Detection of induced triploidy at different ages for larvae of the Japanese pearl oyster, Pinctada fucata martensii, by microfluorometry with DAPI staining

Uchimura, Yuushi; Komaru, Akira; Wada, Katsuhiko T.; Ieyama, Hiroshi; Yamaki, Masaru; Furuta, Hiroyuki

doi:10.1016/0044-8486(89)90246-9

Cited by 20 publications

(3 citation statements)

References 15 publications

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“…Further studies need to examine the ploidy of the spat to verify these results because some authors have reported that the percentage of Table 1 Percentages of triploids in 3-h old embryos for the two experiments (% EXP1) (%EXP2), and percentage of survival for each experiment (% SURl) (%SUR2) estimated 3 h after insemination and after 18 days of culture (% SURl8-1) (% SURl8-2) in the controls and in the different temperatures (T°C), times (in minutes) after insemination (TAI), and shock duration (SHD) treatments (in minutes). T°C 23 23 23 23 23 23 27 27 27 27 27 27 31 31 31 31 31 31 Control Control TAI 10 10 20 20 30 30 10 10 20 20 30 30 10 10 20 20 30 30 1 2 SHD 10 15 10 15 10 15 10 15 10 15 10 15 10 15 10 15 10 polyploid larvae is sometimes reduced during the larval stage in other bivalve species (Uchimura et al 1989). Further studies, including a combination of heat and caffeine (Yamamoto et al 1990) to induce triploidy in A. purpuratus are in progress.…”

Section: Discussionmentioning

confidence: 99%

Induction of triploid embryos by heat shock in the Chilean northern scallopArgopecten purpuratusLamarck, 1819

Toro

Sanhueza

Paredes

et al. 1995

New Zealand Journal of Marine and Freshwater Research

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Triploidy in embryos of the Chilean northern scallop, Argopecten purpuratus Lamarck, 1819, was induced with heat shock. Ploidy levels were assessed by chromosome counting after 3 h from embryos at the 8-16 cell stage of development. The efficiency of induction varied with the time of treatment post-insemination and the temperature of the shock. The duration of the shock showed a positive correlation with the percentage of triploids, the highest of which was obtained using 31°C for 15 min applied 10 min after insemination (66.7 %) but with low percentage of survival embryos after 3 h (26.4 %) and 18 days (11.2 %). The optimal procedure for triploidy induction (62.7 %) and survival after 3 h (62 %) and 18 days of culture (30.5 %) was the heat shock of 31°C for 10 min applied 10 min after fertilisation. We observed no significant difference in growth of shell length between controls and treated larvae after 18 days of culture.

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Section: Discussionmentioning

confidence: 99%

Induction of triploid embryos by heat shock in the Chilean northern scallopArgopecten purpuratusLamarck, 1819

Toro

Sanhueza

Paredes

et al. 1995

New Zealand Journal of Marine and Freshwater Research

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“…Moreover, this article proposes a method for use with embryos, larvae, juvenile and adult preparations which, when combined with image analysis, permits determination of the ploidy ratio of every sample at the earliest stage possible. This method gives an answer to the problem raised by Chalton & Allen (1985) and Uchimura et al (1989) concerning the difficulty of having enough cells at Gl stage to permit ploidy determination in embryos. Mitotic inhibition with aphidicolin allows us to obtain a clear gaussian decomposition of the cell population into diploid and tripioid subpopulations.…”

Section: Discussionmentioning

confidence: 99%

Image analysis: a new method for estimating triploidy in commercial bivalves

Gérard¹,

Naciri²,

Peignon³

et al. 1994

Aquaculture Res

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Abstract. For determination of ploidy levels in bivalves, three techniques are now routinely used: karyological determination, microfluorimetry and flow cytometry. This paper proposes an alternative technique which is based on determination of the optical density (OD) of stained nuclei using image analysis. A karyological and image analysis comparative experiment conducted on juveniles of Crassostrea gigas (Thunberg) and Ruditapes philippinarum (Adam & Reeve) is described and highlights the advantages of the image analysis technique. Initially developed on C. gigas and R. philippinarum, the image analysis method was easily extended to other bivalve species and to different developmental stages, as demonstrated by two experiments conducted on Ostrea edulis (L.). First, percentage of triploids was consistent from the D‐larval to the adult stage of development, despite the use of different preparation techniques depending on the stage examined. Second, a rise in ploidy percentage demonstrated a differential mortality between diploid and triploid animals. Image analysis techniques are cheaper than flow cytometry but more expensive than microfluorimetry. However, image analysis equipment can also be used for algae, eggs and spermatozoa counting, and other purposes such as fluorescence measurements, analysis of autoradiographs and electrophorcsis gels, and measurement of immuno‐histochemical reactions.

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“…In this sense, the fluorometry has been developed so much for static measurements or direct quantification in a single plane or dynamic measurements as in the case of the flow cytometry. In the case of static measurements, special microscopes have been developed to directly measure the fluorescence from cellular samples, with which it has been possible to determine ploidy levels of spermatozoa and polyploidy levels in polyploid organisms 7,8 . In reference to the flow cytometry, probably it is the technique with greater acceptance and diffusion as far as measurement of genomic size 9 .…”

Section: Introductionmentioning

confidence: 99%

Genome size estimation: a new methodology

et al. 2007

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Recently, within the cytogenetic analysis, the evolutionary relations implied in the content of nuclear DNA in plants and animals have received a great attention. The first detailed measurements of the nuclear DNA content were made in the early 40's, several years before Watson and Crick proposed the molecular structure of the DNA. In the following years Hewson Swift developed the concept of "C-value" in reference to the haploid phase of DNA in plants. Later Mirsky and Ris 1 carried out the first systematic study of genomic size in animals, including representatives of the five super classes of vertebrates as well as of some invertebrates. From these preliminary results it became evident that the DNA content varies enormously between the species and that this variation does not bear relation to the intuitive notion from the complexity of the organism. Later, this observation was reaffirmed in the following years as the studies increased on genomic size, thus denominating to this characteristic of the organisms like the "Paradox of the C-value". Few years later along with the no-codification discovery of DNA the paradox was solved, nevertheless, numerous questions remain until nowadays unfinished, taking to denominate this type of studies like the "C-value enigma". In this study, we reported a new method for genome size estimation by quantification of fluorescence fading. We measured the fluorescence intensity each 1600 milliseconds in DAPI-stained nuclei. The estimation of the area under the graph (integral fading) during fading period was related with the genome size.

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Detection of induced triploidy at different ages for larvae of the Japanese pearl oyster, Pinctada fucata martensii, by microfluorometry with DAPI staining

Cited by 20 publications

References 15 publications

Induction of triploid embryos by heat shock in the Chilean northern scallopArgopecten purpuratusLamarck, 1819

Induction of triploid embryos by heat shock in the Chilean northern scallopArgopecten purpuratusLamarck, 1819

Image analysis: a new method for estimating triploidy in commercial bivalves

Genome size estimation: a new methodology

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