Immunolocalization of two different types of bilaterally paired CasBurs neurons per neuromere in crab TGC and POs during late postembryonic development.

<p>(<b>A</b>) CasBurs neurons in the subesophageal and thoracic but not in the abdominal neuromeres of TGC of a postlarva, megalopa. Inset shows a pre-absorption control of an adult TGC. (<b>B</b>) CasBurs neurons in the entire TGC of the 1^st crab. (<b>C–F</b>) Details of neurons in the last thoracic and abdominal neuromeres of the 1^st(<b>C</b>), 3^rd (<b>D</b>), 4^th crab (<b>F</b>). (<b>E</b>, <b>E’</b>) Two distinct types of CasBurs neurons in thoracic segments showing a difference in staining intensity but no size disparity yet. (<b>G</b>) CasBurs neurons in the TGC of a 12^th crab, inset: enlarged cells in the 4^th and the 5^th thoracic neuromeres; note clear disparity in size of the distinct neuron types in abdominal neuromers. (<b>H</b>) Higher magnification of abdominal neuromeres of the 12^th crab’s TGC without apparent size disparity; note the contralateral projection patterns (arrows). (<b>I</b>) Fibers and terminals in the anterior bar of a PO (<b>J</b>) A pair of typically disparate-sized thoracic cell bodies in an adult TGC. <i>Asterisks</i>: foramen of the sternal artery. Whole mounts in all cases; scale bars A, B, C, D, F, I = 100 µm; B inset, E, E’ = 50 µm; H = 200 µm; J = 25 µm.</p

CasBurs drives deposition and hardening of cuticle and hemocyte granulation <i>in vitro</i>.

<p>Semi-thin cross-sections (1 µm) through pieces of dorsal carapace cuticle and hemocytes from control crabs (<i>c</i>; <b>A, D, F, G, I</b>) and after treatment with rCasBurs (<i>tr</i>, <b>B, C, E, H</b>). Control cuticle is thin in stage D₂, but grows rapidly to considerable thickness already visible in ecdysis stage E_1% (<b>D</b>) and even more during stage A_{30 min} (<b>F</b>). Treatment (<i>tr</i>) with rCasBurs, however, clearly stimulates cuticle deposition already in the sensitive stage D₂ (<b>B, C</b>) but also in stage E_1% (<b>E</b>). Hemocytes underneath the hypodermis change from virtually inactive as in E_1% and A_{30 min} controls (<b>G, I</b>) to highly active granulated appearance under rCasBurs treatment readily in stage E_1%. (<b>H</b>). Scale bar: <b>A–C</b> = 25 µm, <b>D</b> and <b>E</b> = 50 µm, <b>F</b> = 100 µm <b>G–I</b> = 10 µm. <i>Arrowheads</i>: setae on the cuticle surface; <i>asterisk</i>: laminated layers of exocuticle.</p

CasBurs synthesis in TGCs and its storage in the POs, and identification of CasBurs heterodimers and CasBurs ß homodimers in POs.

<p>(<b>A</b>) <i>CasBurs α</i> and <i>β</i> (20 ng total RNA tissue equivalents for RT-PCR assay) is expressed only in TGCs but not in ES or brains of a juvenile (∼50 mm CW) and an adult female (F; >125 mm CW) in intermolt, whilst arginine kinase (<i>AK</i>) served as control. (<b>B</b>) About ten times more CasBurs is stored in POs compared with TGCs (means ± SEM pmol/tissue (n = 6) estimated by CasBurs RIA; crab stage 15–16: 70–90 mm carapace width, CW). (<b>C</b>) CasBurs RIA identifies two major peak fractions after RP-HPLC separations of extracts of juvenile and adult POs (Chromatogram shown for juvenile POs only; <i>closed bars</i>: contents of juvenile POs at E_0% stage; <i>open bars</i>: adult females at C₄ stage). (<b>D</b>) Western blot analysis of fractions F17 and F21 obtained from juvenile POs clearly showing the CasBurs ß subunits only in F17 and CasBurs ß and α subunits in F21 (Marker <i>arrowheads</i> at 15 and10 kDa).</p

<i>CasBurs α</i> and <i>ß</i> expressions in TGCs and CasBurs, CCAP and CHH protein contents in TGCs, POs and hemolymph during the molt cycle.

<p>(<b>A</b>) The data of the QRT-PCR assays show copies/µg of TGC total RNA (n = 6–12) of CasBurs α and CasBurs ß after normalization against those of <i>AK</i> (see Suppl. Information for details). <i>Open bars</i>: CasBurs α; <i>closed bars</i>: CasBurs ß. <i>Closed circles</i> represent the ratio of CasBurs ß vs. α. Statistical significance levels at <i>P</i><0.05 (ANOVA, Tukey-Kramer multiple comparison test) as marked with <i>asterisk</i> or different letters for the ratio of CasBurs ß vs. CasBurs α. (<b>B</b>) Changes in the amounts of CasBurs protein in TGC (<i>open bars</i>) and PO (<i>closed bars</i>) during the molt cycle. Note the significant increase in TGC-contents and dramatic decrease in PO-contents during postmolt stage A_{2 hr}. The data are shown as means ± SEM pmol/tissue (n = 8–15). Statistical significance (ANOVA, Tukey-Kramer multiple comparison test, <i>P</i><0.05) is noted with capital letters for Burs in PO and lower case letters for Burs in TGC. (<b>C</b>) CasBurs (<i>closed bars</i>) and CCAP (<i>open bars</i>) contents measured in the same hemolymph samples from various molt stages show amounts close to each other at all sampling points. Concentrations are shown as means ± SEM pM hemolymph (n = 8–15). Statistical significance (ANOVA, Tukey-Kramer multiple comparison test, <i>P</i><0.05) is indicated with different capital letters for Burs and different lower case letters for CCAP. (<b>D</b>) CHH titers of the same hemolymph samples as in C during the molt cycle. Note that significant increases of CHH titers are already beginning during early to mid (E_10%–E_50%) and late (E_100%) ecdysis as indicated by lower case letters at <i>P</i><0.05 (ANOVA, Turkey-Kramer multiple comparison test). Data are presented as means ± SEM pM (n = 8–15).</p

Depressing Antidepressant: Fluoxetine Affects Serotonin Neurons Causing Adverse Reproductive Responses in Daphnia magna

Selective serotonin reuptake inhibitors (SSRIs) are widely used antidepressants. As endocrine disruptive contaminants in the environment, SSRIs affect reproduction in aquatic organisms. In the water flea Daphnia magna, SSRIs increase offspring production in a food ration-dependent manner. At limiting food conditions, females exposed to SSRIs produce more but smaller offspring, which is a maladaptive life-history strategy. We asked whether increased serotonin levels in newly identified serotonin-neurons in the Daphnia brain mediate these effects. We provide strong evidence that exogenous SSRI fluoxetine selectively increases serotonin-immunoreactivity in identified brain neurons under limiting food conditions thereby leading to maladaptive offspring production. Fluoxetine increases serotonin-immuno­reactivity at low food conditions to similar maximal levels as observed under high food conditions and concomitantly enhances offspring production. Sublethal amounts of the neurotoxin 5,7-dihydroxytryptamine known to specifically ablate serotonin-neurons markedly decrease serotonin-immunoreactivity and offspring production, strongly supporting the effect to be serotonin-specific by reversing the reproductive phenotype attained under fluoxetine. Thus, SSRIs impair serotonin-regulation of reproductive investment in a planktonic key organism causing inappropriately increased reproduction with potentially severe ecological impact

Faktory ovlivňující reprodukční ukazatele dojnic ve vybraném chovu holštýnského skotu

The diploma thesis dealt with the analysis, and evaluation of dairy performance and its correlation with reproductive rates of Holstein cattle. Milk cows which were kept in Měcholupy agricultural, Inc., Předslav were analyzed between 2013 - 2015. Th ecompany is located in the foothills of the Bohemian Forest (Šumava). It breeds cattle for milk as well as meat, and pigs. The company management is committed to achieve good levels of milk production. It pays constant attention to this issue, so the average performance has risen since 2013 from 8 726 kilograms of milk to 9 093 kg of milk per cow in 2015. Cows on the 4th and higher lactation reached the highest performance in 2015 9 649 kg of milk. As for the European Union who abolished milk quotas in 2015 and the situation on the milk market is still complicated by banning exports of milk powder to Russia (Russian annexation of the Crimea), the management´s effort has been to produce milk at the lowest possible cost. They keep trying to limit culling as much as possible, so that they solve the problem of mastitis and reproduction at the highest level. Each cow which is treated has an individual approach, as well as the animal husbandry has specific requirements for animal breeding company in terms of quality seed. The results showed that milk cows with other health problems were eliminated the most (38.6% per year from 2013 to 2015) and also the ones with metabolism disorders and problems with hooves. The mastitis issue is dealt with very effectively, so that it is not one of the major problems of the herd (7.24 % for 2013 -2015). Management seeks to eliminate the problem milk cow in the lowest age and conversely on contrary if the animal is healthy, they try to keep it in the herd as long as possible, because such an animal has the greatest economic effect. Reproductive rateis not too satisfactory, but the management has no interest in hormonal control of reproduction. Active search of rutting cows is preferred by nursing staff. They cooperate actively with animal breeders and require a delivery of the insemination doses according to their requirements for each for each co

Additional file 2: of Evolution of neuropeptides in non-pterygote hexapods

Sequence logos of single-copy peptides. The logos show the sequence variability in the single-copy peptides (<30 amino acids) of non-pterygote hexapod lineages determined using WebLogo version 2.8.2 (http://weblogo.berkeley.edu/). For species with two described genes containing different mature neuropeptides, the more conserved sequence was selected. CNMa was not found in Protura and Collembola, while elevenin was not found in Collembola. Pigment-dispersing factor (PDF) was not found in Protura and Diplura. For comparative purposes, the sequence conservation of CAPA-PVK paracopies was added as an example of multiple-copy peptides. A distinct core sequence is obvious in all sequence logos. The N-terminal sequence motifs are variable in a number of peptides, including SIFa, and this variability is similar to that of the N-termini in the different PVK paracopies. (DOC 460 kb