Benha University Faculty of Agriculture Animal Production Dept. EFFECT OF PHOTOPERIOD ON OVARIAN ENDOCRINE FUNCTION IN RABBITS By HUSSEIN MOSTAFA MOHAMMED EL-ZAHER B.Sc. in Agric. Sci. (Animal Production) 1999, Faculty of Agriculture, Zagazig University (Benha Branch) THESIS Submitted in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN ANIMAL PHYSIOLOGY Department of Animal Production Faculty of Agriculture Benha University Egypt 2011 SUPERVISION COMMITTEE EFFECT OF PHOTOPERIOD ON OVARIAN ENDOCRINE FUNCTION IN RABBITS By HUSSEIN MOSTAFA MOHAMMED EL-ZAHER B. Sc. Agric. Sci, (Animal Production) 1999, Faculty of Agriculture, Zagazig University (Benha Branch) THESIS Submitted in partial fulfillment of the Requirement for the degree of MASTER OF SCIENCE IN )ANIMAL PHYSIOLOGY( Under supervision of: ** Prof. Dr. Mohammed, S. Gado …………………………………. Professor of Animal Physiology, Animal Production Dept. Faculty of Agriculture, Benha University. ** Prof. Dr. Gaafar, M. El-Gendi …..……………………………... Professor of Poultry Management, Animal Production Dept. Faculty of Agriculture, Benha University. ** Dr. Magdy Hassan, G. Ishak ….…………………………........ Assoc.Professor of Physiology and Head of Endocrinology Research Unit, Radiobiological Applied Dept. Nuclear Research Center, Atomic Energy Authority. APPROVAL COMMITTE EFFECT OF PHOTOPERIOD ON OVARIAN ENDOCRINE FUNCTION IN RABBITS By HUSSEIN MOSTAFA MOHAMMED EL-ZAHER B. Sc. Agric. Sci. (Animal Production) 1999, Faculty of Agriculture, Zagazig University (Benha Branch) THESIS Submitted in partial fulfillment of the Requirements for the degree of MASTER OF SCIENCE IN ANIMAL PHYSIOLOGY This thesis for M.Sc. degree has been approved by: ** Prof. Dr. Mohammed, S. Gado ………………………………. Professor of Animal Physiology, Animal Production Dept. Faculty of Agriculture, Benha University. ** Prof. Dr. Medhat, H. Khalil ….…………………………… Professor of Animal Physiology, Animal Production Dept. Faculty of Agriculture, Al- Azhar University. ** Prof. Dr. Ahmed, A. Radwn ...………………………..... Professor of Poultry Physiology, Animal Production Dept. Faculty of Agriculture, Benha University. ** Prof. Dr. Gaafar, M. El-Gendi .…………………………... Professor of Poultry Management, Animal Production Dept. Faculty of Agriculture, Benha University. ** Dr. Magdy Hassan G. Ishak …………………………....... Assoc. Professor of Physiology, Head of Endocrinology Research Unit, Radiobiological Applied Dept. Nuclear Research Center, Atomic Energy Authority. Date of Examination: / / 2011. 1 ACKNOWLEDGMENT I am greatly indebted to: Professor Mohamed safwat Abd El-Magied Gado, supervisor of this work and Professor of Animal physiology, Department of Animal Production, Faculty of Agriculture, Benha University for planning the research work, close supervision and valuable comments, without his initiative and constructive advice, this study would not have been possible. I am also thankful for his patient and kindness. Professor Gaafar Mohamed Ibrahim El-Gendi, Professor of Poultry Production , Faculty of Agriculture, Benha University for his helpful and effective support during the lifespan of this work. Dr. Magdy Hassan Gamal Ishak, Assoc. Professor of Physiology and Head of Endocrinology Research Unit, Applied Radiobiological department, Nuclear Research Center, Atomic Energy Authority, Inshas Egypt and Professor Ibrahim Mohamed El-Banna Professor of Physiology, Endocrinology Research Unit, Applied Radiobiological department, Nuclear Research Center, Atomic Energy Authority, Inshas Egypt for suggesting the problem, designing the experiment work, testing the hypothesis, offering all facilities, encouragement, valuable guidance and criticism. TABLE OF CONTENTS Page 1- INTRODUCTION.................................................................................. 1 2- REVIEW OF LITERATURE............................................................... 7 2.1. Overview of reproduction and productivity of female rabbits 7 emphasizing photoperiod effect...................................................... 2.2. Natural Endocrine control of reproduction..................................... 11 2.3. Ovulation induction......................................................................... 17 2.4. Hormonal mechanism during ovulation.......................................... 18 2.5. Neuroendocrine control of ovulation.............................................. 20 2.6. Mechanism of Ovarian Follicle Rupture......................................... 25 2.7. Biosynthesis and biological effect of Melatonin on reproduction............................. 27 2.8. Effect of Photoperiod on Melatonin Biosynthesis.......................... 33 2.9. Pineal gland activity and the role of melatonin in transducing photoperiodic Information............................................................. 38 2.9.1. Pineal gland activity............................................................. 38 2.9.2. Critical features of Melatonin secretion as affected by rhythm..................... 40 2.10. Role of Melatonin in reproduction................................................ 46 2.11. Effect of serotonin in reproduction............................................... 48 2.12. Effect of human chorionic gonadotropin (HCG) on the 50 mechanism of ovulation.............................................................. 2.13. Effect of sexual steroidal hormones on oogenesis and ovulation……...... 52 2.14. Effect of prolactin (PRL) on oogenesis and ovulation.................. 57 2.15. Bromocriptine: an ergoline derivative acts as dopamine agonist used in Hyperprolactinaemia...................................................... 61 2.16. Prostaglandin (PG) function on uterine contraction and ovulation……...... 63 2.17. Effect of indomethacin on the prostaglandin biosynthesis........... 66 3. MATERIALS AND METHODS........................................................... 73 3.1. Grouping of animals and experimental design................................. 73 I 3.2. Hormonal modulators applied.......................................................... 77 3.2.1. Indomethacin....................................................................... 77 3.2.2. Bromocriptine (parlodel cycloset)...................................... 78 3.2.3. Melatonin…………………………………………………. 79 3.2.4. Serotonin………………………………………………..... 80 3.3. Injected materials............................................................................. 81 3.4. Parameters estimated and data collection......................................... 82 3.4.1. Blood samples collection..................................................... 82 3.4.2. Chemicals and reagents used in chemical analysis.............. 82 3.4.3. Progesterone (P4) radioimmunoassay................................. 83 3.4.3.1. Assay buffers....................................................... 84 3.4.3.2. Assay procedure.................................................. 85 4. RESULTS AND DISCUSSIONS.......................................................... 88 4.1. First experiment.............................................................................. 88 4.1.1. Serum progesterone level in untreated does subjected to different lighting regimes applied...................................... 89 4.1.2. Effect of HCG....................................................................... 90 4.1.3. Effect of Melatonin............................................................... 97 4.1.4. Effect of serotonin................................................................. 103 4.1.5. Effect of bromocriptine......................................................... 109 4.1.6. Effect of indomethacin.......................................................... 116 4.2. Second experiment.......................................................................... 122 4.2.1. Serum progesterone level in does treated with HCG............ 122 4.2.2. Serum progesterone level in does treated with HCG in combination with melatonin.............................................. 124 4.2.3. Serum progesterone level in does treated with HCG in combination with serotonin................................................ 130 4.2.4. Serum progesterone level in does treated with HCG in combination with bromocriptine........................................ 136 4.2.5 Serum progesterone level in does treated with HCG in combination with indomethacin......................................... 143 5. CONCLUSION………………………………………………………... 149 6. SUMMARY............................................................................................. 150 II 7. REFERENCES....................................................................................... 160 8. APPENDIX TABLES............................................................................. 212 9. ARABIC SUMMARY............................................................................ ---- III LIST OF TABLES Table Title Page 1.a Design of the first experiment and number of animals 75 subjected to different photoperiods. Design of the second experiment and number of animals 1.b subjected to different photoperiods. 76 Protocol of injected materials during the course of 2 81 experiments Progesterone serum levels (ng/ml) x ±SE in response to injection of HCG vs saline at short darkness photoperiod (8D/16L), equal dark 3 93 and light photoperiod (12D/12L) and long darkness photoperiod (16D/8L). Analysis of variance of data (cumulative area under curve in units) 4 94 collected after injection of HCG vs saline under different photoperiods. Progesterone