Aquaculture 2022

February 28 - March 4, 2022

San Diego, California

DEVELOPMENT OF Aplysia californica EMBRYOS OUTSIDE THE EGG STRAND TO PROVIDE NEW OPTIONS FOR CRYOPRESERVATION?

Allyssa M. Oune*, Jack C. Koch, M. Teresa Gutierrez-Wing, Lucia Arregui,

Rose M. O. Upton, and Terrence R. Tiersch

 

 Aquatic Germplasm and Genetic Resources Center

Louisiana State University Agricultural Center, School of Renewable Natural Resources

 2288 Gourrier Avenue

Baton Rouge, LA 70820

 aoune1@lsu.edu

 



The California Sea Hare (Aplysia californica) is a biomedical model used for studies in molecular neurobiology, electrophysiology, learning, and memory. As with other models, research communit ies often require maintenance of different genetic lines. C ryopreservation has been proposed as one method to preserve these lines. In  many aquaculture species , sperm is cryopreserved, but because A. californica is an internal fertilizer , their sperm is difficult to extract. Thus,  we are investigating if embryos  and larvae  can be cryopreserved instead. During spawning, m ultiple embryos are enclosed within egg capsules which are packaged within a strand (Figure 1A) . The multiple barriers associated with the strand make it difficult to determine how well and how quickly cryoprotectants can penetrate embryos, information that is vital for developing a cryopreservation protocol.  An alternative could be to freeze embryos  or larvae free from the strand.  Egg strands were cut into 1-cm pieces; six  were added to 32 ppt artificial sea water (ASW) , and six were cut open to release embryos into the ASW. Samples were  observed for 16 days at 16°C or  until  >90% of  the  individuals died. Development was assessed and larval stage demographics were counted for each sample. Approximately 87% of embryos  in strands developed normally into the  early veliger stage ,  while only 8% of the free larvae developed into early veligers and of these,  more than half exhibited abnormal development . The remaining  free embryos arrested at an earlier developmental stage, trochophore larvae (Figure 1C). This suggests that the embryos  depend on the environment provided within the strand to undergo normal development . Alt hough  some A. californica embryos can develop outside the strand and capsule, they developed slower and abnormally. Further research to develop ways to culture embryos outside the strand and capsule is needed  to determine whether cryopreserving free larvae would be feasible.