Gene expression in the forebrain of dexamethasone-treated pigs: effects on stress neuropeptides in the hypothalamus and hippocampus and glutamate receptor subunits in the hippocampus

Gene expression studies advance our understanding of the effects of stress and glucocorticoids on brain function and give a new direction to animal welfare research. In this context, the presence of messenger RNA s (m RNA s) for corticotrophin releasing hormone (CRH) and vasopressin (VP) in the porcine hypothalamus has recently been documented. This study investigated the expression of CRH, VP and ionotropic glutamate receptor (iGluR) subunit m RNA s in the brains of pigs treated with the synthetic glucocorticoid dexamethasone (Dex; 5 mg kg(-1)i.v.). In the hypothalamus, VP, but not CRH, m RNA was reduced 3 hours after Dex. In the hippocampus, expression of m RNA s for some iGluR subunits appeared to be differentially regulated 6 hours after Dex. In addition, CRH message was detected in the hippocampus and significantly upregulated in the CA1 region 3 hours after Dex. The relevance of these findings to stress neurobiology of the growing pig is discussed.     

γ-Amino Butyric Acid Control of Arginine Vasopressin Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study aims to ascertain the influence of gamma-amino butyric acid (GABA)(A or B) receptors on arginine vasopressin (AVP) release in vitro and determine whether E(2) modulates GABA-AVP interaction. Within 10 min of ewe killing, saggital midline hypothalamic slices (from the anterior preoptic area to the mediobasal hypothalamus along with the median eminence, 2-mm thick, two per ewe) were dissected, placed in oxygenated minimum essential media (MEM)-alpha at 4 degrees C and within 2 h were singly perifused at 37 degrees C with oxygenated MEM-alpha (pH 7.4; flow rate 0.15 ml/min), either with or without E(2) (24 pg/ml). After 4-h equilibration, 10-min fractions were collected for 4 h interposed with a 10-min exposure at 60 min to a specific GABA(A or B) receptor agonist or antagonist at various doses (0.1-10 mm). GABA(A) (muscimol; no E(2), n = 7 perifusion chambers, with E(2), n = 11) or GABA(B) (baclofen; no E(2), n = 8, with E(2), n = 15) agonists (10 mm) did not influence AVP concentrations. However, AVP release increased (p < 0.05) 20-30 min after exposure to 10 mm GABA(A or B) antagonists (bicuculline, no E(2), n = 7: from 4.6 +/- 0.7 to 33.0 +/- 0.4, with E(2), n = 17: from 11.9 +/- 1.4 to 32.8 +/- 6.0; CGP52432, with E(2), n = 14: from 14.0 +/- 2.6 to 28.8 +/- 3.9 pg/ml). At the end of the collection period, hypothalamic slices responded to KCl (100 mm) with AVP efflux (p < 0.05). GABA(B) but not GABA(A) antagonist-stimulated AVP release was enhanced in the presence of E(2). In summary, AVP release is under the inhibitory influence of GABA input with further potentiation by E(2) through GABA(B) receptors in vitro.     

Effects of grain type, roughage level and monensin level on finishing cattle performance

Interactions among grain type (grain sorghum, corn or wheat), roughage level and monensin level were studied in four feedlot trials using pen-fed crossbred yearling cattle. In Trial 1, cattle fed high-moisture corn (HMC) were more efficient (.1537 vs .1406 for gain/feed; P less than .01) than cattle fed dry-rolled grain sorghum (DRGS). As level (0, 3, 6, 9%) of dietary roughage was increased, feed efficiency (gain/feed) decreased (.1566, .1461, .1479, .1382; linear, P less than .01). In Trial 2, a grain type (DRGS; dry-rolled corn, DRC; dry-rolled wheat, DRW) x roughage level interaction was observed for daily gain and feed efficiency. Feed efficiency (gain/feed) was decreased when roughage was added to diets containing DRC (.1608 vs .1750) or DRGS (.1674 vs .1465), but not to the diet containing DRW (.1664 vs .1607). In trial 3, a grain type x roughage level x monensin level interaction (P less than .08) was observed for feed efficiency. The addition of 27.5 mg of monensin per kilogram of the 0% roughage-DRC diet tended to improve feed efficiency (.1633 vs .1531), but the addition of monensin to the 7.5% roughage-DRC diet tended to depress feed efficiency (.1476 vs .1575). The addition of either roughage (.1493 vs .1420) or monensin (.1500 vs .1413) to the DRW diet improved feed efficiency. In Trial 4, cattle fed a combination of 75% DRW and 25% DRC were more efficient (.1618 vs .1591; P less than .06) than cattle fed DRC. As level of roughage (0, 3.75, 7.5%) increased, feed efficiency decreased linearly (.1645, .1599, .1569; P less than .0001). Monensin had no effect on feed efficiency. The value of feeding roughage and monensin was variable both across grain types and within similar grain types.     

Immunolocalization of Aquaporins 1 and 9 in the Ram Efferent Ducts and Epididymis

Aquaporins (AQPs) are essential membrane protein channels for the transport of water across membranes. Fluid movement in the epididymis is important for modulation of the luminal environment, in which sperm mature and reside. This study was designed to understand the morphology and localization of AQPs in ram efferent ducts (ED) and epididymis. For this purpose, the epididymis of seven animals were removed for histologic and immunohistochemical analyses. AQP1 immunoreactivity was observed in the apex of the ED, and AQP9 was found adjacent to the nuclei of the epithelial cells of the ED. The epithelial lining of ram epididymis is pseudostratified columnar and presents principal, basal, apical and narrow cells. In the initial segment (IS), a moderate reaction for AQP1 was observed in the apical cytoplasm of epithelial cells. An intense reactivity for AQP1 was noted over the microvilli of principal cells and in spermatozoa in the caput. In the corpus and cauda, AQP1 was noted only over the endothelial cells of vascular channels located in intertubular spaces. A weak‐to‐moderate reaction for AQP9 was observed in the nuclei of epithelial cells in the IS, caput and corpus of the epididymis. In the cauda, an intense reaction to AQP9 was observed in the epithelial border. In the IS, caput and corpus, the reactivity for AQP9 differed from those observed in domestic animals. The cauda showed a pattern similar to that previously described. These results indicate that AQPs 1 and 9 have reversed locations and roles in rams, suggesting activity variations related with fluid and solute absorption throughout the epididymis.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Reoviruslike agent in stools: association with infantile diarrhea and development of serologic tests

Reoviruslike particles were visualized by electron microscopy in stool filtrates prepared from stools of infants and young children with severe acute gastroenteritis. Patients who had such particles in their stools and whose paired acute and convalescent serums were tested developed an antibody response to the reoviruslike agent, which was measured by immune electron microscopy and by complement fixation. The reoviruslike agent was antigenically related to the epizootic diarrhea of infant mice virus and the Nebraska calf diarrhea virus.     

The effect of a monensin ruminal delivery device on performance of cattle grazing pasture

A total of 28 trials utilizing 2,262 steers and heifers were conducted to evaluate a sustained monensin2 ruminal delivery device (RDD) on the daily gain of grazing cattle. Five series of trials, with four to eight trials per series within a grazing season, were conducted to evaluate the daily gain response under different environmental conditions and locations. Pastures grazed included both summer (cool- and warm-season grasses and native range) and winter (rye, ryegrass and wheat) growing pastures. The experimental design for all trials in each of five series was a randomized complete block with initial weight or breed as the blocking factor. The treatments compared in series I, III, IV and V (20 trials) were a control and a single RDD, whereas in series II (eight trials), a 2 x 2 factorial was utilized that compared RDD and estradiol 17-beta controlled release implants. In the five series of trials, the monensin RDD increased daily gain .04 (P = .002), .03 (P = .07), .09 (P = .01), .05 (P = .10) and .08 kg (P = .04) compared with the controls. Use of the RDD provided an effective method of achieving consistent monensin delivery and weight gain improvement without requiring the extra labor needed to administer monensin via hand-feeding or self-feeding supplements, blocks or loose mineral.