PHYSIOLOGICAL AND MOLECULAR CHARACTERIZATION OF SOME EGYPTIAN SHEEP BREEDS

PHYSIOLOGICAL AND MOLECULAR CHARACTERIZATION OF SOME

EGYPTIAN SHEEP BREEDS

Journal

Kayali1 I. M., M. A. Rashed 1, M. R. Anous 2, M. A. A. Sallam1 And A. H. Atta1

J. Biol. Chem. Environ. Sci., 2014, Vol. 9 (1): 121-140 www.acepsag.org

1. Dept. of Genetics, 2. Dept of Animal Production, Fac of Agric. Ain Shams

University,cairo,Egypt.

ABSTRACT

Five sheep breeds (Ossimi, Rahmani, Barki, Awassi and Finnish) were used to Physiological screening which revealed that the parameters for fresh semen evaluation were higher in the Ossimi and Finnish rams and low in the Barki and Awassi rams, while Rahmani rams had moderate values. Biochemical analysis for blood plasma proteins succeeded in distinguishing among the tested rams and detecting genetic variation within each breed, but it was not enough to determine the fertility among the rams. While, seminal plasma proteins pattern succeeded to detecting some positive and negative markers to recognized high and low fertility rams of studied breeds. Acr gene, exon5 isolated from sheep rams by two specific primers, according to acrosin gene sequence (Acr) of Ovis canadensis sheep breed. Obtained sequences for Acr gene, exon 5 in the five sheep breeds were compared with each other using BioEdit program. Sequences in Awassi, Barki and Rahmani breeds were identity and were different in Ossimi and Finnish breeds. Eight different nucleotides presented in Finnish breed, while there were five different nucleotides in Ossimi breed. This difference may play important roles in fertility of these rams where Finnish breed have the highest fertility rate. The highest similarity values (1.00) were among Awassi, Barki, and Rahmani breeds, followed by Finnish and Ossimi breeds. While the lowest values (0.98) were between each of Awassi, Barki or Rahmani breeds with Finnish or Ossimi breeds. However, Finnish and Ossimi were closer to each other which in agreement with the semen evaluation. Key words: Rams, Semen, Fertility, Acrosome, PCR, Sequence.


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INTRODUCTION One important element for selection of breeding rams for either

natural mating or artificial insemination relies on semen quality evaluation as reported by Rodríguez-Martínez, (2003). Studied semen characteristics such as semen volume, pH, mean mass activity, initial sperm motility, live sperm count, sperm concentration, total number of sperm in the ejaculate, and overall sperm length was performed by Bhattacharyya et al. (2009) and Davendra et al. (2010). The poorest quality semen collecte during summer with the highest incidence of abnormality. The semen of Saanen and Nubian bucks were characterized by Kamal et al. (2005) who investigated the effect of climate and seasons on the mating behavior and semen characteristics.

Many studies have been carried out on sheep blood protein banding patterns using polyacrylamide gel electrophoresis (SDS-PAGE) techniques Missohou et al., (1999); Tapio et al., (2002); Lovenko, (2002). Wafaa et al. ( 2000) used protein banding pattern variations in the main local Egyptian sheep breeds (Rahmani, Ossimi and Barki) and considered SDS-PAGE as an useful tool for the detection of homogeneity within each breed and generation of specific biochemical markers to identify and differentiate between the three breeds.

In the recent years, several seminal plasma proteins have been identified, isolated and characterized by using SDS-PAGE analysis. Evidence suggests that the protein composition of seminal plasma is varied among species and that some seminal plasma proteins are associated with fertility in various species Wenbin et al., (2009). Ghosh et al. (2008) characterized a 55 kDa protein in Garole sheep (Ovis aries) seminal plasma previously shown to be prevalent in bulls and swine of high fertility. Seminal plasma was partially purified by gel filtration chromatography. 10% SDS-PAGE analysis for their polypeptide profile, revealed a prominent band of 55 kDa including two other minor bands of 69 kDa and 16 kDa.

The acrosome is an unique vesicle surrounding the anterior part of the spermatozoa head and contains a number of hydrolytic enzymes, among which serine proteinase acrosin is the most studied acrosomal enzyme Chen et al., (1991). Acrosin is considered to play an essential role in fertilization, recognition, binding and penetration

J. Biol. Chem. Environ. Sci., 2014, 9 (1), 121-140

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of the zona pellucida of the ovum Klemm et al., (1991). Acrosin is synthesized and stored in the spermacrosome as proacrosin, an inactive zymogene form. During the acrosome reaction, proacrosin is converted into mature acrosin by two part activation process: the liberation of proenzyme segments from C-terminus and cleavage of a peptide bond producing the light and heavy chains Baba et al., (1989). However, penetration of the zona pellucida in acrosin-deficient mice was significantly delayed, and there is ample evidence that acrosin is released during the acrosome reaction, binds the zona pellucida, and is critical for dispersion of acrosomal contents Furlong et al. (2005); Tranter et al. (2000); Urch and Patel (1991).

A previous comparative analysis of Acr sequences from10 mammal species revealed some evidence for adaptive change in this molecule, but P-values were marginal in comparisons of neutral models to selection models Swanson et al., (2003). Furthermore, large divergences between many of the sequences prevented comparison of the most rapidly evolving segment of Acr, exon 5 Adham et al. (1996); Zahn et al. (2002).

This region encodes the carboxy terminus, a proline-rich domain that is not present in other serine proteases and is cleaved during maturation of acrosin Adham et al. (1996); Baba et al. (1989). The function of this region has been debated. Some studies suggest a role in secondary binding of sperm to the zona pellucida Furlong et al. (2005); Mori et al. (1995); Tranter et al. (2000), John and Willie (2007) Comparison of Acr sequences to mitochondrial genes and a nuclear intron show that reproductive genes can be informative phylogenetic markers. In bovid artiodactyls (cattle, sheep, goats, and antelopes), exon 5 of Acr is highly variable, is characterized by a low level of homoplasy, and has a fairly even spread of substitutions across the 3 codon positions. Watanabe et al. (1991) isolated and characterized the genomic region carrying the mouse acrosin gene, including the 5'-flanking sequence; this acrosin gene consists of five exons separated by four introns. Organization of this gene is very similar to those of the genes for other typical serine proteases, except for the phase class of the first intron. However, this study was carried out to characterization of semen’s rams, isolation and characterization of acrosin gene (exon5) in five sheep breeds (Ossimi, Rahmani, Barki, Awassi and Finnish).


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MATERIALS AND METHODS Materials:

The sheep breeds used in the present study was assembled at 2010 from Sakha and Bourg El-Arab Experimental Stations, belonging to Animal Production Research Institute, Agriculture Research Center (ARC). It consisted of 8, 8, 7, 6 and 6 rams at same ages from Barki, Awassi, Finnish, Rahmani and Ossimi breeds, respectively. Awassi, Finnish, Rahmani and Ossimi breeds are recognized by high fertility compare with Barki breed. Semen samples collection:

The fresh semen of each ram was early collected in the morning with the aid of an artificial vagina with inside temperature of 38°C for the 30 sampled animals (six animals from each studied breed) in September month. Blood sample collection:

Blood samples were collected from the jugular using vacutainer glass tubes which containing disodium ethylene diamine tetra acetic acid EDTA (EDTA-Na2) as anticoagulant reagent. Blood plasma was obtained by centrifugation of blood samples at 10000 rpm for 10 minutes at 4°C. Plasma protein (supernatant) was transferred to clean plastic vials and stored at -20°C. The pellet of white cells was immediately stored at -20°C for DNA extraction. Methods:

The following physical characteristics of the fresh semen samples were determined to evaluate their quality. Such as volume, PH value, color, and density were performed. Mass motility:

The wave motion of semen was observed with low power lens (10x), and was graded from 0 to 5 scores according to Chemineau et al. (1991). Progressive motility:

Semen diluted with drops of a 2.9% of sodium citrate solution. Assessments were made with high power lens (40x) using a scale graded from 0 to 5 scores according to Chemineau et al. (1991).


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Percentage of live, dead and abnormality sperms: Using eosin-nigrosin stain, dead sperm were colored by the dye

(pink-red), while live spermatozoa were not stained. Randomly, 100 cells were counted and the recorded number of stained dead sperms could be determined the percentage of dead spermatozoa Evan and Maxwell, (1987). Abnormalities can occur in the head, neck, mid-piece, tail, or any combination of these parts of the sperm cell Salisbury et al. (1978). Sperm concentration:

Sperm cell concentration per ejaculate was calculated with the aid of a haemocytometer according to Loskutoff and Crichton (2001). Percentage of unreacted acrosomes:

The semen were diluted with drops of 2.9% sodium citrate solution on a clean warmed microscope slide to make a smear, then rapidly dried at 37 ºC. The smear was fixed by using immersion in a 5% buffered formal saline (4.5 g sodium chloride + 100 ml formaldehyde + 900 ml water) for 15 minutes. Washed was by added in running tap-water for 15 minutes. The smear was dried and immersion in the buffered Giemsa solution for 90 minutes. The slide were rinsed briefly in a distilled water and dried. Sperms were examined under a microscope. Randomly were counted 100 sperms and recorded number of unreacted acrosome could be determined the percentage of unreacted acrosome Watson, (1975). Semen data analysis:

Semen data analysis was performed using SPSS 15 software. All values were expressed as mean ± standard error of mean (S.E.M). Analysis of variance (ANOVA) with Least Significant Difference (LSD) test was used subsequently for the comparison of mean values and separated the differences among means at a significance level of P < 0.05. SDS-PAGE electrophoresis for blood and seminal plasmaproteins:

Samples were applied to a 15% of polyacrylamide gel. Gel preparation, electrophoretic conditions, staining and destaining of gels were done according to Bollag and Edestein (1994).


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DNA Extraction: DNA was isolated from blood by modification of the method

described by Sambrook et al. (1989). Polymerase chain reaction:

Genomic DNA based analysis of five tested sheep breeds, each individuals sample was used to amplify by two specific primers, forward primer was 5'- TAGGCTCTACCGCCGTGCACA-3' and reverse primer was 5'-AGGCGTGTCGTCCTGGAAGGTC-3'. These primers were designed according to the exon 5 of acrosin gene sequence (ACR) of Ovis canadensis sheep breed found at National Center for Biotechnology Information (NCBI,2013) site with the accession number (EF033138); John and Willie(2007).

PCR reaction was prepared in 10 µl contained: 2 µl DNA, 1 µl for forward primer, 1 µl reverse primer, 1 µl dNTPs, 1 µl MgCl2 , 2 µl 5 X Green GoTaq Flexi buffer,1.9 µl and 0.1 µl Taq polymerase. PCR reaction were achieved in the Techne thermocyclers which programmed as follows: Concerning the first cycle (initial denaturing), the denaturation temperature degree was at 94˚C for 4 min, and for the next 35 cycles, denaturation, annealing and extension temperature degrees were at 94˚ C for 45 sec, at 58˚ C for 1 min and at 72˚ C for 2 min, respectively, and for the last cycle (final extension), the temperature degree was at 72˚ C for 5 min., then stored at 8°C for 10 min. The annealing temperature was 58˚ C in Awassi, Rahmani, Barki and Finnish breeds, while it was 68 C in Ossimi breed. DNA sequencing:

Only one sample of PCR-product for each sheep breed was randomly chosen, cleaned and concentrated using Fermentas (GeneJET PCR Purification Kit #K0702). Cleaned fragments were sequenced through the automated DNA sequencing reaction with specific primers using ABI PRISM Big Dye Terminator Cycle Sequencing Ready Reaction Kit (PE applied Biosystems, Spain), in conjunction with ABI PRISM (310 Genetic Analyzer). Sequences analysis:

The DNA sequences for Acr gene (exon5) which represented the five studied sheep breeds were aligned using BioEdit program through multiple sequence alignment.


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Gels analysis: Gel Analyzer ver. 3 program was used to analyze the gels by

comparing the bands sizes with standard bands (DNA ladder).

RESULTS AND DISCUSSION

Semen evaluation of studied rams breeds: General features of fresh semen samples for sex rams from each

of the studied breeds were evaluated to assess the products fertilizing. This features including semen value of volume, density, color, concentration, mass motility (0-5), progressive motility (0-5), pH, dead (%), abnormality (%) and unreacted acrosome (%) as shown in Fig. 1 and Table 1.

Comparisons and statistical significance for mean values of semen characters in the five studied sheep breeds by ANOVA analysis are listed in Tables (2) and (3). The mean values obtained for density, volume, mass motility, live sperm percentage and sperm concentrations were not significantly different. While the mean values obtained for progressive motility were significantly different between Ossimi and Barki as well as between Ossimi and Finish breeds. Moreover, mean values obtained for abnormal percentage were significantly different between Awassi breed and the other four breeds. In Addition, mean values of unreacted acrosome persentage were only significantly different between Rahmani and Barki breeds.

The highest mean value of progressive motility was in Ossimi breed and the lowest mean value was in Barki breed, while these mean values were convergent in the other breeds. The highest mean value of unreacted acrosome persentage was in Barki breed followed by Awassi breed and the lowest mean value was in Rahmani breed, while the mean values were convergent in the other breed. Moreover, the highest mean value of abnormal percentage was in Awassi breed followed by Barki breed and lowest value was in Finnish breed, while these mean values were convergent in the other breeds. From these results, it could be concluded that Barki and Awassi breeds have low fertility rate. While Ossimi, Finnish, Rahmani breeds have high fertility rate. These results are in agreement with the data obtained from the farm records.

The variation in semen characteristics among the studied rams was in agreement with those of Abdel-Rahman et al. (2000) who


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collected semen during the breeding season from rams by electro ejaculation from two native (Najdi and Naemi) and three imported (Merino, Somalian and Sudanese) sheep breeds. There was a marked reduction in sperm concentration and the percentage live spermatozoa produced by the Somalian (Barbari) and Sudanese (Sawakni) breeds. On the other hand, these two breeds showed the highest percentage of spermatozoal individual motility and the highest concentrations.

Figure (1): Spermatozoa showing Reacted, Unreacted acrosomes

and Normal, abnormal, live and dead sperms in Ossimi, Barki, Rahmani, Awassi and Finnish.

Moreover, Davendra et al. (2010) also compared sperm motion characteristics of adult Malpura and Bharat Merino rams by the computer-aided sperm analysis (CASA) technique. Semen was collected from 8 rams of each breed at the onset of autumn season on 5 occasions at three days interval. The CASA parameters which differed significantly (p<0.05) between the breeds were rapid motile sperm, medium motile sperm, slow motile sperm, linearity, straightness, curvilinear velocity, average path velocity, straight-line velocity, amplitude of lateral head displacement, beat frequency, sperm head area and sperm head elongation were higher in Malpura breed. The semen volume and sperm concentration were higher in Bharat Merino but not significant.


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Table (1): Means and standard errors of theseminalcharacteristics for fresh semen samples of five studied breeds.

Table (2): Comparisons and statistical significance for meanvalues of semen characters in the five studied sheep breeds by ANOVA analysis.

Table (3): Least Significant Difference (LSD) analysis for mean values of progressive motility (%), abnormality (%) andunreacted acrosomes (%) in the five studied sheep breeds


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SDS-PAGE of blood protein (Bulk): SDS-PAGE for blood plasma proteins could be used to detect

some specific markers as illustrated in Fig. (2) and Table (4). One band with a molecular weight of 109 KDa was considered as negative specific markers for Finnish breed which was only absent in it. On the other hand, bands at molecular weight of 61 and 60 KDa could be considered as positive specific markers to recognize Barki and Awassi breeds, respectively. From these results, it could be concluded that the employment of protein banding patterns of blood plasma was very useful tool to identify and differentiate among the studied breeds. Agreement with those of Braend and Tucker (1982), who also differentiated among Indian breeds of goats by using the same techniques. Moreover, Tapio et al. (2002) also differentiate among nine sheep breeds of Finland and North-Western Russia by the same techniques.

Dendrogram tree as shown in Figure (3) illustrate the genetic relationships among the five breeds which divided into two groups. Ossimi, Awassi and Rahmani breeds were clustered in one group, while Finnish and Barki breeds were clustered in the other group. SDS- PAGE of seminal plasma protein (Bulk):

As illustrated in Table (5) and shown in Figure (2), the bands with molecular weights of 283 KDa and 27 KDa were considered as negative specific markers for Rahmani breed because they were only absent in it, while they were presented in Ossimi, Awassi, Barki and Finnish breeds. The band with molecular weight of 167 KDa considered as positive specific markers for Finnish and Ossimi breeds. Moreover, the band with molecular weights 151 KDa was considered as negative specific marker for Finnish and Ossimi breeds, where the rams of these two breeds recognized by their high fertility rate. On the other hand, the bands with molecular weights of 64 KDa and 33 KDa were considered as positive markers for Awassi and Barki breeds, where the rams of these two breeds recognized by their low fertility rate compared with those in the other studied breeds.

From these results, it could be concluded that SDS- seminal plasma protein was useful technique to determine the fertility between the studied breeds, and to detect specific markers for fertility trait among the studied rams. These results were in agreement with Wenbin et al. (2009) who investigated fertility associated proteins in


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ram seminal plasma and the correlation between specific proteins and semen characteristics in sheep.

Dendrogram tree in Figure (3) .shows genetic relationships among the rams of the five breeds based on seminal plasma protein. Finnish and Ossimi breeds were clustered in one group, where the studied rams in these two breeds were high fertility rate, while Awassi and barki breeds were clustered in other group where the studied rams of these two breeds were low fertility rate. Moreover, Rahmani breed was distance from these two groups.

(Bl) (S)

Figure (2): SDS-PAGE of (Bl) blood and (S) for seminal plasma protein banding patterns of bulked sample for the five studied sheep breeds. R: Rahmani, F: Finnish, O: Ossimi, B: Barki, A: Awassi

Table (4): SDS-PAGE of blood protein banding patterns of bulked sample for the five studied sheep breeds.


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Table (5): SDS-PAGE scores of seminal plasma protein banding patterns for the five studied sheep breeds.

(1) (2) Figure (3): Dendrogram tree for the genetic distances among the five

studied breeds based on SDS- PAGE of (1) blood and (2) seminal plasma protein banding patterns.

Detection of acrosin gene, exon 5 in the studied sheep breeds: Blood samples of eight ram individuals from Barki and Awassi

breeds, seven ram individuals from Finnish breed and six ram individuals from Ossimi and Rahmani breeds were randomly taken and DNA was extracted. PCR products resulting by using of the two primers with the applied breeds are presented in Figure (4). The expected band with fragment size of 379 bp was appeared in all tested rams which represented exon 5 in Acr gene.


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Figure (4): PCR products for the Acr gene (exon 5) fragments size

of the Individual samples for Ossimi, Rahmani, Barki, Finnish and Awassi breeds.

Acr gene, exon 5 DNA sequence alignment and similarity matrix: The obtained sequences for Acr gene (exon5) in the five studied

sheep breeds were compared and alignment with each other using BioEdit program as shown in Figure (5). The sequences in Ossimi and Finnish breeds were different from those in Awassi, Barki and Rahmani breeds. As illustrate in Table (6), Eight different nucleotides found in position 175, 211, 212, 243, 247, 264 and 338 in Finnish breed, while there were five different nucleotide in positions 211, 212, 249, 264 and 345 in Ossimi breed. This difference may play important role in fertility variation in those rams where Finnish breed have the highest fertility rate. The results in table (7) showed that the highest similarity values (1.00) were among Awassi, Barki, and Rahmani breeds, followed by Finnish and Ossimi breeds. While the lowest values (0.98) were between each of Awassi, Barki or Rahmani breeds with Finnish or Ossimi breeds


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Table (6): Number and positions of different nucleotides in Acr gene (exon5) sequence for the studied sheep breed.

Table (7): Sequences similarity matrix using pairwise distance among studied sheep breeds.

The dendrogram tree for the five studied sheep breeds based on

the sequence of Acr gene (exon 5) is shown in the Figure 5. The dendrogram was divided into two main clusters, where the first cluster was grouped Finnish and Ossimi breeds together, while the second one was considered Awassi, Barki and Rahmani breeds. However, Finnish and Ossimi were closer to each other in one cluster.

Figure (5): Genetic relationships based on the sequences of Acr gene (exon 5) for rams in the five studied breeds.

The sequences of Acr gene (exon5) of the five studied breeds were compared by alignment with sequences of Acr gene (exon5) of other origins on the data base at the national center for biotechnology information (NCBI) web site using BioEdit program. The sequence was very similar to sequence of Acr gene (exon5) in other mammalians and similarity value was ranged (0.89 – 1.00) which


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mean that Acr gene (exon5) is conserved sequence. This result agreed with John and Willie 2007.

From this study can be concluded that Acr gene (exon5) found in all studied sheep breed with fragment molecular size was 379 bp. This fragment has conservation sequence in these breeds and was identical in Awassi, Rahmani and Barki breed. While, there was different in sequence in Finnsh and Ossimi breeds, this different may play important roles in variation in fertility in these rams. In other hand multiple alignment of Acr gene (exon5) DNA sequence with Acr gene, (exon5) DNA sequence in other organs gives high similarity values.

Figure (5): Multiple alignments of the nucleotide sequences for Acr gene (exon 5) in the rams of the studied breeds.

Conclusion It is concluded from this study that physical screening of fresh

semen used in the present study revealed that the parameter values for fresh semen evaluation were higher in the Ossimi and Finnish rams and low in the Barki and Awassi rams, while Rahmani rams had moderate values. SDS-PAGE for blood and seminal plasma proteins paterns were very useful tools to identify and differentiate among the studied breeds and to detecting some positive and negative markers for the studied breeds. Whereas, the genetic analysis for Acr gene (exon5) showed that this region was found in all rams of studied sheep breeds, this fragment was identical in Awassi, Rahmani and


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Barki breeds, while it was difference in rams of Finnish and Ossimi breeds. This different may play important roles in variation of fertility for these rams.

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التصنيف الفسيولوجى والجزيئى لبعض سالالت األغنام المصرية

، محمود عبد الحفيظ 2، رضا محمد عنوس1، محمد عبد السالم راشد1إبراهيم محمد آيالى 1 و أيمن حنفى عطا1سالم

قسم الوراثة ، آلية الزراعة ، جامعة عين شمس-1

مس قسم اإلنتاج الحيوانى ، آلية الزراعة ، جامعة عين ش-2

ين سالالت روق ب دا لتوضيح الف ة ج ذآور مهم وى لل سائل المن تعتبر دراسة خصائص ال

أثير ه ن إن ل الى ف األغنام المصرية حيث يستخدم الذآر لتلقيح إلناث عديدة وألجيال آثيرة وبالت

ة د دراس ذلك، عن دد ل ل المح ى العام ون ه صوبه تك ة الخ ث أن درج اج حي ى اإلنت ر ف آبي

ام المصرية خصائص السائل المن ذآور بعض سالالت األغن انى و (وى ل األوسيمى و الرحم

وإرتباطها بدرجة الخصوبة المسجلة مسبقا أتضح أن الفرق آان ) البرقى و العواسى و الفينيش

سبة ا بالن يش، أم ى والفين ين ساللة األوسيمى والبرق رق واضحا ب ان الف ة وآ سبة الحيوي فى ن

ع سالالت محل لنسبة التشوهات فى اإلسبرمات فكا ن الفرق واضحا بين العواسى وباقى األرب

ين . الدراسة ا ب رق معنوي ان الف وى فك وان المن ار تفاعل األآروسين فى الحي أما بالنسبة إلختب

سائل SDS-PAGEالبرقى والرحمانى عند إجراء التفريد الكهربى لبروتينات بالزما الدم وال

روتي وذج ب ر نم وى أظه ى المن ى والعواس ن البرق ل م زتين لك زمتين ممي دم ح ا ال ن بالزم

ن ل م ارب آ دم تق ا ال روتين بالزم وذج ب اس نم ى أس ة عل ة الوراثي جرة القراب وأوضحت ش

ى ا ف ى مع يش والبرق ا الفين دة بينم ة واح ى مجموع ا ف انى مع ى والرحم يمى والعواس األوس

سائل الم . مجموعة أخرى زتين آعالمات آما أظهر نموذج بروتين بالزما ال وى حزمتين ممي ن

يش زة للفين البة ممي رى س ة وأخ دة موجب ة واح ة بروتيني ر حزم ط وأظه انى فق البة للرحم س

ا أوضحت زتين لكل من البرقى والعواسى، آم واألوسيمى، باإلضافة لحزمتين موجبتين ممي

و سائل المن ا ال ارب آل شجرة القرابة الوراثية على أساس التفريد الكهربى لبروتين بالزم ى تق

ة ى مجموع ا ف ى مع ا العواسى والبرق دة بينم ة واح ى مجموع ا ف يش مع يمى والفين ن األوس م

ط انى فق ا الرحم ة به م . أخرى ومجموعة ثالث عزل اإلآسون الخامس من جين األآروسين ت

ه وقراءة تتابعة لكل ساللة ومقارنتهم ببعضهما لتحديد الفروق فى القواعد النيتروجينية حيث أن

سالالت محل جين هام وثابت فى آائنات آثيرة جدا فإتضح أنه متساوى الحجم أو الطول فى ال

بينما اإلختالفات فقط آانت فى ثمانى قواعد نيتروجينية موزعة bp 379الدراسة وهى بحجم


140

ةعلى أساس ة الوراثي ا شجرة القراب ه، أم فى أماآن مختلفة بطول الجزء الجينى الذى تم قراءت

ا تتابع النيو آليوتيدات أوضحت تقارب آل من األوسيمى والفينيش معا فى مجموعة واحدة بينم

ة ا فى مجموعة أخرى وأستطاع البحث عمل بصمة وراثي انى مع العواسى والبرقى والرحم

ة ة الهام ز المنطق ذلك تميي ة وآ ت الدراس صرية تح ام الم الالت األغن ذآور س حة ل واض

ا من والمترجمة من جين األآروسين وعزلها و ا يمكنن قرأتها، ومقارنة السالالت ببعضهما مم

.مقارنة السالالت المصرية بالسالالت العالمية

.أسبرمات، ذآور، أآروسين، تفريد آهربى، تتابعات : آلمات دالة

PHYSIOLOGICAL AND MOLECULAR CHARACTERIZATION OF SOME EGYPTIAN SHEEP BREEDS

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