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PubMed2005

Ovulation failure and double ovulation in dairy cattle: risk factors and effects.

Ovulation failure and double ovulation rates were examined in 1917 inseminations performed in high-yielding dairy cows under standard commercial conditions. The ovulation rate was determined 11 days post-insemination by ultrasound detection of at least one corpus luteum in the ovaries. Analyzing the double ovulation and pregnancy rates, the study population consisted only of ovulated cows (n = 1792). Data were analyzed using logistic regression methods. A failure to ovulate was recorded in 125/1917 (6.5%) services: 82/663 (12.4%) during the warm and 43/1254 (3.4%) during the cool period. Based on the odds ratios, the risk of ovulation failure was 3.9 times higher for inseminations performed during the warm period. No significant effects of estrous synchronization, milk production and days in milk at AI, and service and lactation number on ovulation failure were found. Double ovulation was recorded in 277/1792 (15.5%) services: 146 (52.7%) unilateral double ovulations (42.5% left versus 57.5% right); 115 (41.5%) bilateral double ovulations; and 16 (5.8%) triple ovulations. Double ovulation was recorded in 72 (12.4%) and 205 (16.9%) AI during the warm and the cool period, respectively. The percentages of double ovulation for first, second and third or more lactations were 6.7, 16.6 and 25%, respectively. Double ovulation rates for early (less than 90 days), mid- (90-150 days) and late (more than 150 days) lactation periods were 13, 20.7 and 14.2%, respectively. Reaching estrus during the warm period decreased the likelihood of double ovulation by a factor of 0.86; the risk of double ovulation was lower in cows with higher milk production (a 1 kg increase in milk yield led to a 0.97-fold reduced risk of double ovulation); cows in their second and in their third or more lactations showed a likelihood of double ovulation (using the first lactation as reference) increased by factors of 3.4 and 5.6, respectively; and reaching estrus during the early and late lactation period was related to a decreased probability of double ovulation (using the mid-lactation period as reference) by factors of 0.56 and 0.84, respectively. No significant effects of synchronization and service number on the double ovulation rate were found. Pregnancy was recorded in 914/1792 (51%) services: rates of 53.5% (811/1515) were recorded for single ovulations; 37.2% (103/277) for double ovulations: 28.8% (42/146) for unilateral double ovulations; 45.2% (52/115) for bilateral double ovulations; and 56.3% (9/16) for triple ovulations. The likelihood of pregnancy diminished in cows: inseminated during the warm period (by a factor of 0.5); inseminated by one particular bull (by a factor of 0.33); with higher milk production (a 1 kg increase in milk yield decreased the probability of pregnancy by a factor of 0.98); or undergoing unilateral (by a factor of 0.31) and bilateral (by a factor of 0.64) double ovulation. Logistic regression analysis indicated no significant effects of synchronization, days in milk, lactation number and service number on pregnancy rate. Collectively, our results indicate that cows showing estrus in conditions of heat stress had a high risk of ovulation failure. The effect of milk production on double ovulation was negative, whereas lactation number was positively correlated with this factor; the highest incidence of double ovulation occurring during the mid-lactation period.

Animals↗
PubMed1984

Clinical validation of enzymeimmunoassay of human luteinizing hormone (hLH) in the detection of the preovulatory luteinizing hormone (LH) surge in urine.

The preovulatory luteinizing hormone (LH) surge (mean, 60.7 standard error +/- 4.7 mIU/ml) as determined by a solid-phase enzymeimmunoassay in urine has been correlated with clinical parameters in 24 women. In group A, of seven women, the preovulatory LH surge correlated with basal body temperature and cervical mucus. In one of the women in group A, serum levels of pituitary and gonadal hormones confirmed ovulation. In group B, of 17 women, the urinary estrone-3-glucuronide (E1-3-G) peak was either coincident with or preceded the LH surge. The LH surge in all cases occurred 12 to 24 hours prior to follicular rupture, as visualized by real-time sonography. The enzymeimmunoassay for the detection of the preovulatory LH surge is useful in patients for artificial insemination and for aspiration of mature oocytes for in vitro fertilization.

Adult↗
PubMed1990

Immunohistochemical localization of prostaglandin H synthase in the embryo and uterus of the mouse from ovulation through implantation.

Prostaglandins (PGs) in the embryo and endometrium are involved in processes that are important for implantation. Although the presence of PGs (PGE2, PGF2 alpha, PGI2) in decidualized endometrium has been widely reported, less is known about the capacity of the pre-implantation embryo to synthesize PGs. Prostaglandin H (PGH) synthase is necessary for the production of PGs. Using an immunohistochemical method, PGH synthase was localized in the mouse embryo and uterus from superovulation through embryo implantation. No PGH synthase was detected in oocytes at the time of ovulation or in single-cell embryos 1 day post-fertilization (PF). Circular areas of immunostaining became evident in the cytoplasm of blastomeres at the morula stage (day 3 PF). After implantation (day 5 PF), a low level of PGH synthase reactivity was observed in embryonic cells; no PGH synthase was detected in the embryo by day 7 PF. The endometrial glands exhibited maximal immunostaining by day 3 PF, and after implantation, PGH synthase appeared in decidual cells along the border of placentation. Low levels of PGH synthase reactivity were detected in myometrial cells during the period after superovulation through day 7 PF. This is the first demonstration of PGH synthase in the mouse embryo prior to apposition with glandular endometrial epithelium, supporting the hypothesis that the embryo has the potential to produce PGs that may mediate autocrine and/or paracrine responses at the time of nidation.

Animals↗
PubMed1995

Differential constitutive expression of interferon genes in early mouse embryos.

Recent evidence suggests that several processes during mammalian embryogenesis may be regulated by IFNs or IFN-like molecules. With the use of MAPPing, the simultaneous presence of transcripts homologous to IFN-alpha, IFN-beta, IRF-1, and IRF-2 was examined in mouse embryos and in embryonal carcinoma (EC) P19 cells, which are equivalent to epiblast cells of the early postimplantation blastocysts. Transcripts for IFN-alpha, but not for IFN-beta, were detected as maternal transcripts in the ovulated oocyte and persisted over early embryogenesis. IRF-1 transcripts appeared only after the first cell cleavage in the two-cell stage embryo. IRF-2 transcripts were analyzed only in EC P19 cells and were found in both undifferentiated (D-) and differentiated (D+) cells. The IFN-alpha transcripts present in (D-) P19 cells were cloned and the partial cDNA sequences determined. Mu IFN-alpha A and a new Mu IFN-alpha species (Mu IFN-alpha 12) were isolated from (D-) P19 cells. The presence of constitutive IFN-alpha transcripts in early mouse embryos suggests a role for these molecules during embryogenesis.

Animals↗
PubMed1989

The impact of high progesterone levels in the follicular phase of in vitro fertilization (IVF) cycles: a comparative study.

Estrogen (E2) and plasma progesterone (P4) levels are valuable parameters for follicular development in in vitro fertilization (IVF) cycles. Furthermore, the progesterone concentration prior to, during, and following human chorionic gonadotropin (hCG) administration is an important marker for the detection of early luteinization and premature ovulation. The pattern of hormonal profile in relation to the number of oocytes retrieved, fertilized, and cleaved and the fate of the pregnancies achieved were compared in three groups of patients treated by the same protocol. Group I included 22 women who conceived with high progesterone levels on day hCG + 1 (P4, greater than 2.5 ng/ml). Group II included 43 women who conceived with low P4 values (P4, less than 2.5 ng/ml), while group III included 46 patients in whom no pregnancies occurred. A significant decrease in fertilization, cleavage, and pregnancy rates was observed in patients with high progesterone levels on day hCG + 1, compared to those with normal levels. Nevertheless, it is suggested that cycles with high P4 levels in the preovulatory phase should not be canceled, as a fair chance for pregnancy still exists.

Chorionic Gonadotropin↗
PubMed1995

Synchronization of ovulation in dairy cows using PGF2alpha and GnRH.

This paper reports a new method for synchronizing the time of ovulation in cattle using GnRH and PGF(2alpha). In Experiments 1 and 2, lactating dairy cows (n=20) ranging from 36 to 280 d postpartum and dairy heifers (n=24) 14 to 16 mo old were treated with an intramuscular injection of 100 mug GnRH at a random stage of the estrous cycle. Seven d later the cattle received PGF(2alpha) to regress corpora lutea (CL). Lactating cows and heifers received a second injection of 100 mug GnRH 48 and 24 h later, respectively. Lactating cows were artificially inseminated 24 h after the second GnRH injection. Ovarian morphology was monitored daily by trans-rectal ultrasonography from 5 d prior to treatment until ovulation. In Experiment 3, the flexibility in the timing of hormonal injections with this synchronization protocol was evaluated by randomly assigning 66 lactating dairy cows to 3 different treatment groups. Lactating cows received the injection of PGF(2alpha) 48 (Group 1), 24 (Group 2), and 0 h (Group 3) prior to the second injection of GnRH, which was administered at the same time in each group to ensure the second injection of GnRH was given when follicles were at a similar stage of growth. In Experiments 1 and 2, the first injection of GnRH caused ovulation and formation of a new or accessory CL in 18 20 cows and 13 24 heifers. In addition, this injection of GnRH initiated or was coincident with initiation of a new follicular wave in 20 20 lactating cows and 18 24 heifers. Corpora lutea regressed after PGF(2alpha) in 20 20 cows and in 18 24 heifers. All cows and 18 24 heifers ovulated a newly formed dominant follicle between 24 and 32 h after the second injection of GnRH. Ten of 20 cows conceived to the timed artificial insemination. In Experiment 3, the conception rate in Groups 1 and 2 were greater than in Group 3, (55 and 46 % vs 11%, respectively). In summary, this protocol could have a major impact on managing reproduction in lactating dairy cows, because it allows for AI to occur at a known time of ovulation and eliminates the need for detection of estrus.

Journal Article↗
PubMed1998

Immunolocalization of P450C17 in the mare corpus luteum.

Although the mare corpus luteum (CL) is capable of aromatization, the expression of other enzymes involved in estradiol synthesis is not yet clear. This study examined the localization of P450C17 in the mare CL at different stages of its functional development. In ovaries from follicular phase mares P450C17 was localized in the theca cells of ovarian follicles. Following ovulation, no immunostaining for P450C17 was detected in the mature CLs of nonpregnant mares. In pregnant mares, no immunostaining for P450C17 was identified in the corpus luteum prior to secretion of eCG by the feto placental unit at Day 35 of pregnancy. The P450C17 was found to be expressed in CLs retrieved from Day 40 of pregnancy onwards. The changing expression of P450C17 raises the possibility that this may be a regulatory step for estrogen synthesis in the mare ovary.

Animals↗
PubMed1988

PGF2 alpha-induced milk ejection in ewes having cyclic or pregnant corpora lutea.

The production of luteal oxytocin in ewes, resulting from the intrajugular injection of 200 micrograms of PGF2 alpha, could be determined by the increase in intramammary pressure. This simple indirect method of measuring the activity of the corpus luteum enabled easy detection of renewed post-partum ovulation or the onset of pregnancy. The response was monitored every two days between Days 0 and 25, then every 4 days between Days 25 and 59 in: 9 cyclic ewes (group B); 9 cyclic ewes treated with three daily intramuscular injections of 25 mg of Trilostane, a steroid synthesis inhibitor, between Days 7 and 25 (group A); 11 pregnant ewes (group C). Progesterone levels were determined each day from blood sampled in the jugular vein. Trilostane produced a decrease in plasma progesterone, not a total suppression (fig. 3), but did not significantly modify the intramammary pressure variations resulting from PGF2 alpha injections. These were identical in both cyclic and pregnant ewes during the first 15 days: they increased from D0 to D7 and decreased between D12 and D15 (fig. 4). After D15, the increase in intramammary pressure progressively weakened and became 0 at D17 in the cyclic ewes, whereas in the pregnant animals there was a renewed increase in intramammary pressure until D20; this regressed progressively afterwards and disappeared towards D45. This transitory, renewed activity between D15 and D20 might be an indirect or direct result of the message delivered by the embryo to maintain the corpus luteum. Several hypotheses are discussed with a view to explaining this phenomenon.

Animals↗
PubMed2006

Meiotic spindle configuration is differentially influenced by FSH and epidermal growth factor during in vitro maturation of mouse oocytes.

BACKGROUND: To ascertain whether different hormonal treatment protocols could affect metaphase II (MII) spindle morphology, meiotic spindle organization was detected in prepubertal mouse oocytes matured under conditions allowing spontaneous, FSH- or epidermal growth factor (EGF)-dependent meiotic maturation. METHODS: Oocyte-cumulus complexes (OCCs) were matured either spontaneously (control; n=270) or in the presence of hypoxanthine (Hx) plus FSH (n=400) or EGF (n=370). Spindles were detected by immunofluorescence analysis. In vivo ovulated (IVO) oocytes were processed similarly. RESULTS: IVO oocytes displayed spindles underlying the oolemma and with focused poles marked by spots of gamma-tubulin, whereas the majority (89%) of control oocytes had barrel-shaped spindles, positioned away from the oolemma, and with gamma-tubulin distributed along microtubules. Similar configuration/localization was found in 85% of the oocytes matured in vitro in the presence of Hx and FSH. In the presence of Hx-EGF, 35% of the oocytes showed spindles with an IVO-like configuration, although gamma-tubulin was homogeneously distributed throughout microtubules. Independently of spindle shape, 52% of EGF-stimulated oocytes had spindles positioned near the oolemma, in comparison to just 24% of FSH-treated and 13% of control oocytes. CONCLUSIONS: These results indicate that FSH and EGF can differently affect meiotic spindle morphology, and that EGF might be a stronger contributor than FSH to the acquisition of oocyte competence.

Animals↗
PubMed1986

Plasma estradiol, testosterone, and progesterone levels during the ovulatory cycle of the skate (Raja erinacea).

Amounts of estradiol, testosterone, and progesterone in plasma were measured during the reproductive cycle of female Raja erinacea. Estradiol titers correlated directly with follicle size in females undergoing ovarian recrudescence, while highest concentrations were found in females with preovulatory follicles. These data indicate that as follicles grow, their steroidogenic capacity increases. In mature, nonspawning females, titers of estradiol and testosterone varied markedly. Progesterone was not detected in peripheral plasma of skates that did not produce eggs during the observation period. In females producing eggs, estradiol and testosterone predominated during the follicular phase of each spawning cycle. While estradiol and testosterone were elevated, progesterone was not detectable in the peripheral circulation. As ovulation and formation of capsules approached, plasma estradiol and testosterone declined to near baseline levels. Circulating progesterone rose sharply two days before encapsulation of ovulated eggs and remained elevated for only two days. On the day of encapsulation, concentrations of plasma progesterone had fallen to nearly baseline levels. Progesterone titers remained low throughout egg retention and oviposition. These measurements demonstrate that progesterone titers are elevated at specific times during the reproductive cycle of the skate and clearly suggest that progesterone is critically involved in events occurring at ovulation, encapsulation, and possibly oviposition.

Animals↗
PubMed1997

A specific alpha-tubulin is associated with the initiation of parthenogenesis in 'Salmon' wheat lines.

The 'Salmon system' consists of isogenic but alloplasmic wheat lines with either sexual or autonomous embryo development. Using two-dimensional gel electrophoresis these lines have been screened for proteins potentially involved in the initiation of parthenogenesis. A temporally altered expression of the polypeptide 'P 115.1' in the sexual and parthenogenetic 'Salmon' lines seems to be related with the autonomous embryo formation. Around anthesis when most of the egg cells begin the parthenogenetic development, the polypeptide 'P 115.1' was present in ovaries of the parthenogenetic lines but not in ovaries of the sexual line. Moreover, this polypeptide is only expressed in the ovaries of amphidiploid parthenogenetic plants containing differentiated embryo sacs. It is absent from ovaries of the analogous polyhaploid plants, which lack any embryo sac structure within their ovules. Furthermore, the polypeptide was neither detectable in meristematic tissue of root tips nor in leaves. N-terminal amino acid sequencing identified 'P 115.1' as an alpha-tubulin. Thus, 'P 115.1' apparently represents an embryo sac-specific isoform of alpha-tubulin involved in the initiation of embryo development.

Amino Acid Sequence↗
PubMed1984

Human preovulatory follicular fluid, luteinized cells of hyperstimulated preovulatory follicles, and corpus luteum contain placental protein 12.

RIA gel filtration, isoelectric focusing; and immunoperoxidase staining were employed to study the occurrence and physicochemical characteristics of placental protein 12 (PP12) in the human ovary, corpus luteum, and preovulatory follicular fluid. Fluid aspirated from 75 follicles from 22 women hyperstimulated for in vitro fertilization contained 6-230 micrograms/liter PP12-like immunoreactive material. The dose-response curves of follicular fluid PP12, amniotic fluid PP12, and purified human placental PP12 were parallel in the PP12 RIA. In gel filtration, follicular fluid PP12 eluted in the same volume as purified PP12. The isoelectric point of follicular fluid PP12 was 4.9 and that of purified placental PP12 4.6-4.7. A positive correlation was found between follicular fluid estradiol and PP12, progesterone and PP12, and follicular fluid volume and PP12 concentrations. By immunoperoxidase staining, PP12 was not detectable in unstimulated ovarian tissue before ovulation. In hyperstimulated preovulatory follicles biopsied in connection with follicle aspiration, PP12 was found in the granulosa cells which were luteinized (n = 3), whereas in those hyperstimulated follicles (n = 5) with no luteinization, no PP12 was found either. PP12 was seen in all corpora lutea (n = 5) from unstimulated menstrual cycles. These results show that the occurrence of PP12 is not limited to the placenta. The correlation between follicular fluid steroid and PP12 levels and the findings by immunoperoxidase staining suggest that PP12 is related to endocrine phenomena of the ovary, possibly to the luteinization process.

Chromatography, Gel↗
PubMed1998

Mid-follicular phase pulses of inhibin B are absent in polycystic ovarian syndrome and are initiated by successful laparoscopic ovarian diathermy: a possible mechanism regulating emergence of the dominant follicle.

The hypothalamic pulse generator of GnRH is recognized to be central to ovulatory function as evidenced by the anovulation of women with hypogonadotrophic hypogonadism due to Kallmann's syndrome or severe anorexia nervosa. LH is released from the anterior pituitary in pulses, the frequency of which is closely entrained with those of GnRH. In contrast, secretion of FSH is influenced by a number of regulatory molecules, including GnRH, estradiol, inhibin, and activin. The close temporal relationship between changes in levels of inhibin B and FSH in the mid-follicular phase suggests that the release of inhibin B by the preovulatory follicle critically regulates pituitary FSH secretion. Polycystic ovarian syndrome (PCOS) is one of the most common endocrine disorders affecting ovulation, and abnormal ovarian morphology as detected by ultrasonography remains the most sensitive diagnostic marker for this disorder. The etiology of PCOS is unclear, but its effective treatment by both anti-estrogens and by exogenous FSH suggests that a primary disorder of FSH regulation may be central. To investigate the possible role of inhibin B in the pathology of PCOS, serum inhibin B levels were measured in 10 women with PCOS on cycle day 5 of a spontaneous or progestrogen-provoked bleed and compared with levels on cycle day 5 of 10 women with regular ovulatory cycles. The mean serum inhibin B levels in the PCOS patients were significantly higher at 248 (+/- 43.4) pg/mL compared with normal controls, 126 (+/- 18.6) pg/mL (P < 0.01). Ten women with clomiphene resistant PCOS and 5 normal controls consented to undergo serial blood sampling on cycle day 5. Time Series Analysis using a Fourier Transformation to analyze the power spectrum of the data revealed that in normal women there is a distinct periodicity in inhibin B levels with a clear peak detectable every 60-70 min (P < 0.05), whereas in women with ovulatory dysfunction due to PCOS, no such pattern of regular pulsatility was seen. Four women with PCOS whose anovulation was successfully treated with laparoscopic ovarian diathermy (LOD) underwent repeat venous sampling following LOD. Their serum inhibin B levels fell to the upper limit of the normal range (160 +/- 38.5) pg/mL, and pulsatility was initiated. It is possible that inhibin B pulses are being generated directly by the ovary in response to pulses of GnRH in the peripheral circulation, or indirectly in response to FSH pulses arising in the pituitary. The function of inhibin B pulses in the mid-follicular phase of the normal cycle remains to be elucidated, but the absence of the normal pulsatile pattern in women with PCOS, in conjunction with high basal levels of inhibin B arising from the multiple small follicles characteristic of the PCOS ovary, appears to reinforce the development of a large cohort of small, developmentally arrested, and ultimately atretic follicles in these patients. Initiation of normal inhibin B pulsatility by LOD in patients with polycystic ovaries appears to correlate with the post-operative onset of ovular cycles.

Diathermy↗
PubMed2002

Identification and analysis of DYAD: a gene required for meiotic chromosome organisation and female meiotic progression in Arabidopsis.

The dyad mutant of Arabidopsis was previously identified as being defective in female meiosis. We report here the analysis of the DYAD gene. In ovules and anthers DYAD RNA is detected specifically in female and male meiocytes respectively, in premeiotic interphase/meiotic prophase. Analysis of chromosome spreads in female meiocytes showed that in the mutant, chromosomes did not undergo synapsis and formed ten univalents instead of five bivalents. Unlike mutations in AtDMC1 and AtSPO11 which also affect bivalent formation as the univalent chromosomes segregate randomly, the dyad univalents formed an ordered metaphase plate and underwent an equational division. This suggests a requirement for DYAD for chromosome synapsis and centromere configuration in female meiosis. The dyad mutant showed increased and persistent expression of a meiosis-specific marker, pAtDMC1::GUS during female meiosis, indicative of defective meiotic progression. The sequence of the putative protein encoded by DYAD did not reveal strong similarity to other proteins. DYAD is therefore likely to encode a novel protein required for meiotic chromosome organisation and female meiotic progression.

Amino Acid Sequence↗
PubMed2002

Plasma steroid profiles following follicle-stimulating hormone or equine chorionic gonadotropin injection in cows chronically treated with gonadotropin-releasing hormone agonist.

Plasma steroid profiles following follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG) injection were studied in chronically gonadotropin releasing hormone agonist (GnRH-A)-treated cows. Follicular development and irINH secretion were stimulated by FSH or eCG injection. The plasma concentrations of estradiol-17 beta (E(2)) and testosterone (T) were markedly increased following eCG injection. However, significant increases of the plasma E(2) and T concentrations were not detected in FSH-treated cows. Ovulation of developed follicles were depended on the hCG injection in both groups. These results show: 1) Follicular response to an exogenous gonadotropin is still remained, 2) Ovulation of developed follicles is induced by hCG injection and 3) FSH and eCG cause disparate plasma steroid profiles, under the influence of repeated GnRH-A treatment.

Animals↗
PubMed2003

Expression of the uterine Mx protein in cyclic and pregnant cows, gilts, and mares.

Pregnancy and interferon-tau (IFN tau) upregulate uterine Mx gene expression in ewes; however, the only known role for Mx is in the immune response to viral infection. We hypothesize that Mx functions as a conceptus-induced component of the anti-luteolytic mechanism and/or regulator of endometrial secretion or uterine remodeling during early pregnancy. This study was conducted to determine the effects of early pregnancy on uterine Mx expression in domestic farm species with varied mechanisms of pregnancy recognition. Endometrium from cows, gilts, and mares was collected during the first 20 d of the estrous cycle or pregnancy, and total messenger RNA (mRNA) and protein were analyzed for steady-state levels of Mx mRNA and protein. Northern blot analysis of Mx mRNA detected an approximately 2.5 Kb of mRNA in endometrium from each species. In pregnant cows, steady-state levels of Mx mRNA increased 10-fold (P < 0.05) above levels observed in cyclic cows by d 15 to 18. In cyclic gilts, slot blot analysis indicated that endometrial Mx mRNA levels did not change between d 5 and 18 of the cycle. However, in pregnant gilts, Mx levels tended (P = 0.06) to be elevated two-fold on d 16 only, and in situ hybridization indicated that this increase occurred in the stroma. In mares, Mx mRNA was low, but detectable, and did not change between ovulation (d 0) and d 20, regardless of reproductive status. Western blot analysis revealed multiple immunoreactive Mx protein bands in each species. One band was specific to pregnancy in cows. As in ewes, in situ hybridization analysis indicated that Mx mRNA was strongly expressed in the luminal epithelium, stroma, and myometrium by d 18 in cows. However, on d 14 in gilts, Mx was expressed primarily in the stroma, and on d 14 in mares, low levels of Mx expression were confined largely to the luminal epithelium. The uteruses of cows, gilts, and mares express Mx, and expression is upregulated during pregnancy in cows and gilts--animals whose conceptuses secrete interferons during early pregnancy, but that possess different mechanisms for pregnancy recognition.

Animals↗
PubMed2003

[Cloning and characterization of the PTS2 receptor gene (GhPex7) from cotton (Gossypium hirsutum L.)].

Peroxisomal targeting signals (PTS), including PTS1 and PTS2, were proposed to play an important role in introducing proteins into the peroxisomal matrix. Pex7, the PTS2 receptor, is crucial to the import of PTS2 proteins. Based on the sequence of a fragment (F010) recovered from cDNA-AFLP, the full-length cDNA sequence was obtained by RACE and the contig in cotton ESTs, and the coding sequence was further cloned. The GhPex7 cDNA, 1314 bp in length, contained 5 non-coding (77 bp) upstream, 3 complete downstream with polyA signal tail and ORF of 954 bp, which coded for a deduced protein of 317 amino acids. The deduced protein had a predicted MW 35.57 kDa and a pI of 5.603. Homology analysis demonstrated that GhPex7 protein contained three highly conserved domains and three G-bata domains of WD-40 proteins family. The sequence of nucleotides and amino acids shared 83% and 76% identity with known Arabidopsis AtPex7, respectively, and its amino acid sequence shared from 28% to 42% identity with those of Drosophila melanogaster, Saccach, cerevisiae, Mus musculus and Homo sapiens. Southern blotting suggested that at least two copies of GhPex7 gene existed in Gossypium hirstum genome. By Northern blotting and RT-PCR analysis, expression of the GhPex7 was detected in roots, stems, leaves, buds, ovules and fibers, however it was stronger in leaves and stems than in other tissues. At the stage of cotton's ovules and fibers development, RT-PCR analysis also indicated that expression activity of GhPex7 in ovule at 0DPA mutant plant (no-fiber) was stronger than in wild type plant, its expression in wild type fiber at 23DPA was stronger than at 12, 16 DPA too.

Amino Acid Sequence↗
PubMed1991

Evaluation of intra-abdominal vasectomy in llamas and alpacas.

A method of intra-abdominal vasectomy (deferentectomy), using a laparoscopic technique in llamas and alpacas, was evaluated. Food was withheld from 14 animals for 24 hours before sedation and laparoscopy. The ductus deferens was located close to the bladder, and a 2- to 3-cm section of ductus deferens was resected with the aid of a pair of hook scissors. The procedure was completed in 5 to 8 minutes, and animals did not have any adverse effects. Forty-five days after vasectomy, animals were used for detecting sexually receptive females or inducing ovulation during reproductive physiologic studies. The procedure did not impair libido and had no effect on male sexual behavior for many years. The technique is simpler, safer, and faster than the traditional external approach.

Animals↗