The light-dependent daily cycle of ovulation in the oviparous medaka fish, Oryzias latipes (Atherinomorpha: Beloniformes: Adrianichthyidae) artificially pregnant with developing embryos.

In the oviparous medaka fish, Oryzias latipes, mature spermatozoa that were artificially introduced into the ovarian cavity retaining ovulated eggs could internally fertilize these eggs. This enabled us to examine the effect of ovarian gestation on the ovulation cycle. Most freshly ovulated eggs could be internally fertilized in the ovarian cavity. Yet eggs ovulated 24 h after single insemination remained unfertilized in the ovarian cavity. Artificially pregnant females persisted in a daily cycle of ovulation, which occurred shortly before the onset of light under the present reproductive conditions. Females continuously ovulated a certain number of eggs despite ovarian gestation, that is, the presence of embryos within the ovarian cavity. Repeated cycles of ovulation led to crowding in the ovarian cavity because the group of fertilized eggs, with their hardened egg envelope (chorion or zona radiata), plugged the genital orifice. The development of fertilized eggs was retarded and ceased around the initiation stage of blood circulation, but when they were transferred from the ovarian cavity into regular saline, they regained their ability to develop normally up to hatching. These results show that in oviparous female medaka, ovarian gestation exerted little effect on the time of ovulation and the number of ovulated eggs.

Calcium signaling during the early development of medaka and zebrafish.

The ex-utero fertilization and development of the optically clear embryos of teleost fish have long been favorites of developmental biologists. They have, therefore, provided considerable insight with regards to our understanding of embryonic pattern formation and the early development of vertebrates. These attributes have also been most helpful in the visualization of Ca(2+) signaling events that have been reported to accompany many of the fundamental steps and processes that constitute early embryonic development. These include egg activation; segregation of the cytoplasm from the yolk; cytokinesis; axis determination; cellular rearrangement and germ layer establishment; as well as the formation of the first tissue domains. The developing eggs and embryos of medaka (Oryzias latipes) and zebrafish (Danio rerio) have for many decades been a favorite choice of investigators attempting to visualize Ca(2+) signaling events. In this short review, we have attempted to catalog and present a comparative study of the developmental Ca(2+) signals recorded in these most amenable of vertebrate models.

ACETYLCHOLINE AND ACETYLCHOLINESTERASE ACTIVITY IN EARLY EMBRYOS OF THE MEDAKA ORYZIAS LATIPES, A TELEOST.

Acetylcholinesterase (AChE) activity is present in homogenates of medaka embryos during cleavage and epiboly. The levels of AChE activity change little during this period of development and are similar in embryos grown at either 15°C or 25°C. The specific activity of AChE in cells isolated from blastulae is 0.06 mmoles substrate hydrolyzed/min/g protein, a value comparable to that of chicken myoblasts and myotubes in vitro. Acetylcholinesterase activity was detected cytochemically in all cells dissociated from blastulae and gastrulae. In deep blastomeres AChE activity is present nearly throughout the cytoplasm; it is absent from peripheral regions of the cytoplasm which are involved in circus, or limnicolor, movements. Acetylcholine-like activity in extracts of the embryos was assayed using the clam heart ventricle bioassay. The active principle, which caused a decrease in both the frequency and magnitude of ventricular contractions, was inactivated by heating at pH 10 and by incubation at pH 7.0 with commercial AChE. The effect of the active principle on the clam heart was blocked by mytolon chloride, a drug which specifically blocks the effect of acetylcholine on the clam heart. The active principle migrated on Whatman #1 chromatography paper with the same Rf as authentic acetylcholine in three solvent systems. The amount of acetylcholine in blastulae is about 4 picomoles/embryo.