title: Application of a functional marker for the effect of cryoprotectant agents on gorgonian coral (Junceella juncea and J. fragilis) sperm sacs.
abstract: BACKGROUND
  The establishment of coral sperm repositories which retain good post-rewarming viability and fertility play a vital role in species conservation.
  
  
  OBJECTIVE
  This study aimed at obtaining baseline information regarding the effects of cryoprotectant agents (CPAs) on gorgonian coral (Junceella juncea and J. fragilis) sperm sacs.
  
  
  METHODS
  The adenosine triphosphate assay was used to determine the energy level of the gorgonian sperm sacs as an indicator of sperm viability after exposure to cryoprotectants.
  
  
  RESULTS
  The 'no observed effect concentrations' (NOECs) of methanol, dimethyl sulfoxide (DMSO), polypropylene glycol (PG), ethylene glycol (EG) and glycerol for J. juncea sperm sacs were 3 M, 3 M, 1 M, 2 M and 1 M respectively after 20 min exposure; whilst the NOECs for J. fragilis oocytes were 2 M, 3 M, 1 M, 2 M and 2 M, respectively. Methanol and DMSO had the least impact. PG was the most toxic CPA after 10 min exposure. ATP content of J. juncea and J. fragilis sperm sacs did not differ significantly from the control with incubation times of 10-20 min with 2 M EG. However, ATP content dropped significantly after exposing sperm sacs to 2 M EG for 40 min with average values of 2.34 +/- 0.12 and 1.97 +/- 0.48 microg/ml respectively. ATP content for J. juncea and J. fragilis sperm sacs was significantly decreased to 1.79 +/- 0.31 and 2.40 +/- 0.36 microg/ml after 20 min incubation in 2 M PG when compared to the control with 2.98 +/- 0.16 and 4.14 +/- 0.42 microg/ml respectively. Normalized ATP content for sperm sacs of two different gorgonian coral after incubation in methanol, DMSO, PG, EG and glycerol showed that J. juncea sperm sacs were slightly less tolerant to CPAs compared to J. fragilis sperm sacs.
  
  
  CONCLUSIONS
  DMSO or methanol can be considered as efficient CPAs for gorgonian sperm sacs cryopreservation. The ATP luminescence assay provided sensitive and rapid quantification of mitochondrial activity in gorgonian coral sperm sacs. The study on the impact of CPA will contribute to the development of a cryopreservation protocol for coral sperm conservation.
authors.forename: S
authors.initials: S
authors.lastname: Tsai
authors.forename: V
authors.initials: V
authors.lastname: Kuit
authors.forename: Z G
authors.initials: ZG
authors.lastname: Lin
authors.forename: C
authors.initials: C
authors.lastname: Lin
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D037421:Anthozoa / Q000166:cytology / Q000187:drug effects*; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003247:Conservation of Natural Resources; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D008163:Luminescent Measurements; D008297:Male; D000432:Methanol / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011108:Polymers / Q000494:pharmacology; D011409:Propylene Glycols / Q000494:pharmacology; D012662:Semen Preservation / Q000379:methods*; D013076:Sperm Count; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D003451:Cryoprotective Agents; D011108:Polymers; D011409:Propylene Glycols; C012504:polypropylene glycol; D000255:Adenosine Triphosphate; D019855:Ethylene Glycol; D005990:Glycerol; D000432:Methanol; D004121:Dimethyl Sulfoxide