FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Animals (Basel) / Sexual Differentiation and Primordial Germ Cell Distribution...
Animals : an open access journal from MDPI2021; 11(8); 2422; doi: 10.3390/ani11082422

Sexual Differentiation and Primordial Germ Cell Distribution in the Early Horse Fetus.

Abstract: It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy were detected during this time. Tubular structures of the fetal testes were present from day 50 onwards, whereas cell clusters dominated in the fetal ovary. The genital ducts were well-differentiated and presented a lumen in all samples. No sign of mesonephric or paramesonephric duct degeneration was detected. Expression of AMH was strong in the fetal testes but absent in ovaries. Irrespective of sex, primordial germ cells selectively expressed LIN28. Migration of primordial germ cells from the mesonephros to the gonad was detected at 45 days, but not at 60 days of development. Their number and distribution within the gonad were influenced ( < 0.05) by fetal sex. Most primordial germ cells (86.8 ± 3.2% in females and 84.6 ± 4.7% in males) were characterized as pluripotent according to co-localization with CD117. However, only a very small percentage of primordial germ cells were proliferating (7.5 ± 1.7% in females and 3.2 ± 1.2% in males) based on co-localization with Ki67. It can be concluded that gonadal sexual differentiation in the horse occurs asynchronously with regard to sex but already before 45 days of gestation.
Get Full Text
Publication Date: 2021-08-17 PubMed ID: 34438878PubMed Central: PMC8388682DOI: 10.3390/ani11082422Google 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
  • 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 study investigates the development of reproductive organs in horse fetuses during the period of sexual differentiation. It especially focuses on the identification and tracking of primordial germ cells, which are the precursors to both sperm and egg cells.

Objective and Methodology

  • The research aimed to understand and document the growth patterns of the gonads, the primary sexual organs, and the genital ducts in horse fetuses in the period surrounding sexual differentiation. Additionally, it sought to identify and track primordial germ cells.
  • Equine fetuses between 45 and 60 days of gestation were studied with a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. These techniques allowed the researchers to observe detailed structural changes and detect specific cell types and molecules.

Key Findings

  • The size of the fetal gonads increased 23-fold from 45 to 60 days of gestation. Alongside this growth, the researchers noticed a considerable increase in the metanephros volume, an early stage of the kidney.
  • Signs of mesonephros atrophy, or wasting away of an early stage of kidney development, were identified during this timeframe. This suggests that the formation of the gonads and the metanephros is accompanied by the reduction of the mesonephros structure.
  • Different growth patterns were observed in the testes and ovaries. Tubular structures were present in the testes from day 50 onwards, whereas cell clusters dominated in the ovaries.
  • Genital ducts – the structures that will develop into the tubes that carry sperm or eggs – were well-differentiated and exhibited a lumen, or central cavity, in all samples. The researchers found no sign of degeneration of two types of ducts associated with kidney and reproductive organ development, the mesonephric and paramesonephric ducts.

Primordial Germ Cell Observations

  • Primordial germ cells, the precursors to sperm and egg cells, were tracked using their selective expression of LIN28, a protein associated with cell proliferation and development. Their migration from the mesonephros to the gonad was detected at 45 days, but not at 60 days of development. The distribution and count of these cells in the gonad were found to be influenced by the sex of the fetus.
  • A large majority of primordial germ cells showed signs of pluripotency, having the potential to differentiate into various types of cells, according to their co-localization with CD117, a protein associated with stem cells.
  • A small percentage of these cells were found to be proliferating, or multiplying, based on their co-localization with Ki67, a protein associated with cell division.

Conclusion

  • The research concludes that gonadal sexual differentiation in horses occurs asynchronously with regard to sex and begins before 45 days of gestation.

Cite This Article

APA
Scarlet D, Handschuh S, Reichart U, Podico G, Ellerbrock RE, Demyda-Peyrás S, Canisso IF, Walter I, Aurich C. (2021). Sexual Differentiation and Primordial Germ Cell Distribution in the Early Horse Fetus. Animals (Basel), 11(8), 2422. https://doi.org/10.3390/ani11082422

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 11
Issue: 8
PII: 2422

Researcher Affiliations

Scarlet, Dragos
  • Obstetrics, Gynecology and Andrology, Department for Small Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
  • Institute of Veterinary Anatomy and Clinic of Reproductive Medicine, Vetsuisse Faculty Zürich, Winterthurerstrasse 260, 8057 Zürich, Switzerland.
Handschuh, Stephan
  • Vetcore Facility for Research, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
Reichart, Ursula
  • Vetcore Facility for Research, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
Podico, Giorgia
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
  • Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
Ellerbrock, Robyn E
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
  • Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
Demyda-Peyrás, Sebastián
  • Department of Animal Production, School of Veterinary Sciences, National University of La Plata and CONICET CCT-La Plata, Calle 60 and 118 S/N, 1900 La Plata, Argentina.
Canisso, Igor F
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
  • Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
Walter, Ingrid
  • Vetcore Facility for Research, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
  • Institute of Pathology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
Aurich, Christine
  • Center for Artificial Insemination and Embryo Transfer, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 47 references
  1. Sinclair AH, Berta P, Palmer MS, Hawkins JR, Griffiths BL, Smith MJ, Foster JW, Frischauf AM, Lovell-Badge R, Goodfellow PN. A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif.. Nature 1990 Jul 19;346(6281):240-4.
    doi: 10.1038/346240a0pubmed: 1695712google scholar: lookup
  2. Wilhelm D, Palmer S, Koopman P. Sex determination and gonadal development in mammals.. Physiol Rev 2007 Jan;87(1):1-28.
    doi: 10.1152/physrev.00009.2006pubmed: 17237341google scholar: lookup
  3. Ungewitter EK, Yao HH. How to make a gonad: cellular mechanisms governing formation of the testes and ovaries.. Sex Dev 2013;7(1-3):7-20.
    doi: 10.1159/000338612pmc: PMC3474884pubmed: 22614391google scholar: lookup
  4. Gier HT, Marion GB. Development of mammalian testes and genital ducts.. Biol Reprod 1969 Jun;1:Suppl 1:1-23.
    doi: 10.1095/biolreprod1.Supplement_1.1pubmed: 5406326google scholar: lookup
  5. Kerr CL, Hill CM, Blumenthal PD, Gearhart JD. Expression of pluripotent stem cell markers in the human fetal ovary.. Hum Reprod 2008 Mar;23(3):589-99.
    doi: 10.1093/humrep/dem411pubmed: 18203707google scholar: lookup
  6. Mamsen LS, Ernst EH, Borup R, Larsen A, Olesen RH, Ernst E, Anderson RA, Kristensen SG, Andersen CY. Temporal expression pattern of genes during the period of sex differentiation in human embryonic gonads.. Sci Rep 2017 Nov 21;7(1):15961.
    doi: 10.1038/s41598-017-15931-3pmc: PMC5698446pubmed: 29162857google scholar: lookup
  7. McCoard SA, Wise TH, Fahrenkrug SC, Ford JJ. Temporal and spatial localization patterns of Gata4 during porcine gonadogenesis.. Biol Reprod 2001 Aug;65(2):366-74.
    doi: 10.1095/biolreprod65.2.366pubmed: 11466202google scholar: lookup
  8. Allen WR. Fetomaternal interactions and influences during equine pregnancy.. Reproduction 2001 Apr;121(4):513-27.
    doi: 10.1530/rep.0.1210513pubmed: 11277870google scholar: lookup
  9. Canisso IF, Ball BA, Esteller-Vico A, Williams NM, Squires EL, Troedsson MH. Changes in maternal androgens and oestrogens in mares with experimentally-induced ascending placentitis.. Equine Vet J 2017 Mar;49(2):244-249.
    doi: 10.1111/evj.12556pubmed: 26729310google scholar: lookup
  10. Legacki EL, Ball BA, Corbin CJ, Loux SC, Scoggin KE, Stanley SD, Conley AJ. Equine fetal adrenal, gonadal and placental steroidogenesis.. Reproduction 2017 Oct;154(4):445-454.
    doi: 10.1530/REP-17-0239pubmed: 28878092google scholar: lookup
  11. Pashen RL, Sheldrick EL, Allen WR, Flint AP. Dehydroepiandrosterone synthesis by the fetal foal and its importance as an oestrogen precursor.. J Reprod Fertil Suppl 1982;32:389-97.
    pubmed: 6220147
  12. Walt ML, Stabenfeldt GH, Hughes JP, Neely DP, Bradbury R. Development of the equine ovary and ovulation fossa.. J Reprod Fertil Suppl 1979;(27):471-7.
    pubmed: 289826
  13. Barreto RSN, Romagnolli P, Mess AM, Rigoglio NN, Sasahara THC, Simões LS, Fratini P, Matias GSS, Jacob JCF, Gastal EL, Miglino MA. Reproductive system development in male and female horse embryos and fetuses: Gonadal hyperplasia revisited.. Theriogenology 2018 Mar 1;108:118-126.
    doi: 10.1016/j.theriogenology.2017.11.036pubmed: 29207292google scholar: lookup
  14. Aurich C, Schneider J. Sex determination in horses - current status and future perspectives.. Anim Reprod Sci 2014 Apr;146(1-2):34-41.
    doi: 10.1016/j.anireprosci.2014.01.014pubmed: 24598214google scholar: lookup
  15. Kenngott RA, Vermehren M, Ebach K, Sinowatz F. The role of ovarian surface epithelium in folliculogenesis during fetal development of the bovine ovary: a histological and immunohistochemical study.. Sex Dev 2013;7(4):180-95.
    doi: 10.1159/000348881pubmed: 23571709google scholar: lookup
  16. Gall L, De Smedt V, Ruffini S. Co-Expression of Cytokeratins and Vimentin in Sheep Cumulus-Oocyte Complexes. Alteration of Intermediate Filament Distribution by Acrylamide.. Dev. Growth Differ. 1992;34:579–587.
    doi: 10.1111/j.1440-169X.1992.00579.xgoogle scholar: lookup
  17. Kenngott RA, Sauer U, Vermehren M, Sinowatz F. Expression of Intermediate Filaments and Germ Cell Markers in the Developing Bovine Ovary: An Immunohistochemical and Laser-Assisted Microdissection Study.. Cells Tissues Organs 2014;200(2):153-70.
    doi: 10.1159/000369203pubmed: 25999369google scholar: lookup
  18. Martinovic V, Vukusic Pusic T, Restovic I, Bocina I, Filipovic N, Saraga-Babic M, Vukojevic K. Expression of Epithelial and Mesenchymal Differentiation Markers in the Early Human Gonadal Development.. Anat Rec (Hoboken) 2017 Jul;300(7):1315-1326.
    doi: 10.1002/ar.23531pubmed: 27981799google scholar: lookup
  19. Fröjdman K, Ekblom P, Sorokin L, Yagi A, Pelliniemi LJ. Differential distribution of laminin chains in the development and sex differentiation of mouse internal genitalia.. Int J Dev Biol 1995 Apr;39(2):335-44.
    pubmed: 7669546
  20. Fröjdman K, Miner JH, Sanes JR, Pelliniemi LJ, Virtanen I. Sex-specific localization of laminin alpha 5 chain in the differentiating rat testis and ovary.. Differentiation 1999 Mar;64(3):151-9.
    doi: 10.1046/j.1432-0436.1999.6430151.xpubmed: 10234812google scholar: lookup
  21. Terauchi KJ, Shigeta Y, Iguchi T, Sato T. Role of Notch signaling in granulosa cell proliferation and polyovular follicle induction during folliculogenesis in mouse ovary.. Cell Tissue Res 2016 Jul;365(1):197-208.
    doi: 10.1007/s00441-016-2371-4pubmed: 26899251google scholar: lookup
  22. Inomata T, Inoue S, Sugawara H, Kajihara H, Shinomiya T, Wagai I, Ninomiya H, Oshida T, Shirai M, Hashimoto Y. Developmental changes in paramesonephric and mesonephric ducts and the external genitalia in swine fetuses during sexual differentiation.. J Vet Med Sci 1993 Jun;55(3):371-8.
    doi: 10.1292/jvms.55.371pubmed: 8357908google scholar: lookup
  23. Gruenwald P. The relation of the growing müllerian duct to the wolffian duct and its importance for the genesis of malformations.. Anat. Rec. 1941;81:1–19.
    doi: 10.1002/ar.1090810102google scholar: lookup
  24. Kenngott RA, Sinowatz F. Expression and distribution of intermediate-filament proteins and laminin during the development of the bovine Müllerian duct.. Anat Histol Embryol 2008 Jun;37(3):223-30.
    doi: 10.1111/j.1439-0264.2007.00835.xpubmed: 18241300google scholar: lookup
  25. Mullen RD, Behringer RR. Molecular genetics of Müllerian duct formation, regression and differentiation.. Sex Dev 2014;8(5):281-96.
    doi: 10.1159/000364935pmc: PMC4378544pubmed: 25033758google scholar: lookup
  26. Orvis GD, Behringer RR. Cellular mechanisms of Müllerian duct formation in the mouse.. Dev Biol 2007 Jun 15;306(2):493-504.
    doi: 10.1016/j.ydbio.2007.03.027pmc: PMC2730733pubmed: 17467685google scholar: lookup
  27. Childs AJ, Cowan G, Kinnell HL, Anderson RA, Saunders PT. Retinoic Acid signalling and the control of meiotic entry in the human fetal gonad.. PLoS One 2011;6(6):e20249.
    doi: 10.1371/journal.pone.0020249pmc: PMC3108594pubmed: 21674038google scholar: lookup
  28. Houmard B, Small C, Yang L, Naluai-Cecchini T, Cheng E, Hassold T, Griswold M. Global gene expression in the human fetal testis and ovary.. Biol Reprod 2009 Aug;81(2):438-43.
    doi: 10.1095/biolreprod.108.075747pmc: PMC2849816pubmed: 19369649google scholar: lookup
  29. Childs AJ, Kinnell HL, He J, Anderson RA. LIN28 is selectively expressed by primordial and pre-meiotic germ cells in the human fetal ovary.. Stem Cells Dev 2012 Sep 1;21(13):2343-9.
    doi: 10.1089/scd.2011.0730pmc: PMC3424972pubmed: 22296229google scholar: lookup
  30. Gillis AJ, Stoop H, Biermann K, van Gurp RJ, Swartzman E, Cribbes S, Ferlinz A, Shannon M, Oosterhuis JW, Looijenga LH. Expression and interdependencies of pluripotency factors LIN28, OCT3/4, NANOG and SOX2 in human testicular germ cells and tumours of the testis.. Int J Androl 2011 Aug;34(4 Pt 2):e160-74.
    doi: 10.1111/j.1365-2605.2011.01148.xpubmed: 21631526google scholar: lookup
  31. West JA, Viswanathan SR, Yabuuchi A, Cunniff K, Takeuchi A, Park IH, Sero JE, Zhu H, Perez-Atayde A, Frazier AL, Surani MA, Daley GQ. A role for Lin28 in primordial germ-cell development and germ-cell malignancy.. Nature 2009 Aug 13;460(7257):909-13.
    doi: 10.1038/nature08210pmc: PMC2729657pubmed: 19578360google scholar: lookup
  32. Enriquez VA, Cleys ER, Da Silveira JC, Spillman MA, Winger QA, Bouma GJ. High LIN28A Expressing Ovarian Cancer Cells Secrete Exosomes That Induce Invasion and Migration in HEK293 Cells.. Biomed Res Int 2015;2015:701390.
    doi: 10.1155/2015/701390pmc: PMC4637063pubmed: 26583126google scholar: lookup
  33. Aeckerle N, Eildermann K, Drummer C, Ehmcke J, Schweyer S, Lerchl A, Bergmann M, Kliesch S, Gromoll J, Schlatt S, Behr R. The pluripotency factor LIN28 in monkey and human testes: a marker for spermatogonial stem cells?. Mol Hum Reprod 2012 Oct;18(10):477-88.
    doi: 10.1093/molehr/gas025pmc: PMC3457707pubmed: 22689537google scholar: lookup
  34. Curran S, Urven L, Ginther OJ. Distribution of putative primordial germ cells in equine embryos.. Equine Vet J Suppl 1997 Dec;(25):72-6.
    doi: 10.1111/j.2042-3306.1997.tb05105.xpubmed: 9593533google scholar: lookup
  35. Lee G, Jung H, Yoon M. The Lin28 Expression in Stallion Testes.. PLoS One 2016;11(10):e0165011.
    doi: 10.1371/journal.pone.0165011pmc: PMC5087892pubmed: 27798668google scholar: lookup
  36. Jung H, Song H, Yoon M. The KIT is a putative marker for differentiating spermatogonia in stallions.. Anim Reprod Sci 2015 Jan;152:39-46.
    doi: 10.1016/j.anireprosci.2014.11.004pubmed: 25435078google scholar: lookup
  37. Podico G, Canisso IF, Ellerbrock RE, Dias NW, Mercadante VRG, Lima FS. Assessment of peripheral markers and ultrasonographic parameters in pregnant mares receiving intramuscular or intrauterine cloprostenol.. Theriogenology 2020 Jan 15;142:77-84.
    doi: 10.1016/j.theriogenology.2019.09.025pubmed: 31581046google scholar: lookup
  38. Okada CTC, Kaps M, Scarlet D, Handschuh S, Gautier C, Melchert M, Aurich J, Aurich C. Low plasma progestin concentration during the early postovulatory phase impairs equine conceptus development in the late preimplantation phase.. Reprod. Fertil. Dev. 2020;32:1156–1167.
    doi: 10.1071/RDv32n2Ab61google scholar: lookup
  39. Ball BA, Conley AJ, MacLaughlin DT, Grundy SA, Sabeur K, Liu IK. Expression of anti-Müllerian hormone (AMH) in equine granulosa-cell tumors and in normal equine ovaries.. Theriogenology 2008 Oct 1;70(6):968-77.
    doi: 10.1016/j.theriogenology.2008.05.059pubmed: 18599114google scholar: lookup
  40. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A. Fiji: an open-source platform for biological-image analysis.. Nat Methods 2012 Jun 28;9(7):676-82.
    doi: 10.1038/nmeth.2019pmc: PMC3855844pubmed: 22743772google scholar: lookup
  41. Anaya G, Molina A, Valera M, Moreno-Millán M, Azor P, Peral-García P, Demyda-Peyrás S. Sex chromosomal abnormalities associated with equine infertility: validation of a simple molecular screening tool in the Purebred Spanish Horse.. Anim Genet 2017 Aug;48(4):412-419.
    doi: 10.1111/age.12543pubmed: 28224649google scholar: lookup
  42. DeFalco T, Takahashi S, Capel B. Two distinct origins for Leydig cell progenitors in the fetal testis.. Dev Biol 2011 Apr 1;352(1):14-26.
    doi: 10.1016/j.ydbio.2011.01.011pmc: PMC3055913pubmed: 21255566google scholar: lookup
  43. Tilmann C, Capel B. Mesonephric cell migration induces testis cord formation and Sertoli cell differentiation in the mammalian gonad.. Development 1999 Jul;126(13):2883-90.
    doi: 10.1242/dev.126.13.2883pubmed: 10357932google scholar: lookup
  44. Inomata T, Eguchi Y, Nakamura T. Origin of müllerian duct and its later developmental changes in relation to wolffian duct in bovine fetuses.. Zentralbl Veterinarmed A 1989 Mar;36(3):166-74.
    doi: 10.1111/j.1439-0442.1989.tb00717.xpubmed: 2499995google scholar: lookup
  45. Jacob M, Christ B, Jacob HJ, Poelmann RE. The role of fibronectin and laminin in development and migration of the avian Wolffian duct with reference to somitogenesis.. Anat Embryol (Berl) 1991;183(4):385-95.
    doi: 10.1007/BF00196840pubmed: 1867390google scholar: lookup
  46. Kobayashi A, Stewart CA, Wang Y, Fujioka K, Thomas NC, Jamin SP, Behringer RR. β-Catenin is essential for Müllerian duct regression during male sexual differentiation.. Development 2011 May;138(10):1967-75.
    doi: 10.1242/dev.056143pmc: PMC3082302pubmed: 21490063google scholar: lookup
  47. Esteves CL, Sharma R, Dawson L, Taylor SE, Pearson G, Keen JA, McDonald K, Aurich C, Donadeu FX. Expression of putative markers of pluripotency in equine embryonic and adult tissues.. Vet J 2014 Dec;202(3):533-5.
    doi: 10.1016/j.tvjl.2014.08.026pubmed: 25241949google scholar: lookup

Citations

This article has been cited 1 times.
  1. Handschuh S, Glösmann M. Mouse embryo phenotyping using X-ray microCT.. Front Cell Dev Biol 2022;10:949184.
    doi: 10.3389/fcell.2022.949184pubmed: 36187491google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.