Evaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryos

Title:
Evaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryos
Authors:
Cruz, Yolanda P.; Sutherland, Sharon A.; Sutherland, James D.
Abstract:
The enzyme horseradish peroxidase (HRP)is a widely used microinjectable cell marker for studying cell position, lineage, and migration in many kinds of animal embryos. Marked cells are easily identified because they darken when exposed to a chromophore and an HRP substrate such as hydrogen peroxide. This assay, however, requires cytochemical fixation. Thus, when HRP-marked cells need to be identified prior to fixation, visible co-injectants such as dyes and fluorescent substances have been used with HRP. Fluorescent substances have limitations because their excitation could be harmful to the marked cells. Visible but non-fluorescent co-injectants, however, would permit visualization of HRP-marked cells without inflicting such damage. We tested the compatibility of several histological dyes and electrolytic carriers with HRP iontophoresed as a cell marker in 2-cell mouse embryos. The dyes tested were Evans Blue, Cibacron Blue F3GA, Fast Green FCF, and Patent Blue Violet; the electrolytic carriers were KCI, K2SO4, CH3C02K, and KH2PO4. The combination found most useful was Patent Blue Violet in K2SO4. Survival of embryos incubated to the blastocyst stage following injection with HRP + Patent Blue Violet in K2SO4 at the 2-cell stage was significantly greater than that of embryos injected with any other dye. Although the proportion of embryos undergoing the 8-cell-to-morula transition was somewhat decreased by this treatment, the proportion of embryos reaching the blastocyst stage was comparable to that in the uninjected (control) group. Our results indicate that Patent Blue Violet is a useful, HRP-co-injectable dye for short-term cell marking in preimplantation mouse embryos.
Citation:
Cruz, Yolanda P., Sharon A. Sutherland, and James D. Sutherland. 1991. "Evaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryos." International Journal of Developmental Biology 35(1): 57-62.
Publisher:
University of the Basque Country Press
DATE ISSUED:
1991-03
Department:
Biology
Type:
Article
Additional Links:
http://web.a.ebscohost.com/ehost/detail/detail?vid=2&sid=c92df2b6-8917-4146-b50a-b11f8bde95f9%40sessionmgr4003&hid=4109&bdata=JnNpdGU9ZWhvc3QtbGl2ZSZzY29wZT1zaXRl#db=mnh&AN=1868001
PERMANENT LINK:
http://hdl.handle.net/11282/600485

Full metadata record

DC FieldValue Language
dc.contributor.authorCruz, Yolanda P.en
dc.contributor.authorSutherland, Sharon A.en
dc.contributor.authorSutherland, James D.en
dc.date.accessioned2016-03-02T14:54:54Zen
dc.date.available2016-03-02T14:54:54Zen
dc.date.issued1991-03en
dc.identifier.citationCruz, Yolanda P., Sharon A. Sutherland, and James D. Sutherland. 1991. "Evaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryos." International Journal of Developmental Biology 35(1): 57-62.en
dc.identifier.issn0214-6282en
dc.identifier.urihttp://hdl.handle.net/11282/600485en
dc.description.abstractThe enzyme horseradish peroxidase (HRP)is a widely used microinjectable cell marker for studying cell position, lineage, and migration in many kinds of animal embryos. Marked cells are easily identified because they darken when exposed to a chromophore and an HRP substrate such as hydrogen peroxide. This assay, however, requires cytochemical fixation. Thus, when HRP-marked cells need to be identified prior to fixation, visible co-injectants such as dyes and fluorescent substances have been used with HRP. Fluorescent substances have limitations because their excitation could be harmful to the marked cells. Visible but non-fluorescent co-injectants, however, would permit visualization of HRP-marked cells without inflicting such damage. We tested the compatibility of several histological dyes and electrolytic carriers with HRP iontophoresed as a cell marker in 2-cell mouse embryos. The dyes tested were Evans Blue, Cibacron Blue F3GA, Fast Green FCF, and Patent Blue Violet; the electrolytic carriers were KCI, K2SO4, CH3C02K, and KH2PO4. The combination found most useful was Patent Blue Violet in K2SO4. Survival of embryos incubated to the blastocyst stage following injection with HRP + Patent Blue Violet in K2SO4 at the 2-cell stage was significantly greater than that of embryos injected with any other dye. Although the proportion of embryos undergoing the 8-cell-to-morula transition was somewhat decreased by this treatment, the proportion of embryos reaching the blastocyst stage was comparable to that in the uninjected (control) group. Our results indicate that Patent Blue Violet is a useful, HRP-co-injectable dye for short-term cell marking in preimplantation mouse embryos.en
dc.language.isoen_USen
dc.publisherUniversity of the Basque Country Pressen
dc.relation.urlhttp://web.a.ebscohost.com/ehost/detail/detail?vid=2&sid=c92df2b6-8917-4146-b50a-b11f8bde95f9%40sessionmgr4003&hid=4109&bdata=JnNpdGU9ZWhvc3QtbGl2ZSZzY29wZT1zaXRl#db=mnh&AN=1868001en
dc.subject.departmentBiologyen_US
dc.titleEvaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryosen_US
dc.typeArticleen
dc.identifier.journalInternational Journal of Developmental Biologyen
dc.subject.keywordMouse embryosen_US
dc.subject.keywordMicroinjectionen_US
dc.subject.keywordHistological dyesen_US
dc.identifier.volume35en_US
dc.identifier.issue1en_US
dc.identifier.startpage57en_US
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