FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Assist Reprod Genet / Lower blastocyst quality after conventional vs. Piezo ICSI i...
Journal of assisted reproduction and genetics2018; 35(5); 825-840; doi: 10.1007/s10815-018-1174-9

Lower blastocyst quality after conventional vs. Piezo ICSI in the horse reflects delayed sperm component remodeling and oocyte activation.

Abstract: The aim of this study was to evaluate the differential effects of conventional and Piezo-driven ICSI on blastocyst development, and on sperm component remodeling and oocyte activation, in an equine model. Methods: In vitro-matured equine oocytes underwent conventional (Conv) or Piezo ICSI, the latter utilizing fluorocarbon ballast. Blastocyst development was compared between treatments to validate the model. Then, oocytes were fixed at 0, 6, or 18 h after injection, and stained for the sperm tail, acrosome, oocyte cortical granules, and chromatin. These parameters were compared between injection techniques and between sham-injected and sperm-injected oocytes among time periods. Results: Blastocyst rates were 39 and 40%. The nucleus number was lower, and the nuclear fragmentation rate was higher, in blastocysts produced by Conv. Cortical granule loss started at 0H after both sperm and sham injection. The acrosome was present at 0H in both ICSI treatments, and persisted to 18H in significantly more Conv than Piezo oocytes (72 vs. 21%). Sperm head area was unchanged at 6H in Conv but significantly increased at this time in Piezo; correspondingly, at 6H significantly more Conv than Piezo oocytes remained at MII (80 vs. 9.5%). Sham injection did not induce significant meiotic resumption. Conclusions: These data show that Piezo ICSI is associated with more rapid sperm component remodeling and oocyte meiotic resumption after sperm injection than is conventional ICSI, and with higher embryo quality at the blastocyst stage. This suggests that there is value in exploring the Piezo technique, utilized with a non-toxic fluorocarbon ballast, for use in clinical human ICSI.
Get Full Text
Publication Date: 2018-04-10 PubMed ID: 29637506PubMed Central: PMC5984897DOI: 10.1007/s10815-018-1174-9Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Comparative Study
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article is about how Piezo-driven Intracytoplasmic Sperm Injection (ICSI) shows an advantage over conventional ICSI in terms of sperm component remodeling, oocyte activation, and blastocyst quality in horses.

Research Objective

The objective of the study was to compare the impacts of conventional Intracytoplasmic Sperm Injection (ICSI) and Piezo-driven ICSI on blastocyst development, sperm component remodeling, and oocyte activation. The study used the equine model for these assessments.

Methods

  • Equine oocytes that had matured in vitro were subjected to either conventional (Conv) or Piezo ICSI, with the Piezo variant using fluorocarbon ballast.
  • The researchers compared blastocyst development between the two techniques to validate the model.
  • Oocytes were fixed at 0, 6, or 18 hours after injection, then stained for observing the sperm tail, acrosome, oocyte cortical granules, and chromatin.
  • The parameters such as the presence of acrosome, sperm head area, and state of oocyte (whether at Metaphase II – MII stage) at different timings after the injection were compared between the two methods.

Results

  • Both methods yielded comparable rates of blastocysts (39-40%).
  • Nevertheless, the blastocysts resulting from the Conv method had fewer nuclei and a higher nuclear fragmentation rate.
  • The loss of cortical granules started immediately after both sperm and sham injection.
  • Both methods presented the acrosome at the start (0H), but more Conv oocytes retained it up to 18H (72% Conv vs. 21% Piezo).
  • The sperm head area remained constant at 6H in Conv, but it showed a significant increase in Piezo.
  • Consequently, at 6H, a higher number of Conv oocytes remained at MII compared to Piezo.
  • Sham injection didn’t lead to significant meiotic resumption.

Conclusions

  • The study indicates that Piezo ICSI is associated with more rapid sperm component remodeling and oocyte meiotic resumption post-sperm injection than conventional ICSI.
  • The technique also results in higher embryo quality at the blastocyst stage.
  • These points favor exploring the Piezo technique, particularly with the use of a non-toxic fluorocarbon ballast, for clinical human ICSI applications.

Cite This Article

APA
Salgado RM, Brom-de-Luna JG, Resende HL, Canesin HS, Hinrichs K. (2018). Lower blastocyst quality after conventional vs. Piezo ICSI in the horse reflects delayed sperm component remodeling and oocyte activation. J Assist Reprod Genet, 35(5), 825-840. https://doi.org/10.1007/s10815-018-1174-9

Publication

Journal of assisted reproduction and genetics
ISSN: 1573-7330
NlmUniqueID: 9206495
Country: Netherlands
Language: English
Volume: 35
Issue: 5
Pages: 825-840

Researcher Affiliations

Salgado, R M
  • Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, TAMU 4466, College Station, TX, 77843-4466, USA.
Brom-de-Luna, J G
  • Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, TAMU 4466, College Station, TX, 77843-4466, USA.
Resende, H L
  • Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, TAMU 4466, College Station, TX, 77843-4466, USA.
Canesin, H S
  • Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, TAMU 4466, College Station, TX, 77843-4466, USA.
Hinrichs, Katrin
  • Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, TAMU 4466, College Station, TX, 77843-4466, USA. khinrichs@cvm.tamu.edu.

MeSH Terms

  • Acrosome / physiology
  • Animals
  • Blastocyst / cytology
  • Blastocyst / physiology
  • Chromatin
  • Female
  • Horses
  • In Vitro Oocyte Maturation Techniques / veterinary
  • Male
  • Oocytes / cytology
  • Oocytes / physiology
  • Sperm Injections, Intracytoplasmic / methods
  • Sperm Injections, Intracytoplasmic / veterinary
  • Spermatozoa / cytology
  • Spermatozoa / physiology

Conflict of Interest Statement

The authors declare that they have no conflict of interest.

References

This article includes 59 references
  1. Sherins RJ, Thorsell LP, Dorfmann A, Dennison-Lagos L, Calvo LP, Krysa L, Coulam CB, Schulman JD. Intracytoplasmic sperm injection facilitates fertilization even in the most severe forms of male infertility: pregnancy outcome correlates with maternal age and number of eggs available.. Fertil Steril 1995;64:369–375.
    doi: 10.1016/S0015-0282(16)57737-1pubmed: 7615116google scholar: lookup
  2. Salamone DF, Canel NG, Rodriguez MB. Intracytoplasmic sperm injection in domestic and wild mammals.. Reproduction 2017;154:F111–Ff24.
    doi: 10.1530/REP-17-0357pubmed: 29196493google scholar: lookup
  3. Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G. Ovum pick up, intracytoplasmic sperm injection and somatic cell nuclear transfer in cattle, buffalo and horses: from the research laboratory to clinical practice.. Theriogenology 2014;81:138–151.
    doi: 10.1016/j.theriogenology.2013.09.008pubmed: 24274418google scholar: lookup
  4. Dyer S, Chambers GM, de Mouzon J, Nygren KG, Zegers-Hochschild F, Mansour R, Ishihara O, Banker M, Adamson GD. International Committee for Monitoring Assisted Reproductive Technologies world report: assisted reproductive technology 2008, 2009 and 2010.. Hum Reprod 2016;31:1588–1609.
    doi: 10.1093/humrep/dew082pubmed: 27207175google scholar: lookup
  5. Schlegel PN, Girardi SK. Clinical review 87: In vitro fertilization for male factor infertility.. J Clin Endocrinol Metab 1997;82:709–716.
    doi: 10.1210/jcem.82.3.3785pubmed: 9062469google scholar: lookup
  6. Kimura Y, Yanagimachi R. Intracytoplasmic sperm injection in the mouse.. Biol Reprod 1995;52:709–720.
    doi: 10.1095/biolreprod52.4.709pubmed: 7779992google scholar: lookup
  7. Yanagida K, Katayose H, Hirata S, Yazawa H, Hayashi S, Sato A. Influence of sperm immobilization on onset of Ca(2+) oscillations after ICSI.. Hum Reprod 2001;16:148–152.
    doi: 10.1093/humrep/16.1.148pubmed: 11139554google scholar: lookup
  8. Katayama M, Sutovsky P, Yang BS, Cantley T, Rieke A, Farwell R, Oko R, Day BN. Increased disruption of sperm plasma membrane at sperm immobilization promotes dissociation of perinuclear theca from sperm chromatin after intracytoplasmic sperm injection in pigs.. Reproduction 2005;130:907–916.
    doi: 10.1530/rep.1.0680pubmed: 16322550google scholar: lookup
  9. Anzalone DA, Iuso D, Czernik M, Ptak G, Loi P. Plasma membrane and acrosome loss before ICSI is required for sheep embryonic development.. J Assist Reprod Genet 2016;33:757–763.
    doi: 10.1007/s10815-016-0709-1pmc: PMC4889485pubmed: 27059776google scholar: lookup
  10. Lacham-Kaplan O, Trounson A. Intracytoplasmic sperm injection in mice: increased fertilization and development to term after induction of the acrosome reaction.. Hum Reprod 1995;10:2642–2649.
    doi: 10.1093/oxfordjournals.humrep.a135760pubmed: 8567785google scholar: lookup
  11. Morozumi K, Shikano T, Miyazaki S, Yanagimachi R. Simultaneous removal of sperm plasma membrane and acrosome before intracytoplasmic sperm injection improves oocyte activation/embryonic development.. Proc Natl Acad Sci U S A 2006;103:17661–17666.
    doi: 10.1073/pnas.0608183103pmc: PMC1693803pubmed: 17090673google scholar: lookup
  12. Seita Y, Ito J, Kashiwazaki N. Removal of acrosomal membrane from sperm head improves development of rat zygotes derived from intracytoplasmic sperm injection.. J Reprod Dev 2009;55:475–479.
    doi: 10.1262/jrd.20216pubmed: 19444004google scholar: lookup
  13. Zambrano F, Aguila L, Arias ME, Sanchez R, Felmer R. Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100.. Theriogenology 2016;86:1489–1497.
    doi: 10.1016/j.theriogenology.2016.05.007pubmed: 27325573google scholar: lookup
  14. Yanagida K, Hatayose H, Yazawa H, Kimura Y, Konnai K, Sato A. The usefulness of a piezo-micromanipulator in intracytoplasmic sperm injection in humans.. Hum Reprod 1998;14:448–453.
    doi: 10.1093/humrep/14.2.448pubmed: 10099992google scholar: lookup
  15. Takeuchi S, Minoura H, Shibahara T, Shen X, Futamura N, Toyoda N. Comparison of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection into human oocytes.. Gynecol Obstet Investig 2001;52:158–162.
    doi: 10.1159/000052965pubmed: 11598356google scholar: lookup
  16. Hiraoka K, Kitamura S. Clinical efficiency of piezo-ICSI using micropipettes with a wall thickness of 0.625 mum.. J Assist Reprod Genet 2015;32:1827–1833.
    doi: 10.1007/s10815-015-0597-9pmc: PMC4681728pubmed: 26489413google scholar: lookup
  17. Galli C, Vassiliev I, Lagutina I, Galli A, Lazzari G. Bovine embryo development following ICSI: effect of activation, sperm capacitation and pre-treatment with dithiothreitol.. Theriogenology 2003;60:1467–1480.
    doi: 10.1016/S0093-691X(03)00133-Xpubmed: 14519468google scholar: lookup
  18. Garcia-Rosello E, Garcia-Mengual E, Coy P, Alfonso J, Silvestre MA. Intracytoplasmic sperm injection in livestock species: an update.. Reprod Domest Anim 2009;44:143–151.
    doi: 10.1111/j.1439-0531.2007.01018.xpubmed: 18954388google scholar: lookup
  19. Zhou X, Yin M, Jiang W, Jiang M, Li S, Li H, Chen X. Electrical activation of rabbit oocytes increases fertilization and embryo development by intracytoplasmic sperm injection using sperm from deceased male.. J Assist Reprod Genet 2013;30:1605–1610.
    doi: 10.1007/s10815-013-0113-zpmc: PMC3843171pubmed: 24114632google scholar: lookup
  20. Colleoni S, Barbacini S, Necchi D, Duchi R, Lazzari G, Galli C. Application of ovum pick-up, intracytoplasmic sperm injection and embryo culture in equine practice.. Proc Amer Assoc Equine Pract 2007;53:554–559.
  21. Hinrichs K, Choi YH, Norris JD, Love LB, Bedford-Guaus SJ, Hartman DL, Velez IC. 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 2012;241:1070–1074.
    doi: 10.2460/javma.241.8.1070pubmed: 23039983google scholar: lookup
  22. Hinrichs K, Choi YH, Love CC, Spacek S. Use of intracytoplasmic sperm injection and in vitro culture to the blastocyst stage in a commercial equine assisted reproduction program.. J Equine Vet Sci 2014;34:176.
    doi: 10.1016/j.jevs.2013.10.129google scholar: lookup
  23. Hinrichs K, Choi YH, Love LB, Varner DD, Love CC, Walckenaer BE. Chromatin configuration within the germinal vesicle of horse oocytes: changes post mortem and relationship to meiotic and developmental competence.. Biol Reprod 2005;72:1142–1150.
    doi: 10.1095/biolreprod.104.036012pubmed: 15647456google scholar: lookup
  24. Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G. Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.. Anim Reprod Sci 2007;98:39–55.
    doi: 10.1016/j.anireprosci.2006.10.011pubmed: 17101246google scholar: lookup
  25. Altermatt JL, Suh TK, Stokes JE, Carnevale EM. Effects of age and equine follicle-stimulating hormone (eFSH) on collection and viability of equine oocytes assessed by morphology and developmental competency after intracytoplasmic sperm injection (ICSI). Reprod Fertil Dev 2009;21:615–623.
    doi: 10.1071/RD08210pubmed: 19383268google scholar: lookup
  26. Foss R, Ortis H, Hinrichs K. Effect of potential oocyte transport protocols on blastocyst rates after intracytoplasmic sperm injection in the horse.. Equine Vet J 2013;45:39–43.
    doi: 10.1111/evj.12159pubmed: 24304402google scholar: lookup
  27. Smits K, Govaere J, Hoogewijs M, Piepers S, Van Soom A. A pilot comparison of laser-assisted vs piezo drill ICSI for the in vitro production of horse embryos.. Reprod Domest Anim 2012;47:e1–e3.
    doi: 10.1111/j.1439-0531.2011.01814.xpubmed: 21950451google scholar: lookup
  28. Alonso A, Baca Castex C, Ferrante A, Pinto M, Castaneira C, Trasorras V. In vitro equine embryo production using air-dried spermatozoa, with different activation protocols and culture systems.. Andrologia 2015;47:387–394.
    doi: 10.1111/and.12273pubmed: 24684246google scholar: lookup
  29. Dini P, Bogado Pascottini O, Ducheyne K, Hostens M, Daels P. Holding equine oocytes in a commercial embryo-holding medium: new perspective on holding temperature and maturation time.. Theriogenology 2016;86:1361–1368.
    doi: 10.1016/j.theriogenology.2016.04.079pubmed: 27268297google scholar: lookup
  30. Tremoleda JL, Van Haeften T, Stout TA, Colenbrander B, Bevers MM. Cytoskeleton and chromatin reorganization in horse oocytes following intracytoplasmic sperm injection: patterns associated with normal and defective fertilization.. Biol Reprod 2003;69:186–194.
    doi: 10.1095/biolreprod.102.012823pubmed: 12646492google scholar: lookup
  31. Smits K, Govaere J, Peelman LJ, Goossens K, de Graaf DC, Vercauteren D, Vandaele L, Hoogewijs M, Wydooghe E, Stout T, van Soom A. Influence of the uterine environment on the development of in vitro-produced equine embryos.. Reproduction 2012;143:173–181.
    doi: 10.1530/REP-11-0217pubmed: 22089531google scholar: lookup
  32. Carnevale EM. The mare model for follicular maturation and reproductive aging in the woman.. Theriogenology 2008;69:23–30.
    doi: 10.1016/j.theriogenology.2007.09.011pubmed: 17976712google scholar: lookup
  33. Ginther OJ. The mare: a 1000-pound guinea pig for study of the ovulatory follicular wave in women.. Theriogenology 2012;77:818–828.
    doi: 10.1016/j.theriogenology.2011.09.025pubmed: 22115815google scholar: lookup
  34. Jacobson CC, Choi YH, Hayden SS, Hinrichs K. Recovery of mare oocytes on a fixed biweekly schedule, and resulting blastocyst formation after intracytoplasmic sperm injection.. Theriogenology 2010;73:1116–1126.
    doi: 10.1016/j.theriogenology.2010.01.013pubmed: 20202674google scholar: lookup
  35. Choi YH, Love LB, Varner DD, Hinrichs K. Holding immature equine oocytes in the absence of meiotic inhibitors: effect on germinal vesicle chromatin and blastocyst development after intracytoplasmic sperm injection.. Theriogenology 2006;66:955–963.
    doi: 10.1016/j.theriogenology.2006.01.064pubmed: 16574209google scholar: lookup
  36. Martino NA, Dell'Aquila ME, Filioli Uranio M, Rutigliano L, Nicassio M, Lacalandra GM. Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential.. Reprod Biol Endocrinol 2014;12:99.
    doi: 10.1186/1477-7827-12-99pmc: PMC4209075pubmed: 25306508google scholar: lookup
  37. Yoshida M, Cran DG, Pursel VG. Confocal and fluorescence microscopic study using lectins of the distribution of cortical granules during the maturation and fertilization of pig oocytes.. Mol Reprod Dev 1993;36:462–468.
    doi: 10.1002/mrd.1080360409pubmed: 8305208google scholar: lookup
  38. Mortimer D, Curtis EF, Miller RG. Specific labelling by peanut agglutinin of the outer acrosomal membrane of the human spermatozoon.. J Reprod Fertil 1987;81:127–135.
    doi: 10.1530/jrf.0.0810127pubmed: 3118011google scholar: lookup
  39. Ruggeri E, DeLuca KF, Galli C, Lazzari G, DeLuca JG, Stokes JE. Use of confocal microscopy to evaluate equine zygote development after sperm injection of oocytes matured in vivo or in vitro.. Microsc Microanal 2017;23:1197–1206.
    doi: 10.1017/S1431927617012740pmc: PMC5976488pubmed: 29208065google scholar: lookup
  40. Faramarzi A, Khalili MA, Micara G, Agha-Rahimi A. Revealing the secret life of pre-implantation embryos by time-lapse monitoring: a review.. Int J Reprod Biomed (Yazd) 2017;15:257–264.
    doi: 10.29252/ijrm.15.5.257pmc: PMC5510578pubmed: 28744520google scholar: lookup
  41. Gomez-Torres MJ, Ten J, Girela JL, Romero J, Bernabeu R, De Juan J. Sperm immobilized before intracytoplasmic sperm injection undergo ultrastructural damage and acrosomal disruption.. Fertil Steril 2007;88:702–704.
    doi: 10.1016/j.fertnstert.2006.12.063pubmed: 17548082google scholar: lookup
  42. Takeuchi T, Colombero LT, Neri QV, Rosenwaks Z, Palermo GD. Does ICSI require acrosomal disruption? An ultrastructural study.. Hum Reprod 2004;19:114–117.
    doi: 10.1093/humrep/deg511pubmed: 14688168google scholar: lookup
  43. Ramalho-Santos J, Sutovsky P, Simerly C, Oko R, Wessel GM, Hewitson L. ICSI choreography: fate of sperm structures after monospermic rhesus ICSI and first cell cycle implications.. Hum Reprod 2000;15:2610–2620.
    doi: 10.1093/humrep/15.12.2610pubmed: 11098035google scholar: lookup
  44. Katayama M, Koshida M, Miyake M. Fate of the acrosome in ooplasm in pigs after IVF and ICSI.. Hum Reprod 2002;17:2657–2664.
    doi: 10.1093/humrep/17.10.2657pubmed: 12351545google scholar: lookup
  45. Sathananthan AH, Szell A, Ng SC, Kausche A, Lacham-Kaplan O, Trounson A. Is the acrosome reaction a prerequisite for sperm incorporation after intra-cytoplasmic sperm injection (ICSI)?. Reprod Fertil Dev 1997;9:703–709.
    doi: 10.1071/R97028pubmed: 9623490google scholar: lookup
  46. Bourgain C, Nagy ZP, De Zutter H, Van Ranst H, Nogueira D, Van Steirteghem AC. Ultrastructure of gametes after intracytoplasmic sperm injection.. Hum Reprod 1998;13(Suppl 1):107–116.
    doi: 10.1093/humrep/13.suppl_1.107pubmed: 9663775google scholar: lookup
  47. Sutovsky P, Hewitson L, Simerly CR, Tengowski MW, Navara CS, Haavisto A, Schatten G. Intracytoplasmic sperm injection for rhesus monkey fertilization results in unusual chromatin, cytoskeletal, and membrane events, but eventually leads to pronuclear development and sperm aster assembly.. Hum Reprod 1996;11:1703–1712.
    doi: 10.1093/oxfordjournals.humrep.a019473pubmed: 8921120google scholar: lookup
  48. Enders AC, Liu IK, Bowers J, Lantz KC, Schlafke S, Suarez S. The ovulated ovum of the horse: cytology of nonfertilized ova to pronuclear stage ova.. Biol Reprod 1987;37:453–466.
    doi: 10.1095/biolreprod37.2.453pubmed: 3676399google scholar: lookup
  49. Abbott AL, Ducibella T. Calcium and the control of mammalian cortical granule exocytosis.. Front Biosci 2001;6:D792–D806.
    doi: 10.2741/Abbottpubmed: 11438440google scholar: lookup
  50. Liu M. The biology and dynamics of mammalian cortical granules.. Reprod Biol Endocrinol : RB & E 2011;9:149.
    doi: 10.1186/1477-7827-9-149pmc: PMC3228701pubmed: 22088197google scholar: lookup
  51. Bello OD, Cappa AI, de Paola M, Zanetti MN, Fukuda M, Fissore RA, Mayorga LS, Michaut MA. Rab3A, a possible marker of cortical granules, participates in cortical granule exocytosis in mouse eggs.. Exp Cell Res 2016;347:42–51.
    doi: 10.1016/j.yexcr.2016.07.005pmc: PMC6460285pubmed: 27423421google scholar: lookup
  52. Sun FZ, Bradshaw JP, Galli C, Moor RM. Changes in intracellular calcium concentration in bovine oocytes following penetration by spermatozoa.. J Reprod Fertil 1994;101:713–719.
    doi: 10.1530/jrf.0.1010713pubmed: 7966030google scholar: lookup
  53. Fujimoto S, Yoshida N, Fukui T, Amanai M, Isobe T, Itagaki C, Izumi T, Perry ACF. Mammalian phospholipase Cζ induces oocyte activation from the sperm perinuclear matrix.. Dev Biol 2004;274:370–383.
    doi: 10.1016/j.ydbio.2004.07.025pubmed: 15385165google scholar: lookup
  54. Bedford-Guaus SJ, McPartlin LA, Xie J, Westmiller SL, Buffone MG, Roberson MS. Molecular cloning and characterization of phospholipase C zeta in equine sperm and testis reveals species-specific differences in expression of catalytically active protein.. Biol Reprod 2011;85:78–88.
    doi: 10.1095/biolreprod.110.089466pubmed: 21389344google scholar: lookup
  55. Bedford-Guaus SJ, Yoon SY, Fissore RA, Choi YH, Hinrichs K. Microinjection of mouse phospholipase C zeta complementary RNA into mare oocytes induces long-lasting intracellular calcium oscillations and embryonic development.. Reprod Fertil Dev 2008;20:875–883.
    doi: 10.1071/RD08115pubmed: 19007551google scholar: lookup
  56. Betteridge KJ, Eaglesome MD, Mitchell D, Flood PF, Bériault R. Development of horse embryos up to twenty two days after ovulation: observations on fresh specimens.. J Anat 1982;135:191–209.
    pmc: PMC1168142pubmed: 7130052
  57. Hinrichs K, Love CC, Brinsko SP, Choi YH, Varner DD. In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.. Biol Reprod 2002;67:256–262.
    doi: 10.1095/biolreprod67.1.256pubmed: 12080025google scholar: lookup
  58. Bézard J, Magistrini M, Duchamp G, Palmer E. Chronology of equine fertilisation and embryonic development in vivo and in vitro.. Equine Vet J 1989;Supp 8:105–10.
  59. Palmer E, Bézard J, Magistrini M, Duchamp G. In vitro fertilisation in the horse: a retrospective study.. J Reprod Fertil 1991;44(Supp):375–384.
    pubmed: 1795281
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.