 ResearchIn vitro fertilization and artificial activation of eggs of the direct-developing anuran Eleutherodactylus coquiEsteban Toro1,2  and Scott F Michael1  1Department of Tropical Medicine, Box SL-17, Tulane University, New Orleans, LA 70112, USA 2Department of Developmental Biology, Stanford University, Stanford, CA 94045, USA  author email corresponding author email
Reproductive Biology and Endocrinology 2004,
2:60doi:10.1186/1477-7827-2-60 Abstract
Although much is known about the reproductive biology of pond-breeding frogs, there is comparatively little information about terrestrial-breeding anurans, a highly successful and diverse group. This study investigates the activation and in vitro fertilization of eggs of the Puerto Rican coqui frog obtained by hormonally induced ovulation. We report that spontaneous activation occurs in 34% of eggs, probably in response to mechanical stress during oviposition. Artificial activation, as evidenced by the slow block to polyspermy and the onset of zygote division, was elicited both by mechanical stimulation and calcium ionophore exposure in 64% and 83% of the cases, respectively. Finally, one in vitro fertilization protocol showed a 27% success rate, despite the fact that about one third of all unfertilized eggs obtained by hormone injection auto-activate. We expect these findings to aid in the conservation effort of Eleutherodactylus frogs, the largest vertebrate genus. | 
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