Increasing Concentrations of 17β-Estradiol Has Differential Effects on Secretion of Luteinizing Hormone and Follicle-Stimulating Hormone and Amounts of mRNA for Gonadotropin Subunits during the Follicular Phase of the Bovine Estrous Cycle

The hypothesis tested was that 17β-estradiol (E2) would increase amounts of mRNA for α, LHβ, and FSHβ subunits during the follicular phase of the estrous cycle prior to the preovulatory surge of gonadotropins in cows. On Day 16 (Day 0 = estrus) of the estrous cycle, all cows were treated with prostaglandin F2α (PGF2α). Cows served as intact controls (CONT, n = 4) were ovariectomized (OVX, n = 5), or were ovariectomized and administered E2 (OVXE, n = 6) in increasing doses starting at the time of treatment with PGF2α. Cows were bled for 6 h before and for 40 h after PGF2α treatment to characterize pulsatile secretion of LH and FSH. Forty hours after PGF2α treatment, pituitaries were collected for evaluation of amounts of mRNA for α, LHβ, and FSHβ subunits. Amplitude of LH pulses was greater (p \u3c 0.05) in cows from the OVXE than from the CONT group. Concentrations of FSH were greater in cows from both the OVXE and OVX (p \u3c 0.01) groups than from the CONT group. Amounts of mRNA for α and FSHβ subunits were greater (p \u3c 0.01) in pituitaries of cows from the OVX than from the CONT or OVXE groups. Amounts of mRNA for LHP subunit in pituitaries of cows from the OVX group tended to be greater (p \u3c 0.08) than from the CONT group. Cows in the OVXE group tended (p \u3c 0.08) to have greater amounts of mRNA for FSHβ subunit than did CONT cows. Amounts of mRNA for α and LHβ subunits in cows from the OVXE and CONT groups did not differ (p \u3e 0.10). Pituitary weight and content of LH and FSH were not different (p \u3e 0.10) among cows of the different groups. Ovariectomy resulted in enhanced secretion of gonadotropins and increased amounts of mRNA for gonadotropin subunits above values detected in CONT cows. Ovariectomized cows administered E2, in follicular-phase patterns had amounts of mRNA for a and LHβ similar to those in CONT cows even though secretion of LH was enhanced in the OVXE group. We reject our hypothesis and conclude that E2 has a divergent role in regulation of gonadotropins. Release of LH is enhanced by E2, but E2 reduced mRNA for gonadotropin subunits (tended to reduce mRNA for FSHβ subunit) in ovariectomized cows to amounts detected in intact cows during the follicular phase of the bovine estrous cycle

Effects of 17β-Estradiol on Distribution of Pituitary Isoforms of Luteinizing Hormone and Follicle-Stimulating Hormone during the Follicular Phase of the Bovine Estrous Cycle

The objective of this study was to examine the influence of 17β-estradiol (E2) on distribution of LH and FSH isoforms during the follicular phase of the bovine estrous cycle prior to the preovulatory surges of LH and FSH. On Day 16 of the estrous cycle (Day 0 = estrus), intact controls (CONT; n = 4) were treated with prostaglandin F2α (PGF2α) to induce luteal regression and initiation of the follicular phase. Other cows were also treated with PGF2α and either ovariectomized (OVX; n = 5) or ovariectomized and given E2 implants (OVXE; n = 6) to mimic the pattern of increasing E2 concentrations during the follicular phase of the estrous cycle. Pituitaries were collected 40 h after treatment with PGF2α, or ovariectomy (0 h). Aliquots of pituitary extracts were chromatofocused on pH 10.5-4.0 gradients. The LH resolved into thirteen isoforms (designated A-L and S, beginning with the most basic form) while FSH resolved into nine isoforms (designated I-IX, beginning with the most basic form). The percentage of LH as isoform F (elution pH = 9.32 + 0.01) was greater (p \u3c 0.05) in the OVX group (48.5%) than in the OVXE group (45.0% ). LH isoforms I (elution pH = 6.98 ± 0.01) and J (elution pH = 6.48 ± 0.01) were more abundant (p \u3c 0.05) in cows from the OVXE (2.3 and 5.8%, respectively) than the OVX group (1.4 and 3.7%, respectively). Distribution of LH isoforms in cows from the three groups did not differ (p \u3e 0.10). Distribution of FSH isoforms were similar (p \u3e 0.05) among all groups. In summary, removal of the ovary (OVX) resulted in a slight increase in percentage of the basic LH isoform F, while removal of the ovary and administration of E2 (OVXE) in a pattern that mimicked increasing concentrations of E2 during the follicular phase of the estrous cycle resulted in a slight increase in the percentage of acidic LH isoforms (I and J). There was no influence of ovariectomy or treatment with E2 on distribution of FSH isoforms in the pituitary. Thus, gonadotropin heterogeneity does not appear to change significantly during the follicular phase of the bovine estrous cycle

Differential Regulation of Gonadotropin Synthesis and Release in Ovariectomized Ewes after Treatment with a Luteinizing Hormone-Releasing Hormone Antagonist

Our working hypothesis was that synthesis and release of LH, but not FSH, were solely dependent on LHRH. Twenty ovariectomized (OVX) ewes were randomly assigned to one of five treatments (n = 4 per group). Ewes were administered a low (10 μg/kg) or high (100 μg/kg) dose of LHRH antagonist (LHRH-Ant) at 24-h intervals for 3 or 6 days. Control ewes received vehicle (5% mannitol) at 24-h intervals for 6 days. Blood samples were collected every 15 min for 4 h before LHRH-Ant or vehicle and every 2 h during the period of treatment to determine concentrations of LH and FSH. Twenty-four hours after the last treatment with LHRH-Ant or vehicle, anterior pituitaries were collected and divided in half along the midsagittal plane; the number of receptors for LHRH, pituitary content of LH and FSH, and relative amounts of mRNA for α, LHβ, and FSHβ subunits were determined. Concentrations of LH in serum decreased (p \u3c 0.05) from 25.4 ± 4.3 ng/ml before LHRH-Ant to less than 0.5 ng/ml within 4 h after the first treatment of LHRH-Ant and remained low (\u3c 0.5 ng/ml) throughout the study. Serum concentrations of FSH declined gradually during the 3- or 6-day period of treatment with LHRH-Ant, from 37.3 ± 2.4 and 26.5 ± 4.8 ng/ml to 19.9 ± 1.8 and 13.7 ± 2.1 ng/ml, respectively. The magnitude of decline in serum concentrations of LH and FSH did not differ among ewes treated with low or high doses of LHRH-Ant. Pituitary content of LH was not different (p \u3e 0.10) from that in controls, whereas pituitary content of FSH was greater (p \u3c 0.01) in control ewes compared to ewes treated with LHRH-Ant. Receptors for LHRH were nondetectable (\u3c 0.018 x 10-16 mol receptor/μg protein) in pituitaries after 3 or 6 days of treatment with LHRH-Ant (low or high dose). Relative amounts of mRNA for α, LHβ, and FSHβ subunits were lower (p \u3c 0.01) after 6 days of treatment with LHRH-Ant (low or high dose) than after 3 days of treatment with LHRH-Ant (low or high dose). The LHRH was, therefore, required to maintain steady state amounts of mRNA for FSH and LH and to maintain pituitary stores of FSH but not LH. Our data support the hypothesis that differential regulation of LH and FSH release occurs in ewes. While synthesis and release of LH are dependent on LHRH, synthesis but not release of FSH appears to be dependent on LHRH

Twin and family studies reveal strong environmental and weaker genetic cues explaining heritability of eosinophilic esophagitis

Eosinophilic esophagitis (EoE) is a chronic antigen-driven allergic inflammatory disease, likely involving the interplay of genetic and environmental factors, yet their respective contributions to heritability are unknown

Effects of transportation, transport medium and re-housing on Xenopus laevis (Daudin)

Understanding the immediate and longer-term effects of transportation and re-housing in a laboratory species is crucial in order to refine the transfer process, enable the optimal introduction of new animals to a novel environment and to provide a sufficient acclimatisation period before usage. Whilst consideration of animal welfare in most model vertebrate species has received attention, little quantitative evidence exists for the optimal care of the common laboratory amphibian Xenopus laevis. Techniques for the non-invasive welfare assessment of amphibians are also limited and here a non-invasive physiological assay was developed to investigate the impacts of transportation, transport medium and re-housing on X. laevis. First the impacts of transportation and transport medium (water, damp sponge or damp sphagnum moss) were investigated. Transportation caused an increase in waterborne corticosterone regardless of transport medium. Frogs transported in damp sphagnum moss also had a greater decrease in body mass in comparison to frogs not transported, suggesting that this is the least suitable transport medium for X. laevis. Next the prolonged impacts of transportation and re-housing were investigated. Frogs were transported between research facilities with different housing protocols. Samples were collected prior to and immediately following transportation, as well as 1 day, 7 days and 35 days after re-housing. Water-borne corticosterone increased following transportation and remained high for at least 7 days, decreasing to baseline levels by 35 days. Body mass decreased following transportation and remained lower than baseline levels across the entire 35 day observation period. These findings suggest the process of transportation and re-housing is stressful in this species. Together these findings have important relevance for both improving animal welfare and ensuring optimal and efficient scientific research

Bemerkungen über die häusliche Erziehung des Schülers

vom Collegen Dr. GrotjanIn Fraktu

Effect of immunization against LHRH on isoforms of LH in the ovine pituitary

The role of LHRH in modulating intrapituitary LH content as well as the distribution of LH among its isoforms was examined in sheep. Rams (n = 3) and wethers (n = 6) were actively immunized against an LHRH-humanserum globulin conjugate. Pituitaries collected from these animals plus pituitaries from corresponding numbers of nonimmunized rams and wethers were extracted with a buffered saline solution containing protease inhibitors. Immunization markedly reduced total amounts of immunoreactive LH in the pituitary. An aliquot of each pituitary extract was desalted by flow dialysis against water and chromatofocused on a pH 10.5-7.0 gradient. Concentrations of LH in chromatofocusing fractions were determined by radioimmunoassay. LH in pituitary extracts resolved into nine peaks during chromatofocusing which were coded with letters beginning with the most basic isoform. The percentage of LH as the two most basic isoforms, A\u27 and B, was similar (P \u3e 0.05) in all treatment groups. Isoform H constituted a higher percentage (P \u3c 0.05) of the LH in both castrate groups. Nonimmunized wethers had higher percentages of isoforms C, D and E (P \u3c 0.05) and lower percentages (P \u3c 0.05) of the acidic isoforms (coded as peak Z herein) than did other treatment groups. Thus, castration shifted the pattern of intrapituitary isohormones towards the more basic forms. Nonimmunized rams had a higher percentage (P \u3c 0.05) of isoform G than did other groups. Isoform F, the most abundant isoform, was present as a higher percentage (P \u3c 0.05) in immunized rams and wethers than in nonimmunized animals. Hence, ablation of hypothalamic LHRH reaching the pituitary by active immunization not only markedly reduced the quantity of LH in the pituitary, but also altered the distribution of LH among its isoforms yielding a higher percentage of the most abundant isoform F. Hypothalamic LHRH therefore not only increases the quantity of LH in the pituitary but also alters the pattern of intrapituitary isohormones by reducing the percentage present as isoform F. Furthermore, inputs from both the hypothalamus and gonads appear to regulate the distribution of intrapituitary isohormones with hypothalamic influences predominating