Genomic reaction norm models exploiting genotype × environment interaction on sexual precocity indicator traits in Nellore cattle.

Brazilian beef cattle are raised predominantly on pasture in a wide range of environments. In this scenario, genotype by environment (G×E) interaction is an important source of phenotypic variation in the reproductive traits. Hence, the evaluation of G×E interactions for heifer's early pregnancy (HP) and scrotal circumference (SC) traits in Nellore cattle, belonging to three breeding programs, was carried out to determine the animal's sensitivity to the environmental conditions (EC). The dataset consisted of 85 874 records for HP and 151 553 records for SC, from which 1800 heifers and 3343 young bulls were genotyped with the BovineHD BeadChip. Genotypic information for 826 sires was also used in the analyses. EC levels were based on the contemporary group solutions for yearling body weight. Linear reaction norm models (RNM), using pedigree information (RNM_A) or pedigree and genomic information (RNM_H), were used to infer G×E interactions. Two validation schemes were used to assess the predictive ability, with the following training populations: (a) forward scheme-dataset was split based on year of birth from 2008 for HP and from 2011 for SC; and (b) environment-specific scheme-low EC (-3.0 and -1.5) and high EC (1.5 and 3.0). The inclusion of the H matrix in RNM increased the genetic variance of the intercept and slope by 18.55 and 23.00% on average respectively, and provided genetic parameter estimates that were more accurate than those considering pedigree only. The same trend was observed for heritability estimates, which were 0.28-0.56 for SC and 0.26-0.49 for HP, using RNM_H, and 0.26-0.52 for SC and 0.22-0.45 for HP, using RNM_A. The lowest correlation observed between unfavorable (-3.0) and favorable (3.0) EC levels were 0.30 for HP and -0.12 for SC, indicating the presence of G×E interaction. The G×E interaction effect implied differences in animals' genetic merit and re-ranking of animals on different environmental conditions. SNP marker-environment interaction was detected for Nellore sexual precocity indicator traits with changes in effect and variance across EC levels. The RNM_H captured G×E interaction effects better than RNM_A and improved the predictive ability by around 14.04% for SC and 21.31% for HP. Using the forward scheme increased the overall predictive ability for SC (20.55%) and HP (11.06%) compared with the environment-specific scheme. The results suggest that the inclusion of genomic information combined with the pedigree to assess the G×E interaction leads to more accurate variance components and genetic parameter estimates.

Habituation and cross-sensitization of stress-induced hypothalamic-pituitary-adrenal activity: effect of lesions in the paraventricular nucleus of the thalamus or bed nuclei of the stria terminalis.

Habituation of the hypothalamic-pituitary-adrenal (HPA) response to chronic intermittent restraint stress (30 min/day for 15 days) and the cross-sensitization to a heterotypic stress [i.p. lipopolysaccharide (LPS)] were investigated in intact male Sprague Dawley rats, and in rats bearing quinolinic acid lesions to the medial anterior bed nuclei of the stria terminalis (BST) or anterior region of the paraventricular nucleus of the thalamus (PVT). In intact animals, a single period of restraint increased plasma corticosterone levels at 30 min and led to an increase in corticotropin-releasing hormone (CRH) mRNA levels in the PVN at 3 h. LPS had a smaller effect on corticosterone and more variable effect on CRH mRNA. Chronic intermittent restraint stress caused a decrease in body weight and increase in adrenal weights, with concomitant increase in basal corticosterone levels. These animals also displayed marked habituation of the corticosterone and CRH mRNA responses to the homotypic stress of restraint, but no loss of the corticosterone response to the heterotypic stress of LPS and a cross-sensitization of the CRH mRNA response. This pattern of stress responses in control and chronically stressed animals was not significantly affected by lesions to the PVT or BST, two areas which have been implicated in the coping response to stress. Thus, these data provide evidence for independent adaptive mechanisms regulating HPA responses to psychological and immune stressors, but suggest that neither the medial anterior BST nor the anterior PVT participate in the mechanisms of habituation or cross-sensitization.

Immunological stress in rats induces bodily alterations in saline-treated conspecifics.

This work was developed during an investigation on the neuroendocrine-immune interaction in rats immune challenged with sheep red blood cells (SRBC). The structures used for evaluating the immunological response was the direct plaque-forming cells (PFC). An inbred strain of rat was used to overcome the problem of different timings in the peak humoral immune response. Normal rats were injected intraperitoneally with saline or SRBC and were killed 0, 3, 4, 5, and 6 days later. Body and gland weights were recorded, and. serum levels of corticosterone and prolactin were quantified by radioimmunoassay. The hormone levels and gland weights of the saline conspecifics and SRBC-treated rats were found to be similar. When new rats were housed in a separate room and treated with physiological saline, there were again no differences in the body and gland weights or the serum hormone levels between the two home cage control (HCC) groups of animals. Compared with saline conspecifics and SRBC-treated groups, the HCC groups had higher body weights from the third to the sixth day of treatment and had lower gland weights in absolute and relative analysis (pituitary, thyroid, and adrenals) mainly on the fourth and fifth days; thymus weights were highest on the third day. Corticosterone and prolactin levels were significantly lower on the fifth and sixth days, respectively. Because SRBC-treated rats showed a peak direct immune response on the fourth and fifth days and showed peak corticosterone levels on the fifth day after treatment, we conclude that the former animals were under stress and influenced their saline conspecifics through sound or smell. This conclusion agrees with other studies, showing that physically or emotionally stressed rats can influence conspecifics through noise and body odors.

Serum cytokine levels in autoimmune and non-autoimmune hyperthyroid states.

Although the role of interleukin-2 (IL-2) and interferon gamma (gammaIFN) is still poorly understood in hyperthyroid diseases, it is reasonable to assume that these cytokines may be present at higher levels in Graves' disease (GD) than in other primarily non-autoimmune thyroid diseases. In order to look for an easy method to distinguish GD from primarily non-autoimmune causes of hyperthyroidism, we compared 13 healthy individuals with 21 treated and untreated hyperthyroid GD patients and with 19 patients with hyperthyroidism due to other etiologies: 7 cases of multinodular goiter, 5 cases of excessive hormone replacement and 7 cases of amiodarone-associated hyperthyroidism. All patients presented low TSH levels and a dubious clinical thyroid state. We found a good correlation between TSH and serum IL-2 levels (r = 0.56; P<0.01). Serum IL-2 (P<0.01) and gammaIFN (P<0.01) levels were lower in the hyperthyroid group of patients than in control subjects, suggesting a depressed TH1 pattern in the T-cell subset of hyperthyroid patients. GD had normal IL-2 levels, while patients with other forms of thyrotoxicosis presented decreased IL-2 levels (P<0.05). There was no difference between treated and untreated GD patients. We suggest that the direct measurement of serum IL-2 level may help to confirm hyperthyroidism caused by GD.

Serum cytokine levels in autoimmune and non-autoimmune hyperthyroid states

Although the role of interleukin-2 (IL-2) and interferon gamma (yIFN) is still poorly understood in hyperthyroid diseases, it is reasonable to assume that these cytokines may be present at higher levels in Graves' disease (GD) than in other primarily non-autoimmune thyroid diseases. In order to look for an easy method to distinguish GD from primarily non-autoimmune causes of hyperthyroidism, we compared 13 healthy individuals with 21 treated and untreated hyperthyroid GD patients and with 19 patients with hyperthyroidism due to other etiologies: 7 cases of multinodular goiter, 5 cases of excessive hormone replacement and 7 cases of amiodarone-associated hyperthyroidism. All patients presented low TSH levels and a dubious clinical thyroid state. We found a good correlation between TSH and serum IL-2 levels (r = 0.56; P

Serum cytokines in chronic Chagas' disease.

We studied the serum levels of IL-2, IFN-gamma and TNF in different clinical forms of Chagas' disease and in patients clinically compensated and decompensated. Cytokines measured in 91 patients with the chronic form of the disease did not differ from those of 13 normal individuals, suggesting the absence of activation of the TH1 pattern of lymphocyte response. There were no statistical differences among the 17 patients in the indeterminate form of the disease, the patients presenting either early (n = 4) or well-developed signs of cardiomyopathy (n = 62), the digestive (n = 4) or the mixed (n = 4) forms of the disease. Serum TNF was undetectable and IFN-gamma levels did not differ between clinical forms and severities of Chagas' disease. However, we found IL-2 higher levels in the 25 non-controlled patients than in the 66 controlled individuals (p < 0.001). We suggest that IL-2 dosage may be useful as an indicator of the need for more aggressive procedures.

The influence of septal lesions on sodium and water retention induced by Walker 256 tumor.

In the course of studies on the effects of septal area lesions on neuroimmunomodulation and Walker 256 tumor development, it was observed that tumor-induced sodium and water retention was less marked in lesioned than in non-lesioned rats. In the present study possible mechanisms involved in this phenomenon were investigated. The experiments were performed in septal-lesioned (LW; N = 15) and sham-operated (SW; N = 7) 8-week-old male Wistar rats, which received multifocal simultaneous subcutaneous (sc) inoculations of Walker 256 tumor cells about 30 days after the stereotaxic surgery. Control groups (no tumor, sham-operated food-restricted (SFR), N = 7) and lesioned food-restricted (LFR, N = 10) were subjected to a feeding pattern similar to that observed in tumor-bearing animals. Multifocal inoculation of Walker 256 tumor rapidly induces anorexia, which is paradoxically accompanied by an increase in body weight, as a result of renal Na+ and fluid retention. These effects of the tumor were also seen in LW rats, although the rise in fractional sodium balance during the early clinical period was significantly smaller than in SW rats (day 4: SW = 47.6 +/- 6.4% and LW = 13.8 +/- 5.2%; day 5: SW = 57.5 +/- 3.5% and LW = 25.7 +/- 4.8%; day 6: SW = 54.4 +/- 3.8% and LW = 32.1 +/- 4.4%; P < 0.05), suggesting a temporary reduction in tumor-induced sodium retention. In contrast, urine output was significantly reduced in SW rats and increased in LW rats (LW up to -0.85 and SW up to 4.5 ml/100 g body weight), with no change in osmolar excretion. These temporary changes in the tumor's effects on LW rats may reflect a "reversal" of the secondary central antidiuretic response induced by the tumor (from antidiuretic to diuretic).

The influence of septal lesions on sodium and water retention induced by Walker 256 tumor

In the course of studies on the effects of septal area lesions on neuroimmunomodulation and Walker 256 tumor development, it was observed that tumor-induced sodium and water retention was less marked in lesioned than in non-lesioned rats. In the present study possible mechanisms involved in this phenomenon were investigated. The experiments were performed in septal-lesioned (LW; N = 15) and sham-operated (SW; N = 7) 8-week-old male Wistar rats, which received multifocal simultaneous subcutaneous (sc) inoculations of Walker 256 tumor cells about 30 days after the stereotaxic surgery. Control groups (no tumor, sham-operated food-restricted (SFR), N = 7) and lesioned food-restricted (LFR, N = 10) were subjected to a feeding pattern similar to that observed in tumor-bearing animals. Multifocal inoculation of Walker 256 tumor rapidly induces anorexia, which is paradoxically accompanied by an increase in body weight, as a result of renal Na+ and fluid retention. These effects of the tumor were also seen in LW rats, although the rise in fractional sodium balance during the early clinical period was significantly smaller than in SW rats (day 4: SW = 47.6 = 6.4 percent and LW = 13.8 = 5.2 percent; day 5: SW = 57.5 = 3.5 percent and LW = 25.7 = 4.8 percent; day 6: SW = 54.4 = 3.8 percent and LW = 32.1 = 4.4 percent; P<0.05), suggesting a temporary reduction in tumor-induced sodium retention. In contrast, urine output was significantly reduced in SW rats and increased in LW rats (LW up to -0.85 and SW up to 4.5 ml/100 g body weight), with no change in osmolar excretion. These temporary changes in the tumor's effects on LW rats may reflect a "reversal" of the secondary central antidiuretic response induced by the tumor (from antidiuretic to diuretic)

Faster gastric emptying of a liquid meal in rats after hypothalamic dorsomedial nucleus lesion

The effects of dorsomedial hypothalamic (DMH) nucleus lesion on body weight, plasma glucose levels, and the gastric emptying of a liquid meal were investigated in male Wistar rats (170-250 g). DMH lesions were produced stereotaxically by delivering a 2.0 mA current for 20 s through nichrome electrodes (0.3-mm tip exposure). In a second set of experiments, the DMH and the ventromedial hypothalamic (VMH) nucleus were lesioned with a 1.0-mA current for 10 s (0.1-mm tip exposure). The medial hypothalamus (MH) was also lesioned separately using a nichrome electrode (0.3-mm tip exposure) with a 2.0-mA current for 20 s. Gastric emptying was measured following the orogastric infusion of a liquid test meal consisting of physiological saline (0.9 percent NaCl, w/v) plus phenol red dye (6 mg/dl) as a marker. Plasma glucose levels were determined after an 18-h fast before the lesion and on the 7th and 15th postoperative day. Body weight was determined before lesioning and before sacrificing the rats. The DMH-lesioned rats showed a significantly faster (P<0.05) gastric emptying (24.7 percent gastric retention, N = 11) than control (33.0 percent gastric retention, N = 8) and sham-lesioned (33.5 percent gastric retention, N = 12) rats, with a transient hypoglycemia on the 7th postoperative day which returned to normal by the 15th postoperative day. In all cases, weight gain was slower among lesioned rats. Additional experiments using a smaller current to induce lesions confirmed that DNH-lesioned rats had a faster gastric emptying (25.1 percent gastric retention, N =7) than control (33.4 percent gastric retention, N = 17) and VMH-lesioned (34.6 percent gastric retention, N = 7) rats. MH lesions resulted in an even slower gastric emptying (43.7 percent gastric retention, N = 7) than in the latter two groups. We conclude that although DMH lesions reduce weight gain, they do not produce consistent changes in plasma glucose levels. These lesions also promote faster gastric emptying of an inert liquid meal, thus suggesting a role for the DMH in the regulation of gastric motility.

Faster gastric emptying of a liquid meal in rats after hypothalamic dorsomedial nucleus lesion.

The effects of dorsomedial hypothalamic (DMH) nucleus lesion on body weight, plasma glucose levels, and the gastric emptying of a liquid meal were investigated in male Wistar rats (170-250 g). DMH lesions were produced stereotaxically by delivering a 2.0-mA current for 20 s through nichrome electrodes (0.3-mm tip exposure). In a second set of experiments, the DMH and the ventromedial hypothalamic (VMH) nucleus were lesioned with a 1.0-mA current for 10 s (0.1-mm tip exposure). The medial hypothalamus (MH) was also lesioned separately using a nichrome electrode (0.3-mm tip exposure) with a 2.0-mA current for 20 s. Gastric emptying was measured following the orogastric infusion of a liquid test meal consisting of physiological saline (0.9% NaCl, w/v) plus phenol red dye (6 mg/dl) as a marker. Plasma glucose levels were determined after an 18-h fast before the lesion and on the 7th and 15th postoperative day. Body weight was determined before lesioning and before sacrificing the rats. The DMH-lesioned rats showed a significantly faster (P < 0.05) gastric emptying (24.7% gastric retention, N = 11) than control (33.0% gastric retention, N = 8) and sham-lesioned (33.5% gastric retention, N = 12) rats, with a transient hypoglycemia on the 7th postoperative day which returned to normal by the 15th postoperative day. In all cases, weight gain was slower among lesioned rats. Additional experiments using a smaller current to induce lesions confirmed that DMH-lesioned rats had a faster gastric emptying (25.1% gastric retention, N = 7) than control (33.4% gastric retention, N = 17) and VMH-lesioned (34.6% gastric retention, N = 7) rats. MH lesions resulted in an even slower gastric emptying (43.7% gastric retention, N = 7) than in the latter two groups. We conclude that although DMH lesions reduce weight gain, they do not produce consistent changes in plasma glucose levels. These lesions also promote faster gastric emptying of an inert liquid meal, thus suggesting a role for the DMH in the regulation of gastric motility.

Effect of paraventricular nucleus lesion and cold restraint stress on gastric emptying of a liquid meal in rats.

The effects of cold restraint stress on gastric emptying (GE) and the involvement of the hypothalamic paraventricular nucleus (PVN) were investigated in male Wistar rats (200-250 g body weight). Electrolytic lesions were produced stereotaxically in the nucleus by passing a 2.0-mA current for 10 s through stainless steel electrodes. GE was measured by means of a liquid test meal of 5% (w/v) glucose solution plus phenol red (6 mg/dl) dye as marker, given by orogastric infusion. Cold restraint stress induces a significant increase (43.7%, N = 11) in gastric retention of a 5% glucose solution in rats, i.e., a delay in GE of this solution. However, restraint stress alone does not produce any change. Both truncal vagotomy and electrolytic lesion of the PVN completely block the cold restraint-induced delay in GE. However, PVN lesion per se results in a decrease of GE (30.6%, N = 10) when compared to nonoperated controls. In addition, PVN-lesioned rats exposed to cold restraint present a slightly faster GE (14.7%, N = 11) than controls, demonstrating an opposite response to that initially observed without lesion. These data suggest an important role for PVN efferents, probably influencing medullary vagal preganglionic neurons, in the development of this gastric motor impairment under stress conditions.

Effect of paraventricular nucleus lesion and cold restraint stress on gastric emptying of a liquid meal in rats

The effects of cold restraint stress on gastric emptying (GE) and the involvement of the hypothalamic paraventricular nucleus (PVN) were investigated in male Wistar rats (200-250 g body weight). Electrolytic lesions were produced stereotaxically in the nucleus by passing a 2.0-mA current for 10 s through stainless steel electrodes. GE was measured by means of a liquid test meal of 5 per cent (w/v) glucose solution plus phenol red (6 mg/dl) dye as marker, given by orogastric infusion. Cold restraint stress induces a significant increase (43.7 per cent , N = 11) in gastric retention of a 5 per cent glucose solution in rats, i.e., a delay in GE of this solution. However, restraint stress alone does not produce any change. Both truncal vagotomy and electrolytic lesion of the PVN completely block the cold restraint-induced delay in GE. However, PVN lesion per se results in a decrease of GE (30.6 per cent , N = 10) when compared to nonoperated controls. In addition, PVN-lesioned rats exposed to cold restraint present a slightly faster GE (14.7 per cent , N = 11) than controls, demonstrating an opposite response to that initially observed without lesion. These data suggest an important role for PVN efferents, probably influencing medullary vagal preganglionic neurons, in the development of this gastric motor impairment under stress conditions

Thyroid function in hemidecorticate rats

1. Throid function was evaluated in hemidecorticate (HD) and control (C) rats by determining serum T3 and T4 levels and the development of incisors and mandibles and through analysis of various histological features of the thyroid such as follicle size, colloid droplet content and [3H] - glycine uptake by follicular cells. 2. HD animals presented normal levels of circulating T3 but significantly lower T4 levels. 3. There was slight atrophy of the gland in HD animals and fewer colloid droplets were present in the cytoplasm of the follicular cells in this group, indicating a reduction in the breakdown of thyroglobulin. [3H] - glycine uptake by HD indicated that rate of thyroglobulin biosynthesis was not altered in the experimental animals. 4. the growth of mandibles (weight) and incisors (weight and lenght) was reduced in HD compared to the control animals. 5. These results suggest that hemidecortication causes mild hypothyroidism (trophoprivic type) probably by affecting hypothalamic function

Studies on lymphocyte homing in autoimmune prone NZB mice.

Lymphocyte homing patterns in young (3-5 months old) and old (10-12 months old) autoimmune prone NZB mice were investigated by transferring 51Cr labelled lymphoid cells into syngeneic and H-2 compatible allogeneic recipients. We confirmed that non H-2 alloantigens as well as H-2 alloantigens can be important determinants of apparent abnormalities of cellular distribution with the techniques employed. No gross abnormalities of lymphocyte traffic were present in the young NZB mice as compared to the autoimmune resistant strains of mice when syngeneic cells are used. Spleen of older NZB mice appeared to be less attractive to lymph node cells than was the spleen from young NZB mice. Splenocytes of older NZB mice localized significantly more in the liver and less in the lymph nodes as compared with splenocytes from young NZB mice. The mechanism underlying abnormalities of lymphoid cell distribution which feature the autoimmune-prone NZB mice are not yet clear and further studies will be necessary before they can be characterized definitively. Our findings, using syngeneic cells, are in disagreement with those of Zatz and Lance since evidence of abnormal distribution of lymphocytes in young NZB mice were not seen when syngeneic cells were employed.

Regulation of gonadotrophin function in hemidecorticate rats.

The present review focuses on some aspects of the regulation of gonadotrophin (LH and FSH) and prolactin secretion in rats submitted to hemidecortication (HD). The removal of rhinencephalic and cortical structures induces the following changes: Irregular and erratic estrous cycle; Increase in LH and FSH secretion in the afternoon of proestrus and after uni- or bilateral castration; When hormonal replacement therapy was administered to castrated female rats, the controls needed larger doses of estradiol benzoate (E2) to reduce LH levels than the HD animals, which suggests an increase in the sensitivity of the hypothalamus-hypophysis axis to gonadal steroid negative feedback. When HD was performed during the neonatal period (1-3 days old), an earlier onset of puberty and sexual maturation was observed in both female and male HD rats than in controls. During the puberal evolution the HD male rats showed elevation in plasma testosterone levels, an increase in the testicular response to hCG and also in the number of testicular binding sites to LH and FSH at 38-43 days (before the onset of puberty) and to prolactin (after 60 days). The results suggest that after removal of cortical and subcortical structures (HD), with predominant inhibitory influences, the hypothalamus-hypophysis becomes more sensitive to the LH and FSH releasing mechanisms.

Puberty and sexual maturation in hemidecorticate female rats.

The onset of puberty and sexual maturation were studied in female rats that had been hemidecorticated (HD) during the neonatal period (1-3 days old). Hemidecorticate and control rats submitted to photostimulation (14 h light: 10 h dark) had earlier and better synchronized vaginal opening (VO) than rats raised under natural light (12 h light: 12 h dark). HD rats maintained under 14 h light: 10 h dark had vaginal opening 4-5 days earlier than their controls under the same light conditions. Peaks of plasma gonadotrophin (LH) were observed in hemidecorticate animals 10 days (30th day of life) earlier than in the controls (40th day). These LH peaks were preceded by higher adenohypophyseal and uterine weights at 25 and 30 days of age. The estrous cycles observed in adult rats that were hemidecorticated during the neonatal period were similar to that of intact rats. These results suggest that hemidecortication induces early puberty. This effect could be due to accelerated maturation of the ovaries probably due to the unrestricted activity of the medial basal hypothalamus, which in turn might activate the hypothalamic mechanisms that trigger puberty.