西藏当雄牦牛超数排卵及胚胎移植试验

2005年8月初,对西藏当雄10头5～10岁供体母牦牛实施了超数排卵试验,超排选用PGF2α+FSH刺激法和埋置阴道栓Cu-Mate+促卵泡素FSH刺激法,FSH的剂量为8.8～9.0 mg/头.根据超排后供体发情表现、黄体数量及获得胚胎数量及质量评定超排效果,将胚胎鲜胚移植给经同期处理的受体母牦牛.结果为:10头供体牛共获得7枚胚胎,PGF2α+FSH连续递减法得到5枚可用胚胎,Cu-Mate+FSH法没有冲到胚胎,5枚胚胎移植给5头受体后妊娠并出生4头牦犊牛.结论为:PGF2α+FSH连续递减法超排效果较好,胚胎移植取得初步成功.

全放牧婆罗门牛和BMY牛的超数排卵效果及激素水平比较

为了比较婆罗门牛和BMY牛的雌二醇(E2)、孕酮(P4)水平及胚胎生产潜力,并探讨MOET技术在BMY牛扩繁中的应用前景,采用进口激素(日本产FSH 24~36 mg+美国产PG 35 mg)和国产激素(国产FSH 10~12 mg+国产PG 0.8 mg)对全放牧婆罗门牛和BMY牛进行超排,并于超排前(同期发情第7天)、超排开始、超排第3天、超排发情当天和胚胎回收当天采集血液,采用RIA法分析血浆中的E2和P4水平。结果表明,BMY牛的黄体数和获胚(卵)数、可用胚数分别较婆罗门牛高2.09个和0.95、0.36枚,但差异不显著(P>0.05);BMY牛的排卵率较婆罗门牛高18.46%(P<0.01),而可用胚率则较婆罗门牛低2.55%(P<0.01)。BMY牛在5个处理时段的E2和P4平均值分别较婆罗门牛高0.58 pg/mL(P<0.01)和1.44 ng/mL(P>0.05),在各处理时段的P4水平变化规律几乎与婆罗门牛一致,E2/P4值为婆罗门牛的5.16倍。结果提示,BMY母牛的E2和P4水平高于婆罗门牛,在激素水平和繁殖性能方面表现出一定的杂种优势;E2水平低是婆罗门牛发情微弱和排卵率低的主要原因...

肉牛B超活体取卵效果的研究

对肉牛(莫累灰×婆罗门) 利用B 超活体取卵,研究不同年龄和促卵泡素(FSH) 对平均卵泡数、平均卵母细 胞数、平均可利用卵母细胞(COC) 数、卵母细胞回收率、COC 回收率、囊胚率和平均囊胚数的影响。结果表明:肉牛 B 超活体所取卵的数量和质量受年龄的影响,青年牛好于经产牛,经产牛好于老年牛;适量注射FSH 可以提高经产 牛和老年牛的囊胚率和(或) 平均囊胚数。

Amino acid requirements for maturation of rhesus monkey (Macacca mulatta) oocytes in culture

This study evaluated the effects of different amino acid formulations on supporting meiotic and cytoplasmic maturation of rhesus monkey (Macacca mulatta) oocytes in vitro. Five hundred and forty-six cumulus-oocyte complexes (COCs) aspirated from unstimulated adult monkey follicles (greater than or equal to 1000 mum in diameter) were cultured in either modified Connaught Medical Research Laboratories 1066 medium (mCMRL-1066) or in one of eight chemically defined media (modified basic medium 5 supplemented with 5.5 mmol glucose l(-1), 0.003 mmol pantothenic acid l(-1) and different amino acid formulations) as below: (1) modified basic medium 5 (mBM5) containing no amino acid; (2) mBM5 + 0.2 mmol glutamine l(-1); (3) mBM5 + 11 amino acids from hamster embryo culture medium 6 (HECM-6) (11 AA); (4) mBM5 + Eagle's non-essential amino acids (NEA); (5) mBM5 + NEA + 0.2 mmol glutamine l(-1); (6) mBM5 + Eagle's essential amino acids (EA) without glutamine; (7) mBM5 + EA + 0.2 mmol glutamine l(-1); (8) mBM5 + Eagle's 20 amino acids (20 AA) + 0.2 mmol glutamine l(-1); and (9) mCMRL-1066 (control). All media contained FSH, LH, oestradiol and progesterone. After maturation, mature oocytes were subjected to the same fertilization and embryo culture procedures. COCs matured in treatment 5 had greater potential to progress to metaphase II (66%; P < 0.05) than did those in treatments 1 (37.3%), 2 (48.3%)f 3 (41%), 6 (41%) and 9 (43%). Oocytes matured in treatment 8 had the best morula (53%) and blastocyst (18%) developmental responses (P<0.05). The lowest (P<0.05) morula and blastocyst developmental responses were obtained from COCs matured in treatments 1 (0%) and 6 (8%). The other media supported intermediate embryonic development (range 11-38% of morula and blastocyst). These results indicate that the choice of amino acids affects the competence of oocyte maturation and that Eagle's 20 AA with 0.2 mmol glutamine l(-1) is more efficient than the other amino acid formulations for maturation of rhesus monkey oocytes.

Reproductive physiology of ribbon fish (Trichiurus lepturus)

The ribbon fishes ‘of the family Trichiuridac are represented as one of the most important food resources in Indian ocean. High density of the dominant species of ribbon fish (Trichiurus lepturus) in Oman sea and the 'Tillable catch in last yeas (more than 7000 tones per year) makes a trust area for studing their population biolog and stock assessment. As our knowledge on reproductive biology of this species has an important role on their fisheries management, as well as conservation of this stock from decline or over fishing, this research was held to determine some aspects of reproductive physiology of ribbon fish and the effects of environmental factors in gonadal cycle. The goals of the present thesis is to determine some aspects of reproductive physiology such as gonadosomatic index (GSI) , hepatosomatic index (HSI), condition factor (Ko, fecundity, sex ratio, size at first maturity, size at maturity (LM5O) and their relative hormonal & biochemical fluctuations. In this regards annual variation of sex hormones ic. estradiol 17-B, progestron, cortisol, testostrone and gonadotropins FSH (GTH-I) , LH (GTH-ll)I were measured ; gonadal histological studies were done by light & electron micrography. The research was carried out from April 1995 to January 19% in Ras Nleidani in the north part of Oman sea, and the environmental factors such as temperature, salinity, oxygen, rainfall and pH were measured. The effects of these parameters on reproductive cycle and hormonal fluctuationswere discussed by using correlation and principle component analysis (PCA). Female Ribbon fish reproductive strategy shows the same paterns of nonguarder marine teleosts. T. lepturus has more than one spawning season (existance of egges in different size in each month) and therfore it must have asynchronous ovaries and belong to continious spawners. GSI and HSI are good evidences for this type of reproductive patern. The testis of the lobular type , which is typical of most teleosts , is composed of numerous lobules which are separated from each other by a thin layer of fibrous connective tissue. GSI fluctuations revealed prolong- spawning time in males. There is significant increase in 17-13 estradiol. progestrone , cortisol and gonadotropins with maturity and prespawning period of female T lepturus. Plasma concentration of E2 and GTH II incresaed along with water temperature increasing (3300).. Spawning was observed from Nov. 1995 to Apr. 1996 in this species. Progestrone increased significantly with increasing rainfall in this season (P<0.01). Plasma cortisol levels increased with maturation and vitelpgenesis and also with the peak of spawning. From lenght-weight frequency and size distribution in each age groups and also minimum size at first maturity (52a cm) it would he concluded that T. lepturus must be matured at 2 years of age. Serum cholestrol and triglicerides significantly increased when maturation occured in this species. The relationship between alkaline phosphatase activity and hormonal fluctuations with maturity and vitelogenesis were discussed. Proximate compostion (muscle) shows significant variation with spawning period and maturity. Absolute individual fecundity (17420-159150) increased with body length and weight. Ultrastructural observations show dramatic variation in cell membrane (0ocyte membrane), yolk vesicles and, nucleolus dispersal in relation to maturity stages. fluctuations of gonadal hormones were discused in relation with vitelogenesis. Testosterone increased in males from Nov: to Mar. due to environmental impacts and spawning time. Sex ratio in different depth (10-40 m ,80-110 m) shows significnt differences in this ratio for two depths. In 10-40 m depth female shows dominant abundance to male in each months that may be due to their reproductive migration behaviour. The effects of temperature photoperiod and rainfall to maturity and spawning were discussed. According to -pawning period of T. leptunts in our sampling area it could be suggested that ribbon fish fi,theries must be restricted in the peak of spawning seasons (Feb. to Mar.) and in the spawning grounds (under 40 m depths). Other suggestions for population conservation have been mentioned.

猪STCH 基因的Bi2PASA 遗传标记及多态性研究

微粒体应激70 蛋白三磷酸腺苷酶( S TCH) 基因属于应激70 蛋白基因伴侣家族, 在机体免疫反应 和疾病抵抗力等方面起重要作用。根据人和小鼠S TCH 基因的保守序列设计引物, PCR 扩增到猪S TCH 基因第 5 外显子445 bp 片段。序列测定显示, 猪S TCH 基因与人和小鼠S TCH 基因分别具有87113 %和80145 %的同源 性。通过测定和比较中国梅山猪、欧洲约克夏猪及PIC 商品猪的S TCH 基因序列, 发现在猪S TCH 基因编码区 第5 外显子1 050 位点上存在一个单碱基突变位点。利用双向特定等位基因PCR 扩增法(Bi2PASA) 建立了检测 猪S TCH 基因变异的遗传标记, 并用该标记分析了S TCH 基因在中国家猪(梅山猪、荣昌猪和金华猪) 、欧洲家 猪(约克夏猪、大白猪) 、商品猪(PIC 合成系) 以及欧洲野猪的基因频率和多态性。本研究建立的Bi2PASA 遗 传标记和基因变异信息, 将为进一步分析猪S TCH 基因变异与经济性状的相关分析提供基础资料。

幼猴和成年猴颗粒细胞离体下甾体形成的激素调节

用无血清培养技术研究表明: 幼龄和成年恒河猴颗粒细胞在离体下都能对人FSH、环-AMP、FK和GnRH类似物产生反应, 明显增加孕酮和雌激素的产生; hCG能够促进FSH刺激幼猴和年猴颗粒细胞增加孕酮的产生; 在培液中加入DES明显增加FSH刺激孕酮和雌激素产生的效应; 在缺少雌性激素底物的情况下, 猴壳粒细胞也能合成可测量的雌激素。图7参21

双峰驼血浆促卵泡素的放射免疫分析法

用单峰驼促卵泡素标准品(CamFSH),hFSH抗血清和~(125)I-hFSH建立了测定双峰驼血浆FSH的放射免疫分析方法,并通过一系列实验证明,该方法可以用于测定双峰驼血浆FSH。

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Purification, characterization and primary structure of a chymotrypsin inhibitor from Naja atra venom

A chymotrypsin inhibitor, designated NA-CI, was isolated from the venom of the Chinese cobra Naja atra by three-step chromatography. It inhibited bovine (x-chymotrypsin with a K-i of 25 nM. The molecular mass of NA-CI was determined to be 6403.8 Da by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) analysis. The complete amino acid sequence was determined after digestion of S-carboxymethylated inhibitor with Staphylococcus aureus V8 protease and porcine trypsin. NA-CI was a single polypeptide chain composed of 57 amino acid residues. The main contact site with the protease (PI) has a Phe, showing the specificity of the inhibitor. NA-CI shared great similarity with the chymotrypsin inhibitor from Naja naja venom (identities = 89.5%) and other snake venom protease inhibitors. (C) 2003 Elsevier Inc. All rights reserved.

Separation and purification of ovulation-inducing factors in the seminal plasma of the Bactrian camel (Camelus bactrianus)

Ovulation in the Bactrian camel depends upon ovulation-inducing factors in the seminal plasma. The present study was conducted to isolate and purify the bioactive fractions from the seminal plasma of these camels. The seminal plasma was fractionated by anion-exchange chromatography, and six fractions were obtained. The bioactive potential of each fraction was estimated from its effect on rat pituitary tissue cultured in vitro and by the effect of an intramuscular injection of the fraction into female camels in vivo. Both the third fraction (F3) and the fifth fraction (F5) stimulated the release of LH in vitro and in vivo. In addition, female camels ovulated within 48 h after intramuscular injection of F3. However, neither F3 nor F5 had any significant effect on the secretion of FSH, either in vitro or in vivo. When F3 was further fractionated into four subfractions, the third subfraction (F3-3) still stimulated the in vitro release of LH, but not of FSH. An attempt to further purify the ovulation-inducing factors in F3-3 failed owing to the similarity of the molecular characters.

Real-time block flow tracking of atrial septal defect motion in 4D cardiac ultrasound

Real-time cardiac ultrasound allows monitoring the heart motion during intracardiac beating heart procedures. Our application assists atrial septal defect (ASD) closure techniques using real-time 3D ultrasound guidance. One major image processing challenge is the processing of information at high frame rate. We present an optimized block flow technique, which combines the probability-based velocity computation for an entire block with template matching. We propose adapted similarity constraints both from frame to frame, to conserve energy, and globally, to minimize errors. We show tracking results on eight in-vivo 4D datasets acquired from porcine beating-heart procedures. Computing velocity at the block level with an optimized scheme, our technique tracks ASD motion at 41 frames/s. We analyze the errors of motion estimation and retrieve the cardiac cycle in ungated images. © 2007 IEEE.

Molecular characterization of Chinese sturgeon gonadotropins and cellular distribution in pituitaries of mature and immature individuals

Chinese sturgeon (Acipenser sinensis) is a rare and endangered species, and also an important resource for the sturgeon aquaculture industry. To understand molecular characterization of Chinese sturgeon gonadotropins (GTHs), we cloned the full-length cDNAs of gonadotropin subunits common alpha (GTH-alpha), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) from a pituitary cDNA library of mature female. Two subtypes of GTH-alpha were identified. The nucleotide sequences of A. sinensis common alpha I (AsGTH-alpha I), common alpha II (AsGTH-alpha II), FSH beta (AsFSH beta) and LH beta (AsLH beta) subunit cDNAs are 345, 363, 387 and 414 bp in length, and encode mature peptides of 115, 121, 129 and 138 aa, respectively. Then, three polyclonal antibodies were prepared from the in vitro expressed AsGTH-alpha I, AsFSH beta and AsLH beta mature proteins, respectively. Significant expression differences were revealed between immature and mature sturgeon pituitaries. Western blot detection and immunofluoresence localization revealed the existence of three-gonadotropin subunits (AsGTH-alpha, AsFSH beta and AsLH beta) in mature sturgeon pituitaries, but only AsFSH beta was detected in immature individual pituitaries during early stages in the sturgeon life, and obvious difference was observed between males and females. In males, AsFSH beta was expressed in 4-year-old individuals, whereas in females, AsFSH beta was just expressed in 5-year-old individuals. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Identification and characterization of a novel splice variant of gonadotropin alpha subunit in the common carp Cyprinus carpio

In this study, an alternative splicing transcript GtH-alpha 291 was identified by RT-PCR, which is 291 nt and exists not only in the pituitary but also in the ovary in common carp Cyprinus carpio. The analysis of GtH-alpha 291 amino acid sequence by the SignalP server predicted that the 'missing segment' might characterize as a signal peptide. In the secretion experiment, GtH-alpha 357 subunit could be secreted out of HeLa cells while GtH-alpha 291 could not, which confirmed the prediction. Co-immunoprecipitation assay proved that GtH-alpha 291 subunit is able to interact with both FSH-beta and LH-beta as GtH-alpha 357 does. This is the first report concerning an alternative splicing transcript of a GtH alpha subunit. Further studies are necessary to elucidate the specific role of this variant in the regulation of gonadal development and sexual maturation. (c) 2007 The Authors.

Serum concentrations of gonadotropins and steroid hormones of Neophocaena phocaenoides asiaeorientalis in middle and lower regions of the Yangtze River

As a relatively isolated and unique freshwater population, the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) is the most endangered subpopulation of this species. The objective of this study was to improve our understanding of their reproductive endocrinology by measuring (radioimmunoassay) serum concentrations of FSH, LH, estradiol (E-2), progesterone (P-4) and testosterone (T-2) in free-ranging animals. Blood samples were collected from 66 Yangtze finless porpoises (41 males and 25 females) captured in the middle and lower reaches of the Yangtze River. Based on significant variation of serum T-2 concentrations in males with body length (BL) > 138 cm, we inferred that they were mature; in these animals, there were significant seasonal variations in serum T-2 concentrations, with the highest concentrations in March and April (502.0 +/- 319.8 ng/dL, mean +/- S.D.) and the lowest in December (79.4 +/- 83.2 ng/dL). Serum T-2 concentrations positively correlated with serum concentrations of LH and weakly correlated with serum concentrations of E-2, whereas there was a significant negative correlation between serum LH and FSH concentrations in males > 138 cm. The smallest apparently pregnant female porpoise had a BL of 130 cm. Serum P-4 concentrations ranged from 13.2 to 112.4 ng/mL (43.9 +/- 28.3 ng/mL) in pregnant females, and fluctuated under 1.0 ng/mL in non-pregnant females with BL > 130 cm. Serum LH concentrations were significantly higher in non-pregnant females > 130 cm versus those females < 130 cm. To our knowledge, this is the first study of endocrine-related maturity and seasonal breeding characteristics of the Yangtze finless porpoise. (c) 2006 Elsevier Inc. All rights reserved.