The ontogeny of immunoreactive beta-endorphin and beta-lipotropin in the rat testis

We have investigated the ontogeny of immunoreactive beta-endorphin (i-beta E) in the testes of rats from 5 to 150 days of age. i-beta E was measured by RIA in acid extracts of decapsulated testes and characterized by gel filtration chromatography. Significant age-related differences in both the levels and type of i-beta E were observed. Total levels of i-beta E in the testes were very low and barely detectable from 5-20 days of age, but rose sharply in parallel with testes weights from 20-60 days of age; thereafter, no significant changes in i-beta E were found through 150 days of age. Concentrations of i-beta E, expressed in pmol/g testis, fell precipitously between days 5 and 10 and remained relatively constant from 10-150 days. Most of the i-beta E at 5 and 15 days chromatographed like authentic beta-endorphin. However, with the onset of puberty (30-35 days) and during sexual maturation, much of the total i-beta E chromatographed like its' precursor beta-lipotropin (beta LPH). Hypophysectomy decreased the weight and total i-beta E levels of the testes to the same extent without altering the concentrations of i-beta E or the chromatographic pattern of i-beta E. These results indicate that beta E-like and beta LPH-like peptides are present in the rat testis and that age-related changes in both the levels and type of i-beta E correlate with various structural and functional aspects of testicular development.     

Ventilatory control during exercise in calves with artificial hearts

To determine the role of cardiac reflexes in mediating exercise hyperpnea, we investigated ventilatory responses to treadmill exercise in seven calves with artificial hearts and seven controls. In both groups, the ventilatory responses were adequate for the metabolic demands of the exercise; this resulted in regulation of arterial PCO2 and pH despite the absence of cardiac output increase in the implanted group. In this group, there was a small but significant reduction of arterial PO2 by 4 +/- 3 Torr and a rise of blood lactate by 1.1 +/- 1 mmol/l. When cardiac output was experimentally increased in the implanted calves to a level commensurate with that spontaneously occurring in the control calves, ventilation was not affected. However, experimental reductions of cardiac output led to an immediate augmentation of exercise hyperpnea by 4.56 +/- 4.3 l/min and a further significant lactate increase of 1.2 +/- 1.22 mmol/l that was associated with a significant decrease in the exercise O2 consumption (0.32 +/- 0.13 l/min). These observations indicate that neither cardiac nor hemodynamic effects of increased cardiac output constitute an obligatory cause of exercise hyperpnea in the calf.     

Molecular and phylogenetic characterization of pyruvate and 2-ketoisovalerate ferredoxin oxidoreductases from Pyrococcus furiosus and pyruvate ferredoxin oxidoreductase from Thermotoga maritima.

Previous studies have shown that the hyperthermophilic archaeon Pyrococcus furiosus contains four distinct cytoplasmic 2-ketoacid oxidoreductases (ORs) which differ in their substrate specificities, while the hyperthermophilic bacterium Thermotoga maritima contains only one, pyruvate ferredoxin oxidoreductase (POR). These enzymes catalyze the synthesis of the acyl (or aryl) coenzyme A derivative in a thiamine PPi-dependent oxidative decarboxylation reaction with reduction of ferredoxin. We report here on the molecular analysis of the POR (por) and 2-ketoisovalerate ferredoxin oxidoreductase (vor) genes from P. furiosus and of the POR gene from T. maritima, all of which comprise four different subunits. The operon organization for P. furiosus POR and VOR was porG-vorDAB-porDAB, wherein the gamma subunit is shared by the two enzymes. The operon organization for T. maritima POR was porGDAB. The three enzymes were 46 to 53% identical at the amino acid level. Their delta subunits each contained two ferredoxin-type [4Fe-4S] cluster binding motifs (CXXCXXCXXXCP), while their beta subunits each contained four conserved cysteines in addition to a thiamine PPi-binding domain. Amino-terminal sequence comparisons show that POR, VOR, indolepyruvate OR, and 2-ketoglutarate OR of P. furiosus all belong to a phylogenetically homologous OR family. Moreover, the single-subunit pyruvate ORs from mesophilic and moderately thermophilic bacteria and from an amitochondriate eucaryote each contain four domains which are phylogenetically homologous to the four subunits of the hyperthermophilic ORs (27% sequence identity). Three of these subunits are also homologous to the dimeric POR from a mesophilic archaeon, Halobacterium halobium (21% identity). A model is proposed to account for the observed phenotypes based on genomic rearrangements of four ancestral OR subunits.     

Cyclin D1 transgene impedes lymphocyte maturation and collaborates in lymphomagenesis with the myc gene.

Cyclin D1 is the regulatory subunit of certain protein kinases thought to advance the G1 phase of the cell cycle. Deregulated cyclin D1 expression has been implicated in several human neoplasms, most consistently in centrocytic B lymphoma, where the cyclin D1 gene usually has been translocated to an immunoglobulin locus. To determine directly whether constitutive cyclin D1 expression is lymphomagenic, transgenic mice were generated having the cyclin D1 gene linked to an immunoglobulin enhancer. Despite abundant transgene expression, their lymphocytes were normal in cell cycle activity, size and mitogen responsiveness, but young transgenic animals contained fewer mature B- and T-cells. Although spontaneous tumours were infrequent, lymphomagenesis was much more rapid in mice that co-expressed the cyclin D1 transgene and a myc transgene than in mice expressing either transgene alone. Moreover, the spontaneous lymphomas of myc transgenic animals often ectopically expressed the endogenous cyclin D1 gene. These findings indicate that this G1 cyclin can modulate differentiation and collaborate with myc-like genes in oncogenesis.     

Ovarian superstimulatory response relative to follicular wave emergence in heifers

Two experiments were designed to evaluate the responsiveness of beef heifers to superstimulatory treatments administered during the first follicular wave. Heifers were examined daily (Experiment 1) or twice daily (Experiment 2) by ultrasonography to determine the status of follicular wave development and the day of initiation of superstimulatory treatment. Heifers in both experiments were superstimulated with a total dose of 10 ml Folltropin (equivalent to 200 mg of NIH-FSH-P1), divided into 10 equal intramuscular injections over 5 days. On the last day of treatment, heifers received 500 mug of cloprostenol after each injection of Folltropin to induce luteolysis. In the respective groups, superstimulatory treatments were initiated on Day -1, Day 0 (day of ovulation) or Day +1 for Experiment 1, and on Day -1, Day 0, Day +1 or Day +2 for Experiment 2. In Experiment 1, the number of ovulations in each ovary was assessed by ultrasonography and by counting the number of corpora lutea (CL) in each ovary at slaughter. The correlation between both techniques for assessing ovulatory response was high (r= 0.98; P< 0.0001), and there was no significant difference in the mean number of ovulations detected by ultrasound (5.7+/-1.1) versus the mean number of CL counted at slaughter (6.2+/-1.2). In Experiment 1, the mean (+/- SEM) number of CL counted at slaughter in heifers treated on Day -1 (9.4+/-3.8) and Day 0 (7.3+/-1.6) was higher (P< 0.05) than that of heifers treated on Day +1 (0.7+/-0.3). The mean number of follicles >/=7 mm in diameter on the last day of treatment was also higher (P<0.05) in the Day -1 group compared with the Day +1 group; the Day 0 group was intermediate. In Experiment 2, the mean number of ovulations was higher (P< 0.05) in the Day 0 group (18.4+/-3.4) than the Day -1 (9.5+/-2.3), Day +1 (6.7+/-2.2) or Day +2 (6.5+/-2.3) groups. Heifers in the Day -1, and Day 0 groups had more (P< 0.05) follicles >/=7 mm at the end of treatment compared with heifers in the Day +1 or the Day +2 group. The stated hypothesis was supported: exogenous FSH treatment initiated at the time of wave emergence, near the expected time of the endogenous wave-eliciting FSH surge, has a positive effect on the superstimulatory response. A higher superstimulatory response was elicited when treatments were initiated on the day of, or the day before, wave emergence compared with that of later treatments.     

Octopamine receptor subtypes and their modes of action

Octopamine receptor subclasses were first proposed to explain differences in the pharmacological profiles of a range of physiological responses to octopamine obtained in the extensor-tibiae neuromuscular preparation of the locust. Thus, OCTOPAMINE₁ receptors which inhibit an endogenous myogenic rhythm, increase intracellular calcium levels. Also OCTOPAMINE₂ receptors which modulate neuromuscular transmission in this preparation, increase the level of adenylate cyclase activity. The current status of this classification is reviewed by examining the pharmacology of responses to octopamine in a range of preparations. It is concluded that the distinction between OCTOPAMINE₁ and OCTOPAMINE₂ receptor types is still valid, but that OCTOPAMINE₂ receptors exhibit some tissue specific variations. Studies on a clonedDrosophila octopamine/tyramine (phenolamine) receptor are discussed and illustrate many of the difficulties presently encountered in making a definitive classification of octopamine receptors. These include the possibilities that single receptors may activate multiple second messenger systems and that different agonists may differentially couple the same receptor to different second messenger systems.