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The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

Artikel i vetenskaplig tidskrift
Författare Per R. Jonsson
A. L. Wrange
Ulrika Lind
Anna Abramova
Martin Ogemark
Anders Blomberg
Publicerad i Jove-Journal of Visualized Experiments
Nummer/häfte 138
ISSN 1940-087X
Publiceringsår 2018
Publicerad vid Institutionen för marina vetenskaper
Institutionen för kemi och molekylärbiologi
Språk en
Länkar dx.doi.org/10.3791/57825
Ämnesord Environmental Sciences, Issue 138, Barnacle, Crustacean, Balanus (Amphibalanus) improvisus, ostrea-edulis, recruitment, cirripedia, settlement, coatings, adhesion, panels
Ämneskategorier Marin ekologi, Miljövetenskap

Sammanfattning

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly relevant as a model for the studies of osmoregulatory mechanisms because of its extreme tolerance to low salinity. It is also widely used as a model of settling biology, in particular in relation to antifouling research. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae for studies. A protocol for the all-year-round culturing of B. improvisus has been developed and a detailed description of all steps in the production line is outlined (i.e., the establishment of adult cultures on panels, the collection and rearing of barnacle larvae, and the administration of feed for adults and larvae). The description also provides guidance on troubleshooting and discusses critical parameters (e.g., the removal of contamination, the production of high-quality feed, the manpower needed, and the importance of high-quality seawater). Each batch from the culturing system maximally yields roughly 12,000 nauplii and can deliver four batches in a week, so up to almost 50,000 larvae per week can be produced. The method used to culture B. improvisus is, probably, to a large extent also applicable to other marine invertebrates with free-swimming-larvae. Protocols are presented for the dissection of various tissues from adults as well as the production of high-quality RNA for studies on gene expression. It is also described how cultured adults and reared cyprids can be utilized in a wide array of experimental designs for examining gene expression in relation to external factors. The use of cultured barnacles in gene expression is illustrated with studies of possible osmoregulatory roles of Na+/K+ ATPase and aquaporins.

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