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Luteolysis was induced in rats during late pregnancy by fetoplacental removal, and was monitored by the increased activity of luteal 20 alpha-hydroxysteroid dehydrogenase (20 alpha-OHSDH). The extent of enzyme induction over a given length of time varied according to the time of day at which the fetuses and placentae were removed, 11.00 and 21.00 h appearing to give optimal and minimal enzyme activities respectively. The 20 alpha-OHSDH was more readily induced on day 19 than on day 18.

The sustained elevation during Days 12--21 of pregnancy of rat luteal ornithine decarboxylase, which is inducible experimentally by hCG and PGF-2 alpha, was dependent upon intact placentae and oestrogen, but independent of prostaglandin, and was absent at corresponding times of pseudopregnancy. It is suggested that his enzyme activity is a response to an hCG-like hormone of placental origin. Two brief reappearances of enzyme activity on Days 21 and 22 of pregnancy, dependent upon intact placentae and upon both oestrogen and prostaglandin, suggest a pulsatile release of PGF-2 alpha from the uterus or placentae. These experiments illustrate the value of ornithine decarboxylase in seeking unknown hormonal influences upon the corpus luteum.

Seven tk- mutants of herpes simplex virus, type 2 (HSV-2), and three tk- mutants of herpes simplex virus, type 1 (HSV-1), were isolated which did not produce the thymidine kinase (TK) polypeptides but formed smaller polypeptides not seen in wild-type infected cells. Positive TK mRNA selection by hybridization to the cloned tk genes followed by in vitro translation identified the TK polypeptides. Comparisons of the products of partial proteolysis of the polypeptides of four HSV-2 and two HSV-1 tk- mutants to those of the parental TK polypeptides indicated that, in each case, the novel polypeptide was a fragment of the TK polypeptide, showing that these mutants have defects in the structural gene for tk. HSV-2 mutants of this sort have not been previously described. They and the HSV-1 mutants are similar to HSV-1 mutants reported previously. In addition, it was found that TK mRNA was present early in infection but was absent late in infection, suggesting that the shutoff of TK synthesis is due to message degradation. Also, HSV-2 TK mRNA did not hybridize to the cloned HSV-1 tk gene indicating that these genes have extensively diverged.

The mouse major urinary proteins (MUPs) are the products of a multi-gene family of 30-35 genes whose members exhibit diverse tissue specific, developmental, and hormonal controls. Three cDNA clones corresponding to liver MUP mRNAs have been sequenced. Two of the clones (p499, C57BL/6 and p1057, BALB/c) share strong homology whereas a third clone (p199, C57BL/6) has diverged considerably from the others at the nucleic acid (85% homology) and protein (68% homology) levels. The 5' regions of p499 and p199 which show the most sequence divergence were subcloned and shown to hybridize to different liver MUP mRNAs. The p499-5' sequence was expressed in all MUP expressing tissues (liver, lachrymal, submaxillary and mammary) whereas the p199-5' sequence was expressed primarily in the liver and lachrymal. Analysis of liver RNA from mice in different endocrine states indicates that the p499-5' sequence is strongly regulated by thyroxine administration whereas the p199-5' sequence is not. Both sequences appear to be regulated by growth hormone and testosterone. Southern blot analysis of mouse genomic DNA indicates that there are multiple genes homologous to each sequence.

Treatment of pregnant rats with human chorionic gonadotrophin, luteotrophin (luteinizing hormone), luteotrophin-releasing hormone, prostaglandin F2alpha, aminoglutethimide, or by foetoplacental removal or hysterectomy achieved a common multiple-response pattern, namely increased activity of luteal 20alpha-hydroxy steroid dehydrogenase with decreased activity of delta5-3beta-hydroxy steriod dehydrogenase and release of delta4-3-oxo steroids in vitro. 2. Similar effects of foetoplacental removal are noted in pregnant mice. 3. Gonadotrophin induced lower activities of 20alpha-hydroxy steroid dehydrogenase, except at the very end of pregnancy, and partly inhibited the induction caused by foetoplacental removal. 4. The results suggest that existence of a placental factor that restrains these changes until the end of normal pregnancy, which is produced in amounts proportional to the number of placentae and is conveyed to the ovary via the blood. 5. This factor was not replaced by prolactin. 6. It is argued that neither placental lactogen nor pituitary luteotrophin participate in the induction of 20alpha-hydroxy steroid dehydrogenase at late pregnancy in the rat. 7. Aminoglutethimide induced 20alpha-hydroxy steroid dehydrogenase only in late pregnancy. This was partly reversed by progesterone, wholly reversed by progesterone plus oestrogen, and did not involve the pituitary.

Lactose was synthesised within the lumen of purified Golgi membrane vesicles, prepared from lactating rat mammary gland, from externally added glucose and UDP-galactose. An apparent Km of 1.5 mM was shown towards glucose at anomeric equilibrium, but only beta-glucose was utilised. Two apparent Km values, about 17 micro M and 112 micro M, were shown towards UDP-galactose. 5-D-Thioglucose, 6-deoxy-D-glucose and 6-deoxy-6-chloro-D-glucose were alternative substrates, acquiring alpha-lactalbumin dependence when the vesicles were lysed with detergent. Substrates independent of alpha-lactalbumin, or inhibited by it, included a wide range of N-acylated glucosamines as well as phenyl-beta-glucoside. The galactosylation of these by vesicle preparations could be ascribed to a proportion of leaky vesicles. Suitably low concentrations of Triton X-100 activated lactose synthesis by intact vesicles, indicating the membrane as a rate-limiting feature of the system.

Lactose synthesis and fatty acid synthesis in intact lactating-rat mammary gland were measured simultaneously by incorporation of [U-14C]glucose and of both [U-14C]glucose and 3H2O respectively. Both processes were almost abolished by overnight starvation. Self-re-feeding caused recovery of lipogenesis to 100% of normal by 2 h and to 170% by 5 h. Lactose synthesis recovered to 80% of normal by 5 h. Food intubated to starved rats caused partial recovery in 3 h, standard diet favouring lactose synthesis and sugars favouring lipogenesis. Casein and starch were ineffective. Olive oil intubated to fed rats suppressed lipogenesis greatly and lactose synthesis slightly. Paraffin oil or water partly mimicked these effects. Adrenaline (subcutaneous) decreased lipogenesis from glucose, whereas insulin (subcutaneous) caused hypoglycaemia associated with loss of lactose synthesis but unchanged fatty acid synthesis. Streptozotocin and 2-bromo-alpha-ergocryptine (CB-154) impaired lipogenesis but not lactose synthesis. The results are interpreted in terms of competition for intracellular glucose by biosynthetic pathways for lactose and fat, and the possible implications for variations in milk composition are discussed.

1. myo-Inositol was galactosylated by UDP-galactose in the presence of alpha-lactalbumin plus rat mammary Golgi membranes enriched in galactosyltransferase (EC 2.4.1.22). 2. The isolated product migrated on GLC as two peaks of material, apparently identical to galactinol (galactosylinositol) isolated from rat milk. 3. These findings make it likely that in vivo lactose synthetase is responsible for converting inositol into galactinol within the Golgi lumen. 4. This is consistent with the ability of inositol to penetrate the Golgi membrane in vitro, and renders less likely a previously proposed role of beta-galactosidase (EC 3.2.1.23) in mammary galactinol synthesis.

Natural or early weaning of rat litters caused an increased activity of maternal luteal 20alpha-hydroxy steroid dehydrogenase and a decreased release of delta4-3-oxo steroids in vitro. 2. Compound CB-154 (2-bromo-alpha-ergocryptine) caused an increase of 20alpha-hydroxy steroid dehydrogenase activity in mid-lactation but not in early lactation. 3. Prolaction did not prevent these increases in enzyme activity.

1. The activity of 20alpha-hydroxy steroid dehydrogenase in rat ovarian corpora lutea increased at least 50-fold between 2 days before and 2 days after parturition, and then fell gradually during lactation. The activity of 3beta-hydroxy Delta(5)-steroid dehydrogenase decreased by 50% at parturition but remained constant at other times. 2. The 20alpha-hydroxypregn-4-en-3-one/progesterone concentration ratio in the ovary fell tenfold between 1 day before and 1 day after parturition, in contrast with the increase of the ratio for these steroids in plasma. 3. Pregnenolone was metabolized in intact cells or cell-free systems either to pregn-5-ene-3beta,20alpha-diol and then to 20alpha-hydroxypregn-4-en-3-one by 20alpha-hydroxy steroid dehydrogenase and 3beta-hydroxy Delta(5)-steroid dehydrogenase respectively, or directly to progesterone by the latter enzyme. The relative activities of these pathways appeared to reflect the relative amounts of the two enzymes and the concentrations of their respective coenzymes NADPH and NAD(+). 4. From these and other observations it was concluded that the cessation of progesterone secretion, which precedes parturition and lactogenesis at the end of pregnancy, is partly due to the redirected metabolism of pregnenolone away from progesterone and towards 20alpha-hydroxypregn-4-en-3-one as the secreted end product. This is primarily the consequence of the sharp increase in the activity of 20alpha-hydroxy steroid dehydrogenase. This mechanism is super-imposed on the already declining rate of net Delta(4)-steroid release by the ovary. 5. A relationship of these pathways to subcellular compartments of luteal cells is proposed.

1. The presence of palmitoyl-CoA-l-glycerol 1-phosphate palmitoyltransferase (EC2.3.1.15) has been demonstrated in a particulate fraction of baker's yeast. 2. The enzyme has been characterized, and its activity studied as a function of pH and concentration of substrates. 3. Inhibition by thiol poisons and protection by acyl-CoA have been used to obtain information on the active site. 4. By various methods of supplying acyl radicals, the species ;palmitoyl-CoA' has been shown to be the true acyl donor to the transferase.