Lipid class dynamics during development in early life stages of shortbelly rockfish and their application to condition assessment

Lipid class dynamics and the relationship between weight and length were analysed during early life stage (ELS) development in wild populations of shortbelly rockfish Sebastes jordani to investigate their utility in a condition or nutritional status assessment. Analyses from more than 3000 field-captured shortbelly rockfish indicated little variation in weight for a given length and provided limited application in the assessment of condition. The fractionation of total lipids into individual classes revealed trends in lipid metabolism throughout development. These trends were described best using a stage-specific regression model, since Sebastes show defined early life history transitions. Among developmental stages, triacylglycerols (TAG) and polar lipids (PL) were the dominant lipid classes and cholesterol (CHOL), sterol/wax esters, and nonesterified fatty acids were found in lower concentrations. In the preflexion stage, low concentrations and constant levels of TAG along with the combined influence of endogenous and exogenous sources of energy compromised the use of TAG as an index of nutritional status. However, during flexion through juvenile stages, TAG levels increased indicating an accumulation of energy reserves with development, thus providing a valid indicator of nutritional status in later stages. Large fluctuations in PL concentrations during development suggested complex metabolism which may be due to changes in cell volume and cell membrane proliferation with development as well as catabolism. CHOL declined in larval stages, then remained constant with increasing size through the juvenile stage, negating its use as a normalizer for variations of mass in a TAG/CHOL ratio prior to juvenile transformation. This study provides the first documentation of lipid dynamics during early life stages following parturition in a marine viviparous teleost.     

Lipid and fatty acid composition of female gilthead seabream during their reproductive cycle: effects of a diet lacking n-3 HUFA

In female sea bream Sparus aurata fed a control diet (C), ovarian levels of neutral lipids (NL) and polar lipids (PL) remained constant between November and March, while a decrease in NL content was observed in liver and muscle. In the same period, liver PL content increased, while no changes were observed in muscle. Between March and June ovarian NL and PL showed a strong decrease, while NL remained constant in liver and muscle. When fish were fed a diet lacking in n-3 highly unsaturated fatty acids, n-3 HUFA (D), the pattern observed was similar to that found in the fish fed diet C, with the exception of liver NL, which increased between March and June. In general, the changes in fatty acid content, in both groups of fish, were highly influenced by the diet given to the broodstock, although these effects were greater on ovarian NL and PL than on liver and muscle lipids. Despite the fact that gilthead seabream females continue feeding during the spawning season, they probably make use of their liver and muscle reserves during the gonadal maturation process. Furthermore, the fatty acid composition of the broodstock diet was reflected in the body composition, especially in the ovaries.     

Lipids in yolks and neonates of the viviparous lizard Niveoscincus metallicus

Niveoscincus metallicus is a small viviparous skink which provides a substantial amount of yolk to each of its developing embryos although some organic nutrients are also transferred across the placentae. The total amount of lipid present in the yolk of N. metallicus (37% of dry weight) is very much higher than that in the newborns (19% of dry weight), confirming that the yolk is utilised as an energy source during gestation. Triacylglycerols (TAG), which are storage compounds, are the major lipid resource available to the embryos and are present in relatively large amounts in the yolk of N. metallicus. Polar lipids (PL), which form the structural components of membranes, and sterols (ST), which are involved in the synthesis of hormones and vitamins, are also present in the yolk. The proportions of each of these lipid classes differs markedly between yolks and newborns. This may reflect variations in the role played by each lipid class in the provision of nutrients to, and development of, embryos.     

Plasma and ovarian thyroxine levels in relation to sexual maturation and gestation in female Sebastes inermis

Plasma T₄ (L-thyroxine) concentrations in the viviparous rockfish Sebastes inermis showed a peak (119·3±27·8 ng ml⁻¹) during the development of embryos. Ovarian T₄ concentrations increased during vitellogenesis and final maturation and decreased during embryogenesis.However, total T₄ content within the ovary increased continuously up to the larval stages. Plasma oestradiol-17β, (E₂) concentrations peaked at the final maturation stage and then recovered to the level seen during the non-reproductive periods. Plasma testosterone (T) concentrations showed a peak (5·33±1·08 ng ml⁻¹) at the embryo stage and high levels were maintained throughout the gestation period. Plasma T₄ concentrations during the embryo stage and ovarian T₄ content during vitellogenesis were much higher than the levels reported in oviparous fishes. These data suggest that in viviparous rockfish, T₄ may be required for sustaining gestation or embryo development within the maternal body. In addition, the high content of total T₄ in the ovary implies that there probably does exist a maternal-embryo relationship during gestation as well as during vitellogenesis.     

Energy allocation strategy of skipjack tuna Katsuwonus pelamis during their reproductive cycle

The lipid composition of somatic and reproductive tissues was determined for female skipjack tuna Katsuwonus pelamis caught in the western Indian Ocean between latitude 10° N and 20° S and longitude 40° and 70° E. The highest total lipid (TL) contents were in the liver and gonads, with white muscle levels approximately three‐fold lower. Three lipid classes dominated: triacylglycerols (TAG), sterol esters and wax esters (SE‐WE) and phospholipids (PL). Collectively, these accounted for between 70 and 80% of TLs. Changes in lipid concentrations were evaluated over the maturation cycle. Immature fish had the lowest gonad and liver TL levels; concentrations of TL, TAG, SE‐WE and PL accumulated from immature to mature (spawning‐capable) phase, reflecting sustained vitellogenic activity of the liver and a transfer of lipids to developing oocytes from the onset of vitellogenesis. Gonado‐somatic and hepato‐somatic indices were positively correlated with each other and positively related to TL in the gonads and liver. Fulton's condition index and lipid concentrations in muscle did not vary significantly over the maturation cycle; fat content in the main storage tissues was undepleted as the ovary developed. Hence, K. pelamis apparently supports reproduction directly from food intake over the breeding season. In the gonads, reserve lipids (SE‐WE and TAG) and sterols were related to batch fecundity but this was not the case for somatic and hepatic tissues. These results suggest that K. pelamis utilizes an income breeding strategy.     

Transfer of n-3 and n-6 polyunsaturated fatty acids from yolk to embryo during development of the king penguin

This study examines the transfer of lipids from the yolk to the embryo of the king penguin, a seabird with a high dietary intake of n-3 fatty acids. The concentrations of total lipid, triacylglycerol (TAG), and phospholipid (PL) in the yolk decreased by ~80% between days 33 and 55 of development, indicating intensive lipid transfer, whereas the concentration of cholesteryl ester (CE) increased threefold, possibly due to recycling. Total lipid concentration in plasma and liver of the embryo increased by twofold from day 40 to hatching due to the accumulation of CE. Yolk lipids contained high amounts of C(20-22) n-3 fatty acids with 22:6(n-3) forming 4 and 10% of the fatty acid mass in TAG and PL, respectively. Both TAG and PL of plasma and liver contained high proportions of 22:6(n-3) ( approximately 15% in plasma and >20% in liver at day 33); liver PL also contained a high proportion of 20:4(n-6) (14%). Thus both 22:6(n-3) and 20:4(n-6), which are, respectively, abundant and deficient in the yolk, undergo biomagnification during transfer to the embryo.     

Lipid class dynamics and storage depots in juvenile weakfish Cynoscion regalis and their application to condition assessment

Lipid class dynamics, the pattern of change in the primary form and location of lipid stores and their relationship with standard length (L(S) ), were investigated in collections of young-of-the-year weakfish Cynoscion regalis for the purpose of determining the utility of this analysis as an indication of condition. The separation of total lipids into individual classes and the analysis of potential storage depots revealed the general patterns of lipid class dynamics and energy storage in C. regalis during their period of juvenile estuarine residency. Phospholipid and cholesterol exhibited moderate but variable (8·1-40·0 and 1·3-21·5 mg g(-1) , respectively) concentrations across the entire juvenile period and were the predominant lipid classes in juveniles <100 mm L(S) , while wax ester concentrations were low (c. 1 mg g(-1) ) and exhibited the least amount of variability among lipid classes. Triacylglycerols (TAG) and free fatty acids (FFA) exhibited similar dynamics, with relatively low concentrations (<15 mg g(-1) ) in individuals ≤100 mm L(S) . In larger juveniles both TAG and FFA concentrations generally increased rapidly, though there was considerable variability in both measures (0·0-199·7 and 0·0-49·7 mg g(-1) , respectively). Increasing levels of lipids, primarily in the form of TAG, with size indicated an accumulation of energy reserves with growth, thus providing an indication of individual condition for larger juveniles. Separate analysis of liver, viscera and the remaining carcass indicated that liver and viscera did not represent a significant depot of TAG reserves. Analysis of samples derived from whole juvenile C. regalis thus provided an accurate estimate of energy reserves.     

Energetics of early development in the viviparous yellowtail rockfish

Total mean gestation period of all female yellowtail rockfish Sebastes flavidus , from fertilization to parturition, was 29.2 days. Hatching occurred 23 days after fertilization, followed by 6 days of larval incubation. Eggs varied in dry mass and energy content with lipid concentrations contributing the greatest amount of energy per egg. The endogenous energy of the yolk and oil globule declined throughout gestation resulting in limited yolk and oil reserves at parturition.     

Lipid accumulation and utilization by oocytes and eggs of Rhodnius prolixus

Insect eggs must contain the necessary nutrients for embryonic growth. In this article, we investigated the accumulation of triacylglycerol (TAG) in growing oocytes and its utilization during embryonic development. TAG makes up about 60% of the neutral lipids in oocytes and accumulates as oocytes grow, from 2.2 ± 0.1 µg in follicles containing 1.0 mm length oocytes to 10.2 ± 0.8 µg in 2.0 mm length oocytes. Lipophorin (Lp), the hemolymphatic lipoprotein, radioactively labeled in free fatty acid (FFA) or diacylglycerol (DAG), was used to follow the transport of these lipids to the ovary. Radioactivity from both lipid classes accumulated in the oocytes, which was abolished at 4°C. The capacity of the ovary to receive FFA or DAG from Lp varied according to time after a blood meal and reached a maximum around the second day. ³H‐DAG supplied by Lp to the ovaries was used in the synthesis of TAG as, 48 hr after injection, most of the radioactivity was found in TAG (85.7% of labeling in neutral lipids). During embryogenesis, lipid stores were mobilized, and the TAG content decreased from 16.4 ± 2.1 µg/egg on the first day to 10.0 ± 1.3 µg on day 15, just before hatching. Of these, 7.4 ± 0.9 µg were found in the newly emerged nymphs. In unfertilized eggs, the TAG content did not change. Although the TAG content decreased during embryogenesis, the relative lipid composition of the egg did not change. The amount of TAG in the nymph slowly decreased during the days after hatching. © 2011 Wiley Periodicals, Inc.     

Purification and characterization of luteinizing hormone from pituitary glands of rockfish, Sebastes schlegeli

To acquire greater knowledge of the reproductive function of luteinizing hormone (LH) in the viviparous rockfish Sebastes schlegeli, LH from the pituitary glands of mature rockfish was isolated, purified, and localized and its biological activity was characterized. The molecular mass of purified LH was estimated to be approximately 33 kDa, similar to that of known LH. When rockfish LH was purified by reverse-phase high-performance liquid chromatography, its N-terminal amino acid sequences were found to coincide with those of predicted cDNA sequences of rockfish gonadotropin α (ssGTHα) and ssLHβ mature peptides. Immunocytochemical analysis using antisera against ssGTHα (molecular weight [MW], ~14.5 kDa) and ssLHβ (MW, ~18.5 kDa) indicated that the LH-producing cells are mainly distributed throughout the proximal pars distalis and along the periphery of the pars intermedia. Further, in vitro ovarian follicle analysis demonstrated that purified intact rockfish LH significantly enhances E(2) secretion in a dose-dependent manner. This is the first report on the purification and characterization of LH from a viviparous teleost, and these results will enable future research and increase our understanding of the mechanisms underlying the maturation of such fish.     

Excessive ovarian follicular development in pregnant pigtailed macaques (Macaca nemestrina)

To evaluate the status and possible control of ovarian follicular development during pregnancy, circulating levels of estrone (E₁), estradiol-17β (E₂), and follicle-stimulating hormone (FSH) were measured throughout gestation in both intact and ovariectomized pregnant pigtailed monkeys (Macaca nemestrina). From an additional group of pregnant monkeys, ovaries were obtained at late gestation (on day 150 or 159 of pregnancy) for histological studies. Circulating concentrations of E₁ and E₂ increased on day 13 and remained elevated for about 10 days; they then declined and reached low levels on day 32 of gestation. After day 60, there were gradual but smaller increases in estrogen levels to day 140, after which both E₁ and E₂ levels increased significantly, reaching maximum levels (E₁ = 832.2 ± 210.8 pg/ ml; E₂ = 1.66 ± 0.32 ng/ml) at the end of pregnancy. Removal of ovaries on day 35 of gestation did not affect pregnancy or the pattern of estrogen secretion. Serum concentrations of FSH demonstrated only minor fluctuations during pregnancy but were similar to those found during the early follicular phase of cycling pigtailed monkeys investigated in this study. Ovarian histology revealed extensive follicular growth; in addition to the corpus luteum of pregnancy, ovaries were packed with pre-antral, small antral, and medium-sized Graafian follicles. Some of these follicles appeared to be cystic and showed various degree of atresia; their general appearance was similar to the follicles of human females with polycystic ovary syndrome. Our data suggest that FSH may initiate ovarian follicular growth during gestation. High levels of estrogens were incapable of suppressing FSH secretion but may be responsible for the induction of atresia in a large number of follicles in pregnant pigtailed monkeys.     

Branchial placenta in the viviparous teleost Ilyodon whitei (Goodeidae)

Intraluminal gestation, as it occurs in viviparous goodeids, allows a wide diversity of embryo‐maternal metabolic exchanges. The branchial placenta occurs in embryos developing in intraluminal gestation when ovarian folds enter through the operculum, into the branchial chamber. The maternal ovarian folds may extend to the embryonic pharyngeal cavity. A branchial placenta has been observed in few viviparous teleosts, and there are not previous histological analyses. This study analysis the histological structure in the goodeid Ilyodon whitei. The moterno ovarian folds extend through the embryonic operculum and reach near the gills, occupying part of the branchial chamber. These folds extend also into the pharyngeal cavity. In some regions, the epithelia of the ovarian folds and embryo were in apposition, developing a placental structure in which, maternal and embryonic capillaries lie in close proximity. The maternal epithelium has desquamated cells which may enter through the branchial chamber to the pharyngeal cavity and the alimentary tract. The complex processes that occur in the ovaries of viviparous teleosts, and its diverse adaptations for viviparity, as the presence of branchial placenta, are relevant in the study of the evolution of vertebrate viviparity. J. Morphol. 275:1406–1417, 2014. © 2014 Wiley Periodicals, Inc.     

Heterogeneity of the endoplasmic reticulum with respect to lipid synthesis in developing seeds of Brassica napus L.

Studies of the sub-cellular location of storage triacylglycerol (TAG) synthesis in developing embryos of oilseed rape (Brassica napus L.) show that there is heterogeneity of the endoplasmic reticulum (ER) with respect to the enzymes of lipid synthesis. The enzymes of TAG synthesis were detected in two membrane fractions (equilibrium densities 1.05 and 1.10 g· ml^?1) isolated by sucrose-density-gradient centrifugation of homogenates from developing rape embryos. The synthesis of TAG by the lowdensity membranes has not been reported previously and was found in this study because the sucrose density gradients began at only 10% (w/w) sucrose. The pattern of activity of the enzymes involved in the synthesis of TAG in the higher-density fraction closely matched the marker enzymes for the ER; lyso-phosphatidylcholine acyltransferase and cytidine diphosphate-choline:diacylglycerol cholinephosphotransferase. The activity of the ER marker enzymes in the low-density membrane fraction, however, was very much lower when compared to those involved in the synthesis of TAG. Analysis of the lipids extracted from the low-density fraction revealed it contained about 50 mol% TAG compared with 15 mol% in the bulk ER, which may account for the low density of the membranes in this fraction. The possibility that the low-density membranes were the result of contamination of ER by oil bodies was ruled out by the use of oleosins as a marker for oil bodies. It is suggested that the low-density membranes are derived from a domain of the ER which is involved in the formation and secretion of TAG.     

Seasonal dynamics of fatty acid composition in female northern pike (Esox lucius L.)

Seasonal changes in the fatty acid composition of neutral and polar lipids were measured in the ovary, liver, white muscle, and adipopancreatic tissue of northern pike. The role of environmental and physiological factors underlying these changes was evaluated. From late summer (August–September) to winter (January–March), the weight percentage of n-3 polyunsaturated fatty acids (especially 22:6n3) declined significantly in the neutral lipids of all somatic tissues examined. However, large quantities of n-3 polyunsaturated fatty acids accumulated in the recrude cing ovaries during fall and the weight percentage of n-3 polyunsaturated fatty acids in ovary polar lipids also increased significantly. Additionally, the n-3 polyunsaturated fatty acid content of somatic polar lipids increased significantly during fall due to increases in the total polar lipid content of the somatic tissues. This suggests that during fall n-3 polyunsaturated fatty acid are diverted away from somatic neutral lipids and thereby conserved for use in ovary construction and for incorporation into tissue polar lipids. The percentage of n-3 polyunsaturated fatty acid in ovary neutral lipids also declined during fall and early winter, perhaps as an adaptation to conserve these fatty acids for storage in oocyte polar lipids and later incorporation into cellular membranes of the developing embryo. Reductions in the n-3 polyunsaturated fatty acids content of somatic and ovarian neutral lipids during fall were compensated for specifically by increases in the percentage of monounsaturated fatty acids rather than saturated fatty acids. This suggests that the ratio of saturated to unsaturated fatty acids in pike neutral lipid, is regulated physiologically, and hence may influence the physiological functioning of these lipids. During fall and early winter the percentage of saturated fatty acids declined significantly in the polar lipids of all tissues examined. This change was consistent with the known effects of cold acclimation on the fatty acid composition of cellular membranes. As the ovaries were recrudescing from September to January, liver polar lipids exhibited significant decreases in the percentage of total polyunsaturated fatty acids and n-3 polyunsaturated fatty acids and increases in monounsaturated fatty acids, and acquired a fatty acid composition very similar to that of ovary polar lipids. Therefore, seasonal changes in the percentage of polyunsaturated and monounsaturated fatty acids in liver polar lipids probably reflect the liver's role in vitellogenesis rather than the effects of temperature on membrane fatty acid composition. At all times of year, the fatty acid compositions of white muscle and adipopancreatic tissue neutral lipids were very similar, which may indicate a close metabolic relationship between these lipid compartments.Abbreviations AP adipopancreatic - BHT butylated hydroxytoluene - CI confidence interval - EFA essential fatty acids - MUFA monounsaturated fatty acids - NL neutral lipids - PL polar lipids - PUFA polyunsaturated fatty acids - SFA saturated fatty acids     

Proliferation of Oogonia and folliculogenesis in the viviparous teleost Ilyodon whitei (Goodeidae)

Oogonial proliferation in fishes is an essential reproductive strategy to generate new ovarian follicles and is the basis for unlimited oogenesis. The reproductive cycle in viviparous teleosts, besides oogenesis, involves development of embryos inside the ovary, that is, intraovarian gestation. Oogonia are located in the germinal epithelium of the ovary. The germinal epithelium is the surface of ovarian lamellae and, therefore, borders the ovarian lumen. However, activity and seasonality of the germinal epithelium have not been described in any viviparous teleost species regarding oogonial proliferation and folliculogenesis. The goal of this study is to identify the histological features of oogonial proliferation and folliculogenesis during the reproductive cycle of the viviparous goodeid Ilyodon whitei. Ovaries during nongestation and early and late gestation were analyzed. Oogonial proliferation and folliculogenesis in I. whitei, where intraovarian gestation follows the maturation and fertilization of oocytes, do not correspond to the late oogenesis, as was observed in oviparous species, but correspond to late gestation. This observation offers an example of ovarian physiology correlated with viviparous reproduction and provides elements for understanding the regulation of the initiation of processes that ultimately result in the origin of the next generation. These processes include oogonia proliferation and development of the next batch of germ cells into the complex process of intraovarian gestation. J. Morphol. 275:1004–1015, 2014. © 2014 Wiley Periodicals, Inc.     

Inactivation of microsomal triglyceride transfer protein impairs the normal redistribution but not the turnover of newly synthesized glycerolipid in the cytosol, endoplasmic reticulum and Golgi of primary rat hepatocytes.

The requirements for microsomal triglyceride transfer protein (MTP) during the turnover and transfer of glycerolipids from intracellular compartments into secretory very low-density lipoprotein (VLDL) were studied by pre-labelling lipids with [(3)H]glycerol and [(14)C]oleate in primary cultures of rat hepatocytes. The intracellular redistribution of pre-labelled glycerolipids was then compared at the end of subsequent chase periods during which the MTP inhibitor BMS-200150 was either present or absent in the medium. Inhibition of MTP resulted in a decreased output of VLDL triacylglycerol (TAG) and a delayed removal of labelled TAG from the cytosol and from the membranes of the smooth endoplasmic reticulum (SER), the cis- and the trans-Golgi. Inactivation of MTP did not decrease the bulk lipolytic turnover of cellular TAG as reflected by changes in its [(3)H]glycerol:[(14)C]oleate ratios. However, a larger proportion of the resultant TAG fatty acids was re-esterified and remained with the membranes of the various subcellular fractions rather than emerging as VLDL. The effects of BMS-200150 on the pattern of phospholipid (PL) mechanism and redistribution suggested that inhibition of MTP prevented the normal lipolytic transfer of PL-derived fatty acids out of the SER, cis- and trans-Golgi membrane pools. Finally, changes in the (14)C specific radioactivities of the cytosolic and membrane pools of TAG suggested that inhibition of MTP prevented a normal influx of relatively unlabelled fatty acids into these pools during the chase period.     

Large fluxes of fatty acids from membranes to triacylglycerol and back during N-deprivation and recovery in Chlamydomonas

Microalgae accumulate triacylglycerol (TAG) during nutrient deprivation and break it down after nutrient resupply, and these processes involve dramatic shifts in cellular carbon allocation. Due to the importance of algae in the global carbon cycle, and the potential of algal lipids as feedstock for chemical and fuel production, these processes are of both ecophysiological and biotechnological importance. However, the metabolism of TAG is not well understood, particularly the contributions of fatty acids (FAs) from different membrane lipids to TAG accumulation and the fate of TAG FAs during degradation. Here, we used isotopic labeling time course experiments on Chlamydomonas reinhardtii to track FA synthesis and transfer between lipid pools during nitrogen (N)-deprivation and resupply. When cells were labeled before N-deprivation, total levels of label in cellular FAs were unchanged during subsequent N-deprivation and later resupply, despite large fluxes into and out of TAG and membrane lipid pools. Detailed analyses of FA levels and labeling revealed that about one-third of acyl chains accumulating in TAG during N-deprivation derive from preexisting membrane lipids, and in total, at least 45% of TAG FAs passed through membrane lipids at one point. Notably, most acyl chains in membrane lipids during recovery after N-resupply come from TAG. Fluxes of polyunsaturated FAs from plastidic membranes into TAG during N-deprivation were particularly noteworthy. These findings demonstrate a high degree of integration of TAG and membrane lipid metabolism and highlight a role for TAG in storage and supply of membrane lipid components.

In Chlamydomonas, about a third of triacylglycerol (TAG) made during nitrogen deprivation is derived from preexisting membranes, and most membranes made after resupply are derived from TAG.     

Extraembryonic membrane development in a reproductively bimodal lizard, Lacerta (Zootoca) vivipara

Reproductive mode has been remarkably labile among squamate reptiles and the evolutionary transition from oviparity to viviparity commonly has been accompanied by a shift in the pattern of embryonic nutrition. Structural specializations for placental transfer of nutrients during intrauterine gestation are highly diverse and many features of the extraembryonic membranes of viviparous species differ markedly from those of oviparous species. However, because of a high degree of evolutionary divergence between the species used for comparisons it is likely that the observed differences arose secondarily to the evolution of viviparity. We studied development of the extraembryonic membranes and placentation in the reproductively bimodal lizard Lacerta vivipara because the influence of reproductive mode on the structural/functional relationship between mothers and embryos can best be understood by studying the most recent evolutionary events. Lecithotrophic viviparity has evolved recently within this species and, although populations with different reproductive modes are allopatric, oviparous and viviparous forms interbreed in the laboratory and share many life history characteristics. In contrast to prior comparisons between oviparous and viviparous species, we found no differences in ontogeny or structure of the extraembryonic membranes between populations with different reproductive modes within L. vivipara. However, we did confirm conclusions from previous studies that the tertiary envelope of the egg, the eggshell, is much reduced in the viviparous population. These conclusions support a widely accepted model for the evolution of squamate placentation. We also found support for work published nearly 80 years ago that the pattern of development of the yolk sac of L. vivipara is unusual and that a function of a unique structure of squamate development, the yolk cleft, is hematopoiesis. The structure of the yolk sac splanchnopleure of L. vivipara is inconsistent with a commonly accepted model for amniote yolk sac function and we suggest that a long standing hypothesis that cells from the yolk cleft participate in yolk digestion requires further study.     

Peripheral plasma progesterone and utero-ovarian prostaglandin F concentrations in the cow around parturition

The concentration of prostaglandin F in utero-ovarian venous plasma and progesterone in jugular venous plasma were determined by radioimmunoassay in 3 cows over the last 2–3 weeks of gestation. Utero-ovarian prostaglandin F concentrations did not show any consistent pattern in two of three cows until 48–72 h before term when the levels rose sharply from 1 ng/ml to a maximum 4–9 ng/ml during labour. The concentration of progesterone in jugular venous plasma tended to fall gradually over the last 20 days of gestation with a further fall occurring 48-36 h before delivery.In two other cows at around 240 days of gestation the concentration of plasma progesterone in ovarian venous plasma was 50 to 150 times the concentration of progesterone in uterine or jugular venous plasma. It is concluded from these results that the ovaries are the major source of progesterone in cows during late pregnancy. The findings also suggest that prostaglandin F may be the luteolytic factor responsible for the sharp decline in plasma progesterone concentrations over the last 48-36 h preceding parturition.