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Pregnancy-associated glycoprotein (PAG) family localized in chorionic cells within the epitheliochorial/diffuse placenta of the alpaca (Lama pacos)
Authors:Marta MajewskaGrzegorz Panasiewicz  Bozena Szafranska
Institution:Department of Animal Physiology, Faculty of Biology, University of Warmia and Mazury in Olsztyn, Oczapowskiego Str 1A, 10-719 Olsztyn-Kortowo, Poland
Abstract:Pregnancy-associated glycoproteins (PAGs) are abundant embryo-originated products expressed in the pre-placental trophoblast and later in the post-implantational chorionic epithelium of some ungulate species. This paper describes the cellular immunolocalization of the chorionic PAG family in the epitheliochorial placenta type of the alpaca (Lama pacos—Lp), in which the PAGs were named ‘LpPAGs’. Placental Lp sections (5 μm) of different females near mid-pregnancy (150 days post coitum; dpc), advanced pregnancy (244-263 dpc) and late pregnancy (347 dpc) were used for cross-species (heterologous—ht) double fluorescent immunohistochemistry (htdF-IHC). The htdF-IHC was performed with primary rabbit polyvalent anti-porcine PAG polyclonals. The LpPAG immuno-complexes were visualized with secondary goat anti-rabbit immunoglobulins-conjugated with Alexa 488 fluorophore (green), among all nuclei of placental cells stained with propidium iodide (red). This is the first study reporting the immunolocalization of the LpPAG family identified by htdF-IHC at the feto/maternal interface during different pregnancy stages of the alpaca. The most dominant and strongest immune-positive LpPAG signals were found in the well-developed chorionic cell layer. Our htdF-IHC indicated relatively high epitope resemblance to that of the PAGs in camelids and pigs. These data increase our general knowledge of chorionic PAG localization during pregnancy-stage dependent development of the epitheliochorial diffuse placenta type in the alpaca.
Keywords:PAGs  pregnancy-associated glycoproteins  A488  Alexa 488 dye  anti-pPAG-P  anti-porcine PAG polyvalent polyclonals  BNC  binuclear cells  cDNA  complementary DNA  dpc  days post coitum  END  endometrium  fV  fetal vein  fA  fetal artery  GTC  giant trophectoderm cells  H&  E  hematoxylin and eosin staining  htdF-IHC  heterologous double fluorescent immunohistochemistry  ht-ISH  heterologous in situ hybridization  Lp  Lama pacos  LpPAG  PAGs of the alpaca  MNC  mononuclear cells  NRS  normal rabbit serum  RT  room temperature  pPAG  porcine PAG  PI  propidium iodide  TRF  trophoblast  TRD  trophectoderm-chorionic epithelium
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