Regulation of hepatic triacylglycerol synthesis and secretion

Triacylglycerols are the most concentrated storage form of energy for the mammalian organism. These lipids are synthesized and secreted by the liver and serve as a fuel for other tissues. This paper presents a brief review of the regulation of hepatic triacylglycerol synthesis and secretion. Particular attention will be given to the dissociation of the synthesis of triacylglycerols from that of the metabolically closely related nitrogenous phospholipids. Recent evidence is presented which suggests that triacylglycerol synthesis (and secretion) is regulated, at least partially, at the diacylglycerol branchpoint.     

RNA and protein synthesis is required for Ancylostoma caninum larval activation

The developmentally arrested infective larva of hookworms encounters a host-specific signal during invasion that initiates the resumption of suspended developmental pathways. The resumption of development during infection is analogous to recovery from the facultative arrested dauer stage in the free-living nematode Caenorhabditis elegans. Infective larvae of the canine hookworm Ancylostoma caninum resume feeding and secrete molecules important for infection when exposed to a host mimicking signal in vitro. This activation process is a model for the initial steps of the infective process. Dauer recovery requires protein synthesis, but not RNA synthesis in C. elegans. To determine the role of RNA and protein synthesis in hookworm infection, inhibitors of RNA and protein synthesis were tested for their effect on feeding and secretion by A. caninum infective larvae. The RNA synthesis inhibitors α-amanitin and actinomycin D inhibit feeding dose-dependently, with IC(50) values of 30 and 8 μM, respectively. The protein synthesis inhibitors puromycin (IC(50)=110 μM), cycloheximide (IC(50)=50 μM), and anisomycin (IC(50)=200 μM) also displayed dose-dependent inhibition of larval feeding. Significant inhibition of feeding by α-amanitin and anisomycin occurred when the inhibitors were added before 12h of the activation process, but not if the inhibitors were added after 12h. None of the RNA or protein synthesis inhibitors prevented secretion of the activation-associated protein ASP-1, despite nearly complete inhibition of feeding. The results indicate that unlike dauer recovery in C. elegans, de novo gene expression is required for hookworm larval activation, and the critical genes are expressed within 12h of exposure to activating stimuli. However, secretion of infection-associated proteins is independent of gene expression, indicating that the proteins are pre-synthesized and stored for rapid release during the initial stages of infection. The genes that are inhibited represent a subset of those required for the transition to parasitism, and therefore represent interesting targets for further investigation. Furthermore, while dauer recovery provides a useful model for hookworm infection, the differences identified here highlight the importance of exercising caution before making generalizations about parasitic nematodes based on C. elegans biology.     

Estrogen regulates the serum level of phosphorylated prolactin in mice

Phosphorylated prolactin (PPRL) is considered to be the most quantitatively important post-translationally modified form of prolactin (PRL) in rodents. We recently detected two different types of PPRL in the mouse pituitary gland; one was phosphorylated at serine and the other was phosphorylated at serine/threonine. Furthermore, we showed that there are obvious differences in the ratios between PPRLs and non-phosphorylated PRL in the pituitary gland based on age and sex and that estrogen influences PRL phosphorylation at serine in female mice. In the present study, we examined whether estradiol (E2) increases serine PPRL in the male pituitary gland in the same manner as in the female pituitary gland and examined whether PPRL is released into serum. We first determined the relative amounts of intrapituitary PPRLs in male mice under different pharmacological conditions that increased PRL secretion. The results indicated that treatment with E2 increases serine PPRL. We then performed two-dimensional electrophoresis and immunoblotting analysis after immunoprecipitation with anti-mouse PRL antibody using male and female sera under different pharmacological conditions that increased PRL secretion. The results of this experiment indicated that there were PRLs phosphorylated at serine and serine/threonine in the female serum but not in the male serum. The levels of PPRLs in sera were greatly increased with the E2 treatment for both male and female sera. Furthermore, we examined the effect of E2 on PPRL synthesis in cultured male pituitary glands. In this experiment, we observed increased serine PPRL synthesis and stronger immunohistochemical staining of PRL cells with E2 treatment. These findings suggested that serine PPRL synthesis and secretion were influenced by estrogen.     

Tumor necrosis factor-alpha as a possible auto-/paracrine factor affecting estrous cycle in the cat uterus

Tumor Necrosis Factor-alpha (TNFalpha) is a pleiotrophic cytokine, affects either normal or tumor cells, and influences cellular differentiation. TNFalpha role in female reproduction has been proven to be mediated through an influence on prostaglandin (PGs) synthesis and output. To evaluate the possible role of TNFalpha in an auto-/paracrine regulation in the cat uterus, mRNA expression coding for TNFalpha and its receptors (TNFR1 and TNFR2), and TNFalpha protein content at different stages of the estrous cycle were investigated. Additionally, TNFalpha involvement in PG secretion at different stages of the estrous cycle was investigated by in vitro tissue culture. Gene expressions coding for TNFalpha and TNFR1 were the highest at diestrus (P < 0.05). TNFalpha protein expression was the lowest at interestrus (P < 0.05). Nevertheless, TNFR2 was not affected by the estrous stage. TNFalpha at a dose of 1 ng/ml significantly increased PGF2alpha secretion at estrus (P < 0.01) and PGE2 secretion at diestrus (P < 0.001) after 12h incubation. Overall findings indicate that TNFalpha locally produced in the cat's uterus, stimulates PG secretion in an estrous cycle-related manner.     

家蚕4眠期与5龄期后部丝腺细胞蛋白质组成分析

为探讨家蚕丝腺细胞合成分泌蛋白质过程中有关基因的表达调控机理,采用蛋白质双向电泳和图像分析技术,研究了家蚕4眠期和5龄期后部丝腺细胞的蛋白质组成。在4眠家蚕的后部丝腺细胞中检测到425个蛋白斑点,5龄期随着生长发育的进程,可检测到新的特异蛋白斑点,说明不同的发育时期,后部丝腺细胞的蛋白组成是不一致的。5龄前、中期新增加的特异蛋白斑点可能与丝素蛋白的合成、分泌有关,而5龄后期新增加的特异蛋白斑点可能与丝腺组织的形态变化、细胞的分解凋亡有关。     

Stimulation of bovine luteal oxytocin secretion in vitro by a phorbol ester and calcium ionophore

Experiments were conducted to examine the in vitro effects of a phorbol ester and a calcium ionophore on bovine luteal oxytocin (OT) secretion and synthesis and progesterone secretion. Corpora lutea removed from beef heifers on d 8 of an estrous cycle were sliced and incubated for 2 h with .81 nM 12-O-tetradecanoylphorbol-13-acetate (TPA), 1.62 nM TPA or .3 microM calcium ionophore A23187. Both concentrations of TPA increased (P less than .01) OT secretion (ng.g-1.2 h-1; control, 407.1; .81 nM TPA, 494.7; 1.62 nM TPA, 528.1; SE = 21.2). Increased secretion of OT was accompanied by a corresponding increase (P less than .02) in synthesis of the hormone (ng.g-1.2 h-1; control, 368.5; .81 nM TPA, 427.6; 1.62 nM TPA, 492.1; SE = 25.7). Phorbol ester also induced (P less than .025) progesterone secretion (ng.g-1.2 h-1; control, 1,056.2 vs .81 nM TPA, 1,333.3; SE = 86.4). Calcium ionophore increased (P less than .01) OT secretion (ng.g-1.2 h-1; control, 248.9 vs A23187, 327.4; SE = 16) and there was a trend (P = .09) toward increased synthesis of OT in response to the ionophore (control, 124.4 vs A23187, 165.6; SE = 16.4). Because TPA can activate protein kinase C and A23187 increases intracellular calcium, these intracellular constituents probably are involved in promoting secretion of OT and progesterone.     

Oxytocin synthesis and secretion from bovine corpora lutea exposed in vitro to cycloheximide and colchicine

Two experiments were conducted to study the effects of cycloheximide and colchicine on prostaglandin F₂ (PGF₂)-induced secretion and synthesis of oxytocin in bovine luteal tissue in vitro. Corpora lutea were collected from beef heifers on Day 8 of the estrous cycle. In Experiment 1, incorporation of [¹⁴C]-leucine into oxytocin synthesized and secreted by luteal slices after exposure to PGF₂, cycloheximide and cycloheximide plus PGF₂ was examined. In Experiment 2, synthesis and secretion of oxytocin were evaluated in luteal slices incubated with colchicine and PGF₂ alone and in combination. Cycloheximide inhibited incorporation of labeled leucine into luteal proteins by more than 90% and no labeled oxytocin was detected in the media or tissue. Prostaglandin F₂ induced significant secretion of oxytocin that was not inhibited by cycloheximide. Tissue levels of oxytocin after incubation with cycloheximide and/or PGF₂ did not differ and were similar to those of the incubated control. Colchicine alone did not suppress oxytocin secretion and did not alter the ability of PGF₂ to induce significant secretion of this nonapeptide. Tissue concentrations of oxytocin after incubation with colchicine and/or PGF₂ did not differ. These studies indicate that secretion and replenishment of luteal oxytocin in vitro is not contingent upon de novo protein synthesis. Inability of colchicine to suppress oxytocin secretion and synthesis may have been due to the short duration of exposure of luteal tissue to the drug.     

Links between de novo fatty acid synthesis and leptin secretion in bovine adipocytes

Leptin secretion by adipose tissue is involved in many physiological control systems, including those that determine growth, development, body composition, milk production, and reproductive function. In the adipocyte of monogastric animals, malonyl CoA (coenzyme A) seems to link the flux of energy substrates to the control of leptin production. In this study, we tested this for ruminants by examining the effect of cerulenin, an inhibitor of de novo fatty acid synthesis at the step from malonyl CoA to palmitate, on leptin production by cultured bovine adipocytes derived from intermuscular fat. Purified preadipocytes were obtained by the ceiling culture method, and adipogenic media were used to induce their differentiation into adipocytes. We found that leptin concentrations increased significantly with time in culture, and with increases in glucose concentration. Addition of 2-deoxy-D-glucose to the medium, a competitive inhibitor of glucose transport and metabolism, suppressed leptin secretion. In media with high glucose concentrations, cerulenin enhanced leptin secretion. We conclude that, as in monogastrics, malonyl CoA may play a key role in the control of leptin secretion in ruminants.     

Effects of parachlorophenylalanine,quipazine and cyproheptadine on growth hormone and adrenocorticotropin secretion in steers

Adrenergic and perhaps dopaminergic neurons provide inhibitory regulation of growth hormone (GH) secretion in ruminants. This suggests that either serotonergic or other neurons regulate the stimulatory release of GH. The nature of neurotransmitter control of adrenocorticotropin (ACTH) secretion in ruminants has not been determined. Parachlorophenylalanine (PCPA; serotonin synthesis inhibitor), quipazine (serotonin receptor agonist) and cyproheptadine (serotonin receptor antagonist) were utilized in Holstein steers to determine whether serotonin receptors mediate stimulatory actions on GH and ACTH secretion. PCPA (100 mg/kg BW) administered each day at 1900 hr for three successive days did not alter mean GH concentrations, amplitude of GH peaks, nor the number of GH peaks. Likewise, PCPA altered none of these parameters for ACTH. Quipazine injected iv at .1 or .5 mg/kg BW increased plasma GH (P<.05) and ACTH (P<.001) concentrations. There was a dose effect of quipazine on both GH (P<.05) and ACTH (P<.001) secretion. Pretreatment of steers with cyproheptadine (.06 and .6 mg/kg BW) reduced the stimulation of GH by quipazine (P<.0001) and decreased basal GH concentrations (P<.0004). Cyproheptadine at .06 mg/kg BW did not alter quipazine effects on ACTH, however, the higher dose decreased the peak ACTH response (P<.02) to quipazine. Studies with quipazine and cyproheptadine indicated that serotonergic mechanisms are likely involved in the regulation of GH and ACTH secretion in steers.     

家兔胚源性特异蛋白－1的分泌特征及生物活性

经ＳｅｐｈａｄｅｘＧ７５和ＰＡＧＥ分离，以双波长薄层自动扫描仪扫描测定了家兔胚源性特异蛋白－１（ｒＥＳＰ－１）的分泌特性；离体组织培养分析了ｒＥＳＰ－１对子宫蛋白质合成的影响。结果表明，早孕期ｒＥＳＰ－１的分泌呈二次曲线，第３天分泌出现，第４天出现第一峰值，第５天下降，然后迅速增加，最大分泌在第７天，第８天迅速回降。其分泌量随胚胎数（或黄体数）的增加而升高，且受ＦＳＨ－ＬＨ和ＰＭＳＧ－ＬＨ的影响。ｒＥＳＰ－１使子宫细胞ＲＮＡ增加，促进子宫蛋白质合成     

Pituitary effects of steroid hormones on secretion of follicle-stimulating hormone and luteinizing hormone

Steroid hormones have a profound influence on the secretion of the gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH). These effects can occur as a result of steroid hormones modifying the secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus, or a direct effect of steroid hormones on gonadotropin secreting cells in the anterior pituitary gland. With respect to the latter, we have shown that estradiol increases pituitary sensitivity to GnRH by stimulating an increase in expression of the gene encoding the GnRH receptor. Since an estrogen response element (ERE) has not been identified in the GnRH receptor gene, this effect appears to be mediated by estradiol stimulating production of a yet to be identified factor that in turn enhances expression of the GnRH receptor gene. However, the importance of estradiol for enhancing pituitary sensitivity to GnRH during the periovulatory period is questioned because an increase in mRNA for the GnRH receptor precedes the pre-ovulatory rise in circulating concentrations of estradiol. In fact, it appears that the enhanced pituitary sensitivity during the periovulatory period may occur as a result of a decrease in concentrations of progesterone rather than due to an increase in concentrations of estradiol. Estradiol also is capable of altering secretion of FSH and LH in the absence of GnRH. In a recent study utilizing cultured pituitary cells from anestrous ewes, we demonstrated that estradiol induced a dose-dependent increase in secretion of LH, but resulted in a dose-dependent decrease in the secretion of FSH. We hypothesized that the discordant effects on secretion of LH and FSH might arise from estradiol altering the production of some of the intrapituitary factors involved in synthesis and secretion of FSH. To examine this hypothesis, we measured amounts of mRNA for activin B (a factor known to stimulate synthesis of FSH) and follistatin (an activin-binding protein). We found no change in the mRNA for follistatin after treatment of pituitary cells with estradiol, but noted a decrease in the amount of mRNA for activin B. Thus, the inhibitory effect of estradiol on secretion of FSH appears to be mediated by its ability to suppress the expression of the gene encoding activin.     

Distribution of neurosecretion in the supraoptic and paraventricular nuclei of the cow before and following milking]

Distribution of neurosecretion and enzymes in the nuclei was studied by optical microscopy and histochemical methods. The stimulus of milking had no effect on the amount of secretion in the supraoptic nucleus, but synthesis of secretion in the paraventricular nucleus was stimulated by oxytocin liberation associated with milking, and it was still present 60 minutes after milking. It seems that each nuclear region is a functionally independent unit.     

睾丸间质细胞线粒体调控睾酮合成的研究进展

睾丸间质细胞(Leydig cells, LCs)的主要功能是合成和分泌睾酮。在睾丸间质细胞内,以胆固醇为原料,位于线粒体外膜上的类固醇合成急性调节蛋白(steroidogenic acute regulatory protein, StAR)促进胆固醇向线粒体内膜转运,在线粒体内膜胆固醇侧链裂解酶(cholesterol side-chain cleavage cytochrome, P450scc)的催化下生成孕烯醇酮,而后通过光面内质网的羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase, 3β-HSD)和转运蛋白(translocator protein, TSPO)的共同作用合成睾酮。因此,睾丸间质细胞合成和分泌睾酮与线粒体密切相关,线粒体结构和功能的完整性直接影响睾酮的生物合成,而位于线粒体上的StAR和P450scc是睾酮合成的关键调控因子。睾酮能够促进雄性生殖器官发育成熟并维持其功能,对促进蛋白质合成(如肌肉、骨骼及生殖器官的蛋白质合成)具有重要意义。近年来,通过维持线粒体结构完整性和改善线粒体氧化损伤、线粒体生物发生等功能进而促进睾酮的合...     

肾上腺糖皮质激素节律性分泌及主要受控因素

机体在维持内环境稳态和适应外环境的过程中,各种生理机能和代谢活动按照一定的时间顺序发生周而复始的节律变化,称为生物节律。肾上腺糖皮质激素作为机体内维持稳态的一种核心内分泌激素,在维持机体内生命活动发生和营养物质代谢的有序性方面扮演着重要角色。自然状态下,外源受控因素(食物和光照等)通过丘脑的视交叉上核和生物钟之间的复杂的同步反馈调控,使肾上腺糖皮质激素的产生、分泌及生物学作用都具有了节律性特征。本文主要就糖皮质激素的合成、分泌及其内在作用机制的节律性变化特点等方面进行综述,以期为机体内生物节律机制的研究及其对动物生产的指导提供参考。     

Regulation of endometrial prostaglandin F(2alpha) synthesis during luteolysis and early pregnancy in cattle

Luteal regression is caused by a pulsatile release of prostaglandin (PG) F(2alpha) from the uterus in the late luteal phase in most mammals including cattle. Although it has been proposed in ruminants that pulsatile PGF(2alpha) secretion is generated by a positive feedback loop between luteal and/or hypophyseal oxytocin and uterine PGF(2alpha), the bovine endometrium may possess other mechanisms for initiation of luteolytic PGF(2alpha) secretion. It has been recently demonstrated that tumor necrosis factor-alpha (TNF-alpha) stimulates PGF(2alpha) output from bovine endometrial tissue not only during the follicular phase but also during the late luteal phase, suggesting that TNF-alpha is a factor in the initiation of luteolysis in cattle. Furthermore, our recent study has shown that IFN-tau suppresses the action of TNF-alpha on PGF(2alpha) synthesis by the bovine endometrium in vitro, suggesting that IFN-tau plays a luteoprotective role by inhibiting TNF-alpha-induced PGF(2alpha) production in early pregnancy. On the other hand, factors other than oxytocin or TNF-alpha have also been suggested to be involved in the regulation of PGF(2alpha) synthesis by bovine endometrium. The purpose of this review is to summarize our current understanding of the endocrine mechanisms that regulate the timing and pattern of uterine PGF(2alpha) secretion during the estrous cycle and early pregnancy.     

氨基酸供给与内分泌互作调控乳蛋白合成的研究进展

限制性氨基酸研究是泌乳奶牛研究的热点之一,关于其影响乳蛋白合成途径而进行的一系列体内外研究表明:底物效应可能不是主要原因,更重要的是氨基酸及其构成作为信号因子对神经内分泌和细胞内信号通路的影响。本文从乳蛋白合成的影响因素入手,概括了氨基酸供给、激素分泌及二者相互作用对乳蛋白合成的调节,以期为阐明氨基酸供给影响乳蛋白合成的途径提供一定理论参考。     

Effects on functions of ovine blood mononuclear cells for each of several fatty acids at concentrations found in plasma of healthy and ketotic ewes

OBJECTIVE: To assess effects on functions of peripheral blood mononuclear cells (PBMC) obtained from ewes for each of several fatty acids represented in ovine plasma at concentrations mimicking those of ketotic or healthy ewes. SAMPLE POPULATION: Blood samples obtained from 6 Sardinian ewes. PROCEDURE: The PBMC were cultured in media that contained oleic (OA), palmitic (PA), stearic (SA), linoleic (LA), or palmitoleic (POA) acid at concentrations similar to those of ketotic or healthy ewes. Synthesis of DNA was stimulated by use of concanavalin A or pokeweed mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: High concentrations (900, 450, and 225 micromol/L) of OA significantly inhibited DNA synthesis and IgM secretion of PBMC. Conversely, low concentrations (56 or 28 micromol/L) of OA significantly enhanced DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, or 75 micromol/L) significantly inhibited DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, 75, or 38 micromol/L) also significantly inhibited IgM secretion of PBMC. None of the concentrations of LA and POA affected PBMC functions. CONCLUSION AND CLINICAL RELEVANCE: Impaired immunoresponsiveness of ketotic ewes is likely associated with an increase of plasma concentrations of OA, PA, or SA and not with that of LA or POA. At physiologic concentrations, single fatty acids are likely to participate in modulation of immunoresponsiveness by exerting suppressive or stimulatory effects on immune cells.