Generation of a Xenopus laevis F1 albino J strain by genome editing and oocyte host-transfer

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Developmental Biology 426 (2017): 188–193, doi:10.1016/j.ydbio.2016.03.006.Completion of the Xenopus laevis genome sequence from inbred J strain animals has facilitated the generation of germline mutant X. laevis using targeted genome editing. In the last few years, numerous reports have demonstrated that TALENs are able to induce mutations in F0 Xenopus embryos, but none has demonstrated germline transmission of such mutations in X. laevis. In this report we used the oocyte host-transfer method to generate mutations in both tyrosinase homeologs and found highly-penetrant germline mutations; in contrast, embryonic injections yielded few germline mutations. We also compared the distribution of mutations in several F0 somatic tissues and germ cells and found that the majority of mutations in each tissue were different. These results establish that X. laevis J strain animals are very useful for generating germline mutations and that the oocyte host-transfer method is an efficient technique for generating mutations in both homeologs.This work was supported by grants from the NIH (OD010997 and HD084409)

Relazione tecnica sulle attività della campagna oceanografica “Evatir 2016”

La presente relazione tecnica descrive tutte le attività svolte nella Campagna oceanografica “Evatir 2017” condotta a bordo della N/O “G. Dallaporta” (dal 25 luglio al 21 agosto 2016). La Campagna “Evati 2016” è parte integrante del Progetto "Estensione della Campagna acustica Medias (Mediterranean International Acoustic Survey) nelle sub aree geografiche (GSA) 9 (Mar Ligure e Mar Tirreno settentrionale) e 10 (Mar Tirreno centrale e meridionale)", finanziato dal Ministero delle politiche agricole alimentari e forestali (Mipaaf) nell'ambito del Fondo Europeo per gli Affari Marittimi e la Pesca (FEAMP). E’ la sesta campagna rivolta alla valutazione acustica e alla distribuzione spaziale delle popolazioni di piccoli pelagici, insieme allo studio delle condizioni ambientali dell’area di studio. Le specie target del progetto sono state l’acciuga europea (Engraulis encrasicolus) e la sardina (Sardina pilchardus), specie chiave sia a livello commerciale che ecologico. La gestione di tali risorse è abbastanza complessa a causa del loro breve ciclo di vita e dall’ampia oscillazioni inter-annuali nell’abbondanza dello stock, legata al successo o al fallimento del reclutamento annuale. Insieme alle attività di acquisizione acustiche sono stati realizzati campionamenti bilogici, misurazioni, misurazioni di parametri fisico-chimici, rilevamenti di variabili oceanografiche e campionamento di acque (nei Golfi di Salerno, Napoli e Gaeta), monitoraggio della rete da pesca e campionamenti di fitoplancton, zooplancton, solidi sospesi e rilievo delle principali variabili oceanografiche (in prossimità dello stabilimento SOLVAY di Rosignano)

Relazione tecnica sulle attività della Campagna oceanografica “Ancheva 2016”

La presente relazione riporta le attività di ricerca della Campagna oceangrafica “Ancheva 2016”, svolte a bordo della N/O “G. Dallaporta” (nel periodo tra il 4 ed il 26 Luglio 2016) nello Stretto di Sicilia e nel mar Ionio occidentale (GSA 16 e 19) e nelle acque maltesi (GSA 15). Nello specifico, le attività svolte vengono sono di seguito descritte sinteticamente: - Rilevazioni acustiche degli stock di piccoli pelagici con echosounder scientifico Simrad EK60, con trasduttori split beam a scafo; - Campionamenti biologici (di piccoli pelagici) con rete pelagica (volante monobarca), dotata di sistema acustico Simrad ITI per il controllo della geometria della rete (apertura e posizione della rete nella colonna d’acqua); - Campionamenti di tessuti di pesci pelagici (gonadi, fegato e sangue); - Campionamento acqua, in specifiche stazioni, con sonda multiparametrica SEABIRD mod. 9/11 plus (per la misurazione dei parametri fisico-chimici della colonna d’acqua) dotata di bottiglie Niskin (per l’analisi di Nutrienti ed Isotopi di azoto e carbonio); - Campionamento ittioplantonico, mediante con “Bongo 40”, in specifiche stazioni, le cui bocche sono state fissate in alcool, per le analisi degli aminoacidi sulle larve di Engraulis encrasicolus

Relazione tecnica sulle attività della campagna oceanografica “Ancheva 2017”

La presente relazione tecnica descrive le attività svolte nella Campagna oceanografica “Ancheva 2017”, tra il 23 luglio ed il 7 agosto 2017, a bordo della N/O “G. Dallaporta”. Gli echosurvey acquisiti hanno permesso di valutare la biomassa e la distribuzione spaziale dei piccoli pelagici nella piattaforma meridionale della Sicilia, da Marsala a oltre Capo Passero, e nella piattaforma Maltese (GSA 16, FAO sub area 37.2.2). Parallelamente alle suddette attività sono state svolti il campionamento biologico (di piccoli pelagici) con rete pelagica (volante monobarca), il campionamento di tessuti di pesci pelagici (gonadi, fegato e sangue), il campionamento di acqua (per la misurazione dei parametri fisico-chimici della colonna d’acqua e per l’analisi di Nutrienti ed Isotopi di azoto e carbonio) ed, infine, il campionamento ittioplantonico (per le analisi degli aminoacidi sulle larve di Engraulis encrasicolus)

Copernicus Ocean State Report, issue 6

The 6th issue of the Copernicus OSR incorporates a large range of topics for the blue, white and green ocean for all European regional seas, and the global ocean over 1993–2020 with a special focus on 2020

Luteinizing Hormone is an effective replacement for hCG to induce ovulation in Xenopus

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Developmental Biology 426 (2017): 442–448, doi:10.1016/j.ydbio.2016.05.028.Injection of human Chorionic Gonadotropin (hCG) directly into the dorsal lymph sac of Xenopus is a commonly used protocol for induction of ovulation, but recent shortages in the stocks of commercially available hCG as well as lack of a well tested alternative have resulted in frustrating experimental delays in laboratories that predominantly use Xenopus in their research. Mammalian Luteinizing Hormones (LH) share structural similarity, functional equivalency, and bind the same receptor as hCG; this suggests that LH may serve as a good alternative to hCG for promoting ovulation in Xenopus. LH has been found to induce maturation of Xenopus oocytes in vitro, but whether it can be used to induce ovulation in vivo has not been examined. Here we compared the ability of four mammalian LH proteins, bovine (bLH), human (hLH), ovine (oLH), porcine (pLH), to induce ovulation in Xenopus when injected into the dorsal lymph sac of sexually mature females. We find that both ovine and human LH, but not bovine or porcine, are good substitutes for hCG for induction of ovulation in WT and J strain Xenopus laevis and Xenopus tropicalis.This work was supported by a grant from the NIHP40OD010997

Data from: Niche divergence by deep-sea octocorals in the genus Callogorgia across the continental slope of the Gulf of Mexico

Environmental variables that are correlated with depth have been suggested to be among the major forces underlying speciation in the deep sea. This study incorporated phylogenetics and ecological niche models (ENM) to examine whether congeneric species of Callogorgia (Octocorallia: Primnoidae) occupy different ecological niches across the continental slope of the Gulf of Mexico (GoM), and whether this niche divergence could be important in the evolution of these closely related species. Callogorgia americana americana, C. americana delta, and C. gracilis were documented at 13 sites in the GoM (250-1000 m) from specimen collections and extensive video observations. On a first order, these species were separated by depth, with C. gracilis occurring at the shallowest sites, C. a. americana at mid-depths, and C. a. delta at the deepest sites. Callogorgia a. delta was associated with areas of increased seep activity whereas C. gracilis and C. a. americana were associated with narrow, yet warmer, temperature ranges and did not occur near cold seeps. ENM background and identity tests revealed little to no overlap in ecological niches between species. Temporal calibration of the phylogeny revealed the formation of the Isthmus of Panama was a vicariance event that may explain some of the patterns of speciation within this genus. These results elucidate the potential mechanisms for speciation in the deep sea, emphasizing both bathymetric speciation and vicariance events in the evolution of a genus across multiple regions

DESS deconstructed: Is EDTA solely responsible for protection of high molecular weight DNA in this common tissue preservative?

DESS is a formulation widely used to preserve DNA in biological tissue samples. Although it contains three ingredients, dimethyl sulfoxide (DMSO), ethylenediaminetetraacetic acid (EDTA) and sodium chloride (NaCl), it is frequently referred to as a DMSO-based preservative. The effectiveness of DESS has been confirmed for a variety of taxa and tissues, however, to our knowledge, the contributions of each component of DESS to DNA preservation have not been evaluated. To address this question, we stored tissues of three aquatic taxa, Mytilus edulis (blue mussel), Faxonius virilis (virile crayfish) and Alitta virens (clam worm) in DESS, each component of DESS individually and solutions containing all combinations of two components of DESS. After storage at room temperature for intervals ranging from one day to six months, we extracted DNA from each tissue and measured the percentage of high molecular weight (HMW) DNA recovered (%R) and normalized HMW DNA yield (nY). Here, HMW DNA is defined as fragments >10 kb. For comparison, we also measured the %R and nY of HMW DNA from extracts of fresh tissues and those stored in 95% EtOH over the same time intervals. We found that in cases where DESS performed most effectively (yielding ≥ 20%R of HMW DNA), all solutions containing EDTA were as or more effective than DESS. Conversely, in cases where DESS performed more poorly, none of the six DESS-variant storage solutions provided better protection of HMW DNA than DESS. Moreover, for all taxa and storage intervals longer than one day, tissues stored in solutions containing DMSO alone, NaCl alone or DMSO and NaCl in combination resulted in %R and nY of HMW DNA significantly lower than those of fresh tissues. These results indicate that for the taxa, solutions and time intervals examined, only EDTA contributed directly to preservation of high molecular weight DNA

Quattrini_Callogorgia_ENVABUNDATA

Abundances and densities for Callogorgia spp. collected in the Gulf of Mexico, with corresponding site, location, and environmental data

Quattrini_etal_MSH_COI_28S_alignment

Quattrini_etal_MSH_COI_28S_alignmen