广西巴马小型猪GHRH成熟肽真核表达质粒对小鼠生长效应的影响

【目的】探讨广西巴马小型猪促生长激素释放激素（GHRH）成熟肽真核表达质粒对小鼠生长效应的影响,为进一步研究广西巴马小型猪GHRH成熟肽的促生长作用及GHRH基因疫苗的高效制备与安全使用提供参考依据。【方法】利用基因重组技术将广西巴马小型猪GHRH成熟肽序列与p EGFP-N1质粒连接构建重组真核表达质粒,然后按0.5μg/g的剂量肌肉注射经25%蔗糖溶液预处理的小鼠,观察记录小鼠体重变化情况,并分别提取重组质粒注射组小鼠的心脏、肝脏、肾脏及注射部位肌肉DNA,再以PCR检测是否存在基因质粒残留。【结果】以构建的p EGFP-GHRH重组质粒注射小鼠后,其外源GHRH成熟肽DNA可在肌肉中获得表达,但并未整合入小鼠基因组中。虽然重组质粒注射组小鼠的末重与空质粒注射组小鼠的差异不显著（P〉0.05）,但在注射后的10-25 d,重组质粒注射组小鼠体重均显著高于空质粒注射组（P〈0.05）。在相对日增重率方面,注射后的前15 d内,重组质粒注射组小鼠的相对日增重率均高于空质粒注射组。【结论】构建的广西巴马小型猪p EGFP-GHRH重组质粒能在一定时间内促进小鼠生长,即GHRH种属差异性小,在动物中有较高的保守性。     

甲壳动物CHH家族神经激素结构和功能研究进展

甲壳动物主要利用温度、光照周期等外界因子调节其生理状态 ,使它们的生殖活动处于最适条件下 ,来自外界因子的这些信息作用于甲壳动物的中枢神经系统 ,后者将其传递到神经内分泌系统和内分泌系统 ,神经内分泌系统和内分泌系统能够分泌一些促进因子和抑制因子以实施对性腺活动的调控。由于甲壳动物成体的生殖和蜕皮常常交替出现 ,因此 ,神经内分泌系统和内分泌系统的精确调控非常重要。甲壳动物高血糖激素 (ＣＨＨ)家族神经激素是甲壳动物特有的多肽激素 ,它们主要由眼柄的Ｘ -器官窦腺复合体合成 ,它们包括 :甲壳动物高血糖激素 (ＣＨＨ)、…     

显微注射生长激素基因导入中国对虾（Penaeus chinensis）受精卵的研究

本实验采用显微微量注射方法，交羊生长激素基因导入中轻地虾受精卵，受精卵发育到蚤状幼体第三期后采样检测。ＰＣＲ检测结果表明：７个样品共９３尾幼体，有３个样品呈出阳性信号，基因转移比率至少在３％以上。同时斑点杂交结果也证明有两个明显阳性斑点。     

Changes in thyroid hormone levels in eggs and larvae and in iodide uptake by eggs of coho and chinook salmon,Oncorhynchus kisutsch andO. tschawytscha

Developmental profiles of thyroxin (T₄), triiodothyronine (T₃) and radioactive iodide uptake were established for eggs and T₄ and T₃ profiles were established for larvae (whole-body, yolk-only and body-only) of coho and chinook salmon. T₄ and T₃ were consistently present in all samples. In eggs, hormone levels remained fairly constant in all cohorst for at least the first three weeks of incubation, but then fluctuated in both directions in some sample groups. Large increases in T₄ (from 9 ng/g to 245 ng/g) were seen in 1985 chinook eggs 28 days after fertilization. Radioactive iodide uptake (which was used as a possible indicator of thyroxinogenesis) increased at least 10-fold in both 1986 coho and chinook eggs from 23–30 days after fertilization. T₄ (62 ng/g) and T₃ (393 ng/g) were found in the bodies of 28-day-old 1986 chinook embryos. In whole larvae, hormone levels varied depending upon the cohort studied. In general, initial body-only concentrations of both T₄ and T₃ decreased as body weight increased, but before yolksac resorption was completed, both thyroid hormone content and concentration increased (except for chinook T₃). T₄ and T₃ content in larval yolk stayed constant as yolksac size decreased, resulting in increased thyroid hormone concentration in the yolksac. All of these data suggest that the initial source of thyroid hormones in coho and chinook salmon eggs is maternal, but that by approximately 3–4 weeks after fertilization, the developing embryos begin to produce their own thyroid hormones. After hatching, increases in tissue T₄ and T₃ concentration coupled with constant T₄ and T₃ content in diminishing yolksacs suggest that larvae also produce their own thyroid hormones; yolksac content then may reflect both the original maternal hormones and the larva-producted hormones.     

Seasonal changes in circulating growth hormone (GH), hepatic GH-binding and plasma insulin-like growth factor-I immunoreactivity in a marine fish,gilthead sea bream,Sparus aurata

We have studied the seasonal relationship between growth and circulating growth hormone (GH), hepatic GH-binding and plasma insulin-like growth factor-I immunoreactivity in gilthead sea bream,Sparus aurata. The seasonal increase in plasma GH levels preceded by several weeks the summer increase in growth rates. In contrast, a marked increase in hepatic GH-binding with a high degree of endogenous GH occupancy was found during the period of maximum growth which suggests an enhanced sensitivity of liver to GH action. Thus, circulating levels of immunoreactive IGF-I, probably derived from the liver in response to GH action, were positively correlated with growth throughout the experimental period although a consistent relationship between growth and circulating GH was not found. In spite of this, we consider that, in gilthead sea bream, as in several other teleosts, the availability of endogenous GH can limit growth. Thus, under environmental conditions of suboptimal growth, a single intraperitoneal injection of recombinant rainbow trout GH (rtGH) induced over the dose range tested (0.75, 1.5, 3 μg g BW⁻¹) an increase in plasma IGF-I-like immunoreactivity comparable to that seen during the period of maximum growth.     

Hormonal influences on in vitro [35S]-sulfate uptake by gill arches from the goldfish (Carassius auratus L.)

A bioassay for insulin-like growth factor (IGF), based on the in vitro incorporation of [³⁵S]-sulfate into gill arch tissue was used to study the hormonal regulation of proteoglycan synthesis in the goldfish (Carassius auratus). [³⁵S]-sulfate incorporation into gill arch tissue was found to be time-dependent with maximal uptake occurring by 48h, suggesting that proteoglycan synthesis in this tissue was maintained for at least 48h in vitro. The addition of human recombinant IGF-I (IGF-I) to the incubation medium was found to significantly stimulate [³⁵S]-sulfate uptake into the gill arches, whereas bovine growth hormone (GH) was without effect. Porcine insulin was also stimulatory, but results indicate that the effects of porcine insulin and IGF-I may be mediated by a single receptor system. Finally, arches from hypophysectomized fish were significantly less responsive to IGF-I than were arches from sham-operated fish. Furthermore, administration of ovine GH in vivo appeared to increase subsequent responsiveness in vitro. Together, these results provide evidence that the growth-promoting actions of GH in the goldfish may be mediated, at least in part, by a peptide related in structure to mammalian IGF-I.     

Evaluation of commercial catfish, tilapia and salmonid diets for growth promotion of white suckers

In an attempt to identify appropriate diets for use in intensive baitfish culture, the efficacy of three commercial diet formulations, a high-protein, high-lipid, low-carbohydrate diet [a semi-moist salmonid diet (SD)] and two low-protein, low-lipid, high-carbohydrate diets [a catfish diet (CD) and a tilapia diet (TD)] on growth performance of juvenile white suckers, Catostomus commersoni, was examined. All three diets permitted growth during the 16 week trial. The SD promoted highest growth and somatic protein and lipid deposition, whereas the CD and TD formulations promoted markedly higher accumulations of glycogen in the carcass and liver. The high hepatic glycogen content was associated with an elevated hepatosomatic index in the CD- and TD-fed groups. Plasma T3 levels were lower in the CD- and TD-fed groups, possibly a response to the low dietary protein or indicative of dietary insufficiency in these fish. Of the three diets evaluated, the SD was considered to be the best in terms of the growth performance parameters measured, and offers an economical and appropriate diet for intensive culture of juvenile white suckers.     

Influence of growth hormone on the hepatic mixed function oxidase and transferase systems of rainbow trout

The effect of GH treatment on hepatic cytochrome P450 content, aryl hydrocarbon hydroxylase (AHH), aminopyrine-N-demethylase (AND), testosterone hydroxylase, testosterone 5- and 5-reductase, UDP-glucuronyl transferase (UDPGT) and glutathione S-transferase (GST) activities in immature rainbow trout were investigated. Hepatic cytochrome P450 content, AHH and GST activities were measured in both GH implanted and GH injected animals whereas other activities were assayed in GH implanted trout only.GH implants significantly decreased cytochrome P450 content at 15 days compared to the control but no significant effect was observed at 15 or 30 d when GH was injected biweekly. In both cases, AHH activity was significantly decreased by GH treatment compared to the control whereas GST remained unchanged. Compared to the control, GH implanted fish exhibited a pronounced inhibition of AND, a decreased 6 and 16-testosterone hydroxylation, an inhibition of UDPGT with testosterone as substrate and an enhanced 17-testosterone oxidation.     

Calcium currents and exocytosis in single isolated pars intermedia cells from tilapia (Oreochromis mossambicus)

The role of calcium influx through voltage-dependent calcium ion channels in the exocytotic response of single isolated pars intermedia cells from the teleost tilapia (Oreochromis mossambicus) was investigated by means of the whole-cell patch-clamp technique and high resolution electrical measurements. Calcium currents differed from barium currents in several ways: the peak I_Ca was smaller, the current-voltage relationship for calcium attained its maximum at +10 mV instead of 0 mV, and the inward calcium current inactivated more rapidly. Electron micrographs showed that pars intermedia cells possess dense, encored vesicles with an average diameter of 140 nm. Influx of calcium resulted in an increased cell membrane capacitance (C_m) after the depolarizing period, indicating a fast exocytotic response. Comparison with late recordings revealed the presence of a transient in C_m, presumably attributable to movement of ion channel gates. The average increase in C_m was 13.4 fF, suggesting the fusion of at least 23 vesicles with the plasma membrane during the depolarizing pulse. In contrast to calcium, barium did not support significant exocytosis. We conclude that calcium entry through voltage-dependent calcium channels rapidly leads to the exocytosis of secretory vesicles from tilapia pars intermedia cells.     

Evolutionary implications of two rainbow trout growth hormone genes

The primary structures of two rainbow trout growth hormone mRNAs (GH1 and GH2) have been deduced by direct sequencing of their respective cDNA clones and portions of the mRNA. Both GH1 and GH2 mRNA contain open reading frames comprised of 630 nucleotides and encode 210 amino acid residues of which 11 are variant. The translated regions of both mRNA are flanked by a short but rather conserved 5′-end, and a relatively long but highly diverged 3′-end. The differences at translated and 3′-untranslated regions suggest that the GH1 and GH2 mRNA originate from different loci. The GH1 and GH2 mRNA are likely transcribed from two distinct loci which were duplicated during tetraploidization of salmonid genome between 50 to 100 million years ago. The GH2 gene has been isolated and sequenced from a rainbow trout genomic library. This gene spans a region of approximately 4 kilobases. The trout GH gene is comprised of 6 exons and 5 introns, in contrast to 5 exons and 4 introns in mammals. The additional intron in the trout gene interrupts the translated regions that are analogous to the last exon of the mammalian counterpart. The alleged internally repeating sequences in mammalian GH, prolactin (Pr1) and placental lactogen (PL) are not observed in the predicted polypeptide sequence of trout GH. In addition, direct repeats that flank exons I, III and V of mammalian GH, Pr1 and PL genes are absent in trout gene. These findings indicate that the rainbow trout GH gene structure does not support the current hypothesis that internally repeated regions in GH, Pr1 and PL arose from a small primordial gene.