Immunomodulatory effects of excretory/secretory compounds from <i>Contracaecum osculatum</i> larvae in a zebrafish inflammation model

<div><p>Excretory/secretory (ES) compounds isolated from third-stage larvae of the anisakid nematode <i>Contracaecum osculatum</i> parasitizing liver of Baltic cod were investigated for effects on immune gene expression in a zebrafish LPS-induced inflammation model. ES products containing a series of proteins, of which some had enzymatic activity, were injected solely or with LPS. ES proteins alone induced up-regulation of a number of immune-related genes, but generally to a lower degree compared to LPS. When co-injected with LPS, the worm products exacerbated merely expression of five genes affecting Th1, Th2, Th17 and innate responses compared to the LPS-injected group. However, the level of overexpression decreased in an inverse dose-dependent manner. The immune regulating action of <i>C</i>. <i>osculatum</i> ES products is interpreted as an important evolutionary ability of larval parasites in the transport host which makes it less susceptible to host immune responses whereby the probability of reaching the final host is increased.</p></div

Expression of immune genes in zebrafish after exposure to LPS or LPS plus ES compounds.

<p>Fold change ± standard error (fold ± SE) are compared to expression level in LPS-exposed fish. Difference in expression levels is considered significant at P values smaller than 5% and fold changes of at least 2. Asterisk sign: significance of fold changes of each gene in LPS+ES groups compared to the LPS-only group (*P < 0.05; **P <0.01; ***P < 0.001).</p

Enzyme activity of excretory/secretory proteins produced by <i>C</i>. <i>osculatum</i> third-stage larvae.

<p>Enzyme activity of excretory/secretory proteins produced by <i>C</i>. <i>osculatum</i> third-stage larvae.</p

SDS-PAGE of the isolated ES proteins from <i>C</i>. <i>osculatum</i> third-stage larvae; reducing condition, silver staining; Left column: Marker Precision Plus Protein^TM Dual Color Standards; Right column: <i>C</i>. <i>osculatum</i> ES proteins with different molecular weights.

<p>SDS-PAGE of the isolated ES proteins from <i>C</i>. <i>osculatum</i> third-stage larvae; reducing condition, silver staining; Left column: Marker Precision Plus Protein^TM Dual Color Standards; Right column: <i>C</i>. <i>osculatum</i> ES proteins with different molecular weights.</p

Expression of immune genes in zebrafish after exposure to PBS or LPS plus ES compounds.

<p>Fold change in all groups are compared to expression in PBS-injected zebrafish. As the efficiencies of all the assays were within 100% ± 5%, the simplified 2 –ΔΔCq method were used to calculate the relative fold change. Asterisk sign: significance level of fold changes of each gene (*P < 0.05; **P <0.01; ***P < 0.001) and fold changes of at least 2. The elongation factor 1α (<i>EF-1α</i>) was used as the reference gene.</p

Reciprocity in the Developmental Regulation of Aquaporins 1, 3 and 5 during Pregnancy and Lactation in the Rat

Milk secretion involves significant flux of water, driven largely by synthesis of lactose within the Golgi apparatus. It has not been determined whether this flux is simply a passive consequence of the osmotic potential between cytosol and Golgi, or whether it involves regulated flow. Aquaporins (AQPs) are membrane water channels that regulate water flux. AQP1, AQP3 and AQP5 have previously been detected in mammary tissue, but evidence of developmental regulation (altered expression according to the developmental and physiological state of the mammary gland) is lacking and their cellular/subcellular location is not well understood. In this paper we present evidence of developmental regulation of all three of these AQPs. Further, there was evidence of reciprocity since expression of the rather abundant AQP3 and less abundant AQP1 increased significantly from pregnancy into lactation, whereas expression of the least abundant AQP5 decreased. It would be tempting to suggest that AQP3 and AQP1 are involved in the secretion of water into milk. Paradoxically, however, it was AQP5 that demonstrated most evidence of expression located at the apical (secretory) membrane. The possibility is discussed that AQP5 is synthesized during pregnancy as a stable protein that functions to regulate water secretion during lactation. AQP3 was identified primarily at the basal and lateral membranes of the secretory cells, suggesting a possible involvement in regulated uptake of water and glycerol. AQP1 was identified primarily at the capillary and secretory cell cytoplasmic level and may again be more concerned with uptake and hence milk synthesis, rather than secretion. The fact that expression was developmentally regulated supports, but does not prove, a regulatory involvement of AQPs in water flux through the milk secretory cell

Expression of the Small Conductance Ca²⁺-Activated Potassium Channel Subtype 3 (SK3) in Rat Uterus after Stimulation with 17β-Estradiol

<div><p>Preterm births accounts for roughly 9% of all births worldwide and can have detrimental or even lethal consequences for the infant. However to develop new treatment that will lower the rate of preterm births, more knowledge is required on the factors contributing to the contraction and relaxation of the myometrium. The small conductance Ca²⁺-activated potassium channel subtype 3 (SK3) has been identified in the myometrium of several species including humans, mice and rats, but with great inter species variation of the expression pattern and regulation. The aim of this study was to investigate the expression of SK3 in the uterus of rats stimulated with 17β-estradiol and progesterone in order to get an in depth understanding of the rat uterine SK3. Using immunohistochemistry SK3 was localized to the glandular and luminal endometrial lamina epitheliali. Furthermore, a weak signal was observed in the myometrium. Using Western blot the protein level of SK3 was found to increase in uteri from animals treated with 17β-estradiol, an effect that was not reflected at the mRNA level. The levels of mRNA for SK3 were significantly lower in the uterus of 17β-estradiol-treated animals than in the uterus of ovariectomized animals. We conclude that the SK channels are present in the endometrial epithelium, and possibly also in the myometrium of the rat uterus. Furthermore, the hormonal effect on SK3 caused by 17β-estradiol includes divergent regulation at mRNA and protein levels.</p></div

mRNA levels of SK3 in the rat uterus of the different groups of rats.

<p>All data are ratio to β-actin in the same sample and sham values are set to 1. Data are presented as mean ± S.E.M. Asterisk (*) represent significant difference in the different groups of animals (p<0.05). n = 6, 5, 6 and 6 for sham-operated animals, ovariectomized animals (OVX), 17β-estradiol-treated animals (E2) and progesterone-treated animals (P4) respectively.</p