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Background: Seminal plasma of mammals is a complex fluid, which serves as a carrier for the spermatozoa on their journey from the male testes to their target, the oocyte. Seminal plasma contains a variety of biochemical components, some
of which are relatively specific for the regulation of sperm function. The objective of this study was to investigate enzyme
activities aspartate-amino-transferase (AST), alkaline phosphatase (ALP), lactate-dehydrogenase (LDH) and concentrations of macroelements[ sodium (Na+) and potassium (K+) and calcium (Ca2+) ] in seminal plasma. Moreover the present
study was performed to investigate proteins in ram seminal plasma and the correlation between specific proteins and semen
characteristics in cross breed ram.
Materials, Methods & Results: Sixteen crossbred fertile rams (four Baluchi Moghani, four Ghezel Baluchi, four
Ghezel Merino, and four Merino Moghani) were used in this study. Three ejaculates of each animal were collected
during the breeding season and examined spermatozoa with light microscopy for motility, concentration, for dead sperm
and morphology. Seminal plasma was harvested by centrifugation and then subjected to analysis enzyme, mineral composition and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Total protein, AST, ALP, LDH, Na+, K+ and calcium Ca2+
were measured. Spearmans correlation coefficient test was applied to examine the correlation between seminal plasma
protein fractions with all the parameters of the semen. Differences were considered to be statistically significant at P < 0.05.
The highest seminal plasma concentration of Na+, K+ and Ca2+ was recorded in Ghezel Merinos rams. The result showed
that there was a significant negative correlation between Na+ and K+ but small correlation was recorded between Na+ and
Ca2+. The highest plasma concentration of ALP was recorded in Baluchi Moghani ram and the lowest was recorded N
in
Merino Moghani ram (8 1.25 and 3.33 0.01 U/mL, respectively). The highest plasma concentration of LDH and
AST was recorded in Ghezel Baluchi ram (4.68 0.19 U/mL, 19 4 U/mL respectively). Significant correlations were
found between LDH levels and sperm viability. The results obtained indicate that ram seminal plasma protein profile is
characterized by many protein bands with molecular weights, ranging from 8.81 to 95.82 kDa. Five proteins in this study
had quantitative difference between seminal plasma from crossbred rams.
Discussion: The results showed that the relative content of protein bands wasnt significantly different by semen characteristics. Thus this study indicates that the relative content of seminal plasma proteins could not be an essential index
to evaluate ram semen quality. Also increase in the percentage of live and normal sperm corresponded to an increase in
LDH activity in the seminal fluid. It has been stated that LDH plays an important metabolic role in sperm capacitation and
fertilization. This study suggest that the cations Na+ and K+ generally establish the osmotic balance, and seminal plasma
osmolality ultimately plays an important role in the activation sperm cell also potassium ions (K+) are also intracellular
cations, and the concentrations in the seminal fluid may be an indication of sperm plasma membrane integrity. Thus some
enzyme and mineral composition could be an essential index to evaluate cross breed ram semen quality.
Keywords: minerals, rams, SDS-PAGE, seminal plasma, semen quality.
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Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz CORRESPONDENCE: R. Asadpour [r_asadpour@tabrizu.ac.ir Fax: +98 (411) 3357834]. Faculty of Veterinary Medicine, University of Tabriz. PK: 51666-16471, Tabriz, Iran.
R. Asadpour. 2012. Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds.
Acta Scientiae Veterinariae. 40(2): 1027.
INTRODUCTION
The seminal plasma was separated immediately after collection. Fresh semen was centrifuged at
1500 g for 15 min at 5C. The supernatants were
transferred into 1.5 mL eppenddorf of tubes and recentrifuged at 14,000 g for 10 min at 5C to eliminate
the remaining sperm. Seminal plasma was stored at
-20C until used.
R. Asadpour. 2012. Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds.
Acta Scientiae Veterinariae. 40(2): 1027.
Statistical analysis
Seminal plasma compositions were determined according to protocols presented in previous studies: Glucose [26], total protein [27], AST [32], ALP
[23], LDH [2]. Na+, K+ and Ca2+ were also measured
by Micro light1 based on ion-exchanger colorimetric
method [37].
Data analysis was performed using SPSS software program (version 16.0 for Windows). All values
were expressed as Mean standard error of mean
(S.E.M.). Spearmans correlation coefficient test was
applied to examine the correlation between seminal
plasma protein fractions with all the parameters of the
semen. Differences were considered to be statistically
significant at P < 0.05.
RESULTS
Image acquisition
Gel images were analysed to determine molecular weight and relative protein content using the
Total Lab TL120 computer program (v2009, USA).
Table 1. Semen characteristics in fresh ejaculates of cross breed rams with different fertility patterns.
R. Asadpour. 2012. Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds.
Acta Scientiae Veterinariae. 40(2): 1027.
Table 2. Least square means of concentrations of Na+, K+, Ca2+, Alcalin posphatase (ALP), Lactate dehydrogenase (LDH), Aspartate
aminotranferase (AST) and Total protein (TP) in the seminal plasma of cross breed rams.
Different superscript letters in the rows indicate significant differences P < 0.05.
a,b
Figure1. Molecular weight (kDa) of proteins of the seminal plasma (SP), the total
protein fraction (TPF), as revealed by SDSPAGE gel electrophoresis. Gels were
stained with Coomassie Brilliant Blue. Numbers indicate the molecular weight of the
standard in kDa. Lane 1( Merinos Moghani) Lane 2(Merinos Moghani) Lane
3 ( Baluchi Moghani) Lane 4( Baluchi Moghani) Lane 5 (Ghezel Baluchi)
Lane 6 ( Merinous Moghani ) Lane 7(Ghezel Merinous) Lane 8 (Baluchi
Moghani) Lane 9 ( Meronos Moghani Lane 10( Marker).
DISCUSSION
R. Asadpour. 2012. Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds.
Acta Scientiae Veterinariae. 40(2): 1027.
R. Asadpour. 2012. Relationship between Mineral Composition of Seminal Plasma and Semen Quality in Various Ram Breeds.
Acta Scientiae Veterinariae. 40(2): 1027.
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