Respiration rate of Microsetella norvegica collected during James Cook cruise JC087

Respiration of Microsetella norvegica was measured at PAP site during two days, using a UNISENSE microrespiration system and microelectrodes for O2. 10-20 starved Microsetella individuals were carefully placed into 2-ml respiration chambers in filtered sea water, and their respiration was measured for 20 min. The respiration rate was calculated based on the slope of the decrease in oxygen against time in the respiration chamber containing Microsetella, compared to the control where only filtered seawater was present. In total 18 measurements were conducted.

Production, oxygen uptake, and sinking velocity of copepod fecal pellets originated from the central North Sea

Production, oxygen uptake, and sinking velocity of copepod fecal pellets egested by Temora longicornis were measured using a nanoflagellate (Rhodomonas sp.), a diatom (Thalassiosira weissflogii), or a coccolithophorid (Emiliania huxleyi) as food sources. Fecal pellet production varied between 0.8 pellets ind**-1 h**-1 and 3.8 pellets ind**-1 h**-1 and was significantly higher with T. weissflogii than with the other food sources. Average pellet size varied between 2.2 x 10**5 µm**3 and 10.0 x 10**5 µm**3. Using an oxygen microsensor, small-scale oxygen fluxes and microbial respiration rates were measured directly with a spatial resolution of 2 µm at the interface of copepod fecal pellets and the surrounding water. Averaged volume-specific respiration rates were 4.12 fmol O2 µm**-3 d**-1, 2.86 fmol O2 µm**-3 d**-1, and 0.73 fmol O2 µm**-3 d**-1 in pellets produced on Rhodomonas sp., T. weissflogii, and E. huxleyi, respectively. The average carbon-specific respiration rate was 0.15 d**-1 independent on diet (range: 0.08-0.21 d**-1). Because of ballasting of opal and calcite, sinking velocities were significantly higher for pellets produced on T. weissflogii (322 +- 169 m d**-1) and E. huxleyi (200 +- 93 m d**-1) than on Rhodomonas sp. (35 +- 29 m d**-1). Preservation of carbon was estimated to be approximately 10-fold higher in fecal pellets produced when T. longicornis was fed E. huxleyi or T. weissflogii rather than Rhodomonas sp. Our study directly demonstrates that ballast increases the sinking rate of freshly produced copepod fecal pellets but does not protect them from decomposition.

Mesozooplankton biomass data from the Kapisigdlit an inner fjord branch of the Godthaabsfjord system, West Greenland, 2010

Sampling was conducted from March 24 to August 5 2010, in the fjord branch Kapisigdlit located in the inner part of the Godthåbsfjord system, West Greenland. The vessel "Lille Masik" was used during all cruises except on June 17-18 where sampling was done from RV Dana (National Institute for Aquatic Resources, Denmark). A total of 15 cruises (of 1-2 days duration) 7-10 days apart was carried out along a transect composed of 6 stations (St.), spanning the length of the 26 km long fjord branch. St. 1 was located at the mouth of the fjord branch and St. 6 was located at the end of the fjord branch, in the middle of a shallower inner creek . St. 1-4 was covering deeper parts of the fjord, and St. 5 was located on the slope leading up to the shallow inner creek. Mesozooplankton was sampled by vertical net tows using a Hydrobios Multinet (type Mini) equipped with a flow meter and 50 µm mesh nets or a WP-2 net 50 µm mesh size equipped with a non-filtering cod-end. Sampling was conducted at various times of day at the different stations. The nets were hauled with a speed of 0.2-0.3 m s**-1 from 100, 75 and 50 m depth to the surface at St. 2 + 4, 5 and 6, respectively. The content was immediately preserved in buffered formalin (4% final concentration). All samples were analyzed in the Plankton sorting and identification center in Szczecin (www.nmfri.gdynia.pl). Samples containing high numbers of zooplankton were split into subsamples. All copepods and other zooplankton were identified down to lowest possible taxonomic level (approx. 400 per sample), length measured and counted. Copepods were sorted into development stages (nauplii stage 1 - copepodite stage 6) using morphological features and sizes, and up to 10 individuals of each stage was length measured.

Zooplankton carbon budget from Trondheim mesocosm experiment

We measured respiration, egg production and fecal pellet production of five common copepod species, when fed on suspended or aggregated food from two mesocosm, + NP and + NPSi. We hypothetised that calanoid copepods (Temora longicornis, Acartia spp., Centropages spp.) would feed mainly on suspended food, and have low respiration and egestion rates when food was only available as aggregates, while harpacticoids and Oncaea spp. would mainly feed on aggregated food and have low metabolic rates when only suspended food was available. Copepods were collected from the lagoon, and adapted to experimental conditions for 24 h. Food suspension was collected from the mesocosms, and either offered to copepods directly (suspended food) or after rotating in a plankton wheel until most phytoplankton was aggregated together (aggregated food). After 24-h incubation we counted the produced eggs and pellets, and measured copepod respiration using microelectrodes.

Aggregates feeding and pellet production from Microsetella norvegica collected during METEOR cruise M87/1 in Aril 2012

Harpacticoid Microsetella norvegica was fed with 5 concentrations of aggregates, collected from the station 1 (experiment 1) or from station 2 (experiment 2). The aggregates at station 1 were of phytoplankton origin and consisted mainly of Phaeocystis sp. and radiolarians; aggregates at station 2 were detritus collected from deep Mocness tows. M. norvegica was starved in filtered sea water for > 12 h, after which it was incubated together with aggregates for 8 h. After the incubation, pellets were counted and Microsetella and remaining aggregates were counted and measured. Pellet production of M. norvegica reflects feeding so that when pellet production is plotted against aggregate concentration, a functional response can be obtained.

Diurnal egg and pellet production of Calanus finmarchicus collected during METEOR cruise M87/1 in Aril 2012

Egg and pellet production of Calanus finmarchicus was measured at 6-h intervals at all stations during the second leg of the cruise. Calanus was collected at the surface 150-m using a WP2 plankton net, and incubated in chl-max water for 24-h. Each 6 hours females were transferred to a new food solution and eggs and pellets were counted. In the end of the experiment, females were measured for prosome length. The purpose of the exercise was to calculate the minimum carbon consumption of Calanus, and how large proportion of ingestion is egested as fast sinking fecal pellets, and when.

Lenght measurements and carbon biomass of individual zooplankton from METEOR cruise M87/1 in April 2012

Zooplankton samples were taken in five depth strata using a Multinet type Midi, with 50 µm nets. The samples were taken during the second leg only, three times at station 1, two times at station 2 and once at station 3. Zooplankton were identified to species / genus and life-stage, and at least 300 individuals were counted per sample. 10 individuals of each stage / species were measured and the numbers of eggs counted.

Zooplankton egg counts from METEOR cruise M87/1 in April 2012

Zooplankton samples were taken in five depth strata using a Multinet type Midi, with 50 µm nets. The samples were taken during the second leg only, three times at station 1, two times at station 2 and once at station 3. Zooplankton were identified to species / genus and life-stage, and at least 300 individuals were counted per sample. 10 individuals of each stage / species were measured and the numbers of eggs counted.

Zooplankton abundance from METEOR cruise M87/1 in April 2012

Zooplankton samples were taken in five depth strata using a Multinet type Midi, with 50 µm nets. The samples were taken during the second leg only, three times at station 1, two times at station 2 and once at station 3. Zooplankton were identified to species / genus and life-stage, and at least 300 individuals were counted per sample. 10 individuals of each stage / species were measured and the numbers of eggs counted.

Chlorophyll-a in gut from Microsetella norvegica and Oncaea collected during James Cook cruise JC087

Gut chlorophyll of Microsetella norvegica and Oncaea spp. was measured daily at the PAP site from 8th to 14th of June, 2013. Copepods were collected using a WP2 -type net with a mesh size of 50 µm. Immediately after, 40-70 adults and late copepodites of each species (with 2-3 replicate samples) were washed in filtered sea water, placed on CF/F filters and extracted in acetone. Chlorophyll-a on filters was analysed using standard methods.

Vertical distribution and abundance of small zooplankton from the James Cook cruise JC087

Zooplankton samples were collected daily at the PAP site, using a Multinet of the type Midi with 50 µm nets. 5 depth strata (1000-500, 500-300, 300-100, 100-50 and 50-0 m) were collected at each sampling. The samples were preserved in 2% borax bufferred formalin. Zooplankton were identified on a species / genus level including different life-stages and eggs; at least 400 individuals were counted for each sample. When present, 10 individuals of each species and life-stages (for copepods) were measured for their prosome or total length.

The Tabby phenotype is caused by mutation in a mouse homologue of the EDA gene that reveals novel mouse and human exons and encodes a protein (ectodysplasin-A) with collagenous domains

Mouse Tabby (Ta) and X chromosome-linked human EDA share the features of hypoplastic hair, teeth, and eccrine sweat glands. We have cloned the Ta gene and find it to be homologous to the EDA gene. The gene is altered in two Ta alleles with a point mutation or a deletion. The gene is expressed in developing teeth and epidermis; no expression is seen in corresponding tissues from Ta mice. Ta and EDA genes both encode alternatively spliced forms; novel exons now extend the 3′ end of the EDA gene. All transcripts recovered have the same 5′ exon. The longest Ta cDNA encodes a 391-residue transmembrane protein, ectodysplasin-A, containing 19 Gly-Xaa-Yaa repeats. The isoforms of ectodysplasin-A may correlate with differential roles during embryonic development.

Involvement of caspase-dependent activation of cytosolic phospholipase A2 in tumor necrosis factor-induced apoptosis

Tumor necrosis factor (TNF)-induced apoptosis is mediated by caspases, which are cysteine proteases related to interleukin 1β-converting enzyme. We report here that TNF-induced activation of caspases results in the cleavage and activation of cytosolic phospholipase A2 (cPLA2) and that activated cPLA2 contributes to apoptosis. Inhibition of caspases by expression of a cowpox virus-derived inhibitor, CrmA, or by a specific tetrapeptide inhibitor of CPP32/caspase-3, acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO), inhibited TNF-induced activation of cPLA2 and apoptosis. TNF-induced activation of cPLA2 was accompanied by a cleavage of the 100-kDa cPLA2 to a 70-kDa proteolytic fragment. This cleavage was inhibited by Ac-DEVD-CHO in a similar manner as that of poly(ADP)ribose polymerase, a known substrate of CPP32/caspase-3. Interestingly, specific inhibition of cPLA2 enzyme activity by arachidonyl trifluoromethylketone (AACOCF3) partially inhibited TNF-induced apoptosis without inhibition of caspase activity. Thus, our results suggest a novel caspase-dependent activation pathway for cPLA2 during apoptosis and identify cPLA2 as a mediator of TNF-induced cell death acting downstream of caspases.

Folding of Active β-Lactamase in the Yeast Cytoplasm before Translocation into the Endoplasmic Reticulum

Polypeptides targeted to the yeast endoplasmic reticulum (ER) posttranslationally are thought to be kept in the cytoplasm in an unfolded state by Hsp70 chaperones before translocation. We show here that Escherichia coli β-lactamase associated with Hsp70, but adopted a native-like conformation before translocation in living Saccharomyces cerevisiae cells. β-Lactamase is a globular trypsin-resistant molecule in authentic form. For these studies, it was linked to the C terminus of a yeast polypeptide Hsp150Δ, which conferred posttranslational translocation and provided sites for O-glycosylation. We devised conditions to retard translocation of Hsp150Δ-β-lactamase. This enabled us to show by protease protection assays that an unglycosylated precursor was associated with the cytoplasmic surface of isolated microsomes, whereas a glycosylated form resided inside the vesicles. Both proteins were trypsin resistant and had similar β-lactamase activity and Km values for nitrocefin. The enzymatically active cytoplasmic intermediate could be chased into the ER, followed by secretion of the activity to the medium. Productive folding in the cytoplasm occurred in the absence of disulfide formation, whereas in the ER lumen, proper folding required oxidation of the sulfhydryls. This suggests that the polypeptide was refolded in the ER and consequently, at least partially unfolded for translocation.