پایان نامه ارتباط چند شکلی ژن FSHB با صفات تولید مثلی درگاو های نر هلشتاین ایران

فهرست و منابع پایان نامه ارتباط چند شکلی ژن FSHB با صفات تولید مثلی درگاو های نر هلشتاین ایران

فهرست:

چکیده .........................................................................................................................................................1

مقدمه..............................................................................................................................................................3

فصل اول مروری برمنابع. ........................................................................................................................6

1-1 آناتومی دستگاه تولید مثلی گاو نر. ................................................................................................7

1-2 فیزیولوژی تولید مثلی گاو نر. ......................................................................................................8

1-3 عوامل کنترل کننده کیفیت اسپرم. .................................................................................................10

1-3-1 عوامل هورمونی.. .......................................................................................................10

1-3-1-1 GnRH..........................................................................................................................................10

1-3-1-2 گنادوتروپین‌ها و پرولاکتین...........................................................................................................11

1-3-1-3 تستوسترون و آندروژن................................................................................................................12

1-3-1-4 هورمون رشد................................................................................................................................12

1-3-1-5 انسولین ......................................................................................................................................13

1-3-1-6 گلوکورتیکوییدها و آلدسترون ...................................................................................................13

1-3-1-7 هورمون‌های تروییدی و PGF2α.................................................................................................14

1-3-2 عوامل غیرهورمونی موثربرکیفیت اسپرم.....................................................................................................14

1-3-2-1 عوامل ژنتیکی.. ...............................14

1-3-2-1-1 محیطی.. .........................................................................................................................16

1-3-2-1-2 تیم جمع آوری.. ..............................................................................................................16

1-3-2-1-3 فاصله جمع آوری.....................................................................................................................17

1-3-2-1-4 فصل..........................................................................................................................................17

1-3-2-1-5 حرارت.....................................................................................................................................18

1-3-2-1-6  سن دام. .......................................................................................................................19

1-3-2-1-7 تغذیه.........................................................................................................................................20

1-4 ارزیابی اسپرم.......................................................................................................................................21

1-4-1 خصوصیات ظاهری اسپرم............................................................................................................... 22

1-4-2 تحرک اسپرم................................................................................................................................... 22

1-4-3 آکروزوم بی عیب...........................................................................................................................  23

1-4-4 مورفولوژی اسپرم ( شکل اسپرم ).................................................................................................. 23

1-4-5 ارزیابی اسپرم یخ‌زده....................................................................................................................... 24

1-4-6 غلظت اسپرم................................................................................................................................... 25

1-5 صفات تولید مثلی مهم در اصلاح نژاد گاو نر..................................................................................... 26

1-6 رویکردهای مولکولی برای بهبود تولید مثل در گاو............................................................................ 27

1-7 تجزیه و تحلیل ژن  FSH کاندیدا................................................................................ 28

1-8 FSH......................................................................................................................................30

1-8-1 هورمون و تولیدمثل............................................................................................................ 32

1-8-1-2 نقش هورمون FSH  در فعالیت بیضه‌ها.............................................................................. 33

1-8-2 چندشکلی ها:............................................................................................................................ 35

1-8-2 -1 ژن  FSH......................................................................................................................................35

1-8-2-1 -1پلی مورفیسم ایزوفرم های گیرنده FSH....................................................................................36

1-8-2-1-2 رخ دادن جهش های طبیعی از گیرنده FSH..............................................................................37

1-8-2-1-3 تشخیصSNPها در گیرنده FSH...........................................................................................37

1-8-2-1-4 تاثیر SNPها در تولید مثل.........................................................................................................38

1-8-2-1-5 اعمال فیزیولوژی   FSHبا ترکیب بر اسپرم سازی....................................................................39

1-8-2-1-6 پلی مورفیسم گیرندهFSH   وعملکرد بیضه ها..........................................................................42

1-8-2-1-7 چند شکلی در گیرنده FSH در گونه های حیوانی دیگر..........................................................42

فصل دوم مواد و روش.............................................................................................................................45

2-1 روند آزمایش.......................................................................................................................................46

2-2 مواد.....................................................................................................................................................46

2-2-1 حیوانات ........................................................................................................................................46

2-2-2 نمونه برداری..................................................................................................................................48

2-2-3 رکوردهای فنوتیپی.........................................................................................................................48

2-2-4 مواد و وسایل آزمایشگاهی...........................................................................................................48

2-2-4-1 تجهیزات...................................................................................................................................49

2-2-4-2 مواد شیمیایی و کیت‌ها و شرکت سازنده.................................................................................49

2-2-4-3 محلول‌ها و بافرها.....................................................................................................................52

2-2-4-4 آغازگرهای مورد استفاده..........................................................................................................54

2-3 روش‌ها............................................................................................................................................54

2-3-1 صفات مورد بررسی.....................................................................................................................54

2-3-1-1 ارزیابی صفات کیفی اسپرم......................................................................................................54

2-3-1-1-1 حجم اسپرم.........................................................................................................................54

2-3-1-1-2 جمعیت اسپرم در هرمیلی لیتر انزال...................................................................................54

2-3-1-1-3 جمعیت کل در هر انزال....................................................................................................54

2-3-1-1-4 جنبندگی اسپرم تازه و یخ گشایی شده.............................................................................55

2-3-1-1-5 تعداد کل اسپرم زنده قبل انجماد......................................................................................55

2-3-1-1-6 تعداد کل اسپرم زنده بعد از یخ گشایی............................................................................55

2-3-2 روش‌های ژنتیک مولکولی........................................................................................................55

2-3-2-1 استخراج DNA....................................................................................................................55

2-3-2-1-1 استخراج DNA از نمونه‌های خون.................................................................................55

2-3-2-1-2 استخراج DNA از نمونه‌های اسپرم...................................................................................56

2-3-2-1-3 نگهداری DNA استخراج شده..........................................................................................58

2-3-2-1-4 تعیین کمیت و کیفیت DNA استخراج شده......................................................................58

2-3-2-2 واکنش زنجیره‌ای پلی مرز......................................................................................................59

2-3-2-3 تعیین ژنوتیپ.........................................................................................................................60

2-4 تجزیه و تحلیل آماری ..................................................................................................................61

2-4-1 تجزیه و تحلیل ژنوتیپ‌ها..........................................................................................................61

2-4-1-1 محاسبه فراوانی‌های اللی و ژنوتیپی.......................................................................................61

2-4-1-2 آزمون تعادل هاردی-واینبرگ................................................................................................61

2-4-2 تجزیه و تحلیل ارتباط بین ژنهای کاندیدا و صفات مورد بررسی.............................................61

2-4-2-1 مدل آماری برای صفات با توزیع پیوسته...............................................................................62

2-4-2-2 مدل آماری برای صفات با توزیع گسسته..............................................................................62

فصل سوم نتایج و بحث......................................................................................................................64

3-1 خلاصه آماری صفات مورد بررسی ...............................................................................................65

3-2 نتایج تجزیه واریانس عوامل موثر بر کیفیت منی.............................................................................66

3-2-1 فراوانی آللی و ژنوتیپی................................................................................................................68

3-2-1-1-1 بررسی ارتباط جایگاه FSHB ژن هورمون محرک فولیکول با صفات مورد بررسی ...........70

3-2-1-1-2 دلایل احتمالی تاثیر جایگاه  FSHBبر روی صفات مورد ب.............................................73

3-2-2 اثر فصل  اسپرم‌گیری ................................................................................................................75

3-2-3 اثر متقابل فصل اسپرم‌گیری با ژن هورمون محرک فولیکول......................................................79

4- نتیجه گیری......................................................................................................................................84

5- پیشنهادات........................................................................................................................................85

6- منابع.................................................................................................................................................86

7- چکیده انگلیسی............................................................................................................................114

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