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Biology of Reproduction, Vol 14, 163-174, Copyright © 1976 by Society for the Study of Reproduction

Isolation and Translation of Blastokinin mRNA

IRWIN L. LEVEY 1, and JOSEPH C. DANIEL JR. 1

1 Department of Zoology, University of Tennessee, Knoxville, Tennessee 37916


Uterine mRNA synthesis during preimplantation pregnancy of the rabbit was estimated by the assay of the poly(A) content in radiolabeled polysomal RNA. The proportion of newly-synthesized poly(A)-rich RNA in both longterm and rapidly-labeled polysomal RNA increased from estrus to Day 4 of pregnancy, with no further change noted through Day 6.

Uterine RNA preparations were assayed for template activity by: 1) infusion into the uterine lumina of longterm castrate rabbits, and 2) supplementation of a heterologous cell-free translation system derived from wheat embryo. Translation products obtained from both procedures contained a protein with electrophoretic and immunological properties identical to purified blastokinin. Greatest stimulatory activity was observed in poly(A)-rich RNA extracted from day 4 uteri, but was absent in similar preparations from estrous uterus, liver, and ovary.

Note:
ACKNOWLEDGMENTS These investigations were supported by research grant 2 RO HD 06226 from the National Institute of Child Health and Human Development. The wheat embryo cell-free translation system was a generous gift from Dr. Walter Farkas of the University of Tennessee Memorial Research Center and Hospital. We are indebted to Dr. J. J. Ireland for performing the progestin radioimmunoassay, and to Robbie R. Levey for expert technical assistance and help in preparation of this manuscript.

Submitted on February 25, 1975
Accepted on October 3, 1975






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Copyright © 1976 by the Society for the Study of Reproduction.