Chicken primordial germ cells (PGCs) dynamically migrate towards the prospective gonadal area through the germinal crescent region and bloodstream at early embryonic stages. To date, chicken PGCs have been mainly identified by histochemical and immunohistochemical methods or by their morphological characteristics. However, their origin, migration and differentiation are not fully understood because of the lack of specific PGC molecular markers. Here, we have isolated the chicken dead end homologue (CDH) in order to clone its full-length cDNA with an open reading frame of 329 amino acids. The RNA-binding motif present in CDH at amino acids 54–133 was highly homologous to those in the dead end proteins of human, mouse and Xenopus. The temporal and spatial distribution of PGCs was also investigated by in situ hybridization (ISH) on whole-mount embryos with CDH cRNA as a probe. Chicken embryos from stage X to stage 20 were subjected to ISH. The hybridized samples were then sectioned to analyse the translocation of PGCs. CDH-positive cells could be counted from stage X to stage 4, with minimally 30 cells at the blastderm and approximately 260 cells at the germinal crescent. Thus, specific expression of CDH mRNA has been established in chicken PGCs located at the blastderm, germinal crescent and prospective gonadal area by ISH and reverse transcription/polymerase chain reaction. We conclude that isolated CDH is specifically expressed in chicken PGCs during embryogenesis.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Aramaki S, Sato F, Soh T, Yamauchi N, Sakai T, Hattori M-A (2005) Temporal and spatial expression of TGF-β2 in chicken somites during early embryonic development. J Exp Zool 303:323–330
Clawson RC, Domm LV (1963) Developmental changes in glycogen content of primordial germ cells in chick embryo. Proc Soc Exp Biol Med 112:533–537
Clinton M, Haines L, Belloir B, McBride D (2001) Sexing chick embryos: a rapid and simple protocol. Br Poult Sci 42:134–138
Eyal-Giladi H, Kochav S (1976) From cleavage to primitive streak formation: a complementary normal table and a new look at the first stages of the development of the chick. I. General morphology. Dev Biol 49:321–337
Fabian B, Eyal-Giladi H (1981) A SEM study of cell shedding during the formation of the area pellucida in the chick embryo. J Embryol Exp Morphol 64:11–22
Fox M, Damjanov I, Martinez-Hernandez A, Knowles BB, Solter D (1981) Immunohistochemical localization of the early embryonic antigen (SSEA1) in postimplantation mouse embryos and foetal and adult tissues. Dev Biol 83:391–398
Fujimoto T, Ninomiya T, Ukeshima A (1976a) Observations of primordial germ-cells in blood samples from chick-embryo. Dev Biol 49:278–282
Fujimoto T, Ukeshima A, Kiyofuji R (1976b) The origin, migration and morphology of the primordial germ cells in the chick embryo. Anat Rec 185:139–145
Fujioka T, Soh T, Fujihara N, Hattori M-A (2004) Function of TGF-β2 in the growth of chicken primordial germ cells and geminal ridge stroma cells during embryonic development. J Exp Zool 301A:290–296
Ginsburg M (1997) Primordial germ cell development in avians. Poult Sci 76:91–95
Ginsburg M, Eyal-Giladi H (1986) Temporal and spatial aspects of the gradual migration of primordial germ cells from the epiblast into the germinal crescent in the avian embryo. J Embryol Exp Morphol 95:53–71
Ginsburg M, Eyal-Giladi H (1987) Primordial germ cells of the young chick blastderm originate from the central zone of the area pellucida irrespective of the embryo-forming process. Development 101:209–219
Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Karagenc L, Cinnamon Y, Ginsburg M, Petitte JN (1996) Origin of primordial germ cells in the prestreak chick embryo. Dev Genet 19:290–301
Kuwana T, Rogulska T (1999) Migratory mechanisms of chick primordial germ cells toward gonadal anlage. Cell Mol Biol (Noisy-le-grand) 45:725–736
Kuwana T, Maeda-Suga H, Fujimoto T (1986) Attraction of chick primordial germ cells by gonadal anlage in vitro. Anat Rec 215:403–406
Nakabayashi O, Kikuchi H, Kikuchi T, Mizuno S (1998) Differential expression of genes for aromatase and estrogen receptor during the gonadal development in chicken embryos. J Mol Endocrinol 20:193–202
Nieukoop PD, Satasurya LA (1979) Primordial germ cells in the chordates. Embryogenesis and Phylogenesis. Cambridge University Press, Cambridge, pp 118–123
Meyer DB (1960) Application of the periodic acid Schiff technique to whole chick embryos. Stain Technol 35:83–89
Meyer DB (1964) The migration of primordial germ cells in the chick embryo. Dev Biol 10:154–190
Pain B, Clark ME, Shen M, Nakazawa H, Sakurai M, Samarut J, Etches RJ (1996) Long-term in vitro culture and characterisation of avian embryonic stem cells with multiple morphogenetic potentialities. Development 122:2339–2348
Sato F, Kurokawa M, Yamauchi N, Hattori M-A (2006) Gene silencing of myostatin in differentiation of chicken embryonic myoblasts by small interfering RNA. Am J Physiol Cell Physiol 291:C538–C545
Solter D, Knowles BB (1978) Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1). Proc Natl Acad Sci USA 75:5565–5569
Stebler J, Spieler D, Slanchev K, Molyneaux KA, Richter U, Cojocaru V, Tarabykin V, Wylie C, Kessel M, Raz E (2004) Primordial germ cell migration in the chick and mouse embryo: the role of the chemokine SDF-1/CXCL12. Dev Biol 272:351–361
Tajima A, Barbato GF, Kuwana T, Hammerstedt RH (2003) Conservation of a genetically selected broiler line (42L) using cryopreserved circulating primordial germ cells (PGCs) isolated by filtration method. J Poult Sci 40:53–61
Tsuda M, Sasaoka Y, Kiso M, Abe K, Haraguchi S, Kobayashi S, Saga Y (2003) Conserved role of nanos proteins in germ cell development. Science 301:1239–1241
Tsunekawa N, Naito M, Sakai Y, Nishida T, Noce T (2000) Isolation of chicken vasa homolog gene and tracing the origin of primordial germ cells. Development 127:2741–2750
Ukeshima A, Kudo M, Fujimoto T (1987) Relationship between genital ridge formation and settlement site of primordial germ cells in chick embryos. Anat Rec 219:311–314
Vacket L (1962) Some new data concerning the formation of the definitive endoblast in the chick embryo. J Embryol Exp Morphol 10:10–57
Yasuda Y, Tajima A, Fujimoto T, Kuwana T (1992) A method to obtain avian germ-line chimaeras using isolated primordial germ cells. J Reprod Fertil 96:521–528
Yoshizaki G, Sakatani S, Tominaga H, Takeuchi T (2000) Cloning and characterization of a vasa-like gene in rainbow trout and its expression in the germ cell lineage. Mol Reprod Dev 55:364–371
Weidinger G, Stebler J, Slanchev K, Dumstrei K, Wise C, Lovell-Badge R, Thisse C, Thisse B, Raz E (2003) Dead end, a novel vertebrate germ plasm component, is required for zebrafish primordial germ cell migration and survival. Curr Biol 13:1429–1434
Zaccanti F, Vallisneri M, Quaglia A (1990) Early aspects of sex differentiation in the gonads of chick embryos. Differentiation 43:71–80
This research was supported in part by a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Sciences (JSPS; 16380200) to Masa-aki Hattori. Shinya Aramaki was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists (no. 09744).
About this article
Cite this article
Aramaki, S., Sato, F., Kato, T. et al. Molecular cloning and expression of dead end homologue in chicken primordial germ cells. Cell Tissue Res 330, 45–52 (2007). https://doi.org/10.1007/s00441-007-0435-1
- Primordial germ cells
- Cell migration
- In situ hybridization
- Dead end gene