zrps3a, zrpl5a or zrpl11 knockdown heightens vulnerability to alcohol-induced craniofacial deficits in zebrafish embryos.

(A-E) Untreated embryos (A) had normal development of cranial cartilage elements, and these were largely unaffected by treatment with a nonsense morpholino (B) or morpholinos directed against zrps3a (C), zrpl5a (D), or zrpl11 (E). (F-J) Alcohol treatment modestly reduced the size and shape of cranial cartilage elements in otherwise normal embryos. Treatment with a nonsense morpholino (G) did not further worsen cranial development. However, the combination of alcohol with morpholino directed against zrps3a (H), zrpl5a (I), or zrpl11 (J) resulted in ablation of many cranial cartilage elements and reduced ocular size. Embryos are either 4 dpf (A-D, F-I) or 3 dpf (E, J). All views are ventral with equivalent magnification. Abbreviations used: c, cardiac edema; cb, ceratobranchial; ch, ceratohyal; ep, ethmoid plate; m, Meckel’s cartilage.

zrps3a, zrpl5a, or zrpl11 Knockdown Heightens Vulnerability to Alcohol-Induced Apoptosis in 12 hpf Zebrafish Embryos.

(A-E) Untreated embryos (A) had few TUNEL+ cells in the cranial region (arrow). Treatment with a nonsense morpholino (B) caused a modest increase in TUNEL⁺ cells within the cranial region (arrow), as did morpholinos directed against zrps3a (C), zrpl5a (D), or zrpl11 (E). (F-J) Alcohol treatment (F) caused appreciable apoptosis within both cranial and somatic regions. The addition of nonsense morpholino treatment (G) did not further increase TUNEL⁺ cell numbers in alcohol-treated embryos. However, the combination of alcohol with morpholino directed against zrps3a (H), zrpl5a (I), or zrpl11 (J) resulted in higher levels of apoptosis within the cranial region as compared with embryos that received the same morpholino and no alcohol (C-E), or alcohol and no morpholino (F). All views are lateral at equivalent magnification, with rostral at the top and the embryo ‘looking’ left. Arrow indicates the cranial region. (K) Enumeration of TUNEL⁺ cranial cells in alcohol- and morpholino-treated embryos. Values are mean ± S.D. with 10–12 embryos per treatment. ^* Alcohol group differs from no-alcohol control within a treatment at P<0.05. ^† Morpholino-treated differs from its irrelevant-morpholino control at P<0.05. Abbreviation used: Alc, alcohol; C, control; MO, morpholino; y, yolk.