Influence of Inactivation of Tandemly Repeated Pericentromeric DNA Transcription on the Formation of Membraneless Structures at the End of Oocyte Maturation

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Abstract
Tandemly repeated pericentromeric noncoding DNA (TR DNA) makes up approximately 10% of the human genome. Pericentromeric TR DNA includes classical human satellites 1, 2, 3 (HS1, HS2, HS3), which are transcribed in somatic cells. We have previously shown the presence of HS2/HS3 transcripts in late human oogenesis and sequenced them. It has been suggested that the RNPs found may be the site of spatial sequestration of RNA and proteins at the end of human oocytes maturation. The aim of this work was to develop a method for inactivating HS2/HS3 transcripts using antisense oligonucleotides to assess its effect on the size and quantity of DDX4-containing RNPs in late human oogenesis. Inactivation of HS2/HS3 transcription at the end of human oocytes maturation by the microinjection method resulted in a significant decrease of the total HS2/HS3 RNA signal, detected by the method of fluorescence in situ hybridization (FISH). At the same time, an increase in the number of inclusions positive to antibodies to RNA helicase DDX4 was observed. Supposedly, upon inactivation of HS2/HS3 transcription, dissociation of DDX4-containing RNP particles occurred. Such changes of the RNP particles can play a critical role in the development of oocytes and be a cause of the arrest of maturation or the occurrence of pathological syndromes, including those associated with problems of fertilization.

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