Detection and expression of hepatitis B virus X gene in one and two-cell embryos from golden hamster oocytes in vitro fertilized with human spermatozoa carrying HBV DNA.
The main objectives of this study were to introduce motile human sperm carrying Hepatitis B virus (HBV) DNA to golden hamster oocytes in a co-culture environment and to detect the replication and expression of the HBx gene in early embryonic cells. Zona-free hamster oocytes were inseminated with human sperm carrying pBR322-HBV DNA plasmid using the in vitro fertilization (IVF) technique. Both the one- and two-cell stages of early embryonic development were studied. PCR, RT-PCR, and Dot hybridization were performed to observe the HBx gene and its expression in these stages. "Fluorescence in situ hybridization" (FISH) was carried out to confirm the integration of HBV into the pronucleus, nucleus, and the chromosomes of embryos. The results showed that we have the ability to obtain a fertilization rate of 80%. RT-PCR showed that the HBx gene could be expressed in both one- and two-cell stages of embryonic development. The data suggested the possibility of sperm as a vector for the vertical transmission of HBV DNA to the next generation.
Ali
Bahy-Ahmed
BA
Research Center for Reproductive Medicine, Department of Cell Biology and Genetics, Shantou University Medical College, Shantou, Guangdong, China.
Huang
Tian-Hua
TH
Xie
Qing-Dong
QD
D000818:Animals; D006224:Cricetinae; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000166:cytology / Q000821:virology; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics; D006509:Hepatitis B / Q000635:transmission; D006515:Hepatitis B virus / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D018445:Infectious Disease Transmission, Vertical; D008297:Male; D009865:Oocytes / Q000737:chemistry / Q000378:metabolism; D012367:RNA, Viral / Q000032:analysis / Q000378:metabolism; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism / Q000821:virology*; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins
D004279:DNA, Viral; D012367:RNA, Viral; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein