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  • Author: G. P. Aldred x
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G. P. Aldred, P. Fu, R. J. Crawford and R. T. Fernley

ABSTRACT

The primary structure of the sheep renin precursor has been determined from its cDNA sequence. A library of cDNA clones was constructed from adrenalectomized sheep kidney poly(A)+ RNA and screened for sheep renin sequences with a cloned mouse renin cDNA probe. Of the 300 000 clones generated, 24 were hybridization positive and the nucleotide sequences of two of the longest clones were determined. These clones coded for the mature sheep renin protein and the 3′-untranslated sequence but did not extend to the amino-terminal region of preprorenin. Clones corresponding to the 5′ region of renin mRNA were generated by the polymerase chain reaction and their nucleotide sequences determined. The sheep renin precursor consists of 400 amino acids with a putative leader sequence of 14 amino acids and a putative 45 or 53 amino acid prosegment. The mature sheep renin protein has a 73% sequence identity with human renin. Northern analysis demonstrated the presence of renin mRNA in the kidney but not in other tissues in the sheep. While sodium depletion of sheep caused a rise in renin mRNA in the kidney, adrenalectomy also led to a large increase in renal renin mRNA. Southern analysis of genomic DNA suggests that there is only one gene coding for renin in the sheep.

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J. D. Penschow, G. P. Aldred, P. A. Darling, J. Haralambidis, V. E. Hammond, B. H. van Leeuwen, A. J. Mason, H. D. Niall, P. Seeburg and J. P. Coghlan

ABSTRACT

Relative levels of rat ovarian α inhibin (αI) and βA inhibin (βAI) mRNAs were measured during pregnancy by dot-blot hybridization of ovarian poly(A+) RNA. Follicular patterns of αI and βAI expression in contralateral ovaries from the same rats were also studied by hybridization histochemistry. Oligodeoxynucleotide probes specific for porcine αI and βAI were synthesized, 32P end-labelled and used as hybridization probes on dot-blots of ovarian RNA and frozen sections of ovarian tissue from pregnant rats. During pregnancy, levels of αI and βAI mRNAs remained fairly constant from day 7 after mating until parturition and then fell within 16 h post partum. In all ovaries observed, expression of inhibin genes was located in granulosa cells of healthy antral follicles. In general, the strongest signals for αI and βAI mRNAs were obtained in large follicles, with weaker signals in smaller follicles. Follicular patterns of αI and βAI expression during pregnancy were often dissimilar when βI and βAI were compared over a range of follicles. Considerable βI mRNA was detectable in some follicles in which βAI was reduced or undetectable, despite strong signals for both αI and βAI in an adjacent follicle. Essentially, αI mRNA levels were relatively consistent between groups of follicles, whereas βAI levels varied considerably. βAI mRNA was never observed in a follicle in the absence of αI mRNA, indicating that activin production in any follicle occurs in the presence of αI mRNA. Similar patterns of expression were observed in ovaries from pregnant mice. We have shown that expression of αI and βAI inhibin genes is not regulated uniformly within follicles of pregnant rat and mouse ovaries.