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Biomedical subjects

E Gootwine

Publications and source records attributed to E Gootwine.

At least 19 recordsLinked to original sources
PubMed2006

Carrying the FecB (Booroola) mutation is associated with lower birth weight and slower post-weaning growth rate for lambs, as well as a lighter mature bodyweight for ewes.

The present study was conducted in an Assaf flock in which the FecB (Booroola) mutation was segregated to determine whether the FecB mutation affects birthweight and the pre- and post-weaning growth rate of ewe lambs, as well as the mature bodyweight of ewes. Significant differences (P = 0.01) in birthweight (mean +/- s.e.m.) were found between BB ewe lambs (4.03 +/- 0.08 kg) and B+ and ++ ewe lambs (4.16 +/- 0.04 and 4.32 +/- 0.07 kg, respectively), which themselves did not differ significantly (P > 0.05). An FecB-associated maternal effect on the birthweight of ewe lambs was also detected, with the birthweight of lambs born to BB mothers (3.93 +/- 0.08 kg) being significantly (P < 0.0001) different from the birthweight of lambs born to B+ and ++ mothers (4.26 +/- 0.04 and 4.33 +/- 0.07 kg, respectively), which did not differ significantly. The genotypes of the lambs did not affect their preweaning growth rate. However, the post-weaning growth rate of ewe BB lambs (274 +/- 5 g day(-1)) was significantly (P = 0.05) different from the similar (P > 0.05) post-weaning growth rates of B+ and ++ lambs (284 +/- 3 and 290 +/- 4 g day(-1), respectively). The genotype at the FecB locus also affected the mature bodyweight of ewes, with that of BB ewes (67.3 +/- 1.4 kg) being significantly (P < 0.001) different from the similar mature bodyweight of B+ and ++ ewes (70.8 +/- 1.1 and 70.1 +/- 1.7 kg, respectively).

Animals↗
PubMed2004

Placental hormones and fetal-placental development.

Production of growth promoting substances by the placenta is regulated differently from the way production of similar compounds is regulated by maternal organs in various cases. Gene duplication is one of the mechanisms that facilitated the evolution of placental specific endocrine activity. Cattle, sheep and goats, although evolutionarily related, differ significantly from each other in the way their placental growth hormone (GH) and prolactin (PRL)-like hormones have evolved. Cattle carry one copy of the GH gene and there is no evidence yet for expression of that single GH gene copy in the placenta. On the other hand, the ovine GH gene has been duplicated and both oGH copies are expressed in the placenta during early stages of gestation. Prolactin gene duplication in ruminants resulted in the formation of specific placental-expressed prolactin-related genes including the placental lactogen (PL) gene. In homologous state, ovine PL manifests PRL activity, but antagonizes GH activity. Ovine PL activity which can be mediated by PRL receptors or by hetero-dimerization of GH and PRL receptors, provide a novel regulatory mechanism for somatogenic activity dependent on the coexistence of both GH and PRL receptors in the same cells. Another mechanism for specific placental endocrine activity is silencing of the alleles through genetic imprinting. Disruption of genetic imprinting of placental genes has been proposed as one of the explanations for the loss of cloned fetuses generated by somatic cell nuclear transfer.

Animals↗
PubMed2004

Reproductive performance and milk production of Assaf sheep in an intensive management system.

The Assaf breed of dairy sheep, a stabilized cross of the Awassi and East Friesian breeds, has replaced the Awassi as the breed of choice in its country of origin, Israel, and has spread to other Mediterranean countries. In Israel the Assaf breed is managed under an intensive production system involving weaning lambs at birth, rearing them artificially, and milking ewes after parturition. There are several breeding periods in the year when ewes are mated following hormonally synchronized estrus. Records of 18,976 lactations from 5 farms were analyzed to investigate factors that influenced Assaf milk and reproductive performance. Lactation curves were fitted to each lactation, and a range of parameters and calculated values were analyzed. Daily milk yield records also were analyzed to describe a typical Assaf lactation and compared with those of the Awassi breed. Factors affecting age at first lambing also were studied. An average Assaf ewe kept under this intensive management regimen was found to produce 334 L of milk during a 173-d lactation. Mean litter size was 1.57 lambs/ewe lambing, and lambing interval was 272 d. Milk production was affected by litter size, with twin- and triplet-bearing ewes producing approximately 20 L more milk per lactation than single-bearing ewes. Day length was the major environmental variable influencing milk yield. The difference between midsummer and midwinter day lengths accounted for a difference in daily milk yield of 0.44 L in favor of summer. Ewe lambs that were mated for the first time at later ages produced more lambs and more milk due to greater early lactation characteristics. Milk production was found to be negatively associated with subsequent reproductive performance. Comparing these results with those from an earlier study in the Awassi breed, the Assaf was found to produce less milk during a shorter lactation than the Awassi, but its greater litter size made it a more profitable breed.

Age Factors↗
PubMed2001

Large-scale preparation of recombinant ovine prolactin and determination of its in vitro and in vivo activity.

Recombinant bovine Ala-prolactin (PRL) (GenBank Accession No. V00112) in prokaryotic expression plasmid pMON3401 was mutated using a mutagenesis kit, to prepare plasmid encoding ovine PRL (oPRL) (GenBank Accession No. M27057) Escherichia coli cells transformed with this latter plasmid overexpressed large amounts of oPRL upon induction with nalidixic acid. The expressed protein, found in inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding an electrophoretically pure fraction composed of over 98% monomeric protein of the expected molecular mass of approximately 23 kDa. The biological activity of the recombinant oPRL after proper renaturation was evidenced in vitro by its ability to stimulate proliferation of rat lymphoma Nb(2) cells possessing PRL receptors, to stimulate luciferase activity in HEK 293 cells transiently transfected with oPRL receptors, and to induce progesterone secretion in primary cultures of luteal cells obtained from midpregnant ewes. In contrast to ovine growth hormone or ovine placental lactogen, recombinant oPRL had no galactopoietic effect in lactating ewes.

Animals↗
PubMed2000

Active immunization of ewes against ovine placental lactogen increases birth weight of lambs and milk production with no adverse effect on conception rate.

In two experiments, 16 Booroola-Assaf and 35 Assaf ewe-lambs were actively immunized at 5 months of age against recombinant ovine placental lactogen (oPL). At 9 months of age, the ewe-lambs were mated for the first time and then introduced into a frequent mating-system. Anti-oPL antibody titers, reproductive performance, maternal serum levels of oPL during pregnancy, lamb birth weight and milk production of the ewes were followed in the immunized ewes and in their non-immunized control counterparts. All the immunized ewes developed anti-oPL antibodies, which interfered with oPL bioactivity in an in vitro cell proliferation assay. Conception rates did not differ (P>0.05) between immunized and non-immunized ewes. Abundant antibody-bound non-active oPL detected in sera of immunized ewes by western blotting indicated enhanced oPL production by the placenta following immunization. An increase (P<0.02) in serum oPL bioactivity, but not immunoreactivity, was observed in the immunized ewes in late gestation relative to control ewes. The average litter size was 1.83 and 1.32 lambs born per ewe lambing in the first and second experiments, respectively. Average birth weights of lambs born to the immunized ewes were higher (P<0.01) than for lambs born to control ewes by 10, 17 and 39% for those born as singles, twins and triplets, respectively. Immunized ewes produced 19 and 33% more milk (P<0.02) than the control ewes in the first 3.5 months of the first and second lactations, respectively. These findings do not suggest a role for oPL in maternal recognition of pregnancy, but they strongly suggest important roles for oPL in fetal growth and mammogenesis. Immunization of ewes against oPL may thus represent a novel practical technique for enhancing birth weights of lambs born to prolific sheep, as well as milk production by both dairy and mutton ewes.

Animals↗
PubMed1999

Effects of recombinant ovine interferon tau, placental lactogen, and growth hormone on the ovine uterus.

Studies were conducted to determine effects of intrauterine administration of recombinant ovine interferon tau (IFNtau), placental lactogen (PL), and growth hormone (GH) on endometrial function. In the first study, administration of IFNtau to cyclic ewes for one period (Days 11-15) resulted in an interestrous interval (IEI) of approximately 30 days, whereas administration for two periods (Days 11-15 and Days 21-25) extended the IEI to greater than 50 days. Administration of IFNtau from Days 11 to 15 and of PL or GH from Days 21 to 25 failed to extend the IEI more than for IFNtau alone. In the second study, effects of IFNtau, PL, and GH on endometrial differentiation and function were determined in ovariectomized ewes receiving ovarian steroid replacement therapy. Endometrial expression of mRNAs for estrogen receptor (ER), progesterone receptor (PR), and oxytocin receptor (OTR) were not affected by PL or GH treatment; however, uterine milk protein mRNA levels and stratum spongiosum gland density were increased by both PL and GH treatments. Collectively, results indicated that 1) PL and GH do not regulate endometrial PR, ER, and OTR expression or affect corpus luteum life span; 2) down-regulation of epithelial PR expression is requisite for progesterone induction of secretory gene expression in uterine glandular epithelium; 3) effects of PL and GH on endometrial function require IFNtau; and 4) PL and GH regulate endometrial gland proliferation and perhaps differentiated function.

Animals↗
PubMed1998

Establishment and initial characterization of the ovine mammary epithelial cell line NISH.

Analysis of the molecular mechanisms involved in the differentiation and formation of the characteristic three-dimensional structures of the developing mammary gland of the major milk-producing livestock (ducts, end buds, and alveoli) requires in vitro model cell cultures. The few cell lines that have been established from dairy animals do not fully reproduce the entire program of mammary differentiation. Here we present the initial characterization of a unique mammary epithelial cell line derived spontaneously from midpregnant sheep (NISH). These cells form in vitro functional structures resembling ducts, lateral buds, and alveoli that secrete beta-lactoglobulin (BLG) in an ECM (extracellular matrix)-dependent manner. Interestingly, the presence of growth hormone dramatically increased BLG secretion from NISH cells cultured on ECM. It appears that GH is required not only to establish the structural organization but also is continuously needed to maintain BLG expression. Stable transfection of NISH cells with BLG/Human Serum Albumin (HSA) hybrid gene constructs revealed that the relative level of expression was comparable to the in vivo secretion of HSA in transgenic mice carrying these gene sequences. No expression could be detected in cells transfected with hybrid genes carrying either HSA cDNA or the entire HSA gene, and HSA expression was dependent on the presence of intronic sequences. These results demonstrate that NISH cells may prove a useful tool for studying the differentiation and organogenesis of mammary epithelial cells under defined culture conditions. Furthermore, transfected NISH cells may be an alternative for the transgenic mouse model in evaluating the potential of gene constructs to be efficiently expressed in the mammary gland of transgenic farm animals.

Animals↗
PubMed1997

Factors affecting success of embryo collection and transfer in a transgenic goat program.

During a goat transgenic program that took place in Israel from July 1995 to February 1996, Saanen (n = 343) and Nubian x Damascus (n = 378) crossbred goats of mixed ages were used as donors (n = 433) and recipients (n = 288). The effects of season, age, number of surgical procedures, previous hormonal treatments and ovulation rate on the number of microinjectable embryos collected were studied. Likewise, the effects of these parameters on the pregnancy rate as well as the number of embryos transplanted, endogenous progesterone concentrations and exogenous progesterone supplementation were studied in recipient does. Following superovulation with ovine follicle stimulating hormone, 85% of the does responded with 13.6 +/- 5.7 (mean +/- S D) ovulations/doe. Age, month and number of previous hormonal treatments significantly affected the ovulation rate. The average recovery rate was 70%, and it was affected only by the ovulation rate. Pronuclei were visualized in about 30% of the flushed embryos (including unfertilized ova), and those were microinjected with human serum albumin gene construct. About 68% of the injected embryos underwent at least one division during an overnight incubation, and those embryos were transferred, giving about 2.0 transferred embryos per ovulated donor. Of the recipients, 86% responded following synchronization with 3.1 +/- 1.6 (mean +/- S D) ovulations per doe. Breed and month had a significant effect on the ovulation rate. Two or three microinjected embryos were transferred to each recipient, resulting in more than a 40% pregnancy rate during September to November. Lower pregnancy rates were obtained before and after that period. By monitoring plasma progesterone concentrations in the recipients it was found that progesterone concentration was correlated with the ovulation rate. However, the pregnancy rate was not affected by progesterone concentration. During January and February, 30 to 50% of the recipients failed to develop functional corpora lutea (CL) following embryo transfer, which explained the lower pregnancy rate in those months. Of the 86 kids born 4 were transgenic.

Journal Article↗
PubMed1997

Booster immunization with a partially purified citrus tristeza virus (CTV) preparation after priming with recombinant CTV coat protein enhances the binding capacity of capture antibodies by ELISA.

Groups of rabbits and young lambs were immunized subcutaneously and intramuscularly with a recombinant citrus tristeza virus (CTV) coat protein (rCTV-CP) antigen. Three weeks after primary immunization the animals were divided into two groups that were boosted either with rCTV-CP or with a partially purified preparation of CTV particles (ppCTV). Twelve and 15 days after the last injection, the animals were bled and the binding capacity of the antisera for CTV detection was examined for capture antibodies by the indirect ELISA. Considerably higher ELISA titers were obtained from animals that were boosted with ppCTV than with rCP. Boosting with partially purified native antigens after priming with recombinant antigens is expected to extend the applicability of the antisera for detecting other structural and non-structural viral antigens by trapping ELISA.

Animals↗
PubMed1997

The effects of a duplication in the ovine growth hormone (GH) gene on GH expression in the pituitaries of ram lambs from lean and fat-selected sheep lines.

Growth hormone (GH) gene expression was investigated in pituitaries of 14- to 15-month-old ram lambs from flocks selected for high (fat) or low (lean) back fat depth, which were also homozygous for a single GH gene allele, heterozygous or homozygous for a duplication in the GH gene. The pituitaries of lean sheep of all three GH genotypes were significantly heavier than those of fat sheep, but there were no pituitary weight differences between GH genotypes. No significant lean-fat selection line- or GH genotype-specific differences were measured in pituitary GH concentration. However there was a significant increase (P < 0.01) in the total pituitary content of GH in lean compared with fat animals and a significant interaction between GH genotype and lean-fat selection line (P < 0.05) was noted for GH content. No significant differences were measured in the relative concentration of GH mRNA, suggesting that the ratio of GH mRNA per mg total cellular RNA remained constant across lean-fat selection line and GH genotype. We conclude that the pituitary glands of Coopworth sheep selected for low backfat depth (lean) are bigger and have an increased GH content, but appear to contain similar concentrations of GH mRNA and immunoreactive GH as the pituitaries of fat sheep. The presence of the GH gene duplication in sheep has little measurable effect on the expression and storage of GH in the pituitary.

Alleles↗
PubMed1997

The physiological effects of natural variation in growth hormone gene copy number in ram lambs.

The effects of natural variation in the number of copies of the growth hormone (GH) gene on growth parameters, plasma GH profiles, and the response to GHRH challenge were compared in Coopworth ram lambs from selection lines differing in body composition and GH levels. Different genotypes at the GH locus carried two, three, or four copies of the GH gene and GH secretion was studied under ad libitum feeding conditions and in the fasted state. There were no significant effects of GH genotype on any parameters of growth or body composition. Basal serum GH concentration, GH pulse frequency, and GH pulse amplitude differed significantly with selection line and fasting, but did not differ significantly between the GH genotypes. Significant differences of subtle nature were found between the GH genotypes in their responsiveness to GHRH. For the ad libitum-fed Lean selection line animals, the first GHRH challenge resulted in a higher mean maximum response for GH1/GH1 than GH2/GH2 (P < 0.05). Between the first and the second challenges there was a decrease in maximum response for the GH1/GH1 genotype and an increase for the GH2/GH2 genotype (P < 0.05 for GH genotype main effect). The differences between GH genotypes in response to GHRH challenge suggest that polymorphism in the number of GH gene copies in sheep may have physiological implications for the function of the GH axis, which may be manifested in growing lambs only under specific genotype-environment combinations.

Alleles↗
PubMed1993

Plasma immunoreactive inhibin and FSH in prepubertal Assaf and Booroola-Assaf ewe lambs.

Plasma immunoreactive inhibin (i-inhibin) was measured in Assaf and Booroola-Assaf (1/2 Booroola-1/2 Assaf) ewe lambs between the ages of 2 and 15 weeks and correlated with plasma FSH. I-inhibin concentrations were similar in both genotypes and was maximal at 2 weeks of life and decreased thereafter. These levels (mean +/- s.e.) were 1.28 +/- 0.12 and 1.14 +/- 0.10 ng/ml at 2 weeks and 0.55 +/- 0.06 and 0.45 +/- 0.05 ng/ml at 15 weeks of age in Assaf (n = 13) and Booroola-Assaf (n = 13) respectively. In Assaf ewes an inverse correlation between plasma inhibin and FSH was observed at 4 and 15 weeks but not at 2 weeks of age. In Booroola-Assaf ewes, no significant correlation was found at any age studied. At 4 weeks of age, Assaf and Booroola-Assaf ewe lambs had very different ovarian morphology. Assaf lambs had large ovaries with a mean diameter of 1.00 +/- 0.09 cm (mean +/- s.e.) and many preantral and antral follicles present, while Booroola-Assaf had much smaller ovaries (0.66 +/- 0.03 cm, p < 0.05) with only a few antral follicles. It seems likely that the FecB allele from the Boorola Merino influenced the structure of the ovary although the contribution of other Booroola genes cannot be ruled out. In summary, the plasma i-inhibin concentrations decreased between 2 and 15 weeks of age irrespective of genotype. It is speculated that in prepubertal ewe lambs the ratio of bioactive to immunoreactive inhibin varies with age and between the genotypes with different ovarian development.

Aging↗
PubMed1992

Monitoring ovarian response of Booroola Assaf ewe lambs to PMSG, using ultrasonography and serum estradiol.

Ovarian size was determined for 30-day-old ewe lambs treated with pregnant mare serum gonadotropin (PMSG; 1000 IU) using ultrasonography and was compared with direct measurement at laparotomy. Ultrasonography was found to be an accurate and reliable method for estimating ovarian size, and a significantly high correlation was found between the methods (R=0.96, P<0.01). Forty-eight hours following PMSG treatment, serum estradiol levels increased from below detectable levels to a value of 72.8+/-27.4 pg/ml (mean+/-SEM). A high correlation was found also between estradiol levels and ovarian size measured at laparotomy and by ultrasound, with a correlation coefficient of 0.94 and 0.80, respectively. It is suggested that a differential increase in ovarian size and estradiol secretion in immature ewe lambs treated with PMSG may reflect the different stages of ovarian development in individual animals. Noninvasive methods such as ultrasonography and measurements of serum estradiol may be utilized in the estimation of ovarian response.

Journal Article↗
PubMed1992

Differential pituitary response to GnRH in pregnant Booroola-Assaf and Assaf ewes.

Pituitary response to exogenous gonadotrophin-releasing hormone (GnRH) was examined at a late stage of pregnancy in 10 Booroola-Assaf ewes heterozygous at the FecB locus (FecBFec+) and in 11 Assaf ewes that were non-carriers (Fec+Fec+). Basal plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations were similar in the two genotypes. Administration of 100 micrograms of GnRH resulted in a significant increase (P < 0.05) in plasma LH and FSH concentrations in most of the ewes treated. Maximal responses were observed 105-135 min after GnRH treatment. Pituitary responses to GnRH were significantly lower (P < 0.05) in Booroola-Assaf than in Assaf ewes. The decreased pituitary responsiveness observed in FecB gene carriers compared with Fec+Fec+ ewes might be due to differences in the concentrations of ovarian or uterine hormones modulating the release of gonadotrophin. The results suggest that FecB-specific differences can be observed at the pituitary level.

Animals↗