Abstract
In vitro gonad culture systems have proven useful to investigate intrinsic mechanisms of sexual reproduction in animals. Here we describe development of an in vitro culture method for coral ovaries. Mesenterial tissues containing both ovaries and mesenterial filaments were microscopically isolated from the scleractinian coral, Fimbriaphyllia ancora, and culture conditions were optimized. M199 diluted 10× (10% M199, pH 8.1) and supplemented with 25 mM HEPES and the antibiotics, ampicillin, penicillin and streptomycin, supported oocyte survival and maintained the structural integrity of ovaries during short-term culture (~ 6 days). Addition of a commercial antibiotic–antimycotic solution (Anti–Anti) and fetal bovine serum adversely affected ovary maintenance and caused tissue disintegration. Characterization of cultured ovaries showed that there is no difference in cell proliferation of ovarian somatic cells between culture Days 1 and 6. Moreover, the presence of oogonia and expression of a major yolk protein, vitellogenin, were confirmed in ovaries cultured for 6 days. This system will be useful for studying effects of a wide range of substances on coral oogenesis.
Highlights
Sexual reproduction of scleractinian corals is a biological phenomenon that continues to fascinate researchers and the general public
Our recent transcriptome analysis of coral gonads showed that genes encoding growth factors, neuropeptides, and neurotransmitter receptors are expressed in the gonads[20], leading us to hypothesize that coral gametogenesis is regulated by those intrinsic factors
When tissues were maintained in filtered sterile natural seawater (FSW) for 6 days, contours of some oocytes, observed with red fluorescent protein (RFP), became unclear (Fig. 1D,E)
Summary
Sexual reproduction of scleractinian corals is a biological phenomenon that continues to fascinate researchers and the general public. Complex organs corresponding to vertebrate brains, blood vessels, and intestines are lacking in c orals[5,6] Coral germ cells generally develop within specific regions of mesenterial tissues in the p olyp[7] These regions are usually small swellings, conventionally called gonads. Our recent transcriptome analysis of coral gonads showed that genes encoding growth factors, neuropeptides, and neurotransmitter receptors are expressed in the gonads[20], leading us to hypothesize that coral gametogenesis is regulated by those intrinsic factors. To test this hypothesis, functional analysis of genes and proteins is essential. In fish, the mechanism of hormonal regulation during gametogenesis has been
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