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Original Research
9.
Liposome-mediated uptake of exogenous DNA by Sahiwal
cattle spermatozoa -
Thomas V. Babu, M. D. Pratheesh, N. Anandlaxmi ,
K. Amarnath
Vet World. 2012; 5(10): 621-627
doi:
10.5455/vetworld.2012.
621-627
Abstract
Aim:
To investigate the influence of lipofection
treatment and exogenous DNA uptake on the quality of
sahiwal cattle spermatozoa.
Materials and
Methods: Semen collected from sahiwal bulls
(n=7) were evaluated separately for color, volume,
mass activity, concentration, motility and viability
using standard procedures. Pooled sperm samples from
selected bulls (n=3) were transfected with a model
gene construct enhanced green fluorescent protein
(p-EGFP) via lipofection method and confirmed the
genome integration by PCR technique. Furthermore the
effect of transfection on spermatozoa was assessed
based on apotosis, viability and motility.
Results: In the current investigation sahiwal
bulls were selected based on their breeding records
and better semen characteristics. Although the
transfected sperm samples failed to show florescence
under fluorescence microscope, PCR studies confirmed
the successful uptake of the p-EGFP gene in to the
host sperm cell genome. Moreover transfected samples
showed a significant reduction in the viability and
motility without causing any DNA damage induced
apoptosis as demonstrated by DNA Ladder assay.
Keywords: EGFP, lipofection, Sahiwal cattle,
spermatozoa, transfection
