Rick Renaud

Estrogen Metabolism in the Equine Conceptus and Endometrium During Early Pregnancy in Relation to Estrogen Concentrations in Yolk-Sac Fluid

Abstract Because estradiol (E2) production by the early equine conceptus is considered crucial to the establishment of pregnancy, the amounts of E2, estrone (E1), and their sulfates (E2S, E1S) were measured by RIA in yolk-sac fluid of 63 conceptuses collected by transcervical lavage over the period of 11–26 days after ovulation. Amounts increased significantly with age of conceptus, especially for E1S. Then, the metabolism of E2, which may be highly relevant for its action, was examined in the conceptus and endometrium over the period when the conceptus ceases to migrate within the uterus. Eleven conceptuses collected mainly on Days 12, 15, and 18, with endometrial biopsy samples taken immediately thereafter, were used for steroid metabolic studies. Trophoblastic and endometrial tissues were incubated with [3H]-labeled E2 or E1, and with [14C]-E1 in one experiment. Steroids were recovered from the media by solid-phase extraction (SPE) and eluted separately as unconjugated and conjugated fractions. Conjugation increased from Day 12 for the trophoblast (more so by bilaminar than trilaminar tissues on Day 18) and was much greater for endometrium, with almost all as sulfoconjugates. HPLC profiles of free and sulfate fractions were obtained from a gradient of acetonitrile/water. Interconversion (E2 ⇌ E1) by trophoblast varied with development; it favored E2 in older conceptuses, more in bilaminar than trilaminar tissues. Some more polar products were also noted, with loss of tritium seen as [3H]2O at SPE, and confirmed by HPLC in a second system with authentic reference steroids. Almost all radioactivity in the endometrium was present as E2 in both free and sulfate fractions. It was concluded that local metabolism of E2 is quantitatively significant and may play an important role in the actions of the large amounts of estradiol produced by the early equine conceptus.

Free and bound lipids of Neurospora crassa

Abstract Extraction of light and dark grown cells of Neurospora crassa with chloroform-methanol gave a free lipid extract in which the relative amounts and compositions of sterols, fatty acid and carotenoid fractions were determined. Further extraction of the cells with methanolic potassium hydroxide solution liberated a bound lipid fraction from the cells. The levels of the bound lipid fraction were much lower than those of the free lipids but analysis showed that the composition was similar to that of the free lipids.

Isolation of 11β-hydroxylated androgens from testicular vein blood of the mature boar

The isolation of 11 beta-hydroxyandrostenedione and 11 beta-hydroxytestosterone from testicular vein blood of the mature male domestic pig is described. Blood was collected from veins and arteries on the surface of the testes of mature boars. Steroids were extracted from plasma with SEP-PAK C18 cartridges and recovered with acetonitrile. A separation of steroids was made by high performance liquid chromatography (HPLC) using acetonitrile/water (37/63; v/v), and fractions were collected manually with detection at 254 nm. Preliminary identification was based on comparison with the HPLC retention time of an authentic steroid standard. Final characterization was achieved by means of capillary gas chromatography-mass spectrometry (GC-MS). The findings record the first evidence for the secretion of C19-11-hydroxylated steroids by normal testes in a mammalian species.