Ca(V)1 and Ca(V)2 channels engage distinct modes of Ca(2+) signaling to control CREB-dependent gene expression

Activity-dependent gene expression triggered by Ca(2+) entry into neurons is critical for learning and memory, but whether specific sources of Ca(2+) act distinctly or merely supply Ca(2+) to a common pool remains uncertain. Here, we report that both signaling modes coexist and pertain to Ca(V)1 and Ca(V)2 channels, respectively, coupling membrane depolarization to CREB phosphorylation and gene expression. Ca(V)1 channels are advantaged in their voltage-dependent gating and use nanodomain Ca(2+) to drive local CaMKII aggregation and trigger communication with the nucleus. In contrast, Ca(V)2 channels must elevate [Ca(2+)](i) microns away and promote CaMKII aggregation at Ca(V)1 channels. Consequently, Ca(V)2 channels are ~10-fold less effective in signaling to the nucleus than are Ca(V)1 channels for the same bulk [Ca(2+)](i) increase. Furthermore, Ca(V)2-mediated Ca(2+) rises are preferentially curbed by uptake into the endoplasmic reticulum and mitochondria. This source-biased buffering limits the spatial spread of Ca(2+), further attenuating Ca(V)2-mediated gene expression.     

Knocking down Israa,the Zmiz1 intron-nested gene,unveils interrelated T cell activation functions in mouse

We previously reported Israa (immune-system-released activating agent), a novel gene nested in intron 6 of the mouse Zmiz1 gene. Zmiz1 is involved in several functions such as fertility and T cell development and its knockout leads to non-viable embryos. We also reported ISRAA's expression in lymphoid organs, particularly in the thymus CD³⁺ T cells during all developmental stages. In addition, we showed that ISRAA is a binding partner of Fyn and Elf-1 and regulates the expression of T cell activation-related genes in vitro. In this paper, we report the generation and characterization of an Israa^?/? constitutive knockout mouse. The histological study shows that Israa^?/? mice exhibit thymus and spleen hyperplasia. Israa^?/? derived T cells showed increased proliferation compared to the wild-type mice T cells. Moreover, gene expression analysis revealed a set of differentially expressed genes in the knockout and wild-type animals during thymus development (mostly genes of T cell activation pathways). Immunological phenotyping of the thymocytes and splenocytes of Israa^?/- showed no difference with those of the wild-type. Moreover, we observed that knocking out the Zmiz1 intron embedded Israa gene does not affect mice fertility, thus does not disturb this Zmiz1 function. The characterization of the Israa^?/- mouse confirms the role ISRAA plays in the expression regulation of genes involved in T cell activation established in vitro. Taken together, our findings point toward a potential functional interrelation between the intron nested Israa gene and the Zmiz1 host gene in regulating T cell activation. This constitutively Israa^?/? mice can be a good model to study T cell activation and to investigate the relationship between host and intron-nested genes.     

Antidiabetic Drug Sitagliptin with Divalent Transition Metals Manganese and Cobalt: Synthesis,Structure, Characterization Antibacterial and Antioxidative Effects in Liver Tissues

Metals and their complexes have an increasing number of medical applications. Sitagliptin (STG) acts as an antidiabetic drug. Mn(II) and Co(II) complexes were studied and characterized based on physical characterization, FT-IR, DG/TG, XRD, ESM, and TEM. Data revealed that STG acts as a bidentate ligand through the oxygen atom of a carbonyl group and the nitrogen atom of an amino group. Magnetic measurement data revealed that the Mn/STG metal complex has a square planner geometry. The experiment was performed on 40 male albino rats who were divided into four groups: the control group, STG group, group treated with STG/Mn, and group treated with Co/STG. Biomarkers for hepatic enzymes and antioxidants were found in the blood, and hepatic tissue histology was evaluated. STG in combination with Mn and Co administration showed potent protective effects against hepatic biochemical alterations induced by STG alone, as well as suppressing oxidative stress and structural alterations. These complexes prevented any stress and improved hepatic enzymatic levels more than STG alone. The STG/Mn complex was highly effective against Bacillus subtilis and Streptococcus pneumonia, while STG/Co was highly effective against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureas. Therefore, STG combined with Mn and Co produced a synergistic effect against oxidative stress and improved the histological structure of the liver tissues. STG metal complexes with Mn and Co showed the most potential ameliorative antioxidant and hepatoprotective effects.     

A simple method for scanning electron microscope preparation of cells grown in multiwell culture plates

The closeness of wells in multiwell tissue culture plates makes it difficult to remove individual ones without distributing the adjacent wells. Moreover, critical point drying frequently introduces artifact into the culture monolayer grown on plastic substrate. These problems make it difficult to process such cultures for scanning electron microscopy. However, for cell kinetic and correlative morphological studies on primary cultures derived from 7,12-dimethylbenz(alpha)anthracene(DMBA)-induced mammary tumors, we have found that Falcon 24-well tissue culture plates are excellent for maintenance of cells and are convenient to use. By plating the cells in alternating, diagonally disposed wells and while the cells are still in the buffer, individual wells can be cut from a multiwell plate without disturbing the cells in adjacent wells. The isolated wells can be used to carry out scanning electron microscope preparation. The height of the well is reduced with a scalpel prior to critical point drying; the remaining well is useful as a handle in mounting the dried sample to stubs for subsequent coating and viewing. Critical point drying and coating of monolayer samples on Falcon plastic are described. The results do not suggest that any artifacts are introduced in our preparations.     

Annual reproductive cycle and condition index of Mactra corallina (Mollusca: Bivalvia) from the north coast of Tunisia

This work aims to examine the reproductive cycle of the bivalve Mactra corallina inhabiting the Gulf of Tunis (Tunisia). Histological analysis indicated that the reproductive cycle is composed of six stages and was synchronous between sexes. Gonadal development began in January. From April to July, an increase in temperature (from 16 to 28°C) and a rise in chlorophyll a (from 3.25 to 6.35 mg/L) marked the period of sexual maturity in both sexes, which was also confirmed by an elevation in condition index (CI). Partial spawning started in April for males and in May for females, peaking in late summer indicating a prolonged reproductive period. The reproductive cycle of M. corallina finished with an inactive stage during autumn. From the total number of animals used in this study, we identified 104 females (52%) and 96 males (48%) corresponding to a sex ratio 1:0.92 that did not differ significantly from a 1:1 ratio.