The localization of LAP2beta in bovine oocytes after in vitro activation and fertilization.
The present study was designed to clarify the localization of LAP2beta and to compare it with those of lamins A/C and B in bovine oocytes after activation and in vitro fertilization (IVF). After fertilization, LAP2beta was not found until telophase II, and was observed around condensed chromatin after the extrusion of the second polar body, but not in activated oocytes. Although the reaction of LAP2beta was temporally negative or weak on the membrane of the growing small pronuclei, it became strong on the fully grown pronuclei of both activated and fertilized oocytes. Examination of the timing of DNA synthesis using bromodeoxyuridine revealed that the expression of LAP2beta on the pronuclear membrane became strong around the end of the DNA synthesis in both activated and fertilized oocytes. Both male and female pronuclei exhibited the same reactivity to all nuclear proteins examined. It was also shown that LAP2beta first assembled around condensed chromatin, followed by the integration of lamins B and A/C as in somatic cells. LAP2beta staining was maintained on the nuclear membrane of the embryonic cells at interphase until the later stage of preimplantational development. There were no differences between parthenogenetic and fertilized embryos in the expression and localization of LAP2beta from the PN-stage oocyte to the blastocyst. The assembly of LAP2beta was observed around the telophase chromatin of both blastocyst and cumulus cells. Thus, it was shown that the timing of the aggregation of LAP2beta at the second meiosis was different from that in the mitosis of blastocyst and somatic cells. LAP2beta was constantly expressed in the nuclear membrane in in vitro fertilized and parthenogenetic embryos as was lamin B, and lamin A/C was expressed stage-dependently in both types of embryos. Lamin A/C was positive in some inner cell mass cells of parthenogenetic blastocysts, but not those of in vitro fertilized embryos.
Isaji
Mamiko
M
Department of Bioresource and Agrobiosciences, Graduate School of Science and Technology, Kobe University, and Kobe Minori Public Corporation, Kobe City, Japan.
Iwata
Hisataka
H
Harayama
Hiroshi
H
Miyake
Masashi
M
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005307:Fertilization in Vitro; D007315:Insemination, Artificial; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism / Q000502:physiology*
D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; C093413:lamina-associated polypeptide 2