How do maternal androgens and estrogens affect sex determination in reptiles with temperature-dependent sex?

Temperature sex determination (TSD) in reptiles has been studied to elucidate the mechanisms by which temperature is transformed into a biological signal that determines the sex of the embryo. Temperature is thought to trigger signals that alter gene expression and hormone metabolism, which will determine the development of female or male gonads. In this review, we focus on collecting and discussing important and recent information on the role of maternal steroid hormones in sex determination in oviparous reptiles such as crocodiles, turtles, and lizards that possess TSD. In particular, we focus on maternal androgens and estrogens deposited in the egg yolk and their metabolites that could also influence the sex of offspring. Finally, we suggest guidelines for future research to help clarify the link between maternal steroid hormones and offspring sex.

Caiman's fat enriched with n-3 fatty acids: potential food supplement.

Broad-snouted caiman (Caiman latirostris) products (meat, fat and oil) are currently beginning to be valued as a food of special interest due to its high content of n-3 fatty acids. Thus, the objective of this study was to characterize the fats of caiman fed with diets enriched with flaxseeds (Linus usitatissimun) rich in n-3 fatty acids, lignans and antioxidants. Caimans were fed six days a week with: a control diet (C), and a diet enriched with ground flaxseed = 90% C + 10% flaxseed ground (FS), during 30 (FS30) and 60 (FS60) days. Animals fed the flaxseed-enriched diets increased linolenic acid content and reduced the n-6/n-3 ratio of fats relative to controls, and this improvement increased over time. The proportion of eicosapentaenoic acid also increased, but there was no difference at the time the enriched diets were offered. Caiman fat of the FS30 and FS60, showed a decrease in lipoperoxidation (24% and 40%) and reactive oxygen species (44% and 76%) accompanied by an increase in antioxidant systems. Consumption of a flax-enriched diet by caimans increases the content of essential fatty acids and improves the lipoperoxidative status of fat. This provides an enriched fat with potential for the development products for human consumption.

Effect of flaxseed and flaxseed oil supplemented in caiman diet on meat fatty acids.

Increasing polyunsaturated or long-chain fatty acids in meat for human consumption improves both nutritional quality and consumer perception. The increase could occur through the addition of rich sources of omega-3 fatty acids (such as flaxseed or flaxseed oil) to the animal diet. The aim of this study was to evaluate the effects of dietary supplement with two presentations of flax (crushed seeds or oil) on the change of FA content in two cuts of caiman meat (tail and neck). We measured fatty profile in two different caiman meat cuts (neck and tail) from 30 animals (total length 96.7 ± 4.9 cm, snout-vent length 47.8 ± 3 cm, weight 4.2 ± 0.6 kg), raised in individual enclosures, fed three a week for 50 days with crushed chicken head and a dry food formulated for these reptiles in a 70/30 ratio (C, n = 10), control diet with 10% crushed flaxseed (FS, n = 10), and control diet with 10% flaxseed oil (FO, n = 10), while the remaining days animals were fed the control diet. Meats from animals fed both enrichment diet (FS and FO) showed an increase of C18:3n-3 and ΣUFA with respect to control diet. Although both enriched diets raised the levels of C18:3n-3, the neck showed higher values than the tail. We observed that the neck is more susceptible than the tail to be improved by FO, which could suggest that it is more beneficial to consume neck meat. In order to be implemented in caiman farms, flaxseed oil is more expensive than seed, but more effective, easier to manage, and is practical for application on caiman farms.

Optimization of oil extraction from caiman fat. Characterization for use as food supplement.

The leather of Caiman latirostris is highly appreciated in the fashion industry and the meat is valued as an important food but its fat are usually discarded because it has no commercial value. However it is an alternative source of essential fatty acids and could be used for human consumption. The aim was to optimize the oil extraction from Caiman latirostris fat and to carry out the chemical and microbiological characterization for its use as food supplement. The oil obtained by fusion method contains fatty acids with high nutritional quality such as oleic acid (34%), linoleic acid (30%) and α-linolenic acid (2%). The atherogenicity index was 0.29 and the thrombogenicity index 0.47. The presence of mesophilic aerobic bacteria, coliform bacteria, Escherichia coli and Salmonella were not observed, and the oil is stable for 4 months at 25 °C and for at least 8 months in an inert atmosphere at 25 °C.

Maternal conjugated linoleic acid modulates TAG metabolism in adult rat offspring.

Conjugated linoleic acid (CLA) might regulate the lipid depots in liver and adipose tissue. As there is an association between maternal nutrition, fat depots and risk of offspring chronic disease, the aim was to investigate the effect of maternal CLA consumption on TAG regulation and some inflammatory parameters in adult male rat offspring receiving or not receiving CLA. Female Wistar rats were fed control (C) or CLA-supplemented (1 %, w/w) diets during 4 weeks before and throughout pregnancy and lactation. After weaning, male offspring of CLA rats were fed C or CLA diets (CLA/C and CLA/CLA groups, respectively), whereas C male rat offspring were fed a C diet (C/C group) for 9 weeks. Serum TAG levels were increased in the CLA/CLA and CLA/C groups, associated with a reduction of lipoprotein lipase activity and weights of adipose tissue. The liver TAG levels were decreased in the CLA/CLA group, related to a significant reduction of fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) and glucose-6-phosphate dehydrogenase enzyme activities, as well as to the mRNA levels of FAS, ACC, stearoyl-CoA desaturase-1 and sterol regulatory element-binding protein-1c. Even though normal TAG levels were found in the liver of CLA/C rats, a reduction of lipogenesis was also observed. Thus, these results demonstrated a programming effect of CLA on the lipid metabolic pathways leading to a preventive effect on the TAG accretion in adipose tissue and the liver of male rat offspring. This knowledge could be important to develop some dietary strategies leading to a reduced incidence of obesity and fatty acid liver disease in humans.

Effects of trans-fatty acids on liver lipid metabolism in mice fed on diets showing different fatty acid composition.

AIM: Our aim was to investigate the effects of trans-fatty acids (TFA) on liver lipid metabolism in mice fed on experimental diets rich in either oleic or linoleic acid. METHODS: Twenty-two male CF1 mice (22.0 ± 0.1 g) were fed with diets rich in corn oil or olive oil, supplemented or not with TFA (0.75 g TFA/100 g diet), for 4 weeks. Changes in triacylglycerol content, the activity and expression of enzymes involved in lipogenesis and fatty acid oxidation were measured. RESULTS: Supplementation of an olive oil-rich diet with TFA increased liver triacylglycerols, the activity and expression of lipogenic enzymes and sterol regulatory element-binding protein SREBP-1a expression. By contrast, when TFA were added to a corn oil-rich diet, they did not modify these parameters. No significant differences were observed among the experimental groups in the activity and expression of carnitine palmitoyltransferase-Ia, body and liver weights or serum triacylglycerol concentrations. CONCLUSIONS: The effect of TFA on liver fat accumulation depends on the dietary fatty acid composition. Steatosis induced by TFA when included in an olive oil diet (but not in a corn oil diet) was associated with an increased lipogenesis but not with a decreased fatty acid oxidation in animals fed on the olive oil diet. This metabolic change is mediated by SREBP-1a but not by SREBP-1c, and seems to be independent of insulin.

Conjugated linoleic acid reduces hepatic steatosis and restores liver triacylglycerol secretion and the fatty acid profile during protein repletion in rats.

Protein depletion is associated with hepatic steatosis and decreased circulating triacylglycerol (TAG). Since conjugated linoleic acid (CLA) increases lean body mass, protects against muscle catabolism, and modulates lipid metabolism, the aim of this work was to investigate the effects of CLA with two different amounts of dietary fat on the regulation of plasma and hepatic TAG concentration, and its possible connections with changes in fatty acid (FA) profile in plasma, liver and adipose tissue and hepatic oxidative status during protein repletion. Rats were fed a low protein diet (14 days) and then a protein repletion diet (30 days), supplemented or not with CLA, containing 7% (w/w) or 20% (w/w) of fat. Hepatic TAG secretion and removal by muscle and adipose tissue lipoprotein lipase, FA profile and liver oxidative status were evaluated. Protein depletion affected hepatic TAG secretion and peripheral removal, decreasing plasma and increasing liver TAG concentration, whereas protein repletion with CLA improved these abnormalities independently of the amount of dietary fat by increasing hepatic TAG secretion. This prevention in the absence of CLA was not observed. CLA was incorporated in plasma and tissues (adipose > liver > plasma, and c9,t11-CLA > t10,c12-CLA), accompanied by alterations in FA composition, mainly in adipose tissue. The hepatic oxidative stress was overcome by protein repletion. CLA had a beneficial impact on TAG metabolism in protein repleted animals, preventing hepatic steatosis through higher hepatic TAG secretion.

The interactions between the chronic exposure to Aluminum and liver regeneration on bile flow and organic anion transport in rats.

The chronic exposure to Aluminum (Al) may compromise different liver functions, mainly during the hepatic regeneration. The aim of this study is to investigate the interactions between the chronic i.p. exposure to Al and hepatic regeneration (HR) on bile flow and organic anion transport in experimental animals. For this purpose, we studied bile flow and fractional transfer rates for the transport of hepatic organic anions (hepatic uptake, sinusoidal efflux, and canalicular excretion), as well as parameters related with the oxidative stress (OS), on rats chronically treated with Al at 0 and 2 days of HR. The Al treatment and time of HR caused a decrease in the biliary flow and in the hepatic uptake and canalicular excretion constants. In addition, Al and HR increased the lipoperoxidation associated with a reduction of the glutathione content and glutathione peroxidase and catalase enzyme's activities. Since the effects of Al and HR on biliary flow and transport systems were additive, but not on the oxidative status, different mechanisms might be involved on these alterations. Even though the OS may play a key role on the hepatic deleterious effects, there is no unique cause-effect relationship between OS and liver dysfunction in this experimental animal model.

Effects of dietary conjugated linoleic acid at high-fat levels on triacylglycerol regulation in mice.

OBJECTIVE: Our aim was to investigate the effects of dietary conjugated linoleic acid (CLA) at high-fat (HF) levels on parameters related to triacylglycerol (TG) regulation and some potential impacts on liver damage. METHODS: Growing mice were fed a control diet (7% corn oil), an HF diet containing 20% corn oil, or an HF diet containing 3% CLA (HF + CLA) for 30 d. Tissue and organ weights, plasma and tissue TG levels, and parameters related to their regulation were evaluated. Liver oxidative status was also assessed. RESULTS: Dietary CLA showed detrimental and beneficial effects. CLA added to the HF diet caused hepatomegaly (+32%) and exacerbated the hepatic TG accumulation (+168%) observed with the HF diet without inducing liver damage; however, it significantly reduced plasma TG concentrations (-37%) and normalized muscular TG content. An increase in glutathione was associated with total normalization of liver lipid peroxidation. In addition, HF + CLA caused dystrophy of epididymal fat pads, even when the HF diet had increased the adipose tissue mass (30%). The biochemical mechanisms involved in the regulation of lipid levels were related to reduced (-20%) hepatic very low-density lipoprotein-TG secretion and decreased muscle (-35%) and adipose (-49%) tissue contributions to the removal of plasma TG by lipoprotein lipase enzymes. CONCLUSION: Examination of CLA at HF levels showed hepatomegaly and exacerbation of lipid accretion as a negative impact; however, some positive aspects such as hypotriglyceridemia and protection against oxidative stress were also induced. Even the fat reduction is nutritionally important for weight control; the biochemical mechanisms whereby CLA mediates the potential effects could produce undesirable metabolic alterations.

Involvement of oxidative stress in the impairment in biliary secretory function induced by intraperitoneal administration of aluminum to rats.

We have shown that aluminum (Al) induces cholestasis associated with multiple alterations in hepatocellular transporters involved in bile secretory function, like Mrp2. This work aims to investigate whether these harmful effects are mediated by the oxidative stress caused by the metal. For this purpose, the capability of the antioxidant agent, vitamin E, to counteract these alterations was studied in male Wistar rats. Aluminum hydroxide (or saline in controls) was administered ip (27 mg/kg body weight, three times a week, for 90 d). Vitamin E (600 mg/kg body weight) was coadministered, sc. Al increased lipid peroxidation (+50%) and decreased hepatic glutation levels (-43%) and the activity of glutation peroxidase (-50%) and catalase (-88%). Vitamin E counteracted these effects total or partially. Both plasma and hepatic Al levels reached at the end of the treatment were significantly reduced by vitamin E (-40% and -44%, respectively; p<0.05). Al increased 4 times the hepatic apoptotic index, and this effect was fully counteracted by vitamin E. Bile flow was decreased in Altreated rats (-37%) and restored to normality by vitamin E. The antioxidant normalized the hepatic handling of the Mrp2 substrates, rose bengal, and dinitrophenyl-S-glutathione, which was causally associated with restoration of Mrp2 expression. Our data indicate that oxidative stress has a crucial role in cholestasis, apoptotic/necrotic hepatocellular damage, and the impairment in liver transport function induced by Al and that vitamin E counteracts these harmful effects not only by preventing free-radical formation but also by favoring Al disposal.

Biliary secretory function in rats chronically intoxicated with aluminum.

The effects of a chronic aluminum (Al) exposure on biliary secretory function, with special emphasis on hepatic handling of non-bile salt organic anions, was investigated. Male Wistar rats received, intraperitoneally, either 27 mg/kg body weight of Al, as Al hydroxide [Al (+) rats], or the vehicle saline [Al (-) rats] three times a week for 3 months. Serum and hepatic Al levels were increased by the treatment (approximately 9- and 4-fold, respectively). This was associated with enhanced malondialdehyde formation (+110%) and a reduction in GSH content (-17%) and in the activity of the antioxidant enzymes catalase (-84%) and GSH peroxidase (-46%). Bile flow (-23%) and the biliary output of bile salts (-39%), cholesterol (-43%), and proteins (-38%) also decreased. Compartmental analysis of the plasma decay of the model organic anion bromosulphophthalein revealed that sinusoidal uptake and canalicular excretion of the dye were significantly decreased in Al (+) rats (-53 and -43%, respectively). Expression of multidrug resistance-associated protein 2 (Mrp2), the main, multispecific transporter involved in the canalicular excretion of organic anions, was also decreased (-40%), which was associated with a significant decrease in the cumulative biliary excretion of the Mrp2 substrate, dinitrophenyl-S-glutathione (-50%). These results show that chronic Al exposure leads to oxidative stress, cholestasis, and impairment of the hepatic handling of organic anions by decreasing both sinusoidal uptake and canalicular excretion. The alteration of the latter process seems to be causally related to impairment of Mrp2 expression. We have addressed some possible mechanisms involved in these deleterious effects.