Worm Breeder's Gazette 13(2): 12b (February 1, 1994)
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
We have used an in situ hybridization protocol to explore the distribution of mRNAs in germlines and embryos (Evans et al., manuscript submitted). This protocol yielded strong staining for glp-1 mRNA with very little background; glp-1 mRNA was detected throughout the distal arm of the germline, in oocytes, and in all cells of 1-8 cell embryos. After the 8-cell stage, glp-1 mRNA disappears from all cells but persists in P3 and P4 ,consistent with observations made by Geraldine Seydoux (WBG 13(1): 25). We have also detected endogenous mRNAs for fem-3 and lag-2 ,and exogenous lacZ RNA that had been injected into the gonad. For the sex-determination gene fem-3 ,mRNA distribution was similar to glp-1 ;RNA was detected in the germline (distal arm and oocytes) and in 1-8 cell embryos. For lag-2 ,mRNA was detected in the distal tip cell, but not in the germline (see Henderson et al. abstract in this WBG). Following the last worm meeting, we received several requests for the protocol. Since then, we've made some modifications. In some cases, decreasing the hybridization time to 10-12 h may significantly improve sensitivity in addition to making the procedure shorter. Anyone wanting further information should call or e-mail tomevans@macc.wisc.edu (note that this is a different e-mail address from the last directory). Also, if anyone has made any improvements on the procedure or any interesting discoveries with it, we would love to hear from you.