Abstract
This work was designed to evaluate the ovarian follicular development, oocytes morphology, methods of oocytes reterival, and the effect of different in vitro maturation (IVM) media on cumulus cell expansion and nuclear maturation of Jennies oocytes. Experiment 1, the number of small (<6 mm), medium (6 to 9 mm) and large size (>10 mm) ovarian follicles was recorded. Cumulus-oocyte-complexes (COCs) were reterived and classified into 4 Grades based on their cumulus-cells investment and the homogenous of the ooplasm. In Experiment 2, COCs were recovered by using 18-G, 20-G needle or slicing and scraping of ovarian follicles to determine the number and morphology of the recovered COCs. In Experiment 3, Grade A and B COCs were IVM in DMEM-HG, DMEM-LG, DMEM-F12, TCM199, TCM199-F12 or CR1aa media supplemented with 10 % FCS + 10 μg FSH/mL + 10 IU hCG/mL + 50 μg/mL gentamicin. Maturation was performed for 36 h at 38.5 °C under 5 % CO2 in humidified air. After IVM, cumulus cell expansion and oocytes nuclear canfiguration were determined. An average of 6.40 ± 0.26 follicles was recorded per Jenny ovary, representing 3.37 ± 0.46, 1.89 ± 0.14 and 1.14 ± 0.16, for the small, medium and large size follicles, respectively. Oocyte recovery was higher (P < 0.05) in large and medium size follicle than in the small one (62 %, 60 % and 45.1 %, respectively). Small size follicles produced higher (P < 0.05) percentage of Grade A COCs than large or medium size follicles. A higher number of oocytes was recovered by slincing and scraping of follicles (4.86 ± 0.67), then aspiration of follicles using 18-G needle (3.14 ± 0.36 COCs/ovary, P < 0.05). Aspiration using 18-G needle or slicing and scraping of follicles using produced a significantly higher (P < 0.05) percentage of Grade A COCs compared to aspiration of follicles using 20-G needle (56.6 %, 46.7 % and 32.0 %, respectively, P < 0.05). IVM of COCs in CR1aa and TCM 199-F12 media significantly increased (P < 0.05) Grade 3 cumulus-cell expansion compared with TCM199, DMEM-F12, DMEM-LG and DMEM-HG (65.5 % and 64.0 %, 52.8 %, 32.1 %, 0.0 % and 7.4 %, respectively). The proportion of IVM oocytes reaching the M II stage was significantly higher (P < 0.05) for oocytes matured in TCM199-F12 or CR1aa media than TCM199, DMEM-HG, DMEM-LG, DMEM-F12 (69.1 % and 62.2 %, 55.7 %, 45.8 %, 39.0 % and 40.7 %, respectively). The proportion of degenerated oocytes IVM in TCM199-F12 (10.3 %), CR1aa (11.3 %) or TCM199 (13.1 %) was lower (P < 0.05) than that matured in DMEM-HG, DMEM-LG or DMEM-F12 media (23.7 %, 29.3 % and 22.9 %, respectively). Conclusion: Slicing and scraping or aspiration of follicles using 18-G needle increased the number and percentage of Grade A Jennies oocytes. TCM199-F12, CR1aa and TCM199 medi are more suitable for IVM of Jenny oocytes by promoting cumulus cells expansion and nuclear maturation to M II stage.
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References
Alm H, Torner H (1994) In vitro maturation of horse oocytes. Theriogenology 42:345–9
Alm H, Torner H, Kanitz W, Becker F, Hinrichs K (1997) Comparison of different methods for the recovery of horse oocytes. Equine Vet J Suppl 25:47–50
Bézard J, Mekarska A, Goudet G, Duchamp G, Palmer E (1997) Timing of in vivo maturation of equine preovulatory oocytes and competence for in vitro maturation of immature oocytes collected simultaneously. Equine Vet J Suppl 25:38–42
Blanchard TL, Taylor TS, Love CL (1999) Estrous cycle characteristics and response to estrus synchronization in mammoth asses (Equus assinus Americans). Theriogeneology 52:827–834
Brück I, Greve T, Hyttel P (1999) Morphology of the oocyte-follicular connection in the mare. Anat Embryol (Berl) 199:21–8
Choi YH, Hochi S, Braun J, Oguri N (1993) In vitro maturation of equine oocytes collected by aspiration and additional slicing of ovaries. Theriogenology 40:959–966
Choi YH, Hochi S, Braun J, Sato K, Oguri N (1997) In vitro maturation of equine oocytes collected by follicle aspiration and by the slicing of ovaries. Theriogenology 40:959–966
Choi YH, Chung YG, Seidel GE Jr, Squires EL (2001) Developmental capacity of equine oocytes matured and cultured in equine trophoblast-conditioned media. Theriogenology 56:329–339
Choi YH, Love LB, Varner DD, Hinrichs K (2004) Factors affecting developmental competence of equine oocytes after intracytoplasmic sperm injection. Reprod 127:187–194
Colleoni S, Luciano AM, Gandolfi F (2004) Cumulus-oocyte communications in the horse: role of the breeding season and of the maturation medium. Reprod Domest Anim 39(2):70–5
Dell’Aquila ME, Masterson M, Maritato F, Hinrichs K (2001) Influence of oocyte collection technique on initial chromatin configuration, meiotic competence, and male pronucleus formation after intracytoplasmic sperm injection (ICSI) of equine oocytes. Mol Reprod Dev 60:79–88
Dell’Aquila ME, Cho YS, Minoia P, Traina V, Lacalandra GM, Maritato F (1997) Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in-vitro fertilization or intracytoplasmic sperm injection. Hum Reprod 12:2766–72
Derar RJ, Hussein HA (2011) Ovarian Follicular Dynamics during the Estrous Cycle in Jennies in Upper Egypt. Veterinary Medicine International Volume 2011, Article ID 860518, 6 pages doi:10.4061/2011/860518
Driancourt MA, Prunier A, Palmer E, Mariana JC (1983) Seasonal effects on ovarian follicular development in pony mares. Reprod Nutr Dev 23:207–215
Erice I, Gil L, Josa A, Echegaray A, Martinez F, Espinosa E (1998) Effect of mare’s age and recovery methods on the recovery rate of equine follicular oocytes for IVM procedures. Theriogenology 49:735–41
Farin CE, Rodriguez KF, Alexander JE, Hockney JE, Herrick JR, Kennedy-Stoskopf S (2007) The role of transcription in EGF- and FSH-mediated oocyte maturation in vitro. Anim Reprod Sci 98(1–2):97–112
Fernandes CB, Peres KR, Alvarenga MA, Landim-Alvarenga FC (2006) The use of transmission electron microscopy and oocyte transfer to evaluate in vitro maturation of equine oocytes in different culture conditions. J Equine Vet Sci 26:159–167
Fernando P, Starkey P (2004) Donkeys and Development: Socio-Economic Aspects of Donkey Use in Africa, in: Fielding, D. and Starkey, P. (Eds). Donkeys, People and Development. A resource book in the Animal Traction Network for Eastern and Southern Africa (ATNESA). ACP-EU Technical Centre for Agricultural and Rural Cooperation (CTA)
Galli C, Maclellan LJ, Crotti G, Turini P, Ponderato N, Duchi R, Merlo B, Mari G, Barbacini S, Lazzari G (2002) Development of equine oocytes matured in vitro in different media and fertilised by ICSI. Theriogenology 57:718
Galli C, Colleoni S, Turini P, Crotti G, Dieci C, Lodde V, Luciano AM, Lazzari G (2014) Holding equine oocytes at room temperature for 18 h prior to in vitro maturation maintains their developmental competence. J Equine Vet Sci 3:174–175
Gambini A, Jarazo J, Karlanian F, De Stéfano A, Salamone DF (2014) Effect of collection-maturation interval time and pregnancy status of donor mares on oocyte developmental competence in horse cloning. J Anim Sci 92:561–7
Ginther OJ, Scraba ST, Bergfelt DR (1987) Reproducitve seasonality of the jenny. Theriogenology 27:587–592
Goudet G, Bézard J, Duchamp G, Gérard N, Palmer E (1997) Equine oocyte competence for nuclear and cytoplasmic in vitro maturation: effect of follicle size and hormonal environment. Biol Reprod 57:232–45
Goudet G, Bézard J, Belin F, Duchamp G, Palmer E, Gérard N (1998) Oocyte competence for in vitro maturation is associated with histone H kinase activity and is influenced by estrous cycle stage in the mare. Biol Reprod 59:456–462
Guignot F, Bezard J, Palmer E (1999) Effect of time during transport of excised mare ovaries on oocyte recovery rate and quality after in vitro maturation. Theriogenology 52:757–66
Hamano S, Kuwayama M (1993) In vitro fertilization and development of bovine oocytes recovered from the ovaries of individual donors: A comparison between the cutting and aspiration method. Theriogenology 39:703–712
Hinrichs K (1997) Cumulus expansion, chromatin configuration and meiotic competence in horse oocytes: a new hypothesis. Equine Vet J Suppl 25:43–6
Hinrichs K (2010) The equine oocyte: factors affecting meiotic and developmental competence. Mol Reprod Dev 77:651–661
Hinrichs K (2013) Assisted reproduction techniques in the horse. Reprod Fert Dev 25:80–93
Hinrichs K, Schmidt AL (2000) Meiotic competence in horse oocytes: interactions among chromatin configuration, follicle size, cumulus morphology, and season. Mol Reprod 62:1402–1408
Hinrichs K, Williams KA (1997) Relationships among oocyte-cumulus morphology, follicular atresia, initial chromatin configuration, and oocyte meiotic competence in the horse. Biol Reprod 57:77–84
Hinrichs K, Choi YH, Love LB, Varner DD, Love CC, Walckenaer BE (2005) Chromatin configuration within the germinal vesicle of horse oocytes: changes post mortem and relationship to meiotic and developmental competence. Biol Reprod 72:1142–50
Hinrichs K, Choi YH, Norris JD, Love LB, Bedford-Guaus SJ, Hartman DL, Velez IC (2012) Evaluation of foal production following intracytoplasmic sperm injection and blastocyst culture of oocytes from ovaries collected immediately before euthanasia or after death of mares under field conditions. J Am Vet Med Assoc 241:1070–4
Kanitz W, Alm H, Becker F, Nürnberg G, Kurth J, Hinrichs K (2000) Repeated follicle aspiration in mares: consequences for follicle growth and oocyte quality. J Reprod Fertil Suppl 56:463–472
Kebede H, Lemma A, Negussie H (2012) Ultrasonographic studies on ovarian dynamics and associated estrus manifestations of Jennies under controlled management, Ethiopia. Trop Anim Health Prod 44:1965–70
Kugler V., Grunenfelder H.P., Broxham E., 2008. Donkey Breeds in Europe: Inventory, Description, Need for Action, Conservation; Report 2007/2008. St. Gallen, Switzerland: Monitoring Institute for Rare Breeds and Seeds in Europe. http://www.save foundation.net/pdf/donkey.
Lemma A, Bekana M, Schwartz HJ, Hildebrandt T (2006) The effect of body condition on ovarian activity of free ranging tropical Jennies (Equus asinus). J Vet Med A 53:1–4
Li-jiang G, Chang-lin X, Rui-qing S, Jing-he T, Ming-jiu L (2001) Ovarian follicular and hormonal dynamics in Dezhou Jenny during estrous cycle monitored by real time ultrasonography. Acta Vet Zootech Sin 42:1256–1263
Mlodawska W, Okólski A (1997) Equine oocyte-cumulus morphology as affected by follicular size. Equine Vet J Suppl 25:43–46
Pereira GR, Lorenzo PL, Carneiro GF, Ball BA, Bilodeau-Goeseels S, Kastelic J, Pegoraro LM, Pimentel CA, Esteller-Vico A, Illera JC, Granado GS, Casey P, Liu IK (2013) The involvement of growth hormone in equine oocyte maturation, receptor localization and steroid production by cumulus-oocyte complexes in vitro. Res Vet Sci 95:667–74
Rambags BPB, van Boxtel DCJ, Tharasanit T, Lenstra JA, Colebrander B, Stout TAE (2005) Oocyte mitochondrial degeneration during reproductive ageing in the mare. Havemeyer foundation. Monogr Ser 18:25–7
Rosenkrans CF Jr, Zeng GQ, McNamara GT, Schoff PK, First NL (1993) Development of bovine embryos in vitro as affected by energy substrates. Biol Reprod 49:459–462
Shabpareh V, Squires EL, Seidel GE Jr, Jasko DJ (1993) Methods for collecting and maturing equine oocytes in vitro. Theriogenology 40:1161–1175
Sosnowski J, Lechniak D, Brzozowska M, Switoński M (1997) Cytogenetic analysis of horse oocytes matured in vitro for different periods of time. Reprod Nutr Dev 37:63–8
Starkey P (1997) “Donkey Work”, in Elisabeth Svendsen (ed.), The Professional Handbook of the Donkey, 3rd edition. London: Whittet Books (1997). ISBN 978-1-873580-37-0. pp.183–206
Torner H, Alm H, Kanitz W, Goellnitz K, Becker F, Poehland R, Bruessow KP, Tuchscherer A (2007) Effect of initial cumulus morphology on meiotic dynamic and status of mitochondria in horse oocytes during IVM. Reprod Domest Anim 42:176–83
Willis P, Caudle AB, Fayrer-Hosken RA (1991) Equine oocyte in vitro maturation: influences of sera, time and hormones. Mol Reprod Dev 30:360–368
Zhao G, Wub K, Cuib L, Zhaob L, Liub Y, Tanc X, Zhoub H (2011) In vitro maturation and artificial activation of donkey oocytes. Theriogenology 76:700–704
Acknowledgments
This paper was funded by the National Research Center (NRC) of Egypt (in-house project 9040207). The authors, therefore, acknowledge with thanks the technical and financial support for NRC.
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Abdoon A.S., Abdelrahman H.A. and Kandil O.M. design the work, performed all experiments Sherif I. Shawki assist in the practical work. Abdoon write the manuscript and all the authors read and approved the final manuscript.
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Abdoon, A.S.S., Abdel-Rahman, H.A., Shawki, S.M. et al. Influence of follicle size, methods of retrieval on oocytes yield and morphology in Egyptian Jennies ovaries with special reference to maturation rate in vitro. Vet Res Commun 38, 287–295 (2014). https://doi.org/10.1007/s11259-014-9617-y
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DOI: https://doi.org/10.1007/s11259-014-9617-y