Differential effect of components of the extracellular matrix on differentiation and apoptosis

AbstractBackground: Epithelial cells are closely associated with a basement membrane, but the intimate relationships that affect growth, differentiation and survival remain enigmatic. We have previously reported that granulosa cells adjacent to the basement membrane of the ovarian follicle have a higher degree of differentiation compared with cells located distal to the basement membrane. By contrast, granulosa cells distal to the basement membrane are the first to undergo apoptosis during follicular atresia. Moreover, growth of granulosa cells in vitro on a naturally produced basement-membrane-like extracellular matrix (ECM) enhances progesterone production and the cellular response to gonadotropic hormones by an undefined mechanism.Results: To investigate the effect of the ECM on granulosa cell differentiation and death, primary granulosa cells were cultured on ECMs that lacked or contained bFGF (basic fibroblast growth factor). These otherwise identical ECMs were deposited by HR9 mouse endodermal cells, which do not synthesize bFGF, or by HR9 cells transfected with the bFGF gene. Both ECMs provided protection against apoptosis in serum-free medium, but only the bFGF-containing ECM maintained expression of the steroidogenic P450scc enzyme system and the production of progesterone. Moreover, culturing the cells on this ECM enhanced the expression of the 30 kDa steroid acute regulatory protein which plays a key role in steroid hormone biosynthesis. Laminin, but not fibronectin, was able to replace the ECM in protecting the cells from apoptosis; but not in maintaining steroidogenesis, whereas bFGF was able to enhance steroidogenesis without protecting the cells against apoptosis. Cells cultured on both ECMs or laminin had a well-developed actin cytoskeleton compared with cells cultured on non-coated dishes, which underwent apoptosis.Conclusions: Cellular responses to ECM are mediated by the combined action of macromolecular constituents and regulatory molecules, such as bFGF, that are sequestered and stored in the ECM. ECM or laminin protects against cell death by interacting with specific integrin receptors and maintaining a well-developed actin cytoskeleton. ECM-bound bFGF provides differentiation signals for granulosa cells, which are in intimate contact with the ECM. Thus, a clear distinction can be made between the survival activity and the differentiation stimulus exerted by the ECM on epithelial cells

The effect of dietary taurine and its potential biosynthesis on juvenile grey mullet (Mugil cephalus) performance

The grey mullet is a catadromous species that spawns in the taurine-rich seawater environment, followed by the young fish generally migrating to less saline, low taurine waters during the larva-juvenile transition. Consequently, this study aimed to (1) determines whether there is a dietary taurine requirement in juvenile grey mullet for enhanced growth and (2) the potential for taurine biosynthesis. The experimental system consisted of sixteen 400-l V-tanks, where filtered, UV-treated ambient seawater (40 ‰) entered the bottom of the tanks at a rate of 7 tank exchanges/day. This allowed the testing of four 1 mm pelleted diets (0, 0.5, 1.0, and 2.0% taurine DW diet) in replicates of 4 tanks/treatment for 58 days. Grey mullet juveniles demonstrated (P<0.05) a specific requirement for a 0.5% taurine DW diet for improved growth. Fish fed the taurine diets displayed populations with a markedly (P < 0.05) higher average number of surviving fish (23.4±1.1) of moderately sized (10- 20 g) cohorts than smaller (< 10 g) individuals (12.5±1.1). In contrast, the fish fed the taurine control (0% taurine) exhibited similar average numbers of small and moderate sized fish (18.0±3.6-20.0±4.1). Dietary taurine accumulated highly (P<0.05) in the muscles in a dose dependent manner but less so (P<0.05) in eyes, and liver. The gene expression of liver cysteine sulfinic acid decarboxylase (CSD) exhibited an upregulation (P<0.05) with taurine diets from 0 to 1% but was down regulated (P<0.05) in fish fed the 2% taurine DW diet

The effect of algal turbidity on larval performance and the ontogeny of digestive enzymes in the grey mullet (Mugil cephalus)

A study comprised of two trials determined the effects of water turbidity produced by live microalgae and inert clay particles on the larval rearing of grey mullet (Mugil cephalus). Trial 1 evaluated the effect of microalgae produced water turbidity on grey mullet larval performance and digestive tract (DT) enzyme ontogeny. Two microalgae (Nannochloropsis oculata and Isochrysis galbana) water turbidity levels (0.76 and 1.20 NTU, respectively) and a non-microalgae control (0.26 NTU) were investigated on 2 to 23 dph grey mullet larvae. The higher turbidity (1.2 NTU) larvae (5 dph) consumed markedly (P < .05) more rotifers than other treatment fish, independently of the microalgae type. There was no clear effect of the turbidity treatments on DT enzyme ontogeny. However, in all treatments lipase and alkaline proteases appeared to be modulated by the diet. Alkaline phosphatase activity was ca. 8 times higher and α-amylase activity increased 5.3 times in 79 dph fish compared to 40 dph individuals. The ratio of alkaline phosphatase and leucine-alanine aminopeptidase indicated gut maturation occurred around 61 dph. Trial 2 compared the most effective N.occulata produced turbidity level (1.2 NTU) with the identical water turbidity produced by inert clay on larval performance. M. cephalus larvae exposed to high algal turbidity demonstrated superior performance (P < .05), in terms of rotifer ingestion, dry weight gain and survival, compared to cohorts reared under the clay treatment and the lower microalgae produced turbidity. These findings suggested that water algal turbidity is not the dominant factor determining improved grey mullet larval performance.info:eu-repo/semantics/acceptedVersio

Physiological changes in the spawning gilthead seabream, Sparus aurata, succeeding the removal of males

The physiological effects triggered in females by the removal of males from a group of spawning fish were examined in the multiple batch spawner, the gilthead seabream, Sparus aurata. One week after the removal of males, a large portion of the oocytes underwent atresia, and sporadic release of low quality eggs continued at low frequency over a period of seven weeks. The transcript levels of the three native gonadotropin releasing hormone (GnRH) forms, salmon (s)GnRH, seabream (sb)GnRH, and chicken (c)GnRH-II, and the two βGtH subunits were measured. Brain mRNA levels for all three GnRHs and pituitary βLH mRNA levels significantly declined in the females as a result of removing the males compared to females that were maintained with males. Pituitary βFSH mRNA levels showed the opposite trend and were significantly higher in females that were separated from males. Circulating levels of LH, testosterone, estradiol, 17α,20β-dihydroxy-4-pregnen-3-one, and 17α,20β,21-trihydroxy-4-pregnen-3-one all declined in the group of females without males. These results imply the existence of an endocrine response to socio-sexual stimuli during the reproductive process in the gilthead seabream. J. Exp. Zool. 292:555–564, 2002. © 2002 Wiley-Liss, Inc

Cyclic ADPR and calcium signaling in sea bream (Sparus aurata) egg fertilization.

The cell egg is in a state of quiescence and only after its fusion with the sperm, a series of pre-programmed metabolic processes will be activated, culminating with embryonic development. The egg/sperm fusion induces a transitory increase of Ca(2+) in the cytoplasm, which is responsible for the activation of both precocious and late reactions. The release of Ca(2+) occurs by stimulation of the ionic specific channels. In addition to IP(3), a new Ca-release inducer was recently evidenced, cyclic ADP ribose (cADPR), in some invertebrates and mammals. Here, we report the first evidence of the cADPR presence in fish. Our data also demonstrate that in the sea bream egg, cADPR is involved in the fertilization process; in fact, its level increases after the entrance of the sperm. By in vitro experiments, it was shown that cADPR induces a release of Ca(2+) in the egg homogenate, indicating that in sea bream, the increase of cADPR can induce an intracellular Ca(2+) release. Since cADPR is a product of NAD(+) metabolism, the activity of several enzymes involved in the NAD(+) metabolism was investigated. Sea bream eggs are pelagic and only floating eggs after insemination develop into viable embryos. In the present work, NAD(+) metabolism was studied in both types of egg. All the tested enzymes showed similar specific activity in both floating and sinking eggs. In the latter, cADPR was not detectable and the nucleotides content was significantly lower, evidencing a scarce energetic charge in sinking eggs

Decreased melanoma CSF-1 secretion by Cannabigerol treatment reprograms regulatory myeloid cells and reduces tumor progression

ABSTRACTDuring solid tumor progression, the tumor microenvironment (TME) evolves into a highly immunosuppressive milieu. Key players in the immunosuppressive environment are regulatory myeloid cells, including myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs), which are recruited and activated via tumor-secreted cytokines such as colony-stimulating factor 1 (CSF-1). Therefore, the depletion of tumor-secreted cytokines is a leading anticancer strategy. Here, we found that CSF-1 secretion by melanoma cells is decreased following treatment with Cannabis extracts. Cannabigerol (CBG) was identified as the bioactive cannabinoid responsible for the effects. Conditioned media from cells treated with pure CBG or the high-CBG extract reduced the expansion and macrophage transition of the monocytic-MDSC subpopulation. Treated MO-MDSCs also expressed lower levels of iNOS, leading to restored CD8+ T-cell activation. Tumor-bearing mice treated with CBG presented reduced tumor progression, lower TAM frequencies and reduced TAM/M1 ratio. A combination of CBG and αPD-L1 was more effective in reducing tumor progression, enhancing survival and increasing the infiltration of activated cytotoxic T-cells than each treatment separately. We show a novel mechanism for CBG in modulating the TME and enhancing immune checkpoint blockade therapy, underlining its promising therapeutic potential for the treatment of a variety of tumors with elevated CSF-1 expression