Alkaline phosphatase expression in ascidian egg fragments and andromerogons

Alkaline phosphatase (AP) activity is expressed in the endodermal cell lineage of ascidian embryos beginning at gastrulation. AP expression is resistant to levels of actinomycin D which completely suppress the appearance of other tissue-specific enzyme and morphological markers including acetylcholinesterase (AchE), a larval muscle enzyme whose expression requires embryonic transcription. The resistance of AP expression to actinomycin D has led to the proposal that AP may be expressed independent of embryonic transcription by the translational activation of maternal AP mRNA. To test this hypothesis, AP expression was examined in fragments of unfertilized and fertilized Styela plicata eggs by histochemical methods. As expected, nucleate fragments from fertilized eggs developed into larvae which exhibited AP activity in their endodermal cells and AchE activity in their muscle cells. In contrast, anucleate fragments from fertilized eggs, cultured until controls reached the larval stage, did not develop AP or AchE activity. The lack of AP activity was unrelated to the absence of cleavage or to the ooplasmic composition of the anucleate fragments. Anucleate fragments from unfertilized eggs were also AP negative, unless they were inseminated, after which they often developed to the larval stage as andromerogons and exhibited AP activity in their endodermal cells. The development of endodermal AP in andromerogons suggests that the factors responsible for AP expression are not localized in or attached to the maternal nucleus. In summary, the results suggest that AP expression requires nuclear events and is not determined exclusively by maternal cytoplasmic factors such as preformed mRNA.