Veratrum californicum was responsible for large losses of sheep grazing high mountain ranges in central Idaho in the 1950s. Veratrum induces various
birth defects including the cyclopic-type craniofacial defect (monkey-faced lambs) that is specifically induced in lambs after pregnant ewes grazed the plant on the 14th day of gestation. The steroidal
alkaloids cyclopamine (1) and
jervine (2) were isolated from Veratrum and shown to be primarily responsible for the malformations.
Cyclopamine (1) and
jervine (2) are potent
teratogens that inhibit Sonic hedgehog (Shh) signaling during gastrulation-stage embryonic development, producing
cyclopia and
holoprosencephaly. Although losses to the sheep industry from Veratrum are now relatively infrequent, occasional incidents of toxicoses and craniofacial malformations are still reported in sheep and other species. However, the benefits to biomedical research using
cyclopamine (1) as a tool to study human diseases have greatly expanded. A competitive inhibition
enzyme-linked
immunosorbent assay (ELISA) to detect and measure
cyclopamine (1) and
jervine (2) was developed using polyclonal
antibodies produced in ewes. The limits of detection of the assay were 90.0 and 22.7 pg for
cyclopamine (1) and
jervine (2), respectively. This assay was used for the detection and measurement of
cyclopamine (1) spiked into sheep blood. The simple extraction-ELISA methods developed in this study demonstrate the potential of using these techniques for the rapid screening of
biological samples to detect the presence and concentration of
cyclopamine (1) and
jervine (2) and will be beneficial to pharmacological studies and livestock diagnostics.