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Rhodamine 123 as a cytoplasmic stain for mammalian zygotes
1Department of Animal Science North Carolina State University Raleigh, NC, 27695-7621 USA.
Abstract:
A mitochondrion-specific fluorescent dye, rhodamine 123, stains the cytoplasm of ova from mice, rabbits, sheep, cattle and pigs. Mouse zygotes stained with rhodamine 123 are often observed with areas of negative stain where the pronuclei are located. However, such areas of negative staining are not observed in zygotes from rabbits, sheep or cattle. In vitro viability of mouse zygotes stained with rhodamine 123 is high if observed with fluorescent microscopy for 5 min or less. Observation for more than 5 min results in a significant effect on embryo developmental potential in vitro.
Insights
Rhodamine 123, a fluorescent dye, stains ova cytoplasm in various mammals. Mouse zygotes show unique negative staining at pronuclei, impacting embryo development if observed for over five minutes.
Area of Science:
- Reproductive biology
- Cell biology
- Developmental biology
Background:
- Mitochondrion-specific fluorescent dyes are crucial for visualizing cellular structures.
- Rhodamine 123 is a widely used dye for mitochondrial staining in oocytes and zygotes.
- Differential staining patterns in zygotes across species may indicate variations in pronuclear development or dye interaction.
Purpose of the Study:
- To investigate the staining characteristics of rhodamine 123 in the cytoplasm of ova from different mammalian species.
- To examine the effect of rhodamine 123 staining on the in vitro viability and developmental potential of mouse zygotes.
Main Methods:
- Ova from mice, rabbits, sheep, cattle, and pigs were stained with rhodamine 123.
- Mouse zygotes were observed under fluorescent microscopy to note staining patterns, particularly around pronuclei.
- In vitro viability assays were conducted on stained mouse zygotes with varying observation times.
Main Results:
- Rhodamine 123 successfully stained the cytoplasm of ova across all tested species.
- Mouse zygotes exhibited distinct areas of negative staining at the pronuclei, unlike zygotes from rabbits, sheep, or cattle.
- In vitro viability of mouse zygotes remained high with observation times of 5 minutes or less, but was significantly reduced with longer observation periods.
Conclusions:
- Rhodamine 123 is a reliable stain for mammalian ova cytoplasm, with species-specific variations in zygote staining patterns.
- Prolonged observation of rhodamine 123-stained mouse zygotes negatively impacts their in vitro developmental potential, highlighting the importance of minimizing exposure time.
