SEPARATION OF X AND Y-BEARING BUFFALO FROZEN SPERMATOZOA USING GRADIENT MEDIUM AND EVALUATION BY FLUORESCENCE IN-SITU HYBRIDIZATION

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RASTEGARNIA A.R.; AFSHANI M.; EFTEKHARI YAZDI P.

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Journal: Journal of Veterinary Research Year:2011 | Volume:67 | Issue:1 Page(s): 35-42

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Title

SEPARATION OF X AND Y-BEARING BUFFALO FROZEN SPERMATOZOA USING GRADIENT MEDIUM AND EVALUATION BY FLUORESCENCE IN-SITU HYBRIDIZATION

Author(s)

RASTEGARNIA A.R. | AFSHANI M. | EFTEKHARI YAZDI P. | Issue Writer Certificate

Keywords

SPERMQ2

SEX PRESELECTIONQ3

ALLGRAD GRADIENTQ2

BUFFALOQ1

Abstract

BACKGROUNDS: SPERM sexing as one of the important ways for pre-selection of offspring along with artificial insemination, has the potential to considerably improve animal breeding and the efficiency of dairy and meat production. The discrepancy between X- and Y- chromosome bearing SPERM is the basis of this procedure.OBJECTIVES:The aim of this study was to evaluate a discontinuous SPERM isolation medium ALLGRAD GRADIENT in separation of BUFFALO SPERMatozoa according to sex chromosomes by fluorescence in-situ hybridation (FISH).METHODS: Afourlayer discontinuous ALLGRAD GRADIENT, 65%-95% with 10% differences between each subsequent layer were prepared in gradient separation for post thaw X- and Y-bearing SPERMatozoa. Following centrifugation, SPERM from the top and bottom fraction were aspirated and fixed on a slide. The FISH procedure was performed by using X and Ybovine specific DNAprobes. At least 400 SPERMatozoa were scored for each sample. The proportions of X- and Y-bearing SPERMatozoa were determined by the presence of red or green fluorescent signals.RESULTS: There was no significant difference in the percentage of SPERMatozoa with the specific signals of X and Y chromosomes in the control groups. After separation, in treatment groups, the percentage of Xbearing SPERMatozoa in the bottom layer exceeded that of the one in the top layer (78.3 vs. 21.7) (p<0.05). Adversely, in all treatment groups the significant difference between the frequencies of Y-bearing SPERMatozoa in the top layers was evident (75.5vs.24.5) (p<0.05).CONCLUSIONS: This study showed that using four-step discontinuous gradient by Allgrad isolation medium was a reliable method for the separation of X- and Y-bearing frozen-thawed BUFFALO SPERMatozoa and can be more expedient for IVF in BUFFALO.

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APA: Copy

RASTEGARNIA, A.R., AFSHANI, M., & EFTEKHARI YAZDI, P.. (2011). SEPARATION OF X AND Y-BEARING BUFFALO FROZEN SPERMATOZOA USING GRADIENT MEDIUM AND EVALUATION BY FLUORESCENCE IN-SITU HYBRIDIZATION. JOURNAL OF VETERINARY RESEARCH, 67(1), 35-42. SID. https://sid.ir/paper/35284/en

Vancouver: Copy

RASTEGARNIA A.R., AFSHANI M., EFTEKHARI YAZDI P.. SEPARATION OF X AND Y-BEARING BUFFALO FROZEN SPERMATOZOA USING GRADIENT MEDIUM AND EVALUATION BY FLUORESCENCE IN-SITU HYBRIDIZATION. JOURNAL OF VETERINARY RESEARCH[Internet]. 2011;67(1):35-42. Available from: https://sid.ir/paper/35284/en

IEEE: Copy

A.R. RASTEGARNIA, M. AFSHANI, and P. EFTEKHARI YAZDI, “SEPARATION OF X AND Y-BEARING BUFFALO FROZEN SPERMATOZOA USING GRADIENT MEDIUM AND EVALUATION BY FLUORESCENCE IN-SITU HYBRIDIZATION,” JOURNAL OF VETERINARY RESEARCH, vol. 67, no. 1, pp. 35–42, 2011, [Online]. Available: https://sid.ir/paper/35284/en

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