Dynamic changes in plasma concentrations of gonadotropins, inhibin, estradiol-17beta and progesterone in cows with ultrasound-guided follicular aspiration.

To elucidate the effects of ultrasound-guided transvaginal follicular aspiration, plasma concentrations of FSH, LH, inhibin, estradiol-17beta and progesterone, and folliculogenesis were examined in Holstein cows. Four clinically healthy cows with regular estrous cycles were scanned by ultrasound per rectum once a week for 9 weeks before the commencement of follicular aspiration. All visible follicles were divided into 3 categories based on their sizes (2 < or = small < 5 mm; 5 < or = medium < 10 mm, large > or = 10 mm). The follicular aspiration was started at random during the estrous cycle and conducted under epidural anesthesia induced with 5 ml of 2% lidocaine once a week for 6 weeks. The average number of total visible follicles > or = 2 mm in diameter at 7 days after aspiration (21.7 +/- 7.4, n = 24) was similar to that before starting aspiration (26.7 +/- 10.5, n = 36). Plasma inhibin and estradiol-17beta declined and fell to a trough on 1.5 days and returned to pre-aspiration values by 5 days after aspiration. Plasma concentrations of FSH increased and reached peak levels between 1 and 1.5 days after aspirations. Plasma concentrations of LH also increased and reached peak levels between 0.5 and 1.5 days after aspirations. Both plasma FSH and LH had returned to pre-aspiration levels by 5 days after aspirations. Plasma concentrations of progesterone did not change with the follicular aspiration. These results demonstrate that follicular aspiration decreases plasma concentrations of inhibin and estradiol-17beta, which in turn leads to a rise in plasma concentrations of FSH and LH. It is suggested that marked increases in plasma concentrations of FSH and LH after the aspiration stimulate the development and maturation of a new cohort of follicles within one week in cows.

Binding of [35S]GTPgammaS stimulated by (+)-pentazocine sigma receptor agonist, is abundant in the guinea pig spleen.

Here we measured sigma receptor agonist, [3H](+)-pentazocine binding and (+)-pentazocine-stimulated [35S]GTPgammaS binding throughout brain regions and peripheral organs of mice and guinea pigs to investigate the distribution of G protein-coupled sigma receptors. There was no significant correlationship between both distributions, in which the [3H](+)-pentazocine binding is highest in the liver of each species, while the [35S]GTP-gammaS binding is highest in the guinea pig spleen. The agonist-stimulated [35S]GTPgammaS binding in the spleen was also confirmed by in situ autoradiography using sections. Thus it is suggested that there are at least two subtypes, metabotropic and nonmetabotropic sigma receptors, and the former ones are abundant in the guinea pig spleen.

Dose-related opposite modulation by nociceptin/orphanin FQ of substance P nociception in the nociceptors and spinal cord.

We previously reported that the intraplantar (i.pl.) application of nociceptin/orphanin FQ (N/OFQ) at extremely low doses elicited a nociception through a substance P (SP) release from nociceptor endings. In the present study, the nociception induced by SP (and N/OFQ) was abolished by intrathecal (i.t.) injection of neurokinin(1) (SP receptor) antagonist, suggesting the involvement of the stimulation of nociceptive primary SP neuron and SP release into spinal synapses. On the other hand, similar low doses of N/OFQ (i.t.) exerted nociceptive responses, characterized by scratching, biting, and licking, and these responses were blocked by an neurokinin(1) antagonist (i.t.) or capsaicin pretreatment or in tachykinin 1 gene knockout mice (tac1(-/-) mice), suggesting that N/OFQ receptor (NOR) also exists on the spinal terminals of SP neurons. When wide ranges of N/OFQ doses were used, a typical bell-shaped dose-response relationship was observed in both peripheral and central nociception tests. Furthermore, N/OFQ (1 nmol) administered i.pl. blocked SP (i.pl.)-induced flexor responses, which were abolished by pertussis toxin pretreatment or in NOR gene knockout (NOR(-/-)) mice. On the other hand, N/OFQ administered i.t. blocked SP (i.t.)-induced scratching, biting, and licking in capsaicin-pretreated and tac1(-/-) mice, and this antinociception was abolished in NOR(-/-) mice. All these findings suggest that N/OFQ has biphasic actions depending on doses in the nociceptors and spinal synapses and has postsynaptic antinociceptive actions in spinal cord by modulating SP signaling.

Parallel stimulations of in vitro and in situ [35S]GTPgammaS binding by endomorphin 1 and DAMGO in mouse brains.

Metabotropic activities of endomorphin 1, a candidate for endogenous mu-opioid receptor ligands, were examined in comparison with the actions of [D-Ala2, N-Me-Phe4, Gly5ol]-enkephalin/DAMGO, a well-known synthetic mu-opioid agonist. Endomorphin 1 stimulated [35S]GTPgammaS binding to synaptic membranes from the mouse amygdala in a naloxone-reversible manner. DAMGO had the same effect in such preparations. In in situ [35S]GTP-gammaS binding experiments using brain sections, both endomorphin 1 and DAMGO similarly stimulated this binding in specific cellular locations throughout the brain regions. These findings strongly support the view that endomorphin 1 selectively acts on a mu-opioid receptor.

Characterization of nociceptin-stimulated in situ [35S]GTPgammaS binding in comparison with opioid agonist-stimulated ones in brain regions of the mice.

We studied the characterization of receptor-mediated G protein activity by nociceptin throughout brain regions, using in situ GTPgammaS binding autoradiography. Nociceptin-stimulated GTPgammaS binding was markedly observed in amygdala, hippocampal pyramidal cell layers, temporal and entorhinal cortex, infralimbic organ, anterior olfactory nucleus, and rostral part of thalamus. These nociceptin-stimulated activities were not affected by naloxone, naltrindol nor norbinaltorphimine which completely blocked mu-, delta- or kappa-opioid agonist-stimulated GTPgammaS binding, respectively. In addition, the distribution of nociceptin-stimulated activities throughout brain regions was found to be different from such opioid receptor-mediated ones.

Viability of frozen-thawed bovine embryos bisected in sucrose: A preliminary report.

A method for producing identical twin calves is described in which Day 7 frozen-thawed bovine embryos in 12.5% sucrose solution were bisected using a fine microsurgical blade. The resulting bisected embryos were transferred nonsurgically to the uterine horn ipsilateral to the corpus luteum of synchronous recipients (+/-1 d), two bisected embryos per recipient. The pregnancy rate when both halves remained in the same zona pellucida was 50% (5 10 ); the pregnancy rate was 1 5 for morulae and 4 5 for blastocysts. The pregnancy rate for unfrozen morulae bisected in PBS and transferred without zona pellucida was 27% (4 15 ). The in vitro survival rate of embryos bisected in 12.5% sucrose when both halves remained in the original zona pellucida was 82% (18 22 ), which was higher than when embryos were bisected in PBS (53%, 9 17 ).

Production of indirect hemolysin by Yersinia enterocolitica and its properties.

Hemolysis was demonstrated on blood agar by adding lecithin from strains of biovar 1 Yersinia enterocolitica. Hemolytic activity was also observed in culture filtrates containing lecithin. No hemolytic activity was detected from any blood agar or culture filtrate without lecithin or after incubation at 37 degrees C. Crude indirect hemolysin was prepared by ammonium sulfate fractionation from culture filtrates, and two enzymes, phospholipase A and lipase, were fractionated by gel filtration. Hemolytic substances were recognized as lysolecithin and fatty acid when lecithin wad decomposed by phospholipase A and lipase, respectively.