Neuronal precursor cell-expressed developmentally down-regulated 4-1 (NEDD4-1) controls the sorting of newly synthesized Ca(V)1.2 calcium channels

Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) proteins are ubiquitin ligases, which attach ubiquitin moieties to their target proteins, a post-translational modification that is most commonly associated with protein degradation. Nedd4 ubiquitin ligases have been shown to down-regulate both potassium and sodium channels. In this study, we investigated whether Nedd4 ubiquitin ligases also regulate Ca(v) calcium channels. We expressed three Nedd4 family members, Nedd4-1, Nedd4-2, and WWP2, together with Ca(v)1.2 channels in tsA-201 cells. We found that Nedd4-1 dramatically decreased Ca(v) whole-cell currents, whereas Nedd4-2 and WWP2 failed to regulate the current. Surface biotinylation assays revealed that Nedd4-1 decreased the number of channels inserted at the plasma membrane. Western blots also showed a concomitant decrease in the total expression of the channels. Surprisingly, however, neither the Ca(v) pore-forming α1 subunit nor the associated Ca(v)β and Ca(v)α(2)δ subunits were ubiquitylated by Nedd4-1. The proteasome inhibitor MG132 prevented the degradation of Ca(v) channels, whereas monodansylcadaverine and chloroquine partially antagonized the Nedd4-1-induced regulation of Ca(v) currents. Remarkably, the effect of Nedd4-1 was fully prevented by brefeldin A. These data suggest that Nedd4-1 promotes the sorting of newly synthesized Ca(v) channels for degradation by both the proteasome and the lysosome. Most importantly, Nedd4-1-induced regulation required the co-expression of Ca(v)β subunits, known to antagonize the retention of the channels in the endoplasmic reticulum. Altogether, our results suggest that Nedd4-1 interferes with the chaperon role of Ca(v)β at the endoplasmic reticulum/Golgi level to prevent the delivery of Ca(v) channels at the plasma membrane.

Detrital Zircon Evidence for Non-Laurentian Provenance, Mesoproterozoic (ca. 1490-1450 Ma) Deposition and Orogenesis in a Reconstructed Orogenic Belt, Northern New Mexico, USA: Defining the Picuris Orogeny

Detrital zircon and igneous zircon U-Pb ages are reported from Proterozoic metamorphic rocks in northern New Mexico. These data give new insight into the provenance and depositional age of a >3-km-thick metasedimentary succession and help resolve the timing of orogenesis within an area of overlapping accretionary orogens and thermal events related to the Proterozoic tectonic evolution of southwest Laurentia. Three samples from the Paleoproterozoic Vadito Group yield narrow, unimodal detrital zircon age spectra with peak ages near 1710 Ma. Igneous rocks that intrude the Vadito Group include the Cerro Alto metadacite, the Picuris Pueblo granite, and the Penasco quartz monzonite and yield crystallization ages of 1710 +/- 10 Ma, 1699 +/- 3 Ma, and 1450 +/- 10 Ma, respectively. Within the overlying Hondo Group, a metamorphosed tuff layer from the Pilar Formation yields an age of 1488 +/- 6 Ma and represents the first direct depositional age constraint on any part of the Proterozoic metasedimentary succession in northern New Mexico. Detrital zircon from the overlying Piedra Lumbre Formation yield a minimum age peak of 1475 Ma, and similar to 60 grains (similar to 25%) yield ages between 1500 Ma and 1600 Ma, possibly representing non-Laurentian detritus originating from Australia and/or Antarctica. Detrital zircons from the basal metaconglomerate and the middle quartzite member of the Marquenas Formation yield minimum age peaks of 1472 Ma and 1471 Ma, consistent with earlier results. We interpret the onset of ca. 1490-1450 Ma deposition followed by tectonic burial, regional Al2SiO5 triple-point metamorphism, and ductile deformation at depths of 12-18 km to reflect a Mesoproterozoic contractional orogenic event, possibly related to the final suturing of the Mazatzal crustal province to the southern margin of Laurentia. We propose to call this event the Picuris orogeny.

Association of the (CA)n repeat polymorphism of insulin-like growth factor-I and -202 A/C IGF-binding protein-3 promoter polymorphism with adult height in patients with severe growth hormone deficiency

A number of mathematical models for predicting growth and final height outcome have been proposed to enable the clinician to 'individualize' growth-promoting treatment. However, despite optimizing these models, many patients with isolated growth hormone deficiency (IGHD) do not reach their target height. The aim of this study was to analyse the impact of polymorphic genotypes [CA repeat promoter polymorphism of insulin-like growth factor-I (IGF-I) and the -202 A/C promoter polymorphism of IGF-Binding Protein-3 (IGFBP-3)] on variable growth factors as well as final height in severe IGHD following GH treatment. DESIGN, PATIENTS AND CONTROLS: One hundred seventy eight (IGF-I) and 167 (IGFBP-3) subjects with severe growth retardation because of IGHD were studied. In addition, the various genotypes were also studied in a healthy control group of 211 subjects.

Detrital Zircon Evidence for Mixing of Mazatzal Province Age Detritus With Yavapai Age (ca. 1700-1740 Ma) and Older Detritus in the ca. 1650 Ma Mazatzal Province of Central New Mexico, USA

Preliminary detrital zircon age distributions from Mazatzal crustal province quartzite and schist exposed in the Manzano Mountains and Pedernal Hills of central New Mexico are consistent with a mixture of detritus from Mazatzal age (ca. 1650 Ma), Yavapai age (ca. 1720 Ma.), and older sources. A quartzite sample from the Blue Springs Formation in the Manzano Mountains yielding 67 concordant grain analyses shows two dominant age peaks of 1737 Ma and 1791 Ma with a minimum peak age of 1652 Ma. Quartzite and micaceous quartzite samples from near Pedernal Peak give unimodal peak ages of ca. 1695 Ma and 1738 Ma with minimum detrital zircon ages of ca. 1625 Ma and 1680 Ma, respectively. A schist sample from the southern exposures of the Pedernal Hills area gives a unimodal peak age of 1680 Ma with a minimum age of ca. 1635 Ma. Minor amounts of older detritus (>1800 Ma) possibly reflect Trans-Hudson, Wyoming, Mojave Province, and older Archean sources and aid in locating potential source terrains for these detrital zircon. The Blue Springs Formation metarhyolite from near the top of the Proterozoic section in the Manzano Mountains yields 71 concordant grains that show a preliminary U-Pb zircon crystallization age of 1621 ¿ 5 Ma, which provides a minimum age constraint for deposition in the Manzano Mountains. Normalized probability plots from this study are similar to previously reported age distributions in the Burro and San Andres Mountains in southern New Mexico and suggest that Yavapai Province age detritus was deposited and intermingled with Mazatzal Province age detritus across much of the Mazatzal crustal province in New Mexico. This data shows that the tectonic evolution of southwestern Laurentia is associated with multiple orogenic events. Regional metamorphism and deformation in the area must postdate the Mazatzal Orogeny and ca. 1610 Ma ¿ 1620 Ma rhyolite crystallization and is attributed to the Mesoproterozoic ca. 1400 ¿ 1480 Ma Picuris Orogeny.

Agonist specific L-type Ca(2+)-current stimulation in ventricular myocytes by a novel steroid-like compound

In cardiac muscle the amplitude of Ca(2+) transients can be increased by enhancing Ca(2+) influx. Among the processes leading to increased Ca(2+) influx, agonists of the L-type Ca(2+)-channel can play an important role. Known pharmacological Ca(2+)-channel agonists act on different binding sites on the channel protein, which may lead not only to enhanced peak currents, but also to distinct changes in other biophysical characteristics of the current. In this study, membrane currents were recorded with the patch-clamp technique in the whole-cell configuration in guinea pig isolated ventricular myocytes in combination with confocal fluorescence Ca(2+) imaging techniques and a variety of pharmacological tools. Testing a new positive inotropic steroid-like compound, we found that it increased the L-type Ca(2+)-current by 2.5-fold by shifting the voltage-dependence of activation by 20.2 mV towards negative potentials. The dose-response relationship revealed two vastly different affinities (EC(50(high-affinity))=4.5+/-1.7 nM, EC(50(low-affinity))=8.0+/-1.1 microM) exhibiting differential pharmacological interactions with three classes of Ca(2+)-current antagonists, suggesting more than one binding site on the channel protein. Therefore, we identified and characterized a novel positive inotropic compound (F90927) as a member of a new class of Ca(2+)-channel agonists exhibiting unique features, which set it apart from other presently known L-type Ca(2+)-channel agonists.