects in control of apoptotic pathways may contribute to a variety of diseases including cancer, autoimmune and neurodegenerative conditions and AIDS. While many components of the regulatory network controlling apoptosis have been defined, the mechanisms of action and patterns of interaction of these factors remain controversial. This article summarizes some of the known aspects of signaling pathways involved in apoptosis.     

Examination of solidification pathways and the liquidus surface in the Nb-Ti-Al system

The solidification pathways, subsequent solid-state transformations, and the liquidus surface in the Nb-Ti-Al system have been examined as part of a larger investigation of phase equilibria in Nb-Ti-Al intermetallic alloys. Fifteen alloys ranging in composition from 15 to 40 at. pct Al, with Nb to Ti ratios of 4:1, 2:1, 1.5:1, 1:1, and 1:1.5, were prepared by arc melting and the as-cast microstructures were characterized by optical microscopy (OM), microhardness, X-ray diffraction (XRD), differential thermal analysis (DTA), backscattered electron imaging (BSEI), electron probe microanalysis (EPMA), and transmission electron microscopy (TEM). The results indicate that the range of primary β solidification is much wider than that indicated in previously reported liquidus surfaces, both experimental and calculated. Differential thermal analysis has identified the existence of a β to σ+γ transformation in three alloys where it was previously thought not to exist; confirmation was provided by high-temperature vacuum heat treatments in the single-phase β region followed by rapid quenching. The location of the boundary between the β, σ, and δ primary solidification fields has been redefined. A massive β → δ transformation, which was observed in the cast microstructure of a Nb-25Ti-25Al alloy, was repeatable through cooling following homogenization. A β → δ+σ eutectoid-like transformation in the 25 at. pct Al alloys, was detected by DTA and evaluated through microstructural analysis of heat-treated samples. Trends in the β phase with variations in composition were established for both lattice parameters and microhardness. As a result of this wider extent of the primary β solidification field, a greater possibility exists for microstructural control through thermal processing for alloys consisting of either σ+γ, β+σ, or β+δ phases. An erratum to this article is available at .     

Mutations in the nucleotide-binding sites of P-glycoprotein that affect substrate specificity modulate substrate-induced adenosine triphosphatase activity

The amino- and carboxy-terminal nucleotide-binding domains (NBD1 and NBD2) of P-glycoprotein (P-gp) share over 80% sequence identity. Almost all of NBD1 can be exchanged by corresponding NBD2 segments with no significant loss of function, except for a small segment around the Walker B motif. Within this segment, we identified two sets of residues [ERGA --> DKGT (522-525) and T578C] that, when replaced by their NBD2 counterparts, cause dramatic alterations of the substrate specificity of the protein [Beaudet, L., and Gros, P. (1995) J. Biol. Chem. 270, 17159-17170]. We wished to gain insight into the molecular basis of this defect. For this, we overexpressed the wild-type mouse Mdr3 and variants bearing single or double mutations at these positions in the yeast Pichia pastoris. P-gp-specific ATPase activity was measured in yeast plasma membrane preparations after detergent solubilization and reconstitution in Escherichia coli proteoliposomes. P-gp proteoliposomes from P. pastoris showed a strong verapamil- and valinomycin-stimulated ATPase activity, with characteristics (KM, Vmax) similar to those measured in mammalian cells. Mutations did not appear to affect the KM for Mg2+ATP ( approximately 0.4 mM), but maximum velocity (Vmax) of the drug-stimulated ATPase activity was severely affected in a substrate/modulator-specific fashion. Indeed, all mutants showed complete loss of verapamil-induced ATPase, while all retained at least some degree of valinomycin-induced ATPase activity. Photolabeling studies with [125I]iodoarylazidoprazosin, including competition with MDR drugs and modulators, suggested that drug binding was not affected in the mutants. The altered drug resistance profiles of the ERGA --> DKGT(522-525) and T578C mutants in vivo, together with the observed alterations in substrate-induced ATPase activity of these proteins, suggest that the residues involved may form part of a signal pathway between the membrane regions (substrate binding) and the ATP binding sites.     

Cytosolic Ca2+ acts by two separate pathways to modulate the supply of release-competent vesicles in chromaffin cells

Recovery from depletion of the readily releasable pool of vesicles (RRP) in adrenal chromaffin cells was studied at differing basal [Ca2+]i or following protein kinase C (PKC) activation by phorbol esters. Following depletion, the pool size was estimated at varied times from cell capacitance jumps in response to paired depolarizations. The experimentally observed RRP recovery time course and steady-state size could be predicted from the measured [Ca2+]i signal assuming a Michaelis-Menten-type regulation of the vesicle supply by Ca2+. An elevated recruitment activity was observed at increased [Ca2+]i even when protein kinase C was blocked, but maximum effects could be obtained only after stimulation of PKC by phorbol esters or by prolonged elevations in [Ca2+]i. We suggest that, in chromaffin cells, elevated cytosolic Ca2+ modulates exocytotic plasticity via PKC-dependent and -independent pathways.     

Craniofacial pain syndromes that mimic temporomandibular joint disorders

Undiagnosed pain in the face, head and neck is very frustrating to all parties involved: the patient, the family of the patient, and the clinician. Simple pains may be treated with ease; however, chronic pain is a different matter. Often, patients consult numerous doctors, clinics, and hospitals in their search of pain relief. The patient may then undergo extensive and at times, unnecessary invasive treatment with the hope that the relentless agony of their suffering will be relieved. The purpose of this article is primarily one of education. Physicians and dentists alike encounter these unfortunate sufferers and yet, are confounded by the multitude of symptoms and complaints. Therefore, it was felt that an article such as this was needed. Many concepts presented within are either new or revised. It will be interesting to the reader just how similar many of these pain syndromes appear. Yet, when a systematic diagnostic approach is attempted, it will become clear just how different these syndromes are as well.     

TOR: a new orphan receptor expressed in the thymus that can modulate retinoid and thyroid hormone signals

Vitamin A and other fat-soluble hormones and vitamins have important roles as modulators of essential biological processes such as homeostasis, development, differentiation, and oncogenesis and also as regulators of the immune system. The active form of vitamin A, retinoic acid, as well as vitamin D3 and thyroid hormones exert their actions by binding to specific nuclear receptors that represent one subfamily of the steroid/thyroid hormone receptor superfamily. To identify new members of the retinoid/thyroid hormone receptor subfamily that could play a role in the immune system, a screening of a T cell cDNA library was performed using a retinoid X receptor probe. A clone was isolated encoding a novel nuclear receptor expressed mainly in the thymus and T cell lines. This new receptor, TOR (thymus orphan receptor), is most closely related in both its DNA-binding domain and ligand-binding domain, 90% and 53%, respectively, to ROR alpha/RZR alpha and clusters with these two receptors and RZR beta in a phylogenetic tree, when both the DNA-binding domain and the ligand-binding domain sequences of nuclear receptors are compared. Thus, TOR is part of a subgroup of receptors, one of which has recently been reported to be activated by melatonin. TOR binds specifically to a direct repeat of the half-site sequence 5'-AGGTCA-3' with a four- or five-nucleotide spacer, DNA sequences that also serve as binding sites for thyroid hormone (TR), and retinoic acid receptors (RAR). In transient transfection experiments TOR does not activate a reporter gene carrying these sequences in the absence or the presence of any known nuclear receptor ligands. TOR, however, is able to repress TR and RAR activity on DR-4-TREs or DR-5-RAREs, respectively. Therefore, our data suggest that TOR, similar to COUP-TF, can negatively regulate retinoic acid and thyroid hormone signals. However, the response elements recognized by TOR and COUP-TF differ as do the expression patterns of these receptors. Thus, one important role of TOR could be to modulate retinoid and thyroid hormone signals in the thymus.     

Neuroanatomy in Rett syndrome: cerebral cortex and posterior fossa

Rett syndrome (RS), a neurodevelopmental disorder of unknown etiology occurring almost exclusively in females, is characterized by autistic-like behavior, motor dysfunction, loss of language skills, dementia, and microcephaly. This study is a follow-up and extension of a previously reported neuroimaging study of patients with RS. We replicated previously reported findings with a larger patient population, and the volumetric MRI analysis was extended to include an analysis of neuroanatomy of the posterior fossa. Twenty girls with RS were compared with individually age- and gender-matched normal controls. Patients with RS showed global reduction in gray- and white-matter volumes. The prefrontal, posterior-frontal, and anterior-temporal regions showed the largest bilateral decrease in gray-matter volume, whereas white-matter volume was uniformly reduced throughout the brain. We found confirmation for the preferential reduction in caudate nucleus volume. However, we observed no preferential reduction in midbrain volume despite a preferential reduction in the midsagittal area of this region. We also present an individual case comparison between monozygotic twins discordant for RS.