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First published online February 1, 2009

RNA Interference of the Period Gene Affects the Rhythm of Sperm Release in Moths

Abstract

The period (per) gene is 1 of the core elements of the circadian clock mechanism in animals from insects to mammals. In clock cells of Drosophila melanogaster, per mRNA and PER protein oscillate in daily cycles. Consistent with the molecular clock model, PER moves to cell nuclei and acts as a repressor of positive clock elements. Homologs of per are known in many insects; however, specific roles of per in generating output rhythms are not known for most species. The aim of this article was to determine whether per is functionally involved in the circadian rhythm of sperm release in the moth, Spodoptera littoralis. In this species, as in other moths, rhythmic release of sperm bundles from the testis into the upper vas deferens occurs only in the evening, and this rhythm continues in the isolated reproductive system. S. littoralis was used to investigate the expression of per mRNA and protein in the 2 types of cells involved in sperm release: the cyst cells surrounding sperm bundles in the testes, and the barrier cells separating testicular follicles from the vas deferens. In cyst cells, PER showed a nuclear rhythm in light/dark (LD) cycles but was constitutively cytoplasmic in constant darkness (DD). In barrier cells, nuclear cycling of PER was observed in both LD and DD. To determine the role of PER in rhythmic sperm release in moths, testes-sperm duct complexes were treated in vitro with double-stranded fragments of per mRNA (dsRNA). This treatment significantly lowered per mRNA and protein in cyst cells and barrier cells and caused a delay of sperm release. These data demonstrate that a molecular oscillator involving the period gene plays an essential role in the regulation of rhythmic sperm release in this species.

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Article first published online: February 1, 2009
Issue published: February 2009

Keywords

  1. Spodoptera littoralis
  2. circadian clock
  3. RNAi
  4. period
  5. sperm release

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PubMed: 19150927

Authors

Affiliations

Joanna Kotwica
Department of Animal Physiology, Faculty of Biology, University of Warsaw, Warsaw, Poland
Piotr Bebas
Department of Animal Physiology, Faculty of Biology, University of Warsaw, Warsaw, Poland
Barbara O. Gvakharia
Department of Zoology, Oregon State University, Corvallis, OR, USA, Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331-2194
Jadwiga M. Giebultowicz
Department of Zoology, Oregon State University, Corvallis, OR, USA, [email protected]

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