Evaluation Of Bull Semen Quality In Relation To Fertility Associated Proteins, Lipid Peroxidation And In Vitro Sperm Characters
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Date
2011
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Tamil Nadu Veterinary and Animal Sciences University
Abstract
Fresh semen samples were collected from 22 breeding bulls to screen for the
presence of fertility associated proteins. Seminal plasma proteins revealed 15
numbers of protein bands. The fertility related proteins such as 15/14 kDa and 28
kDa were present in all the 22 bulls (100 %), while 26 kDa protein was present in 18
bulls (81.82 %) and 55 kDa protein was present in 14 bulls (63.3 %). SDS- PAGE of
sperm membrane protein revealed a total of 14 protein bands. Proteins related with
bull fertility such as 15/14 kDa protein was observed in all the 22 bulls (100 %), 28
kDa protein was present in 21 bulls (95.45 %), 26 kDa protein was present in 14
bulls (63.64 %) and 55 kDa protein was present in only 11 bulls (50.00 %).
Heparin binding proteins of the seminal plasma revealed 7 protein bands, the
fertility related proteins such as 15/14 kDa protein was present in all the 22 bulls
(100 %) and 28 kDa protein was present in 18 bulls (81.82 %). 10 protein bands
were observed in the heparin binding proteins of the sperm membrane. 15/14 kDa
fertility related protein was present in all the bulls, while 28 kDa protein was present
in 11 bulls (50.00 %). Based on the presence of 28 kDa heparin binding protein insperm membrane, bulls in the present study were categorized into group I bulls
which were positive for the protein and group II bulls negative for the protein.
To study the effect of oxidative stress and role of fertility associated proteins
on in vitro sperm characters, frozen thawed semen of the bulls evaluated for their in
vitro sperm characters after treating with H 2 O 2 and with fertility associated proteins.
In group I bulls, frozen thawed semen samples treated with 25μg of fertility
associated protein had significantly (P < 0.01) lower level of MDA than the control
at 60 min ((1.86 ± 0.17 vs 2.67 ± 0.19), at 120 min (2.25 ± 0.19 vs 2.79 ± 0.24) and
at 180 min (2.81 ± 0.26 vs 3.77 ± 0.41).In group IIbulls, semen samples treated with
fertility associated protein also had significantly lower level of MDA than the
control at 60 min (2.03 ± 0.12 vs 3.04 ± 0.15), at 120 min (2.55 ± 0.13 vs 3.61 ±
0.28) and at 180 min (3.16 ± 0.14 vs 4.84 ± 0.46).In H 2 O 2 treatment, bulls in group I
had significantly (P < 0.05) lower MDA level than group II bulls at 30 min of
incubation (2.85± 0.16 vs 3.26 ± 0.12) and at 60 min of incubation (3.04 ± 0.16 vs
3.51 ± 0.12).
Bulls in group I when treated with fertility associated proteinhad
significantly more number of sperm cells with intact plasma membrane than the
control at 120 min (36.02 ± 2.10 vs 28.22 ± 3.10) and at 180 min (26.69 ± 2.06 vs
15.27 ± 2.11) of incubation. Similarly, bulls in group II when treated with fertility
associated protein had significantly more number of sperm cells with intact plasma
membrane than the control at 120 min (32.50 ± 3.03 vs 25.96 ± 3.32) and at 180 min
(23.36 ± 2.17 vs 15.06 ± 2.16) of incubation.In H 2 O 2 treated semen samples, group I
bulls had significantly (P < 0.01) more number of sperm cells with intact plasma
membrane than the group II bulls at 10 min (45.75 ± 3.44 vs 31.79 ± 3.34) and at 30
min(26.03 ± 2.53 vs 17.54 ± 1.59) post thaw incubation periods.
Bulls in group I and II did not differ with each other in the per cent of sperm
cells with HMMP at 10 min of incubation with H 2 O 2 (15.30 ± 0.92 vs 14.86 ± 0.77).
When the semen samples were treated with fertility associated proteins, bulls in
group I had significantly (P< 0.01) more per cent of sperm cells with HMMP than
the group II during different points of incubation; 60 min (17.51 ± 0.90 vs 13.57 ±0.85), at 120 min (14.54 ± 0.58 vs 9.58 ± 0.76) and at 180 min (10.54 ± 0.55 vs 7.49
± 0.89) incubation periods.
In H 2 O 2 treated samples, group I bulls had significantly (P < 0.05) more
number of DNAintact sperm cells than the group II bulls at 60 min of incubation
(94.01 ± 0.51 vs 92.26 ± 0.56). In H 2 O 2 treatment, group I bulls had significantly (P
< 0.01) less number of apoptotic cells (10.53 ± 1.35 vs 13.90 ± 1.52)than group II at
30 min of incubation. When the samples were treated with fertility associated
protein, group I bulls had significantly (P <0.05) less number of apoptotic cells than
group II during incubation.
Significant reductions in motility and velocity parameters were observed
during incubation in control as well as in fertility associated protein treated samples.
Treatment with fertility associated protein caused significant reductions in motility
parameters when compared to the untreated control.
In group Ibulls, the per cent of sperm cells with B pattern in heparin and
heparin with fertility associated proteins treated groups were significantly (P < 0.01)
higher than the control group at 60 min (62.37 ± 1.92, 63.24 ± 1.55 and 50.86 ±
1.82), at 120 min (69.79 ± 1.97, 69.78 ± 1.41 and 53.10 ± 1.76) and at 180 min post
thaw incubation (74.48 ± 1.98, 74.29 ± 1.33 and 54.79 ± 2.08). But B pattern cells in
heparin treated group and heparin with fertility associated proteins treated group did
not differ significantly at any point of incubation.
When the bulls were ranked by analyzing the in vitro sperm characters
during incubation from immediate post thaw to 180 min in the frozen thawed semen
samples by Duncan Multiple Range Test, bulls in the group I had 5 bulls each in
rank 1 and rank 2, while only one bull was categorized under rank 3. Group II bulls
had 4 bulls each in rank 1 and 2 and 3 bulls in rank 3.
Description
Keywords
Bull semen, fertility associated proteins, lipid peroxidation, sperm characters