Emerging perspectives in store-operated Ca2+ entry: roles of Orai, Stim and TRP

Depletion of intracellular Ca2+ stores induces Ca2+ influx across the plasma membrane through store-operated channels (SOCs). This store-operated Ca2+ influx is important for the replenishment of the Ca2+ stores, and is also involved in many signaling processes by virtue of the ability of intracellular Ca2+ to act as a second messenger. For many years, the molecular identities of particular SOCs, as well as the signaling mechanisms by which these channels are activated, have been elusive. Recently, however, the mammalian proteins STIM1 and Orai1 were shown to be necessary for the activation of store-operated Ca2+ entry in a variety of mammalian cells. Here we present molecular, pharmacological, and electrophysiological properties of SOCs, with particular focus on the roles that STIM1 and Orai1 may play in the signaling processes that regulate various pathways of store-operated entry.     

EMG sensor location: Does it influence the ability to detect differences in muscle contraction conditions?

Even though it is well known that electromyography (EMG) characteristics are influenced by electrode placement it is common to use a single pair of sensors per muscle for EMG. This study was designed to determine if the ability to distinguish between contraction conditions was influenced by sensor location. Subjects (n = 10; 27+/-5.3 years; 82+/-13.4 kg; 178+/-7.1 cm) completed six elbow flexor conditions: three isometric contraction intensities (100% maximum effort, 80%, 50%) and three isotonic contraction intensities (heavy weight, 80% and 50% of the weight). Three pairs of electrodes were placed centrally, medially and laterally on the biceps brachii belly in line with the muscle fibers. Isometric contractions were held for 5s, with the middle 3 s analyzed. Isotonic exercises included five repetitions of elbow flexion-extension, with the middle three repetitions analyzed. Average EMG (EMG(AVG)), root mean square EMG (EMG(RMS)) and mean power frequency (MPF) were calculated for each extracted data set. Dependent variables were analyzed using 2 (contraction type) x 3 (intensity) repeated measures ANOVAs per sensor. EMG(AVG) was influenced by the interaction between contraction type and intensity for all sensors (p < 0.05). EMG(RMS) as well as MPF were influenced by the interaction between contraction type and intensity for the lateral and central leads (p < 0.05) but not the medial leads (p > 0.05). Different conclusions could have been reached from the same experiment due to different sensor locations. These differences were primarily related to comparing contraction types (i.e., isotonic vs. isometric).     

Thermographic mapping of the skin surface of the head in bald-headed male subjects

Thermal patterns on the skin are related to skin blood perfusion. While knowledge concerning thermal distribution patterns of the face are well established, the thermal distribution patterns of the skin surface of other aspects of the head has only been investigated once previously in a study employing low resolution thermographic equipment [van Dulken, H., van Voss, S.F.C.H, 1971. The normal thermographic patterns of the human head. Acta Neurochir. 23, 247–58.] The purpose of the study was to re-examine the thermal distribution patterns of the surface of the entire head in 12 healthy bald headed male subjects (mean age 39±7.8 (SD) years) using a high definition infrared (IR) thermographic camera. The thermal distribution patterns were monitored using dynamic infrared thermography (DIRT) before, during and after a 2 min local skin cooling procedure with a pair of fans. Infrared thermal images (thermograms) were taken of the superior, anterior, lateral and posterior aspects of the head. The thermal patterns in general followed the anatomical location of the main superficial arteries and veins of the head as described in anatomy books although there were some variations, especially with regard to the forehead and top of the head (superior aspect), ranging from an asymmetrical distribution pattern to a lack of a clear thermal pattern. The thermal patterns in the left and right lateral thermograms in each individual were relatively symmetrical. During the rewarming period the helix, auricular lobule of the auricles and the nose were relatively cold. The frontal thermograms were characterized by cool nasal and cheek areas and a clear warm area surrounding the eyes, especially around the inner canthus.     

Loopholes in the regulation of invasive species: genetic identifications identify mislabeling of prohibited aquarium plants

Numerous invasive aquatic species introductions can be traced to the aquarium trade. Many potentially harmful aquarium species may be difficult to identify based on morphology alone. As such, some prohibited or invasive species may be available for purchase if they are mislabeled as species without restrictions. Here we compare molecular identifications to internet vendors’ identifications for accessions of a popular genus of aquarium plants that are difficult to distinguish morphologically (Myriophyllum; watermilfoils). Specifically, we identified the extensive mislabeling of M. heterophyllum—an invasive species in the northeastern and western US. Furthermore, genotypes of M. heterophyllum found in our aquarium survey have also been found in invasive populations, suggesting their potential introduction through escape from aquaria, water gardens, or nurseries. Two additional taxa were sold under incorrect names. Finally, our survey revealed that Myriophyllum taxa present in the aquarium trade generally have poorly known distributions and ecologies, and therefore their invasive potential is unknown. Our study confirms that molecular identification methods can provide a valuable tool to survey commercial pathways for potentially harmful species that are otherwise difficult to identify.     

SP-SR interneurones: a novel class of neurones of the CA2 region of the hippocampus

The CA2 region of the hippocampus has distinctive properties and inputs and may be linked with the pathology of specific psychiatric and neurological disorders. It is, therefore, important to understand CA2 circuitry and its involvement in the circuitry of the hippocampus. Properties of CA2 basket cells have been reported. However, other classes of CA2 interneurones with cell bodies located in stratum pyramidale remained to be described. In this study, the unusual axonal arbors of a novel subclass of dendrite-preferring CA2 interneurones whose somata are located in the pyramidal cell layer was revealed following intracellular recordings and biocytin labeling. One to four apical dendrites emerged from the soma, branched in stratum radiatum (SR) forming a tuft, but rarely penetrated stratum lacunosum-moleculare (SLM). One or two basal dendrites branched close to the soma, the branches extended through stratum oriens (SO) and often reached the alveus. Unlike CA2 bistratified cells, the axons of these cells arborized almost exclusively in SR with few, if any, branches extending to stratum pyramidale (SP), SO, or SLM. These interneurones again, unlike bistratified cells, were immunonegative for parvalbumin and cholecystokinin. Electrophysiologically, they were similar to some CA2 basket and bistratified cells in that they presented a "sag" in response to hyperpolarizing current injections and displayed spike frequency adaptation. They targeted the apical dendrites of neighboring CA2 pyramidal cells and received inputs from them.     

Multi-Tissue Microarray Analysis Identifies a Molecular Signature of Regeneration

The inability to functionally repair tissues that are lost as a consequence of disease or injury remains a significant challenge for regenerative medicine. The molecular and cellular processes involved in complete restoration of tissue architecture and function are expected to be complex and remain largely unknown. Unlike humans, certain salamanders can completely regenerate injured tissues and lost appendages without scar formation. A parsimonious hypothesis would predict that all of these regenerative activities are regulated, at least in part, by a common set of genes. To test this hypothesis and identify genes that might control conserved regenerative processes, we performed a comprehensive microarray analysis of the early regenerative response in five regeneration-competent tissues from the newt Notophthalmus viridescens. Consistent with this hypothesis, we established a molecular signature for regeneration that consists of common genes or gene family members that exhibit dynamic differential regulation during regeneration in multiple tissue types. These genes include members of the matrix metalloproteinase family and its regulators, extracellular matrix components, genes involved in controlling cytoskeleton dynamics, and a variety of immune response factors. Gene Ontology term enrichment analysis validated and supported their functional activities in conserved regenerative processes. Surprisingly, dendrogram clustering and RadViz classification also revealed that each regenerative tissue had its own unique temporal expression profile, pointing to an inherent tissue-specific regenerative gene program. These new findings demand a reconsideration of how we conceptualize regenerative processes and how we devise new strategies for regenerative medicine.     

ATP7B expression in human breast epithelial cells is mediated by lactational hormones.

A role for the copper transporter, ATP7B, in secretion of copper from the human breast into milk has previously not been reported, although it is known that the murine ortholog of ATP7B facilitates copper secretion in the mouse mammary gland. We show here that ATP7B is expressed in luminal epithelial cells in both the resting and lactating human breast, where it has a perinuclear localization in resting epithelial cells and a diffuse location in lactating tissue. ATP7B protein was present in a different subset of vesicles from those containing milk proteins and did not overlap with Menkes ATPase, ATP-7A, except in the perinuclear region of cells. In the cultured human mammary line, PMC42-LA, treatment with lactational hormones induced a redistribution of ATP7B from a perinuclear region to a region adjacent, but not coincident with, the apical plasma membrane. Trafficking of ATP7B was copper dependent, suggesting that the hormone-induced redistribution of ATP7A was mediated through an increase in intracellular copper. Radioactive copper ((64)Cu) studies using polarized PMC42-LA cells that overexpressed mAtp7B protein showed that this transporter facilitates copper efflux from the apical surface of the cells. In summary, our results are consistent with an important function of ATP7B in the secretion of copper from the human mammary gland.