Changes of sex hormone-binding globulin/SHBG expression in the hypothalamo-hypophyseal system of rats during pregnancy, parturition and lactation.

Sex hormone-binding globulin (SHBG) is expressed in hypothalamic magnocellular neurons. High co-localization rates of SHBG with oxytocin have been observed in the hypothalamus, indicating that SHBG plays a role in pregnancy, parturition and lactation. Further studies have shown that hypothalamic SHBG expression is malleable to changing steroid conditions. In this study, we have examined SHBG levels in the supraoptic and paraventricular hypothalamic nuclei and in the posterior pituitary lobe of late pregnant, parturient and early lactating rats by IN SITU hybridization, immunocytochemistry, and ELISA. Immunocytochemical and biochemical analysis showed that the SHBG levels increased during late pregnancy in hypothalamic nuclei. During parturition, SHBG levels fell in the magnocellular nuclei but increased in the posterior pituitary lobe. SHBG levels increase again during lactation. At day six of lactation, there was no significant difference in SHBG levels compared to normal cycling female rats, which served as control in this study. IN SITU hybridization showed increased SHBG mRNA signal during late pregnancy. The highest SHBG expression was observed during parturition. Our data indicate that hypothalamic SHBG expression changes during pregnancy, parturition and lactation, parallel to ovarian steroid and co-localized OT levels. This may in part be linked to known steroid actions on synthesis and secretion of magnocellular hypothalamic peptide hormones, important for the control of parturition and lactation.     

Cyclic nucleotides, adenylate cyclase, and cyclic AMP phosphodiesterase in mammary glands from pregnant and lactating mice.

In the mammary glands of mice, levels of cyclic AMP increased during pregnancy and then fell precipitously following parturition. In contrast, levels of cyclic GMP fell during the gestation period and then rose rapidly during the early days of lactation. Adenylate cyclase and cyclic AMP hohsphodiesterase activities were elevated during the pregnancy and lactation periods.     

Prolactin and the dietary protein/carbohydrate ratio regulate the expression of SNAT2 amino acid transporter in the mammary gland during lactation

The sodium coupled neutral amino acid transporter 2 (SNAT2/SAT2/ATA2) is expressed in the mammary gland (MG) and plays an important role in the uptake of alanine and glutamine which are the most abundant amino acids transported into this tissue during lactation. Thus, the aim of this study was to assess the amount and localization of SNAT2 before delivery and during lactation in rat MG, and to evaluate whether prolactin and the dietary protein/carbohydrate ratio might influence SNAT2 expression in the MG, liver and adipose tissue during lactation. Our results showed that SNAT2 protein abundance in the MG increased during lactation and this increase was maintained along this period, while 24 h after weaning it tended to decrease. To study the effect of prolactin on SNAT2 expression, we incubated MG explants or T47D cells transfected with the SNAT2 promoter with prolactin, and we observed in both studies an increase in the SNAT2 expression or promoter activity. Consumption of a high-protein/low carbohydrate diet increased prolactin concentration, with a concomitant increase in SNAT2 expression not only in the MG during lactation, but also in the liver and adipose tissue. There was a correlation between SNAT2 expression and serum prolactin levels depending on the amount of dietary protein/carbohydrate ratio consumed. These findings suggest that prolactin actively supports lactation providing amino acids to the gland through SNAT2 for the synthesis of milk proteins.     

Influence of the metabolic state during lactation on milk production in modern sows

Selection for prolificacy in sows has resulted in higher metabolic demands during lactation. In addition, modern sows have an increased genetic merit for leanness. Consequently, sow metabolism during lactation has changed, possibly affecting milk production and litter weight gain. The aim of this study was to investigate the effect of lactational feed intake on milk production and relations between mobilization of body tissues (adipose tissue or skeletal muscle) and milk production in modern sows with a different lactational feed intake. A total of 36 primiparous sows were used, which were either full-fed (6.5 kg/day) or restricted-fed (3.25 kg/day) during the last 2 weeks of a 24-day lactation. Restricted-fed sows had a lower milk fat percentage at weaning and a lower litter weight gain and estimated milk fat and protein production in the last week of lactation. Next, several relations between sow body condition (loss) and milk production variables were identified. Sow BW, loin muscle depth and backfat depth at parturition were positively related to milk fat production in the last week of lactation. In addition, milk fat production was related to the backfat depth loss while milk protein production was related to the loin muscle depth loss during lactation. Backfat depth and loin muscle depth at parturition were positively related to lactational backfat depth loss or muscle depth loss, respectively. Together, results suggest that sows which have more available resources during lactation, either from a higher amount of body tissues at parturition or from an increased feed intake during lactation, direct more energy toward milk production to support a higher litter weight gain. In addition, results show that the type of milk nutrients that sows produce (i.e. milk fat or milk protein) is highly related to the type of body tissues that are mobilized during lactation. Interestingly, relations between sow body condition and milk production were all independent of feed level during lactation. Sow management strategies to increase milk production and litter growth in modern sows may focus on improving sow body condition at the start of lactation or increasing feed intake during lactation.     

Changes in maternal taurine levels in response to pregnancy and lactation

Taurine levels in various tissues and fluids of female rats were measured throughout pregnancy and lactation. The taurine concentration of liver markedly increased at days 19 and 21 of pregnancy to 188% of levels for nonpregnant, nonlactating control rats and then fell rapidly after delivery to reach only 30% of the control level by 3 days postpartum. Muscle and heart taurine concentrations were significantly negatively correlated with liver taurine levels. Brain taurine levels were low at days 14, 19 and 21 of pregnancy and day 14 of lactation. Urinary excretion of taurine decreased to 32% of control levels at day 21 of pregnancy and was negatively correlated with the hepatic taurine concentration over the course of pregnancy and lactation. The ratio of glycine- to taurine-conjugated bile acids was strongly negatively correlated with the hepatic taurine concentration. The milk taurine level was positively correlated with hepatic taurine concentration during lactation. The hepatic taurine pool appears to increase just before parturition and to rapidly decrease during the first few days of lactation when high levels of taurine are secreted in the milk. Our data suggests that the accumulation of taurine in the liver may be related to both a decreased renal clearance of taurine and a shifting of tauring from other tissues to the liver and that this enlarged pool of hepatic taurine may serve as a source of taurine for secretion in the early milk.     

Dietary protein restriction during lactation in primiparous sows with different live weights at farrowing: I. Consequences on sow metabolic status and litter growth

The hypothesis that the restriction of dietary protein during lactation has different impacts on sow metabolic status and milk production according to body weight was evaluated. From 5-months of age until farrowing, the gilts were fed to achieve body weights of 180 or 240 kg at farrowing. At this time, 38 sows were assigned to one of three groups: " 180 kg" sows not restricted in dietary protein during lactation (180CP); "180 kg" restricted in protein (180LP), or "240 kg" sows restricted in protein (240LP). Catheters were fitted in the jugular vein of 24 sows and serial blood samples were collected 1 d before and 1 d after weaning. Amongst the protein-restricted animals, heavy sows (240LP) had a smaller appetite than light sows in early lactation, resulting in lower energy and protein intakes in the 240LP than in the 180LP sows. Body protein losses were 8, 11 and 13.5% of calculated body protein mass at farrowing in the 180CP, 180LP and 240LP sows, respectively. At the end of lactation, IGF-I concentrations were lower in the 180LP than in the sows from the other groups, probably because of the uncoupling between GH and IGF-I secretions. Low IGF-I concentrations likely promote lean tissue mobilization. Glucose and insulin profiles suggested an insulin resistance state in the 240LP sows compared with the 180LP sows, which may explain, at least in part, the lower feed intake and body reserve mobilization in these sows. Plasma pre- and post-prandial concentrations of amino acids in late lactation differed among the three treatment groups. Throughout lactation, litters from the 180LP and 240LP sows had a slower growth rate than litters from sows which were not restricted, suggesting that endogenous protein mobilization throughout lactation does not completely compensate for a low protein intake.     

Lactation during hibernation in wild black bears: effects on plasma amino acids and nitrogen metabolites.

This study examined the seasonal and reproductive influences on individual plasma amino acid concentrations and nitrogen metabolites in a black bear population (Ontario, Canada). During hibernation, 11 of 23 plasma amino acids were significantly higher (13%-108%) in lactating than in nonlactating females, without an alteration in plasma total protein or total essential or nonessential amino acid levels. The greatest changes were observed in glutamine, arginine, and glycine levels. Plasma urea, urea/creatinine, and ammonia levels were significantly lower in hibernating compared with active female bears, but lactation had no effect on these parameters. Taken together these results show that lactation during hibernation is an additional metabolic challenge that results in increased mobilization of individual plasma amino acids and no accumulation of nitrogen end products, underlining the remarkable efficiency of amino acid and urea recycling in denning female black bears.     

饲料蛋白水平对湘华鲮幼鱼生长性能、体成分及血清生化指标的影响

为研究饲料中不同蛋白水平对湘华鲮[Sinilabeo decorus Tungting (Nichols)]幼鱼形体指标、体成分及血清生化指标的影响, 配制蛋白水平为32.57%、37.58%、42.76%、47.83%和52.22%的5种等脂饲料。选取初始质量为(14.40±1.08) g的湘华鲮幼鱼450尾, 随机分成5组, 每组3个重复, 每个重复30尾, 进行为期60d的饲养实验。结果表明: (1)饲料不同蛋白水平对湘华鲮的成活率和饲料系数无显著影响(P>0.05); 蛋白质效率随饲料蛋白水平的升高显著降低(P<0.05); 52.22%蛋白组的增重率和特定生长率均显著低于32.57%和42.76%蛋白组(P<0.05)。(2)随着饲料蛋白水平升高, 湘华鲮肥满度呈降低趋势, 且52.22%蛋白组显著低于其他组(P<0.05); 肝体比呈先降后升的趋势, 且37.58%蛋白组显著低于32.57%、47.83%和52.22%蛋白组(P<0.05); 肠体比呈降低趋势, 且52.22%蛋白组显著低于32.57%蛋白组(P<0.05)。(3)湘华鲮肌肉粗蛋白质含量随着饲料蛋白水平升高而增大, 且52.22%蛋白组显著大于32.57%和37.58%蛋白组(P<0.05), 42.76%蛋白组显著大于32.57%蛋白组(P<0.05); 饲料蛋白水平对全鱼的水分、粗脂肪、粗蛋白质、灰分及肌肉的水分和粗脂肪含量没有显著影响(P>0.05)。(4)肌肉中均检测出17种氨基酸(除色氨酸), 其中52.22%蛋白组的必需氨基酸总和(ΣEAA)、非必需氨基酸总和(ΣNEAA)、鲜味氨基酸总和(ΣDAA)及氨基酸总和(ΣTAA)显著大于32.57%蛋白组(P<0.05)。(5)不同蛋白组间甘油三酯(TG)和极低密度脂蛋白(VLDL)水平均表现为37.58%<32.57%<42.76%<47.83%<52.22%, 且52.22%蛋白组显著高于37.58%蛋白组(P<0.05)。综合生长性能、形体指标、体成分及血清生化指标, 初步认定湘华鲮幼鱼最适蛋白质水平为37.58%—42.76%。通过特定生长率(SGR)和饲料蛋白水平的折线法分析表明, 湘华鲮幼鱼达到最大生长时饲料蛋白质水平为42.91%。     

Longitudinal Profiling of the Tissue-Specific Expression of Genes Related with Insulin Sensitivity in Dairy Cows during Lactation Focusing on Different Fat Depots

In dairy cows the milk associated energy output in early lactation exceeds the input via voluntary feed intake. To spare glucose for mammary lactose synthesis, peripheral insulin sensitivity (IS) is reduced and fat mobilization is stimulated. For these processes a link between IS and the endocrine functions of adipose tissue (AT) is likely; we thus aimed to characterise the mRNA expression from bovine AT derived proteins and receptors that are related to IS according to the literature in metabolically active tissues plus systemic IS throughout lactation. Conjugated linoleic acids (CLA) reduce milk fat thus decreasing the milk drain of energy and potentially dampening lipolysis, but may also affect IS. Subcutaneous (s.c.) AT and liver from pluriparous cows receiving either control fat or CLA supplement (100 g/day from 1 to 182 days in milk each) were biopsied covering week −3 to 36 relative to parturition. In an additional trial with primiparous cows treated analogously and slaughtered on days in milk 1, 42 or 105, samples from liver, udder, skeletal muscle and 3 visceral and 3 s.c. AT were obtained and assayed for mRNA abundance of adiponectin, its receptors, leptin, leptin receptor, PPARγ, PPARγ2, IL-6, and TNF-α. In pluriparous animals, the mRNA abundance of most of the target genes decreased after parturition in s.c. AT but increased in liver. In primiparous cows, AT depot specific differences were mostly related to retroperitoneal AT; adiponectin receptor 1 and TNF-α were affected predominantly. CLA effects in primiparous cows were largely limited to decreased PPARγ2 mRNA abundance in udder tissue. In pluriparous cows, insulin secretion was increased by CLA resulting in decreased systemic IS but without consistent changes in tissue target mRNA abundance. The temporal gene expression profiles from the adipokines and related receptors support their coactive function in adapting to the needs of lactation.     

MMP-2 expression precedes the final ripening process of the bovine cervix

Collagen is denatured in the gradual cervical ripening process during late pregnancy, already before the onset of final cervical ripening at parturition. Matrix Metallo Proteinases (MMPs) might be responsible for this process. To investigate the presence and potential function of MMPs at the different stages of the ripening process, serial cervical biopsies were obtained from 10 cows at Days 185 and 275 of pregnancy (approximately 5 days before calving), at parturition and at 30 days after parturition. The mRNA and protein expression of MMP-1, MMP-2, and MMP-9 and of the tissue inhibitors of MMPs (TIMP)-1 and TIMP-2 were semi-quantitatively determined using RT-PCR, respectively, zymography, Westernblot, and ELISA techniques and the localization of MMP-2 protein and presence of granulocytes by immunohistochemistry and Luna staining. At parturition compared to 185 days pregnancy the MMP-1 protein expression and the numbers of granulocytes were significantly increased by 3 and 26-fold respectively. MMP-2 mRNA and protein expression had already increased 2.5 (P < 0.05) and twofold (P < 0.05) at 5 days before parturition, prior to final ripening. At that time, MMP-2 was present in smooth muscle cells and extra cellular matrix. TIMP-1 mRNA expression was significantly increased at parturition and TIMP-2 mRNA expression peaked at 5 days before parturition. The increased expression of MMP-2 at 5 days before parturition, suggests that in the cow MMP-2 is responsible for collagen denaturation in the last part of gradual cervical ripening, while MMP-1 and MMP-9 are only active during the final cervical ripening process at parturition.     

Up-regulation of phosphatidylinositol 3-kinase and glucose transporter 4 in muscle of rats subjected to maternal undernutrition

Early postnatal nutrition has been associated with the long-term effects on glucose homeostasis in adulthood. To elucidate the molecular mechanisms by which undernutrition during early life leads to changes in insulin sensitivity, we investigated the insulin signaling in skeletal muscle of rats during development. Offspring of dams fed with either protein-free or normal diets during the first 10 days of lactation were studied from lactation period until adulthood. Early maternal undernutrition impaired secretion of insulin but maintained normal blood glucose levels until adulthood. Insulin receptor (IR) activation after insulin stimulation was decreased during the period of protein restriction. In addition, glucose uptake, insulin receptor substrate 1 (IRS-1) phosphorylation and glucose transporter 4 (GLUT-4) translocation in muscle were reduced in response to insulin during suckling. In contrast, non- or insulin-stimulated glucose uptake and GLUT-4 translocation were found significantly increased in muscle of adult offspring. Finally, basal association of phosphatidylinositol 3-kinase (PI3-kinase) with IRS-1 was increased and was highly stimulated by insulin in muscle from adult rats. Our findings suggest that early postnatal undernutrition increases insulin sensitivity in adulthood, which appears to be directly related to changes in critical steps required for glucose metabolism.     

Involvement of skeletal muscle protein, glycogen, and fat metabolism in the adaptation on early lactation of dairy cows

During early lactation, high-yielding dairy cows cannot consume enough feed to meet nutrient requirements. As a consequence, animals drop into negative energy balance and mobilize body reserves including muscle protein and glycogen for milk production, direct oxidation, and hepatic gluconeogenesis. To examine which muscle metabolic processes contribute to the adaptation during early lactation, six German Holstein cows were blood sampled and muscle biopsied throughout the periparturient period. From pregnancy to lactation, the free plasma amino acid pattern imbalanced and plasma glucose decreased. Several muscle amino acids, as well as total muscle protein, fat, and glycogen, and the expression of glucose transporter-4 were reduced within the first 4 weeks of lactation. The 2-DE and MALDI-TOF-MS analysis identified 43 differentially expressed muscle protein spots throughout the periparturient period. In early lactation, expression of cytoskeletal proteins and enzymes involved in glycogen synthesis and in the TCA cycle was decreased, whereas proteins related to glycolysis, fatty acid degradation, lactate, and ATP production were increased. On the basis of these results, we propose a model in which the muscle breakdown in early lactation provides substrates for milk production by a decoupled Cori cycle favoring hepatic gluconeogenesis and by interfering with feed intake signaling.     

Expression of the estrogen receptor in blood neutrophils of dairy cows during the periparturient period

During the period around parturition, cows experience an increased susceptibility to inflammatory disorders in the mammary gland and uterus. This increased susceptibility has been correlated with a decreased functionality of neutrophils, major components in the innate immune defence. As sex steroid levels vary extensively in the period around parturition, an influence of these changes on the functionality of neutrophils has been suggested. Indeed, it has been shown that 17beta-estradiol affects some functions of bovine neutrophils. In spite of these observations, receptors for 17beta-estradiol have not yet been demonstrated in these cells. The investigation of the presence of estrogen receptors in bovine neutrophils was therefore the main objective of this study. The expression of estrogen receptors was evaluated at the protein level by flow cytometry, and at the mRNA level by polymerase chain reaction. A clear positive signal was obtained using flow cytometry for the estrogen receptor protein in bovine neutrophils. Further discrimination between the estrogen receptor subtypes alpha and beta revealed the expression of the estrogen receptor beta, whereas for the estrogen receptor alpha no reproducible positive signal could be obtained with the available antibodies. Both subtypes were found at the mRNA level. Subsequently, the estrogen receptor protein expression level in neutrophils obtained from cows in early lactation was compared with those from cows in late pregnancy. Additionally, the influence of endogenous 17beta-estradiol and progesterone levels was assessed. No difference was found for the estrogen receptor protein expression in neutrophils from cows in early lactation compared with late gestation neither were the endogenous 17beta-estradiol and progesterone levels correlated with the protein expression.     

骨骼肌钝挫伤恢复过程中HIF-1α、AMPKα2表达的改变

目的：观测大鼠骨骼肌钝挫伤（SMBI）恢复进程中腺苷酸活化蛋白激酶α2（AMPKα2）、缺氧诱导因子-1α（HIF-1α）的表达变化,探讨SMBI恢复可能生物学机制。方法：雄性Wistar大鼠48只,随机选取6只作为正常对照组;另42只大鼠重物砸伤后肢小腿建立钝挫伤模型后分7组（n=6）,造模后各组分别在伤后12 h、2 d、5 d、7 d、10 d、15 d、30 d取材,检测小腿三头肌HIF-1α、AMPKα2表达的变化。结果：损伤后12 h HIF-1α、AMPKα2表达均明显升高,在伤后15 d开始回落接近正常;HIF-1α、AMPKα2表达的峰值出现在伤后2 d内,伤后5 d后表达量开始下降;除HIF-1α mRNA在伤后2 d组表达出现峰值外,其余时间点二者mRNA与蛋白表达时程变化基本一致。结论：SMBI后,HIF-1α、AMPKα2可能通过参与调解缺氧适应、肌细胞再生、能量代偿起到促进伤后恢复的作用。     

Chronic Maternal Vitamin B12 Restriction Induced Changes in Body Composition & Glucose Metabolism in the Wistar Rat Offspring Are Partly Correctable by Rehabilitation

Maternal under-nutrition increases the risk of developing metabolic diseases. We studied the effects of chronic maternal dietary vitamin B12 restriction on lean body mass (LBM), fat free mass (FFM), muscle function, glucose tolerance and metabolism in Wistar rat offspring. Prevention/reversibility of changes by rehabilitating restricted mothers from conception or parturition and their offspring from weaning was assessed. Female weaning Wistar rats (n = 30) were fed ad libitum for 12 weeks, a control diet (n = 6) or the same with 40% restriction of vitamin B12 (B12R) (n = 24); after confirming deficiency, were mated with control males. Six each of pregnant B12R dams were rehabilitated from conception and parturition and their offspring weaned to control diet. While offspring of six B12R dams were weaned to control diet, those of the remaining six B12R dams continued on B12R diet. Biochemical parameters and body composition were determined in dams before mating and in male offspring at 3, 6, 9 and 12 months of their age. Dietary vitamin B12 restriction increased body weight but decreased LBM% and FFM% but not the percent of tissue associated fat (TAF%) in dams. Maternal B12R decreased LBM% and FFM% in the male offspring, but their TAF%, basal and insulin stimulated glucose uptake by diaphragm were unaltered. At 12 months age, B12R offspring had higher (than controls) fasting plasma glucose, insulin, HOMA-IR and impaired glucose tolerance. Their hepatic gluconeogenic enzyme activities were increased. B12R offspring had increased oxidative stress and decreased antioxidant status. Changes in body composition, glucose metabolism and stress were reversed by rehabilitating B12R dams from conception, whereas rehabilitation from parturition and weaning corrected them partially, highlighting the importance of vitamin B12 during pregnancy and lactation on growth, muscle development, glucose tolerance and metabolism in the offspring.     

Southern elephant seal,Mirounga leonina: composition of milk during lactation

An analysis of milk constituents during various stages of lactation in the southern elephant seal Mirounga leonina was carried out. Forty-six milk samples were taken from 30 females throughout lactation during 1985, 1987, 1990 and 1991 on Stranger Point, King George Island, South Shetland Islands. Total nitrogen (TN), non-protein nitrogen (NPN), sugar, fat, ash and water were measured, and from some of these data true protein and energy content were calculated. The results showed a high degree of variation in water and fat concentrations among samples at different stages of lactation. During the first 20 days the fat content of milk increased from about 12 to approximately 52%, while water content fell from 70 to 33%. The composition of milk changes rapidly during the first days post-partum. Protein, minerals and sugar appear to remain stable after the fourth day of lactation. Milk samples contain significant levels of sugars; thin layer chromatography indicates the presence of lactose and glucose together with other unidentified components. There is evidence of a striking change in composition of the milk in the later part of lactation; the progressive increase in the fat:water ratio is abruptly reversed just prior to weaning.