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Sexing murine embryos with an indirect immunofluorescence assay using phage antibody B9-Fab against SDM antigen.

Wang N, Yuan A, Ma J, Deng Z, Xue L - J. Vet. Med. Sci. (2015)

Bottom Line: In the present study, novel anti-SDM B9-Fab recombinant clones (obtained by chain shuffling of an A8 original clone) were used to detect SDM antigens on murine embryos.Overall, 43.5% (60/138) were classified as presumptive males and 56.5% (78/138) as presumptive females, with 85.0 and 88.5% of these, respectively, confirmed as correct predictions (based on PCR analysis of a male-specific [Sry] sequence).We concluded that the anti-SDM B9-Fab molecule had potential for non-invasive, technically simple immunological sexing of mammalian embryos.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, PR China.

ABSTRACT
The use of serologically detectable male (SDM; also called H-Y) antigens to identify male embryos may be limited by the source of anti-SDM antibody. In the present study, novel anti-SDM B9-Fab recombinant clones (obtained by chain shuffling of an A8 original clone) were used to detect SDM antigens on murine embryos. Murine morulae and blastocysts (n=138) were flushed from the oviducts of Kunming mice and incubated with anti-SDM B9-Fab for 30 min at 37°C. With an indirect immunofluorescence assay, the membrane and inner cell mass had bright green fluorescence (presumptive males). Overall, 43.5% (60/138) were classified as presumptive males and 56.5% (78/138) as presumptive females, with 85.0 and 88.5% of these, respectively, confirmed as correct predictions (based on PCR analysis of a male-specific [Sry] sequence). We concluded that the anti-SDM B9-Fab molecule had potential for non-invasive, technically simple immunological sexing of mammalian embryos.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescent detection of SDM antigen on murine embryos by B9-Fab. Fluorescenceand optical images of 8-cell embryos (A and B), morulae (D and E) and expandedblastocysts (G and H) after being detected with an indirect immunofluorescence assayinduced by B9-Fab (classified as males). Also, note the optical image of an 8-cellembryo (C), morula (F) and expanded blastocyst (I) before treatment with B9-Fab.Fluorescence and optical images of morulae (J and K), and early blastocysts (M and N)after being detected in an indirect immunofluorescence assay induced by B9-Fab(classified as females). Optical images of a morula (L) and an early blastocyst (O)before treatment with B9-Fab.
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fig_001: Immunofluorescent detection of SDM antigen on murine embryos by B9-Fab. Fluorescenceand optical images of 8-cell embryos (A and B), morulae (D and E) and expandedblastocysts (G and H) after being detected with an indirect immunofluorescence assayinduced by B9-Fab (classified as males). Also, note the optical image of an 8-cellembryo (C), morula (F) and expanded blastocyst (I) before treatment with B9-Fab.Fluorescence and optical images of morulae (J and K), and early blastocysts (M and N)after being detected in an indirect immunofluorescence assay induced by B9-Fab(classified as females). Optical images of a morula (L) and an early blastocyst (O)before treatment with B9-Fab.

Mentions: The membrane and inner cell mass of morulae and blastocysts (presumptive males) had brightgreen fluorescence (Fig. 1Fig. 1.


Sexing murine embryos with an indirect immunofluorescence assay using phage antibody B9-Fab against SDM antigen.

Wang N, Yuan A, Ma J, Deng Z, Xue L - J. Vet. Med. Sci. (2015)

Immunofluorescent detection of SDM antigen on murine embryos by B9-Fab. Fluorescenceand optical images of 8-cell embryos (A and B), morulae (D and E) and expandedblastocysts (G and H) after being detected with an indirect immunofluorescence assayinduced by B9-Fab (classified as males). Also, note the optical image of an 8-cellembryo (C), morula (F) and expanded blastocyst (I) before treatment with B9-Fab.Fluorescence and optical images of morulae (J and K), and early blastocysts (M and N)after being detected in an indirect immunofluorescence assay induced by B9-Fab(classified as females). Optical images of a morula (L) and an early blastocyst (O)before treatment with B9-Fab.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488409&req=5

fig_001: Immunofluorescent detection of SDM antigen on murine embryos by B9-Fab. Fluorescenceand optical images of 8-cell embryos (A and B), morulae (D and E) and expandedblastocysts (G and H) after being detected with an indirect immunofluorescence assayinduced by B9-Fab (classified as males). Also, note the optical image of an 8-cellembryo (C), morula (F) and expanded blastocyst (I) before treatment with B9-Fab.Fluorescence and optical images of morulae (J and K), and early blastocysts (M and N)after being detected in an indirect immunofluorescence assay induced by B9-Fab(classified as females). Optical images of a morula (L) and an early blastocyst (O)before treatment with B9-Fab.
Mentions: The membrane and inner cell mass of morulae and blastocysts (presumptive males) had brightgreen fluorescence (Fig. 1Fig. 1.

Bottom Line: In the present study, novel anti-SDM B9-Fab recombinant clones (obtained by chain shuffling of an A8 original clone) were used to detect SDM antigens on murine embryos.Overall, 43.5% (60/138) were classified as presumptive males and 56.5% (78/138) as presumptive females, with 85.0 and 88.5% of these, respectively, confirmed as correct predictions (based on PCR analysis of a male-specific [Sry] sequence).We concluded that the anti-SDM B9-Fab molecule had potential for non-invasive, technically simple immunological sexing of mammalian embryos.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, PR China.

ABSTRACT
The use of serologically detectable male (SDM; also called H-Y) antigens to identify male embryos may be limited by the source of anti-SDM antibody. In the present study, novel anti-SDM B9-Fab recombinant clones (obtained by chain shuffling of an A8 original clone) were used to detect SDM antigens on murine embryos. Murine morulae and blastocysts (n=138) were flushed from the oviducts of Kunming mice and incubated with anti-SDM B9-Fab for 30 min at 37°C. With an indirect immunofluorescence assay, the membrane and inner cell mass had bright green fluorescence (presumptive males). Overall, 43.5% (60/138) were classified as presumptive males and 56.5% (78/138) as presumptive females, with 85.0 and 88.5% of these, respectively, confirmed as correct predictions (based on PCR analysis of a male-specific [Sry] sequence). We concluded that the anti-SDM B9-Fab molecule had potential for non-invasive, technically simple immunological sexing of mammalian embryos.

No MeSH data available.


Related in: MedlinePlus