Jugular Plasma Research Articles

Many peptides that are present in the external zone of the median eminence are not secreted into the hypophysial portal blood of sheep.

Apart from the well recognized factors that are produced by the hypothalamus and secreted into hypophysial portal blood to regulate pituitary function, there is a range of neuropeptides that are present in the median eminence and could be secreted to serve a modulatory function. In this study we have collected hypophysial portal blood and jugular venous blood from sheep in an attempt to identify which of these putative modulatory peptides might be secreted from the median eminence. We have measured neuropeptide Y (NPY), substance P (SP), galanin (GAL), neurokinin A (NKA), peptide histidine isoleucine (PHI), vasoactive intestinal peptide (VIP), neurotensin (NT) and cholecystokinin (CCK). We also examined the sheep median eminence using immunohistochemistry for NPY, SP and GAL and determined degradation profiles of NPY, SP, GAL and NKA in portal and jugular plasma. In no instance did we find that levels of the above peptides were consistently higher in portal blood than in peripheral blood. In some cases levels of peptide were lower in portal plasma e.g. for NPY (6/10 sheep). In one experimental series SP levels in portal plasma were significantly (p < 0.05) lower than levels in jugular plasma but this was not found in another experimental series. Galanin levels were significantly (p < 0.01) lower in portal plasma compared to levels in jugular plasma. We conducted in vitro studies to determine whether or not the above peptides are selectively degraded in portal blood but were unable to show any differences between the rates of degradation in portal and jugular plasma. Immunohistochemistry revealed projections into the external zone of the median eminence for NPY, GAL and SP. This study shows that none of the above peptides are secreted into the hypophysial portal blood of sheep. For some peptides e.g. GAL, enzymes from the endothelial cells of the portal vessels may enhance degradation. Projections into the external zone of the median eminence of neuronal systems containing these peptides may serve to modulate the secretion of the well recognized release and inhibiting factors by acting on the neurosecretory terminals.

Plasma concentrations of 13,14-dihydro- 15-ketoprostaglandin F2α in mares during uterine involution

Summary Twelve mares were allowed to foal naturally, after which they were monitored to study uterine involution. Starting on day 3 after parturition, the internal genital tract was examined per rectum manually and ultrasonographically every other day for changes in uterine characteristics and ovarian activity. By day 5, gravid and nongravid uterine horns were similar in size, and by day 7, uterine fluid was absent. On day 7 after parturition, endometrial biopsy samples were obtained for histologic evaluation, and uterine swab specimens were obtained for microbiologic culture. Uterine swab specimens from 10 of 12 mares had slight bacterial growth. The uteri of 8 of the 12 mares were histologically involuted by day 7. All mares ovulated 7 to 12 days after parturition. Concentrations of 13,14-dihydro-15-keto-PGF2α(pgfm) were measured in jugular plasma samples obtained daily for 21 days after parturition. Concentrations of pgfm were low by the day after parturition, and there was no significant correlation between uterine involution and pgfm concentrations in these mares’. All 12 mares were bred at the first estrus after parturition, and 9 became pregnant.

Jugular blood plasma acetylcholinesterase (AchE) activity during oestrus in dairy cattle

263 Ramachandra, S.G., Prakash, N. and Narayana. K., 1992. Jugular blood plasma acetylcholinesterase (AchE) activity during oestrus in dairy cattle. Anim. Reprod. Sci .• 27: 263-267. Blood plasma acetylcholinesterase (AchE) activity was monitored in blood samples collected every 2 h before and after prostaglandin-induced oestrus in dairy cows. A maximum AchE activity of 138 ± 31 enzyme units mlI was found at 2 h after the onset of oestrus and there were peak activities of 112± \3 enzyme units ml' and 112± 7 enzyme units ml' at 8 and 24 h. respectively. There was a follicle stimulating hormone (FSH) surge at 8 h after the onset of oestrus. Preceding the initial peak AchE activity. there was a peak oestradiol concentration.

Effect of prostaglandin F2 α ( PGF2 α) on sources of progesterone and pregnancy in intact, ovariectomized and hysterectomized 90–100 day pregnant ewes

A single dose of 8 or 16 mg of PGF 2 α per 58 kg body weight was injected intramuscular into intact, ovariectomized or hysterectomized 90–100 day pregnant sheep in three separate experiments. Both doses of PGF 2 α decreased the weights of the corpora lutea ( P≤0.05) and the concentration of progesterone in ovarian venous plasma at 72 hr ( P≤0.05) compared to the 0 hr sample within treatment groups and to control ewes at 72 hr in intact and hysterectomized pregnant ewes. In hysterectomized pregnant ewes, progesterone in jugular plasma declined ( P≤0.05) from 0 to 72 hr but never fell below 4 mg/ml and this decrease in progesterone after 8 or 16 mg PGF 2 α was greater than in control hysterectomized ewes ( P≤0.05). There was a significant decrease in progesterone over time in jugular or uterine venous plasma in the presence of absence of the ovaries in 90–100 day pregnant ewes ( P≤0.05) but the profiles of progesterone were not different between vehicle and PGF 2 α -treated ewes ( P≥0.05). Uterine venous progesterone never declined below 30 ng/ml in the presence or absence of the ovaries and there was a significant quadratic increase ( P≤0.05) in uterine venous progesterone toward the end of the 72 hr sampling period indicating an increase in steroidogenic activity of the placenta. PGF 2 α did not affect the number of abortions in intact or ovariectomized pregnant ewes ( P>0.05). Thus, the corpus luteum of sheep at 90–100 days of pregnancy is functional and responsive to PGF 2 α , placentomes are functional but do not appear to be responsive to the doses of PGF 2 α tested and PGF 2 α was not an abortifacient over the 72 hr treatment period.

Distribution and characterization of peptidylglycine alpha-amidating monooxygenase activity in the ovine brain and hypothalamo-pituitary axis.

The production of alpha-amidated peptide hormones from their glycine-extended precursors is catalyzed by the specific enzyme peptidylglycine alpha-amidating monooxygenase (PAM). In the present study, the distribution and subcellular localization of PAM activity in the sheep brain was examined and compared with known sites of amidated peptide synthesis and release. Of the brain regions assayed, the preoptic anterior and medial basal areas of the hypothalamus contained the greatest concentration of amidating activity. Lower concentrations (greater than 3-fold less) were found in the anterior and neurointermediate pituitary, median eminence, cerebral cortex, hippocampus, pons-medulla, and brainstem. Very low amounts of activity were present in the cerebellum and pineal gland. In most tissues tested, PAM activity was 40-75% higher in the membrane-associated fraction than in the soluble fraction. In the hypothalamus, affinity constants were identical for both membrane-associated and soluble fractions, and ranged from 12.3-13.3 microM. Maximal velocity was higher in the membrane fraction (4.7-4.8 pmol/microgram.h) than in the soluble fraction (2.6-2.9 pmol/microgram/h). Levels of amidating activity in hypophysial-portal and jugular plasma were similar and were 20- to 25-fold lower than in hypothalamic extracts. Insulin-induced hypoglycemia did not significantly alter PAM levels in portal or peripheral plasma, suggesting that amidating activity is not released during this stress. These results indicate that the hypothalamus is the richest source of amidating activity in the sheep brain, and suggest that amidation of neurohypophysial and hypothalamic releasing peptides may occur before axonal transport, given the much lower levels in median eminence, neurointermediate pituitary, and portal plasma.

Increased angiotensin-(1-7) in hypophysial-portal plasma of conscious sheep.

Studies were undertaken to characterize angiotensin peptides in hypophysial-portal blood of conscious sheep and to determine whether the median eminence (ME) secretes angiotensin peptides into the hypophysial-portal circulation. Simultaneous measurements of angiotensin peptides in jugular and hypophysial-portal plasma were performed in 6 sheep. Cerebrospinal fluid (CSF) was collected and data for hypophysial-portal plasma were corrected for CSF contamination. Angiotensin peptides were also measured in extracts of sheep ME. In a separate group of 4 sheep, simultaneous measurements of angiotensin peptides in arterial and jugular plasma were performed. Using high performance liquid chromatography-based radioimmunoassays, 8 angiotensin peptides were measured: Ang-(1-7), Ang II, Ang-(1-9), Ang I, Ang-(2-7), Ang III, Ang-(2-9), and Ang-(2-10). Renin, angiotensinogen and prolyl endopeptidase were also measured. No differences in angiotensin peptide levels in arterial and jugular plasma were observed. Angiotensin peptide levels in hypophysial-portal plasma were similar to those in jugular plasma, except for Ang-(1-7), the levels of which were 5-fold higher in hypophysial-portal plasma, and Ang I, for which the levels in hypophysial-portal plasma were 46% of the jugular levels. Renin and angiotensinogen levels were similar in arterial, jugular, and hypophysial-portal plasma. Angiotensin peptide contents of sheep ME were less than 16 fmol/ME. However, the prolyl endopeptidase content of sheep ME was 430-fold higher than plasma levels. The low levels of angiotensin peptides in sheep ME indicate that it does not secrete these peptides into the hypophysial-portal circulation. Rather, the high level of prolyl endopeptidase in ME is consistent with region-specific metabolism of Ang I delivered to the ME by arterial blood, generating increased levels of Ang-(1-7) in hypophysial portal plasma. The increased levels of Ang-(1-7) in hypophysial-portal plasma may play a role in regulation of pituitary function.

A potentially new metabolic pathway: ethyl esterification of acitretin.

1. The acidic retinoid, acitretin, was esterified to etretinate (ethyl ester) by rat and human liver 12,000 g supernatant. The amount of etretinate formed was increased by adding ethanol to the rat preparation. 2. This esterification almost certainly involves enzymic catalysis, and the amounts of etretinate formed were increased by the use of fresh rat liver. 3. Co-administration of acitretin and ethanol to rats resulted in a maximum plasma concentration of etretinate at approximately 1 h after dosing. Secondary maxima were induced by administering ethanol alone at 5 and 8 h after dosing with acitretin. 4. Comparison of acitretin and etretinate concentrations in rat portal and jugular vein plasma after ethanol administration indicated that the ester was formed mainly systematically, rather than during absorption. 5. The results of our study in the rat could indicate that the presence of etretinate in plasma of some patients being treated with acitretin may result from the intake of alcohol.

The form and function of the Leydig cells in hypophysectomized rams treated with pituitary extract when spermatogenesis is disrupted by heating the testes

The morphology and in-vivo function of the Leydig cells were studied in rams when spermatogenesis had been disrupted by a single exposure of the testes 20 days earlier to a temperature of about 42 degrees C for 45 min. To avoid complications due to changed negative feedback from the testes to the pituitary with consequent changes in the degree of gonadotrophic stimulation, ten of the animals (five heated and five unheated) were surgically hypophysectomized when the testes were heated and then treated twice daily with pituitary extract. Six intact rams (three heated and three unheated) were also studied. The heat-affected testes were about half the size of the unheated testes, and blood plasma flow was closely related to testis weight. There were no differences in the testosterone concentrations in spermatic venous blood, testicular lymph or rete testis fluid, or in oestradiol in spermatic venous plasma from heated or unheated testes. Consequently, testosterone secretion by the heat-affected testes was markedly reduced, and the concentrations in jugular blood were also lower in the heat-affected rams than in controls. The volume of the interstitial tissue was less in absolute terms in the heat-affected rams, but it made up a greater fraction of the testes. The absolute volume of the blood plus lymph vessels, and their fraction of the interstitial tissue were lower in the heat-affected testes, although there was no effect on their volume as a fraction of the whole testis. The heat-affected testes of the hormone-treated rams had fewer Leydig cells, but each cell was larger; no equivalent difference was found in the intact rams. However, the dose of pituitary extract chosen was somewhat excessive, as there were higher than normal concentrations of FSH, LH and testosterone in jugular blood plasma, of testosterone and oestradiol in testicular venous blood plasma and of testosterone in rete testis fluid in the hormone-treated hypophysectomized rams. The testes of the unheated hypophysectomized rams increased in size by about 20% during treatment with pituitary extract, although testicular blood plasma flow was lower per unit weight of testis. The absolute volume of each Leydig cell and the total volume in absolute terms and as a fraction of the interstitial tissue was greater in the hormone-treated than in the untreated rams, but not the volume as a fraction of the whole testis. The total number of Leydig cells was higher in the hormone-treated unheated rams than in all the other rams taken together.(ABSTRACT TRUNCATED AT 400 WORDS)

Catheterization of the caudal vena cava via the lateral saphenous vein in the ewe, cow, and gilt: an alternative to utero-ovarian and medial coccygeal vein catheters2

Current methods for obtaining venous blood from the reproductive organs of livestock often have a low rate of success or involve intensive surgical procedures that may impair ovarian function. Therefore, the caudal vena cava was catheterized via the lateral saphenous vein to determine the feasibility of using this method for chronic sampling of blood draining from the reproductive organs of ewes (n = 6), cows (n = 6), and gilts (n = 7). Blood samples were collected at 2-cm (ewes and gilts) or 5-cm (cows) intervals during insertion of catheters. Correct placement, defined as the position at which plasma concentrations of progesterone or estrogen were at least threefold greater than in jugular venous plasma, varied among species and among animals within species. It seemed, however, that a majority of catheters would be placed correctly if secured at 48 to 52 cm in ewes, 52 cm in gilts, and 90 to 100 cm in cows. Saphenous vein catheters were secured for sequential sampling of vena caval blood during the follicular phase of ewes (n = 25), cows (n = 4), and gilts (n = 5). Catheters remained patent for the duration of sampling in all individuals. Concentrations of estrogen in jugular and vena caval plasma were correlated (ewe P less than .0003; cow P less than .0001; gilt P less than .0001). Profiles of progesterone and estrogen revealed an episodic pattern of secretion in vena caval but not jugular plasma. Catheterization of the vena cava via the saphenous vein is a relatively simple and noninvasive method for obtaining blood containing uterine and ovarian hormones before their metabolism.

Duodenal Rapeseed Oil Infusion in Early and Midlactation Cows. 3. Plasma Hormones and Mammary Apparent Uptake of Metabolites

Rapeseed oil was infused continuously (1.0 to 1.1 kg/d) into the duodenum of Holstein × Friesian multiparous cows during the first 3 wk of lactation (oil treatment, 6 cows versus 6 controls, early lactation trial) or after 100 d of lactation (midlactation trial, 9 cows in a crossover design).In the midlactation trial, plasma glucose, 3-hydroxybutyrate, and free glycerol were not affected by oil infusion. Postprandial plasma NEFA were higher in oil-infused than in control cows; plasma triglycerides, phospholipids, and cholesterol were significantly increased in oil treatment. Plasma insulin was lower and somatotropin higher in oil-infused cows, whereas insulin-like growth factor-I and triiodothyronine were not affected.During the early lactation trial, there were few significant effects of oil infusion on measured plasma metabolites and hormones. Preprandial glucose and NEFA were lower in oil treatment during wk 2, preprandial phospholipids were higher in wk 1 and 3, and free cholesterol was higher in wk 1.Responses of plasma glucose and NEFA to insulin challenge were not clearly affected by oil treatment during either trial. Differences in plasma triglycerides between jugular and mammary veins were higher in oil treatment in both trials. Oil infusion did not affect jugular-mammary differences of other metabolites except for a slight increase in cholesterol esters in midlactation. The relationships between jugular-mammary differences and jugular plasma concentrations showed that up to 24, 27, 54, 17% of jugular glucose, NEFA, triglycerides, and 3-hydroxybutyrate apparently were taken up by the mammary gland.

DEVELOPMENT OF A TWO-SITE IMMUNORADIOMETRIC ASSAY FOR DIMERIC INHIBIN USING ANTIBODIES AGAINST CHEMICALLY SYNTHESIZED FRAGMENTS OF THE α AND β SUBUNIT

A two-site (liquid-phase) immunoradiometric assay (IRMA) for dimeric inhibin has been developed using antibodies raised against synthetic peptide sequences corresponding to the N-terminus (1-32) of the alpha subunit and the C-terminal region (82-114) of the beta A subunit of Mr approximately 30,000 human inhibin. Highly-purified Mr 32,000 bovine inhibin (standard) gave a dilution curve parallel to those for bovine follicular fluid (bFF), human (h)FF and rat ovary extract. Whilst the assay detected both Mr 56,000 and 32,000 inhibin forms in bFF, little reaction with higher Mr forms was evident. Cross-reaction of 'free' inhibin subunit (Mr 25,000 form) and recombinant human activin A in the IRMA were minimal (less than 0.1 and less than 2% respectively). Although the detection limit of the IRMA (approximately 50 pg/tube) was similar to that of several reported radioimmunoassays (RIA), the IRMA was unable to detect dimeric inhibin in jugular or utero-ovarian vein plasma of heifers. Similarly, when assayed by IRMA, bFF, hFF and rat ovary extract contained 8-58 times less inhibin than was indicated by RIA. These observations are consistent with earlier evidence that the ovary secretes a substantial excess of 'free' inhibin alpha subunit and that this material reaches the peripheral circulation. Surprisingly, however, the inhibin contents of bFF, hFF and rat ovary extract determined by in vitro bioassay were 8-23 times greater than the corresponding IRMA values, being similar to those derived by RIA.(ABSTRACT TRUNCATED AT 250 WORDS)

Utilization of Medium-Chain Triglycerides by Neonatal Piglets: Chain Length of Even- and Odd-Carbon Fatty Acids and Apparent Digestion/Absorption and Hepatic Metabolism

Jugular plasma concentrations of medium-chain fatty acids (MCFA) and whole blood concentrations of D-beta-hydroxybutyrate (BHBA) were measured in 1-d-old pigs at 0, 1, 2, 4 and 8 h after forcefeeding 12 mL of one of four triglycerides: tri-7:0, 8:0, 9:0 or 10:0. Both BHBA and MCFA were highest at 1 h. The MCFA measured at 1 h decreased (P less than 0.01) with increasing chain length. The BHBA was not different in pigs given tri-7:0, 8:0 or 9:0 but was lower (P less than 0.05) for pigs given tri-10:0. Isolated hepatocytes converted [1-14C] C7 or C9 to CO2 and acid soluble products more than 40% faster than cells given 8:0 or 10:0 and consumed 7% more O2. Even- and odd-MCFA were oxidized faster (P less than 0.01) than 18:1 (n-9). Theoretical calculations from fatty acid oxidation accounted for 96 to 142% of measured O2 consumption for the various fatty acids. In all instances, L-carnitine had no effect. Appearance of 14C in lipid products increased progressively with chain length from 7:0 to 18:1 (n-9). Collectively, these data suggest that changes in chain length within the medium-chain family may dramatically influence the rate and extent of digestion and/or absorption and metabolism of medium-chain triglycerides by neonates. This may be a consequence of increased hydrophobicity with increasing chain length or, for odd-chain fatty acids, a reflection of anaplerotic carbon effects from propionyl-CoA metabolism.

Changes in Rat Luteal Ultrastructure and P45Oscc mRNA and Protein Content after in Vivo Treatment with a Gonadotropin-Releasing Hormone Agonist1

Previous studies have demonstrated that plasma progesterone levels decrease in pregnant rats treated in vivo with a gonadotropin-releasing hormone agonist (GnRH-Ag), without changes in testosterone or estradiol levels in ovarian vein plasma. The objective of this study was to determine the loci of GnRH-Ag disruption of progesterone synthesis by examining luteal mitochondria, lipid droplets, cellular composition, and P450 side-chain cleavage (P450scc) enzyme and mRNA content in the pregnant rat. On Day 7 or 11 of pregnancy, osmotic minipumps containing GnRH-Ag were implanted into 5-7 rats. Sham operations were performed on 5-6 controls at each time period. Five micrograms per day of GnRH-Ag were released for about 24 h, after which corpora lutea and jugular vein plasma were collected. The corpora lutea were prepared for microscopy or analyzed for P450scc enzyme and mRNA content. Plasma progesterone levels were measured by RIA. In those rats treated with GnRH-Ag, progesterone levels had decreased, and within the luteal cells, there was an increase in the number of lipid droplets and a decrease in the number of tubular cristae within the mitochondria. Concomitantly, P450scc enzyme and mRNA content decreased on both Day 8 and Day 12 of pregnancy. Also, GnRH-Ag treatment decreased the ratio of large to small steroidogenic luteal cells on Day 8 of pregnancy, but did not alter cellular ratios on Day 12 of pregnancy. These observations suggest that treatment with GnRH-Ag inhibits progesterone synthesis by decreasing the amount of P450scc mRNA and enzyme content, which may alter the mitochondrial cristae structure on Day 8 and Day 12 of pregnancy. The reduction in tubular cristae and P450scc enzyme in the mitochondria may account for the increase in lipid droplets, as less cholesterol is converted to pregnenolone. An additional mechanism of inhibition may be the reduction in the number of large steroidogenic luteal cells, which appear to be the major source of progesterone in the rat corpus luteum on Day 8 of pregnancy.

Vulvar electrical impedance in periparturient cows and its relation to plasma progesterone, oestradiol-17β and PGFM

In previous studies we described the vulvar electrical impedance (VEI) - which refers to tissue hydration - during the oestrous cycle of cows. The objectives of the present study were to: (1) describe VEI changes around the time of calving; and (2) determine the relationship between periparturient changes in VEI and jugular plasma concentrations of oestradiol-17β (E 2), progesterone (P 4), and 13, 14-dihydro-15-keto PGE 2α (PGFM). Cylindrical bipolar electrodes were implanted inside the vulvar labiae of seven Israeli-Friesian cows. From day 245 of gestation through 15 days postpartum, VEI was measured, once daily in the morning, by means of a Hewlett-Packard multifrequency impedance analyser. Jugular blood samples for hormone determination were collected immediately after each VEI measurement. VEI decreased from approximately 150 Ω 27 days before parturition (day of calving = day 0) to 120 Ω on day − 10. The % prepartal VEI decrease was greater at current frequencies < 10 kHz than at > 10 kHz. This indicated that the decrease was caused mainly by an increase in the extracellular compartment. During that period P 4 decreased from ∼ I 1 ng/ml to 6 ng/ml and E 2 increased from 50 pg/ml to 150 pg/ml. VEI remained low until parturition, while P 4 continued to decrease to < 1 ng/ml and E 2 continued to increase to ∼ 600 pg/ml at calving. There was no significant prepartal relationship between PGFM and VEI. VEI increased postpartum from 120 Ω on day 1 postpartum to 155 Ω on day 11 and remained high thereafter. A negative linear relationship between PGFM and VEI was found during the postpartum period. The prepartal decrease of VEI occurred when the E 2:P 4 (pg/ml:ng/ml) ratio was ≥ 10:1. Measuring VEI may become a tool for (1) studying the dynamics of hormone action on the hydration of target tissues; (2) determining impending calving; and (3) monitoring the recovery of genital tissue after calving.

Ovine follicular fluid suppresses the ovarian secretion of androgens, oestradiol and inhibin

An experiment was conducted to examine the effect of steroid-free ovine follicular fluid (oFF) on ovarian hormone secretion. Eight Merino x Finnish Landrace ewes in which the left ovary and vascular pedicle had been autotransplanted to a site in the neck were studied during the breeding season. Luteal regression was induced in all animals by injection of cloprostenol (100 micrograms, i.m.) on day 10 of the luteal phase. Four of the eight animals were treated with steroid-free oFF (3 ml, s.c.) in the early follicular phase, 24 and 36 h after injection of cloprostenol. Samples of both ovarian and jugular venous blood were collected at 4-h intervals from 20 h before until 96 h after injection of cloprostenol. Ovarian and jugular venous blood samples were also collected at 10-min intervals from 48 to 52 h after injection of cloprostenol to investigate the pattern of pulsatile secretion of ovarian hormones. Samples were assayed for oestradiol, androstenedione, testosterone and inhibin and the ovarian secretion rates calculated. Both injections of oFF resulted in a fourfold increase in the concentration of inhibin in jugular venous plasma within 4-8 h of administration (P less than 0.01) with concentrations remaining increased (P less than 0.05) until 56 h after cloprostenol (32 h after the first oFF injection). Following oFF injection there was a profound (100%; P less than 0.001) and prolonged decrease in the peripheral concentration of FSH until 60 h after cloprostenol at which time the concentration of FSH increased five- to sixfold (P less than 0.001) to a peak lasting 24 h. In contrast to FSH, the concentration of LH in jugular venous plasma rose immediately following oFF treatment and continued to increase, exhibiting a profile similar to that described for FSH. No preovulatory LH surge was detected in any of the oFF-treated ewes while untreated ewes had an LH surge within 58.0 +/- 1.2 (S.E.M.) h. Within 8 h of the first injection of oFF the ovarian secretion rate of oestradiol, androstenedione and inhibin began to decline to reach a nadir of less than 1 ng/min within 32-36 h (56-60 h after cloprostenol; P less than 0.01). Testosterone secretion, already barely detectable, did not change significantly following injection of oFF but remained low for 36 h following oFF and did not exhibit the increase observed over this period in controls. After injection of oFF the episodic secretion of oestradiol, androstenedione, testosterone and inhibin was markedly suppressed in spite of numerous pulses of LH.(ABSTRACT TRUNCATED AT 400 WORDS)

Purification, partial characterization, and development of a specific radioimmunoassay for goat placental lactogen

Placental lactogen (PL) was isolated from goat cotyledonary tissue by a combination of mild alkaline extraction, anion and cation exchange chromatography, chromatofocussing and molecular filtration. The product, enriched 15,000-fold from the initial extract, was homogeneous when examined by SDS-gel electrophoresis (Mr 22,500) and isoelectricfocussing indicated a pI of 8.35 with a trace contaminant of pI 8.0. When assessed by relative binding activity in radioreceptor assays (RRA), goat PL exhibited somatotropic activity equivalent to 2.2 units/mg dry weight and lactogenic activity equivalent to 28.5 units/mg. A radioimmunoassay (RIA) for goat PL is described that is highly sensitive (190 pg/tube) and has acceptable repeatability within and between assays (6 and 13%, respectively). The assay is not affected by goat pituitary extracts or partly purified goat growth hormone and prolactin. Despite the marked increase in sensitivity of the RIA over that previously available when goat PL was measured by RRA, the hormone was not detected in jugular plasma of goats before Day 44 of pregnancy; concentrations increased thereafter and highest levels were measured during the last third of pregnancy in animals bearing triplets. Measurements by RIA are in general agreement with those obtained earlier in several studies in which RRAs were used. The hormone was detected in amniotic fluid. Maternal concentrations of goat PL declined before parturition and were undetectable by 18 h post partum.

Factors affecting the voluntary intake of food by sheep

The site where osmotically active substances act to depress food intake was determined in sheep. After 5.5 h of food deprivation, solutions of sodium chloride or polyethylene glycol-200 (PEG-200) were added to either the reticulo-rumen or the abomasum. The sheep were then immediately offered pelleted lucerne (Medicago sativa). Water was withheld during the first 60 min of feeding but was available from 60 to 90 min. There was a linear inhibition in food intake in the first 10 min after loading 2.37, 6.25, 12.5, 25.0 or 50.0 g NaCl into the rumen according to a 5 x 5 Latin square design (P = 0.0001). The intake reduction was 3.49 g food/g NaCl. An osmotic load of PEG-200 equivalent to 50 g NaCl also significantly inhibited food intake in the first 10 min of the meal compared with a control treatment. The inhibition of food intake after loading 55 g NaCl into the rumen was not affected by injecting lidocaine hydrochloride into the reticulum immediately before NaCl loading. NaCl injected into the abomasum did not significantly affect food intake in the first 10 min of feeding even though the tonicity of abomasal digesta was increased to unphysiological levels. There was no consistent relationship between food intake and the change in the tonicity of jugular plasma following solute loading and drinking. The sensing site of hypertonicity was localized to the wall of the reticulo-rumen where neuronal receptors appear to be capable of detecting osmotic pressure within the physiological range to depress food intake. These receptors should be identified and characterized because of their possible significance in limiting food intake by ruminants.

Myometrial Activity and Plasma Progesterone and Oxytocin Concentrations in Cycling and Early-Pregnant Ewes1

Myometrial activity and plasma progesterone (P) and oxytocin (OT) were measured in early pregnant (n = 5) and cycling (n = 5) ewes. Electromyography (EMG) leads and jugular and inferior vena cava (IVC) catheters were surgically placed in ewes about 1 wk before data collection. When ewes returned to estrus, they were bred to either an intact or vasectomized ram. Continuous EMG data were collected, and blood samples were collected twice daily from day of estrus (Day 0) until Day 18. Ewes bred with an intact ram were checked surgically for pregnancy on Day 20. Computerized, quantitative analysis of EMG events showed no difference in signal from the right to left uterine horns, and no differences between pregnant and cycling ewes (p less than 0.05) until Days 14-18 when nonpregnant ewes returned to estrus and had increased EMG activity. The mean number of EMG events 180-900 s in length decreased in pregnant ewes, but this difference was not significant (p less than 0.05). Jugular plasma progesterone (P) levels confirmed corpus luteum (CL) formation in all ewes, and no differences in P between pregnant and nonpregnant ewes were measured until Days 14-18, when cycling ewes underwent luteolysis and pregnant ewes maintained CL. IVC plasma oxytocin concentrations were increased in pregnant ewes compared to concentrations in nonpregnant ewes on Days 5-13 (p less than 0.05), and the difference was largest at Day 6 (means +/- SEM pg/ml: pregnant = 68.7 +/- 13.9, nonpregnant = 30.9 +/- 19.9).(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of gestation on jugular plasma thyroxine levels in Murrah buffalo and Karan Swiss cows

Plasma samples from 106 pregnant Karan Swiss (Brown Swiss x Sahiwal) cows and 108 Murrah buffalo were tested for thyroxine (T(4)) levels to determine the relationship between the hormonal changes and advancing pregnancy in the two species. All samples were collected within 2 months (January and February) to avoid seasonal interference on T(4) levels. In pregnant cows, the concentration of T(4) increased sharply during the first trimester, reaching a peak at the third month of gestation followed by a gradual decline until the last month of pregnancy. In pregnant buffalo, peripheral plasma T(4) levels fluctuated slightly throughout pregnancy without exhibiting a specific trend. Statistical analysis revealed that the magnitude of T(4) levels was significantly lower in buffalo (P < 0.01) than in the cows throughout pregnancy and that the hormonal patterns of the two species were significantly different (P < 0.05) during gestation. It was hypothesized from this study that T(4) requirements for the fetal buffalo calf may be lower than that for the fetal cattle calf since the buffalo gestation period is a month longer and the metabolic rate lower vis-a-vis the cow.

Parotid secretion patterns during meals and their relationships to the tonicity of body fluids and to gastrin and pancreatic polypeptide in sheep

The flow pattern of unilateral parotid saliva in sheep was compared when a total of 800 g lucerne (Medicago sativa) hay was offered as one, two, four or eight distinct meals. These patterns were related to changes in the tonicity of rumen fluid and plasma and to plasma concentrations of gastrin and pancreatic polypeptide. Sheep having ad lib access to hay overnight were offered fresh hay from 08.00 to 09.00 hours and were then given one, two, four, or eight meals of fresh hay according to a schedule such that the mean deprivation period was 6.5 h for each meal frequency-size. Neither the peak in saliva flow rate nor the time of this peak differed among the different meal sizes. The flow rate decreased rapidly after reaching a maximum at 3.2 min into the meal. After 7 min of eating, the tonicity of plasma and rumen fluid had increased by only 2.2 and 8.2 mosmol/kg respectively. These increases would not cause the rapid decline in parotid flow observed after 3.2 min of eating. There was no postprandial change in the concentration of gastrin in jugular plasma. However, it did increase significantly (P = 0.0043) from 16 to 4 min before eating commenced. There was a postprandial peak in plasma pancreatic polypeptide concentration after 4.5 min of eating. However, the parotid flow rate remained low after the concentration of this peptide returned to prefeeding levels. The rapid decrease in parotid secretion rate observed early in the meal may be due to subsiding central excitation rather than to an inhibitory factor limiting production.