The spatial distribution of maternal mRNA is determined by a cortical cytoskeletal domain in Chaetopterus eggs

Abstract

Messenger RNA molecules are localized in the cortical region of eggs and unevenly segregated to the embryonic cells during early development of the annelid Chaetopterus. The egg cortex is enriched in two organelles, ectoplasmic spherules and associated structures, which are similar in appearance to nuage. The physical basis of cortical mRNA localization was examined in stratified eggs and in eggs extracted with the nonionic detergent Nonidet P-40 (NP-40). The cortical organelles were displaced to the most centrifugal zone of stratified eggs. In situ hybridization with poly(U) or cloned DNA probes showed that a large proportion of the poly(A) +RNA, histone mRNA, and actin mRNA molecules was also displaced to the centrifugal zone. Extraction with NP-40 revealed a detergent-insoluble cytoskeletal domain (CD) in the egg cortex which contained the remnants of ectoplasmic spherules and nuage embedded in a fibrous network. Although most of the total protein and RNA was extracted by NP-40, a large proportion of the poly(A) +RNA, histone mRNA, and actin mRNA molecules was retained in the CD. In situ hybridization of stratified eggs extracted with NP-40 indicated that the CD, with its associated organelles and mRNA molecules, is displaced to the centrifugal zone as a unit. The results suggest that the tenacious association of mRNA molecules with the cortical CD may be responsible for maternal mRNA localization during early development.