Association of single nucleotide polymorphisms in the growth hormone and growth hormone receptor genes with blood serum insulin-like growth factor I concentration and growth traits in Angus cattle

This study was conducted to identify polymorphisms in the promoter and coding regions of the bovine growth hormone and growth hormone receptor genes and to study association of polymorphisms identified in these genes with growth traits and serum insulin-like growth factor-I (IGF-I) concentration. The denaturing gradient gel electrophoresis method and sequencing were utilized to identify three new single nucleotide polymorphisms in the promoter region of the growth hormone gene in Angus cattle. Polymerase chain reaction-based restriction fragment length polymorphism procedures were developed for rapid determination of the single nucleotide polymorphism genotypes in the growth hormone and the growth hormone receptor genes among Angus calves from lines divergently selected for high or low blood serum IGF-I concentration. The IGF-I concentration and growth traits were analyzed using animal models. The single nucleotide polymorphism in the promoter region of the growth hormone receptor gene was associated with serum IGF-I concentration on d 42 of the postweaning test and with mean IGF-I concentration. The associated effects of the markers need to be verified in other populations.     

Reproductive performance of bulls divergently selected on the basis of blood serum insulin-like growth factor I concentration.

The objectives of this study were to examine differences in scrotal circumference, sperm motility, and percentage of normal sperm cells between two lines of Angus beef cattle divergently selected for blood serum IGF-I concentration. Data were obtained from an ongoing experiment involving 100 spring-calving (50 high and 50 low line) and 100 fall-calving (50 high and 50 low line) purebred Angus cows. Scrotal circumference, percentage of motile sperm cells, and percentage of normal sperm cells did not differ between high and low IGF-I line yearling bulls (P = .79, .50, and .56, respectively). The IGF-I concentrations measured at d 28, 42, and 56 of the postweaning test are abbreviated as IGF28, IGF42, and IGF56, respectively. Coefficients for the quadratic regression of scrotal circumference on IGF28 and IGF42 tended to be negative (P = .07 and .08, respectively), as did the coefficient for the quadratic regression of the percentage of motile sperm cells on IGF42 (P = .08). The coefficient for the linear regression of percentage of normal sperm cells on IGF28 was positive (P = .02). The coefficient for the quadratic regression of percentage of normal sperm cells on IGF56 was negative (P = .04). Coefficients for the quadratic regression of scrotal circumference and percentage of normal sperm cells on mean IGF-I concentrations were negative and important (P = .04 and .08, respectively). Thus, scrotal circumference and percentage of normal sperm cells are related to blood serum IGF-I concentration in yearling Angus bulls.     

Cloning and characterization of a genomic probe for malignant catarrhal fever virus

A genomic probe specific for malignant catarrhal fever (MCF) virus was cloned by using purified viral DNA from MCF-virus strain WCll. Restriction endonuclease analysis of the purified viral DNA was used to identify the cloned viral genomic fragment. Dot blot hybridization by use of the genomic probe (pRP-5) indicated that the probe hybridized specifically with WCll-MCF virus, as well as with one other isolate of MCF-associated herpesvirus. Hybridization also was observed to a non-MCF virus strain of bovine herpesvirus.     

Insulin and insulin-like growth factor-I stimulate DNA synthesis in bovine mammary tissue in vitro

Effects of insulin and insulin-like growth factor I (IGF-I) on [3H]thymidine incorporation, in vitro, by mammary tissue slices obtained from prepartum and lactating cows were investigated. Both insulin and IGF-I induced up to a 10-fold increase in [3H]thymidine incorporation in the mammary slices cultured in serum-free media. The effect of insulin-stimulated [3H]thymidine incorporation occurred at a threshold of greater than 1.75 pmol/ml and appeared to reach maximum at greater than 8.8 nmol/ml. The response to IGF-I occurred at greater than 6.5 pmol/ml and reached the equivalent of maximal insulin-stimulated incorporation at 39 pmol/ml. No synergistic or additive effects were observed between these two factors. The in vitro response took 3 to 4 d to reach maximum and was inhibited by cytarabine. Mammary tissue obtained from lactating cows incorporated more [3H]thymidine per microgram DNA in response to insulin (175 pmol/ml) than mammary tissue from pregnant cows. Culture of mammary tissue slices with growth hormone, cortisol, prolactin, or triiodothyronine showed no stimulation of [3H]thymidine incorporation over control. Autoradiography of the cultured lactating tissue showed incorporation of [3H]thymidine by 51, 24 and 29% of the ductal epithelial, secretory alveolar epithelial and myoepithelial cells, respectively. All alveolar epithelial cells that incorporated [3H]thymidine contained secretory products. Among nonsecretory cells, 25 and 28% of the fibroblasts and white blood cells, respectively, were labeled. Insulin-like growth factor I, but not bovine somatotropin, stimulated [3H]thymidine uptake into DNA in lactating bovine mammary tissue. Thus, our data support the concept that bovine somatotropin acts through IGF-I to increase DNA synthesis in mammary cells.     

Impact of blood serum insulin-like growth factor I on platelet-activating factor in bull spermatozoa

The objective of this study was to examine differences in platelet-activating factor [1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine; PAF] in spermatozoa between two lines of Angus beef cattle divergently selected for blood serum insulin-like growth factor I (IGF-I) concentration. Endogenous lipids were extracted from the spermatozoa and endogenous PAF content was determined by radioimmunoassay. The amount of PAF detected in spermatozoa obtained from high IGF-I bulls (n = 8) ranged from 0.145 to 3.571 pM/10(6) cells. The level of PAF extracted from spermatozoa obtained from low IGF-I- bulls (n = 5) ranged from 0.001 to 1.024 pM/10(6) cells. Polynomial regression analysis revealed a significant cubic relationship (R(2) = 0.374; F = 6.292; P < 0.05) between spermatozoa PAF content and blood serum IGF-I concentration. Spermatozoa-derived PAF levels (mean +/- SEM) were significantly higher (P < 0.05) in the high IGF-I group (1.90 +/- 0.39 pM/10(6) cells) than in the low IGF-I group (0.59 +/- 0.20 pM/10(6) cells). High IGF-I bulls have a greater than three-fold higher PAF content in their spermatozoa than low IGF-I bulls. The data demonstrate that not only is PAF present in bull spermatozoa but that levels are significantly higher in individuals with high serum IGF-I concentrations.     

Influence of diet on basal and growth hormone-stimulated plasma concentrations of IGF-I in beef cattle

The effects of nutrition on plasma concentrations of insulin-like growth factor-I (IGF-I) were characterized in steers under basal conditions and following single i.m. injection of bovine growth hormone (bGH, .1 mg/kg BW). Nutritional effects on IGF-I were studied in three trials. In all trials steers were individually fed and penned Angus or Hereford x Angus (280 kg). In the first trial, two diets (LPLE1: 8% CP and 1.96 Mcal ME/kg, 4.5 kg.hd-1.d-1; MPHE1: 11% CP, 2.67 Mcal ME/kg, 6.5 kg.hd-1.d-1) were fed (n = 5/diet). Plasma IGF-I concentrations averaged 74 (LPLE1) and 152 (MPHE1) ng/ml (P less than .02). Following bGH injection, IGF-I increased to peak concentrations between 12 and 24 h (averaging 105 and 208 ng/ml at peak for LPLE and MPLE, respectively, P less than .01). In the second trial, steers were fed diets composed of 8, 11 or 14% CP and 1.96 or 2.67 Mcal ME/kg dry matter (6.35 kg.hd-1.d-1 in a factorial arrangement for 84 d, n = 4/diet). Within the low ME diet groups, plasma IGF-I was similar in steers fed 11 and 14% CP but greater at these two CP levels than in steers fed 8% CP (P less than .05). Within the high ME diet groups, plasma IGF-I increased linearly with CP (P less than .01). In the third trial, steers were fed diets to result in a negative N status. Insulin-like growth factor-I was lower (P less than .02) during feed restriction than when steers were full-fed. The IGF-I response to bGH was diminished or absent in underfed steers (P less than .01). These data are interpreted to suggest that diet composition and intake affect plasma concentrations of IGF-I in steers. In cattle, CP may be the primary nutritional determinant of basal IGF-I, but the IGF-I response to CP may be affected by the available ME. Undernutrition can attenuate the IGF-I response to GH and uncouple the regulation of IGF-I normally ascribed to GH.     

The influence of tropical adaptation on plasma concentrations of insulin-like growth factor-I in purebred and crossbred beef cattle

In an effort to determine whether tropical adaptation influences circulating concentrations of the growth-related hormone IGF-I, 3-breed diallel matings were conducted using temperate Bos taurus (Angus), tropical Bos indicus (Brahman), and tropical Bos taurus (Romosinuano). Purebred Angus, Braham, and Romosinuano and crossbred Angus-Braham, Angus-Romosinuano, and Braham-Romosinuano heifers and steers were evaluated in 2 separate calf crops from 2003 and 2004. Blood samples were obtained from 10 heifers of each breed group (n = 90) for each year at weaning and on d 0 and 84 of postweaning trials. Samples were also taken from 10 steers of each breed group (n = 90) at weaning and on d 0 and 60 of individual finishing phase feeding trials for each year. Concentrations of IGF-I were determined by RIA. Analyses included effects of sire breed, dam breed, year of record, the age of the dam of the calf in years, and interactions. Age of calf in days was investigated as a linear and quadratic covariate. Separate analyses were conducted for steers and heifers. The direct effect of Angus was to reduce (P < 0.03) heifer concentrations of IGF-I at d 84 and in the repeated measures analysis. In the repeated measures analysis, the direct effect of Romosinuano was to increase concentrations of IGF-I (P = 0.01). Relative to the temperate Bos taurus breed, plasma concentrations of IGF-I were numerically greater in male and female tropically adapted breed groups.     

Detection of infectious laryngotracheitis virus infected cells with cloned DNA probes.

A genomic library of infectious laryngotracheitis virus (ILTV) DNA BamH1 fragments was prepared and two cloned fragments were evaluated for their potential as probes for the detection of ILTV infected cells. The virus was purified by a modified sucrose density gradient procedure for the isolation of pure ILTV DNA. A genomic library was constructed using BamH1-digested ILTV DNA and pGEM7 as a vector. A 1.1 kb cloned BamH1 fragment of ILTV DNA was tested in a slot or dot blot assay for the detection of ILTV infected cells. The limit of detection for this probe was at least 0.12 ng of pure ILTV DNA. The probe was able to identify both chicken embryo liver (CELi) cells and choriallantoic membranes infected with ILTV. Chicken embryo liver cells infected with several field isolates and a vaccine strain of ILTV were positive by dot blot analysis using this probe. Some qualitative differences in the degree of hybridization to cells infected by different ILTV isolates were observed. Uninfected cells and cells infected with fowlpox virus, turkey herpesvirus, Marek's disease virus or Newcastle disease virus were negative by the same assay. Compared with the 1.1 kb fragment, a larger 6 kb cloned BamH1 fragment of ILTV DNA showed a stronger hybridization signal to DNA from ILTV infected cells.     

Diagnosis of malignant catarrhal fever by PCR using formalin-fixed, paraffin-embedded tissues.

A previously described polymerase chain reaction (PCR) assay (amplification of a 238-bp fragment of ovine herpesvirus 2 [OHV-2] genomic DNA) for diagnosis of sheep-associated malignant catarrhal fever (MCF) was adapted for use on formalin-fixed, paraffin-embedded tissues. Variables affecting its use were examined. Archived tissues from cattle, white-tailed deer (Odocoileus virginianus), and bison (Bison bison) diagnosed with MCF by clinical signs or histologic lesions were obtained from 2 veterinary diagnostic laboratories. Tissues from healthy animals and from animals diagnosed with other common bovine viral diseases were examined as controls. A total of 86 blocks from 37 suspect MCF cases were examined. Forty-one blocks from healthy animals and animals with unrelated viral diseases were examined as controls. The assay was specific for sheep-associated MCF and did not yield false-positive signals from healthy animals or from cases of infectious bovine rhinotracheitis, bovine virus diarrhea, mucosal disease, or parainfluenza-3 virus infection. A wide variety of tissues were suitable substrates, including spleen, lymph node, intestine, brain, lung, and kidney. Extracted DNA provided a more suitable target than did unextracted tissue lysate. The highest levels of viral DNA were present in lymphoid organs and intestine, but the data indicate that in acute clinical cases, most organs contain sufficient viral DNA to serve as a suitable diagnostic specimen. Fixation of 0.5-cm3 blocks of tissue in 10% neutral buffered formalin was deleterious to the target DNA, and PCR signals progressively diminished after fixation for >45 days. Detection of genomic DNA of OHV-2 by PCR was successful for archived tissues that were 15 years old.     

凋亡素基因的克隆

为了探索应用凋亡素杀死肿瘤细胞的可能性,用ＰＣＲ方法扩增了鸡贫血病毒（ｃｈｉｃｋｅｎａｎｅｍｉａｖｉｒｕｓ,ＣＡＶ）的ＶＰ３基因（凋亡素）片段,然后将该片段重组于ｐＵＣ１９载体,并进行了限制性内切酶鉴定与序列分析,证实该片段与鸡贫血病毒Ｃｕｘ－１株的ＶＰ３ＤＮＡ序列一致,该ＤＮＡ全长３６３ｂｐ,编码１２１个氨基酸。     

大肠杆菌免疫刺激DNA的克隆及其对抗体产生的影响

利用聚合酶链式反应（Polymerase Chain Reaction,PCR）从大肠杆菌基因组中扩增免疫刺激基因，并把扩增的基因片段克隆和pGEM－T载体。PstI酶切结果表明重组质粒（rP）均含有扩增的基因片段；rP1与rP2是相同的重组质粒。利用酶联免疫吸附检测了重组质粒对小鼠产生抗体的影响，结果显示：重组质粒1与牛血清白蛋白（BSA），免疫小鼠能产生较同的抗体水平，而重组质粒3则对小鼠产生的BSA抗体水平没有影响。对重组质粒1的序列分析,克隆的基因片段中富含CpG序列，并含有数个具有强烈免疫刺激作用的RRCGYY（如：AACGTT）序列。本研究为基因免疫机理的深入研究和基因免疫佐剂的开发奠定了基础。     

Association of a genetic marker with blood serum insulin-like growth factor-I concentration and growth traits in Angus cattle

The objective of this research was to evaluate a biallelic genetic marker identified in the first promoter region of the bovine IGF-I gene. The point mutation was identified as a T-to-C transition by sequencing the polymorphic fragments. A PCR-RFLP procedure was developed for determining the marker genotypes. Marker genotypes were determined for 760 Angus calves from divergent lines that were created by selection for high or low serum IGF-I concentration (allele A: 63.9%, B: 36.1%). Data were analyzed using the multiple-trait derivative-free restricted maximum likelihood computer programs with animal models. The full animal model included fixed effects of marker genotype, birth year, season of birth, sex, age of dam, and selection line; random effects of animal, maternal genetic, and maternal permanent environmental effects; and a covariate for age of calf. Traits analyzed included blood serum IGF-I concentrations on d 28, 42, and 56 of the postweaning test, mean IGF-I concentration, birth weight, weaning weight, on-test weight, off-test weight, off-test hip height, postweaning gain, and weight gain during the 20-d period immediately after weaning. Results from the analysis across selection lines showed a significant association of the BB genotype with higher weight gain during the first 20 d after weaning and a slight dominance effect of the marker on postweaning gain. Analysis within the low IGF-I line also showed a significant association of the BB genotype with higher weight gain during the first 20 d after weaning and with on-test weight, although analysis within the high IGF-I line did not show any significant association. The associated effects of the marker need to be verified in other cattle populations.     

用RAPD技术筛选中国荷斯坦牛产奶量性状遗传标记

选用320条10碱基随机引物在由36头高产（305d产奶量〉8500kg）和32头低产（305d产奶量〈5600kg）中国荷斯坦牛所组成的2个DNA池间进行RAPD-PCR扩增，从中筛选出稳定性好、带型清晰且在2个DNA池间有明显差异的RAPD引物24条。用筛选到的24条引物对所有个体进行PCR检测，得到了5个与奶牛产奶量性状相关的RAPD标记，并对其中4个进行了克隆测序和SCAR标记转化以及生物信息学分析。结果表明：SCAR标记yield—s139与产奶量密切相关，应用电子克隆方法已将该标记由921bp延伸到2141bp，经同源性比较发现，此标记对应于奶牛基因组中LINEs家族中的一个重复元件，推测该元件附近可能存在与产奶量性状相关的QTLs或主效基因。     

Secretory patterns of growth hormone and insulin-like growth factor-I during peripubertal period in intact and castrate male cattle

Fifteen Angus bulls and 15 Angus steers 9 months of age and 275 kg of body weight were bled at 20-min intervals over a 6-hr period and serum GH and IGF-I concentrations were measured by RIA. There were no differences between bulls and steers in the mean GH concentration, pulse frequency and amplitude when analyzed by the computer program PULSAR. Mean IGF-I concentration was not different between the two sex phenotypes, nor was there a significant correlation between the integrated IGF-I and GH concentrations. Subsequently, five bulls and five steers were selected from the 30 animals, full-fed a diet for growth in individual pens for 3 months and bled at 15-min intervals over a 24-hr period. Bulls tended to show a greater weight gain and feed conversion efficiency (P<.10) than steers during the 3-month period. Serum GH concentrations had a pulsatile pattern in all animals with no apparent diurnal rhythm during the 24-hr bleeding. Although mean GH concentration was not different between the two sex phenotypes, bulls tended to have lower baseline levels (P<.10) and greater peak amplitudes than steers. Serum IGF-I concentrations fluctuated within a two-fold concentration range, with no obvious pulsatility similar to that of GH. Mean IGF-I concentrations of each of the 10 animals were correlated with mean peak GH amplitudes (r = .79), but not with mean GH. These results suggest that gonadal hormone(s) modulates the GH secretory pattern and increases IGF-I secretion which may be related to the greater growth rate of bulls compared with steers.     

Canine thyrotropin beta-subunit gene: cloning and expression in Escherichia coli, generation of monoclonal antibodies, and transient expression in the Chinese hamster ovary cells

The gene encoding the mature β subunit of canine thyroid stimulating hormone (cTSHβ) was cloned, sequenced and expressed in Escherichia coli and in Chinese hamster ovary (CHO) cells, and monoclonal antibodies against the recombinant cTSHβ purified from E. coli were generated. The gene fragment that encodes mature TSHβ was cloned from the canine genomic DNA by direct polymerase chain reaction (PCR) using primers that were designed based on the consensus sequences from other species. The resulting 891 basepairs (bp) of genomic DNA consisted of two coding exons of the canine TSHβ gene and an intron of 450 bp. The two exons, which encode the mature cTSHβ subunit, was joined together by an overlap PCR and was expressed in E. coli as 6×His-tagged protein. The purified recombinant cTSHβ with a molecular weight of about 15 kDa was recognized by the polyclonal antibodies prepared against the native canine TSH in Western blot. Monoclonal antibodies were raised against the purified cTSHβ and subsequently characterized. For transient expression in CHO cells that are permanently transfected with the bovine common gene, a 60-oligonucleotide signal peptide coding sequence was added to the 5′ end of the cTSHβ gene before it was cloned into the mammalian expression vector pRSV and used to transfect CHO cells. The medium from these transfected cells, presumably containing the bovine and canine TSHβ in heterodimeric confirmation, exhibited TSH bioactivity as indicated by the stimulation of cAMP production in the cultured FRTL-5 thyrocytes.     

DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies

Genomic DNA extracted from bovine mummified tissue is valuable material for detection of some genes that may contribute to fetal abnormalities. In this study bovine genomic DNA was extracted from the hardened tissue samples of ten bovine mummified fetuses. The amount of genomic DNA extracted from 2 g of the mummified tissues by the phenol/chloroform-ethanol method was low (less than 4 microg/ml) for all samples. The extracted DNA was then amplified by the GenomiPhi DNA amplification system. After amplification, the amount of DNA was increased to more than 100 microg/ml for all samples. This amplification system was shown to be a good tool for amplifying the genomic DNA of the mummified fetuses. The amplified genomic DNA was used for testing the mummies for Factor XI gene deficiency, an autosomal recessive deficiency involved in the early stages of the intrinsic blood coagulation pathway. Exon 12 of the Factor XI gene of the mummies was amplified by PCR. Two of the ten mummified fetuses were heterozygous for the Factor XI gene as indicated by the presence of two amplified DNA fragments of 320 bp and 244 bp. Factor XI deficiency has already been described in Holstein cattle. However, no report is available for bovine fetus. In this study, DNA was extracted and amplified from the bovine mummified fetuses, and the samples were successfully tested for Factor XI gene deficiency in the mummies.     

Effects of flax supplementation and a combined trenbolone acetate and estradiol implant on circulating insulin-like growth factor-I and muscle insulin-like growth factor-I messenger RNA levels in beef cattle

We evaluated effects of a 5% (dry matter basis) ground flaxseed supplement (flax) and a trenbolone acetate and estradiol-17beta implant, Revalor-S, on circulating IGF-I and muscle IGF-I messenger RNA (mRNA). Sixteen crossbred yearling steers (initial BW = 397 kg) were assigned randomly to one of four treatments: 1) flax/implant; 2) nonflax/implant; 3) flax/nonimplant; and 4) nonflax/nonimplant. Serum was harvested from blood collected on d 0 (before implant or flax addition), 14, and 28, and used in subsequent analyses of circulating IGF-I. Biopsy samples (0.5 g) were obtained from the longissimus muscle on d 0, 14, and 28. Total RNA was isolated from the muscle samples, and real-time quantitative-PCR was used to assess relative differences in IGF-I mRNA. Flax supplementation had no effect (P > 0.10) on circulating IGF-I concentrations. Following implantation, sera from implanted steers had 52 and 84% greater (P < 0.05) IGF-I concentrations than sera from nonimplanted steers on d 14 and 28, respectively. On d 28, local muscle IGF-I mRNA levels increased 2.4-fold (P < 0.01) in biopsy samples obtained from implanted compared with nonimplanted steers. Muscle biopsy samples from nonflax cattle had 4.4-fold higher (P < 0.01) levels of IGF-I mRNA than those from flax cattle on d 28. To determine whether a component of flax, alpha-linolenic acid (alphaLA), was directly responsible for IGF-I mRNA down-regulation, we incubated primary cultures of bovine satellite cells, from implanted and nonimplanted steers, in two concentrations of alphaLA (10 nM and 1 microM). An implant x dose interaction (P < 0.05) was observed for IGF-I mRNA concentrations in bovine satellite cells cultured for 72 h with alphaLA. Satellite cells from nonimplanted steers had similar (P > 0.10) IGF-I mRNA concentration regardless of the level of alphaLA exposure; however, satellite cells from implanted steers exposed to 10 nM and 1 microM alphaLA had 2.5- and 2.0-fold greater IGF-I mRNA levels, respectively, than cells from implanted steers that were not exposed to alphaLA (P < 0.05). Administration of a Revalor-S implant increased circulating IGF-I and local muscle IGF-I mRNA concentrations in finishing cattle. However, muscle IGF-I mRNA levels were decreased by flax supplementation. Muscle cell culture experiments suggested that alphaLA was not responsible for the IGF-I mRNA down-regulation.     

Changes in metabolites, metabolic hormones, and luteinizing hormone before puberty in Angus, Braford, Charolais, and Simmental heifers

We determined changes in insulin, glucose, free fatty acids (FFA), growth hormone (GH), insulin-like growth factor I (IGF-I) and LH before puberty in Angus, Braford, Charolais, and Simmental heifers. Our primary objective was to identify metabolites and metabolic hormones that serve as metabolic cues for onset of puberty. Angus (n = 12). Braford (n = 7), Charolais (n = 9), and Simmental (n = 7) heifers were assigned at weaning (289 +/- 25 d of age; 264 +/- 23 kg) to open-sided pens with slotted floors, and they were fed a corn silage-concentrate diet formulated to provide gains of .91 kg/d. Puberty was defined as the 1st d (d 0) that serum progesterone (determined in blood samples collected at weekly intervals) exceeded 1 ng/ml. Blood samples were collected before and after feeding at 15-min intervals for 8 h at 21-d intervals before puberty in a subsample of heifers (at least five per breed). Angus and Simmental heifers weighed less and were younger (P less than .05) at puberty than Charolais and Braford heifers. Serum FFA before feeding and frequency of LH release increased (P less than .05) from d-40 +/- 3 to d-17 +/- 3 in all breeds. Conversely, concentrations of insulin were greater (P less than .05) at -40 than at -17 d from puberty in Angus, but not in Braford, Charolais, or Simmental heifers. Frequency of GH release was greater at d -40 than at d -17 in Angus heifers; however, in Braford and Charolais heifers frequency of GH release was greater at d -17 than at d -40. Concentrations of IGF-I (measured every 2 wk) increased linearly (P less than .07) from d -56 to 0 from puberty in Angus but not in other breeds. In conclusion, frequency of LH release and concentrations of FFA increased before puberty in all breeds; however, consistent changes in other metabolites and hormones were observed only in Angus heifers.     

Effect of exogenous estradiol on plasma concentrations of somatotropin, insulin-like growth factor-I, insulin-like growth factor binding protein activity, and metabolites in ovariectomized angus and braham cows

To determine the effect of breed and estradiol-17β on selected hormones and metabolites, ovariectomized (3 mo) Angus (n = 14) and Brahman (n = 12) cows were paired by age and body weight and randomly assigned as either nonimplanted controls (CON) or implanted with estradiol (E2) for 45 d. After Day 7 and through Day 42, plasma concentration of somatotropin was greater for E2 than CON cows (treatment X day, P < 0.05). During an intensive blood sampling on Day 36, E2 cows tended (P < 0.10) to have greater somatotropin pulse amplitudes than CON cows, but other parameters of somatotropin release were not affected (P > 0.10) by E2 treatment. The effect of breed was apparent on Day 36 as Brahman cows had greater (P < 0.05) somatotropin pulse amplitude, basal secretion, and mean concentration than Angus cows. Overall, plasma concentration of IGF-I was greater (P < 0.01) for E2 than CON cows (158.3 vs. 104.2 ng/ml) and was greater for Brahman than Angus cows (164.1 vs. 98.4 ng/ml). However, there was a trend (P < 0.10) for a treatment X breed X day interaction for IGF-I (i.e., the magnitude of increase in IGF-I concentration was greater in E2-Angus than E2-Brahman cows). After Day 7 and through Day 42, total plasma IGF binding protein (IGFBP) activity was greater (P < 0.01) for E2 than CON cows. Ligand blotting revealed at least five forms of IGFBP activity, and E2 cows had greater (P < 0.05) binding activity of IGFBP-3 and the 30- and 32-kDa IGFBP than CON cows. Brahman cows had greater (P < 0.05) IGFBP-3 and the 32-kDa IGFBP than Angus cows. After Day 14 and through Day 42, concentration of urea nitrogen (PUN) was greater (P < 0.001) for CON than E2 cows (treatment X day, P < 0.001). Brahman had greater (P < 0.01) PUN than Angus cows (16.6 vs. 14.2 mg/dl). Plasma concentration of glucose was greater (P < 0.01) for E2 than CON cows (78.9 vs. 76.4 mg/dl) but was not affected (P > 0.10) by breed. In summary, these data suggest that some, but not all, of the positive effects of estradiol on peripheral concentration of IGF-I and IGFBP activity can be attributed to increased somatotropin. Moreover, breed influenced basal and E2-induced secretion of somatotropin and IGF-I such that differences between Brahman and Angus cows in plasma IGF-I concentrations were abated within 3 wk of estradiol implantation. Thus, breed influences the metabolite and hormonal response of cattle to estrogenic implants.