Muscle-bound primordial stem cells give rise to myofiber-associated myogenic and non-myogenic progenitors

Myofiber cultures give rise to myogenic as well as to non-myogenic cells. Whether these myofiber-associated non-myogenic cells develop from resident stem cells that possess mesenchymal plasticity or from other stem cells such as mesenchymal stem cells (MSCs) remain unsolved. To address this question, we applied a method for reconstructing cell lineage trees from somatic mutations to MSCs and myogenic and non-myogenic cells from individual myofibers that were cultured at clonal density.Our analyses show that (i) in addition to myogenic progenitors, myofibers also harbor non-myogenic progenitors of a distinct, yet close, lineage; (ii) myofiber-associated non-myogenic and myogenic cells share the same muscle-bound primordial stem cells of a lineage distinct from bone marrow MSCs; (iii) these muscle-bound primordial stem-cells first part to individual muscles and then differentiate into myogenic and non-myogenic stem cells.     

Static mechanical stretching accelerates lipid production in 3T3-L1 adipocytes by activating the MEK signaling pathway

Understanding mechanotransduction in adipocytes is important for research of obesity and related diseases. We cultured 3T3-L1 preadipocytes on elastic substrata and applied static tensile strains of 12% to the substrata while inducing differentiation. Using an image processing method, we monitored lipid production for a period of 3-4 wk. The ratio of %-lipid area per field of view (FOV) in the stretched over nonstretched cultures was significantly greater than unity (P < 0.05), reaching ～1.8 on average starting from experimental day ～10. The superior coverage of the FOV by lipids in the stretched cultures was due to significantly greater sizes of lipid droplets (LDs) with respect to nonstretched cultures, starting from experimental day ～10 (P < 0.05), and due to significantly more LDs per cell between days ～10 and ～17 (P < 0.05). The statically stretched cells also differentiated significantly faster than the nonstretched cells within the first ～10 days (P < 0.05). Adding peroxisome proliferator-activated receptor-γ (PPARγ) antagonist did not change these trends, as the %-lipid area per FOV in the stretched cultures that received this treatment was still significantly greater than in the nonstretched cultures without the PPARγ antagonist (14.44 ± 1.96% vs. 10.21 ± 3%; P < 0.05). Hence, the accelerated adipogenesis in the stretched cultures was not mediated through PPARγ. Nonetheless, inhibiting the MEK/MAPK signaling pathway reduced the extent of adipogenesis in the stretched cultures (13.53 ± 5.63%), bringing it to the baseline level of the nonstretched cultures without the MEK inhibitor (10.21 ± 3.07%). Our results hence demonstrate that differentiation of adipocytes can be enhanced by sustained stretching, which activates the MEK signaling pathway.     

Reproductive Cycle and Developmental Processes During Embryogenesis of Clavularia hamra (Cnidaria,Octocorallia)

Clavularia hamra Gohar, 1948 is a common octocoral on the reefs of the Gulf of Eilat (northern Red Sea). Reproductive biology of C. hamra was studied for two years. Direct observations of spawning were conducted in situ and in aquaria. Cleavage of eggs and further embryo metamorphosis into mature planulae were examined by scanning electron microscopy and histological sections. Clavularia hamra is dioecious. Young oocytes appear annually in September, gradually grow in size and attain maturity within 11 months. The main spawning event of the population is highly synchronized, occurring on a single night each year between the last quarter and the new moon in mid summer. The released orange eggs are held together by mucus and remain attached amongst the polyps on the outside of the female colonies. Twelve hours after spawning, the young embryos have blastomeres of equal size with numerous microvilli on their external surface. Due to unequal cleavage, bizarre embryos are also formed. By 48 h a blastopore is visible, indicating that a gastrula is developing. Eight days after spawning mature planulae are observed. The external mode of embryo development on the surface of the parent colony reduces dispersal of the planulae. However, this reproductive feature enhances formation of locally dense populations of C. hamra, with distinct habitat preferences.     

Space partitioning by stony corals soft corals and benthic algae on the coral reefs of the northern Gulf of Eilat (Red Sea)

The major faunistic and floristic components occupying space on the coral reefs of the northern Gulf of Eilat (Red Sea) are stony corals, soft corals and benthic algae. The percent living coverage of the three components and the relative abundance of the different species of each component were studied by line transects, on the reef flats and the upper forereef zones of nine localities. A wider and higher range of living coverage values of stony corals were recorded at the upper fore-reef zones (18.30–49.09%) compared with the reef flats (5.50–31.66%) at the different stations. The most abundant stony corals on the reef flats areCyphastrea microphthalma, Stylophora pistillata, Favia favus, Porites lutea, Platygyra lamellina and the hydrozoanMillepora dichotoma. The fire coralM. dichotoma dominates the upper fore-reef zone in most of the stations. The average percent living coverage of soft corals on the reef flats ranged between 0.20 and 17.06%, and on the upper fore-reef zones between 1.68 and 15.13%. Seventy percent of the total living coverage of the soft coral community is contributed by 2 to 3 species. They tend to form large monospecific carpets, such as those composed ofSinularia sp.,Sarcophyton glaucum andLobophytum pauciflorum. The common benthic algae on the coral reef studied occur as turfs or macroscopic noncalcareous algae. They play a significant role in occupying space, especially on the reef flats. The most abundant algae recorded in all localities are the turfsSphacelaria tribuloides, Jania sp. and the macroscopic non-calcareous algaeTurbinaria elatensis andColpomenia sinuosa. Comparison between reef flats and upper fore-reef zones, in terms of average living cover of stony corals, shows that the variation among the reef flats is grater than the variation among the upper fore-reef zones. However, there is no significant variation in the average living coverage of soft corals between these two zones. Annual living-coverage values of algae on the reef flats are significantly higher than those of the upper fore-reef zones. Extremely low tides occurring periodically but unpredictably at Eilat cause mass mortality of the benthic communities on the reef flats reopening new spaces for settlement. The coexistence of stony corals, soft corals and algae on the reef ecosystem is due to different biological properties of each component. Opportunistic life histories of certain stony corals and most algae enable quick colonization of newly opened spaces. Lack of predators, high tolerance against abiotic factors and ability to form large aggregates of colonies are suggested as possible factors supporting the existence of soft corals in shallow water. Biological factors such as competition, predation and grazing pressure play an increasingly important role in controlling space utilization by the components studied with the advancement of succession.     

SVEP1 expression is regulated in estrogen-dependent manner

The SVEP1 protein comprises modules related to the selectin super family and other motifs found in cell surface molecules. Earlier, we demonstrated that SVEP1 is expressed in osteogenic cells both in vivo and in vitro; in the current study we elaborate on the regulation of SVEP1 by 17beta-Estradiol (17betaE2). SVEP1 message is expressed in vivo by bone marrow cells of sham-operated rats, but not in estrogen-depleted ovariectomized (OVX) rats. We demonstrated that 17betaE2 treatment increases the level of the SVEP1 expression in cultured osteoblasts. SVEP1 was identified also in breast carcinoma (BC) cells known to reside in bone when metastasized from the primary tumor. SVEP1 expression was demonstrated by immunohistochemistry and fluorescence-activated cell sorting (FACS) on various BC cell lines. The chromatin immunoprecipitation (ChIP) assay was applied to analyze the estrogen receptor (ER) binding to the putative SVEP1 promoter. We demonstrated that treatment with 17betaE2 or ICI 182,780 affects this binding and regulates the mRNA and protein levels of SVEP1 in BC cells. We propose that SVEP1 may serve as a useful biomarker for studying the mechanism of cells interactions within the local microenvironment affected by estrogen.     

MS-KIF18A, a kinesin, is associated with estrogen receptor

The study of MS-KIF18A kinesin protein is focused on its cellular distribution and association with a cargo protein. Indirect immunofluorescence (IF) analyzed the intracellular distribution of endogenous MS-KIF18A and the transfected enhanced green fluorescence protein (eGFP)-MS-KIF18A in osteogenic cells. In both cases, the proteins were localized at the plasma membrane, cytosol, and nucleus. Bioinformatics analysis suggested interactions between MS-KIF18A and estrogen receptor (ERalpha) which were further elucidated by immunoprecipitation (IP). We identified interaction between endogenous MS-KIF18A with 66 and 46 kDa isoforms of ERalpha in MBA-15 cells. Moreover, MS-KIF18A and 66 kDa ERalpha complex has been demonstrated between ectopically expressed proteins in COS-7 cells. We have shown that anti-MS-KIF18A antibody immunoprecipitated the ERalpha and pERK in cells challenged with 17beta-estrogen (17beta-E2). The hormone activation induced mitogen-activated protein kinases (MAPK) pathway and increased p-ERK. The activation was interfered when cells were pre-treated with either ICI-182,780 or MAPK inhibitor PD98059 prior the challenge with 17beta-E2 that resulted in a decrease in association between MS-KIF18A and p-ERK1/2. The obtained results suggest a role for the proteins in a non-genomic response of MBA-15 cells challenged with 17beta-E2. This study presents a novel interaction between MS-KIF18A and ER that may have important physiological and pharmacological implications for estrogen action in various cells.