Sex Reversal in Zebrafish fancl Mutants Is Caused by Tp53-Mediated Germ Cell Apoptosis

The molecular genetic mechanisms of sex determination are not known for most vertebrates, including zebrafish. We identified a mutation in the zebrafish fancl gene that causes homozygous mutants to develop as fertile males due to female-to-male sex reversal. Fancl is a member of the Fanconi Anemia/BRCA DNA repair pathway. Experiments showed that zebrafish fancl was expressed in developing germ cells in bipotential gonads at the critical time of sexual fate determination. Caspase-3 immunoassays revealed increased germ cell apoptosis in fancl mutants that compromised oocyte survival. In the absence of oocytes surviving through meiosis, somatic cells of mutant gonads did not maintain expression of the ovary gene cyp19a1a and did not down-regulate expression of the early testis gene amh; consequently, gonads masculinized and became testes. Remarkably, results showed that the introduction of a tp53 (p53) mutation into fancl mutants rescued the sex-reversal phenotype by reducing germ cell apoptosis and, thus, allowed fancl mutants to become fertile females. Our results show that Fancl function is not essential for spermatogonia and oogonia to become sperm or mature oocytes, but instead suggest that Fancl function is involved in the survival of developing oocytes through meiosis. This work reveals that Tp53-mediated germ cell apoptosis induces sex reversal after the mutation of a DNA–repair pathway gene by compromising the survival of oocytes and suggests the existence of an oocyte-derived signal that biases gonad fate towards the female developmental pathway and thereby controls zebrafish sex determination

Pragmatic skills predict online counterfactual comprehension:Evidence from the N400

Counterfactual thought allows people to consider alternative worlds they know to be false. Communicating these thoughts through language poses a social-communicative challenge because listeners typically expect a speaker to produce true utterances, but counterfactuals per definition convey information that is false. Listeners must therefore incorporate overt linguistic cues (subjunctive mood, such as in If I loved you then) in a rapid way to infer the intended counterfactual meaning. The present EEG study focused on the comprehension of such counterfactual antecedents and investigated if pragmatic ability—the ability to apply knowledge of the social-communicative use of language in daily life—predicts the online generation of counterfactual worlds. This yielded two novel findings: (1) Words that are consistent with factual knowledge incur a semantic processing cost, as reflected in larger N400 amplitude, in counterfactual antecedents compared to hypothetical antecedents (If sweets were/are made of sugar). We take this to suggest that counterfactuality is quickly incorporated during language comprehension and reduces online expectations based on factual knowledge. (2) Individual scores on the Autism Quotient Communication subscale modulated this effect, suggesting that individuals who are better at understanding the communicative intentions of other people are more likely to reduce knowledge-based expectations in counterfactuals. These results are the first demonstration of the real-time pragmatic processes involved in creating possible worlds

Karyotype variation in aminoethylcysteine resistant cell and callus cultures and regenerated plants of a dihaploid potato (Solanum tuberosum)

Karyotypic studies on cell suspensions and calli of an S-(2-aminoethyl)cysteine resistant cell line of the interdihaploid potato H2578 (2n=24) revealed a high degree of variation in the number of chromosomes (33–217) and dicentric chromosomes (0–8). The suspensions also exhibited megachromosomes and fused chromosomes. Differential staining showed that in suspensions dicentrics survived mitotic cycles mainly by parallel separation of the chromatids during anaphase and hardly by the breakage-fusion-bridge cycle. Two phenotypes of plantlets regenerated, each with 34 or 35 chromosomes with gross structural mutations and with the triploid amount of DNA. Chromosomal variation was related to the degree of tissue organizatio

Nuclear DNA amounts in European Callitriche species (Callitrichaceae)

Nuclear DNA amounts were determined for nine species, one subspecies and one hybrid of the European Callitrichaceae. The 2C DNA values ranged from 1 -82 pg to 8-30 pg due to variation in chromosome numbers (2n = 6-38) and loss (maximally 40-9%) or amplification (maximally 28-5%) of the DNA amount. The results are discussed in relation to phylogeny

Initial acytokinesis during leaf protoplast culture of dihaploid and tetraploid Solanum tuberosum and diploid S. bulbocastanum

Leaf protoplasts of dihaploid (2n=2x=24) and tetraploid (2n=4x=48) Solanum tuberosum, rr, and diploid S. bulbocastanum (2n=2x=24) were cultured in liquid medium. The cultures were studied for early karyological changes during their development. Giemsa staining of spread preparations revealed extremely low percentages of protoplasts developing into calli with the parental chromosome number, and high percentages of acytokinetic cells. The nuclear divisions within a cell were synchronous which allowed the occurrence of spindle interaction, resulting in nuclear poly- and aneuploidization. Although polyploidization was also found in uninucleate cells, a major increase in the formation of true-to-type calli would certainly be established by the improvement of early cross wall formation

Behaviour of chromosomes in potato leaf tissue cultured <i>in vitro</i> as studied by BrdC-Giemsa labelling

Cells of leaf explants of a monohaploid potato (Solanum tuberosum) were stimulated to mitosis on a medium with 5-bromodeoxycytidine during a period of 7 days. The cells cycled with mono- or diplochromosomes which showed differential staining of the sister chromatids and sister chromatid exchanges by the fluorescent plus Giemsa technique after two rounds of BrdC incorporation. Through the staining pattern the course of the first three cell cycles could be traced and the duration of the cycles estimated. Polyploidisation was enhanced by selective stimulation of polyploid cells and by endoreduplication of G2-phase cells. The percentage of polyploid mitoses increased from 10 to 7