TY - JOUR
T1 - A scalable culturing system for the marine annelid Platynereis dumerilii
AU - Kuehn, Emily
AU - Stockinger, Alexander W.
AU - Girard, Jerome
AU - Raible, Florian
AU - Duygu Özpolat, B.
N1 - Funding Information:
BDO received funding from Hibbitt Startup Funds. FR received funding from projects P30035, and I2972, by Austrian Science Fund (FWF). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank John Carr, Kate Dever, Lindsey DeMelo and the rest of the MRC staff at the MBL for their help in building the shelving setup and culture husbandry; we thank colleagues working on P. dumerilii for their helpful suggestions; and Guillaume Balavoine for providing the P. dumerilii larvae to start our cultures. We also thank two reviewers and the editor for their helpful suggestions that improved this manuscript.
Publisher Copyright:
© 2019 Kuehn et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Platynereis dumerilii is a marine segmented worm (annelid) with externally fertilized embryos and it can be cultured for the full life cycle in the laboratory. The accessibility of embryos and larvae combined with the breadth of the established molecular and functional techniques has made P. dumerilii an attractive model for studying development, cell lineages, cell type evolution, reproduction, regeneration, the nervous system, and behavior. Traditionally, these worms have been kept in rooms dedicated for their culture. This allows for the regulation of temperature and light cycles, which is critical to synchronizing sexual maturation. However, regulating the conditions of a whole room has limitations, especially if experiments require being able to change culturing conditions. Here we present scalable and flexible culture methods that provide ability to control the environmental conditions, and have a multi-purpose culture space. We provide a closed setup shelving design with proper light conditions necessary for P. dumerilii to mature. We also implemented a standardized method of feeding P. dumerilii cultures with powdered spirulina which relieves the ambiguity associated with using frozen spinach, and helps standardize nutrition conditions across experiments and across different labs. By using these methods, we were able to raise mature P. dumerilii, capable of spawning and producing viable embryos for experimentation and replenishing culture populations. These methods will allow for the further accessibility of P. dumerilii as a model system, and they can be adapted for other aquatic organisms.
AB - Platynereis dumerilii is a marine segmented worm (annelid) with externally fertilized embryos and it can be cultured for the full life cycle in the laboratory. The accessibility of embryos and larvae combined with the breadth of the established molecular and functional techniques has made P. dumerilii an attractive model for studying development, cell lineages, cell type evolution, reproduction, regeneration, the nervous system, and behavior. Traditionally, these worms have been kept in rooms dedicated for their culture. This allows for the regulation of temperature and light cycles, which is critical to synchronizing sexual maturation. However, regulating the conditions of a whole room has limitations, especially if experiments require being able to change culturing conditions. Here we present scalable and flexible culture methods that provide ability to control the environmental conditions, and have a multi-purpose culture space. We provide a closed setup shelving design with proper light conditions necessary for P. dumerilii to mature. We also implemented a standardized method of feeding P. dumerilii cultures with powdered spirulina which relieves the ambiguity associated with using frozen spinach, and helps standardize nutrition conditions across experiments and across different labs. By using these methods, we were able to raise mature P. dumerilii, capable of spawning and producing viable embryos for experimentation and replenishing culture populations. These methods will allow for the further accessibility of P. dumerilii as a model system, and they can be adapted for other aquatic organisms.
UR - http://www.scopus.com/inward/record.url?scp=85076242012&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0226156
DO - 10.1371/journal.pone.0226156
M3 - Article
C2 - 31805142
AN - SCOPUS:85076242012
SN - 1932-6203
VL - 14
JO - PloS one
JF - PloS one
IS - 12
M1 - e0226156
ER -