The control values were measured using a chamber filled with MFSW
4) with 40 ml of MFSW
containing 3 g of sucrose and 10 ml of 25% glutaraldehyde.
Abbreviations: AG, apical ganglion; AN, apical nerve; MFSW
, Millipore-filtered seawater; SCP, small cardioactive peptide.
To enhance embryonic development, either horse serum (HS) (Sigma) or bovine serum albumin (BSA) (ICN) was added to the MFSW culture.
5% BSA/ MFSW in plastic petri dishes unlined by an agarose cushion fall to undergo chorion expansion and die after 3 or 4 days.
Ten larvae were exposed to the substrate in MFSW either in the presence or absence of 0.
Formaldehyde was freshly prepared by dissolving paraformaldehyde (EM grade, Ted Pella, CA) in MFSW, adjusting the pH to 7.
Gastrulating embryos were fixed with 1% glutaraldehyde dissolved in MFSW
at room temperature for 2 h.
Sperm concentrate could be stored in the refrigerator in sealed Eppendorf vials for 1 or 2 h or used immediately by gently drawing several drops into a Pasteur pipette and adding them to a vial containing 10 ml of MFSW
to activate them.