Sucar Sofia, Moore Ginger L, Ard Melissa E, Ring Brian C
Department of Biology, Valdosta State University, Georgia 31698 Agrobiotechnology Laboratory, National Institute for Agricultural Technology (INTA), Balcarce 7620, Buenos Aires, Argentina.
Department of Biology, Valdosta State University, Georgia 31698 Department of Neurosurgery, University of Florida, McKnight Brain Institute, Gainesville, Florida 32610.
G3 (Bethesda). 2016 Apr 7;6(4):1107-19. doi: 10.1534/g3.115.022475.
The mangrove killifish, Kryptolebias marmoratus, is unique among vertebrates due to its self-fertilizing mode of reproduction involving an ovotestis. As a result, it constitutes a simplistic and desirable vertebrate model for developmental genetics as it is easily maintained, reaches sexual maturity in about 100 days, and provides a manageable number of relatively clear embryos. After the establishment and characterization of an initial mutagenesis pilot screen using N-ethyl-N-nitrosourea, a three-generation genetic screen was performed to confirm zygotic mutant allele heritability and simultaneously score for homozygous recessive mutant sterile F2 fish. From a total of 307 F2 fish screened, 10 were found to be 1° males, 16 were sterile, 92 wild-type, and the remaining 189, carriers of zygotic recessive alleles. These carriers produced 25% progeny exhibiting several zygotic phenotypes similar to those previously described in zebrafish and in the aforementioned pilot screen, as expected. Interestingly, new phenotypes such as golden yolk, no trunk, and short tail were observed. The siblings of sterile F2 mutants were used to produce an F3 generation in order to confirm familial sterility. Out of the 284 F3 fish belonging to 10 previously identified sterile families, 12 were found to be 1° males, 69 were wild-type, 83 sterile, and 120 were classified as */+ (either wild-type or carriers) with undefined genotypes. This screen provides proof of principle that K. marmoratus is a powerful vertebrate model for developmental genetics and can be used to identify mutations affecting fertility.
红树鳉（Kryptolebias marmoratus）在脊椎动物中独一无二，因为它具有涉及卵睾的自体受精繁殖模式。因此，它构成了一个简单且理想的发育遗传学脊椎动物模型，因为它易于饲养，大约100天就能达到性成熟，并且能提供数量可控、相对清晰的胚胎。在用N-乙基-N-亚硝基脲建立并表征了初始诱变预筛选之后，进行了三代遗传筛选，以确认合子突变等位基因的遗传性，并同时对纯合隐性突变不育F2代鱼进行评分。在总共筛选的307条F2代鱼中，发现10条为初级雄鱼，16条不育，92条为野生型，其余189条为合子隐性等位基因的携带者。正如预期的那样，这些携带者产生了25%的后代，表现出几种与先前在斑马鱼和上述预筛选中描述的合子表型相似的表型。有趣的是，还观察到了新的表型，如金色卵黄、无躯干和短尾。不育F2代突变体的同胞被用来产生F3代，以确认家族性不育。在属于10个先前确定的不育家族的284条F3代鱼中，发现12条为初级雄鱼，69条为野生型，83条不育，120条被归类为*/+（野生型或携带者），基因型未定义。该筛选提供了原理证明，即红树鳉是发育遗传学的一个强大的脊椎动物模型，可用于识别影响生育力的突变。
