Food restriction induced thyroid changes and their reversal after refeeding in female rats and their pups

In the present study, two groups of pregnant female rats were submitted to food restriction (24 h fast versus 24 h diet intake) from the 14th day of pregnancy until either the 14th day (group B) or the 4th day after parturition (group C). All pups and their mothers were sacrificed on day 14 after delivery. The body weight of the 14-day-old pups (group B) was 46% less than the controls (group A). Free thyroxine and free triiodothyronine levels in the plasma were reduced by 44 and 16% in pups and by 20 and 36% in their mothers, respectively. These reductions were correlated with a decrease in thyroid iodine content of the pups (-50%) and their mothers (-24%). Radioiodine uptake (131I) by the thyroid gland of pups was significantly increased by 27%. Plasma TSH levels were decreased by 38% in pups and by 44% in dams. Morphological changes in thyroid glands were observed in energy restricted dams and in their pups. Some of follicles in pups were empty. Moroever in dams, we noted the presence of peripheral resorbed vacuoles, sign of thyroid hyperactivity. After a refeeding (group C) period of ten days, total recovery occurred in plasma thyroid hormone levels (FT4 and FT3) and in thyroid iodine contents of pups in spite of a partial recovery of body weights and plasma TSH levels. In dams, a partial recovery occurred in plasma thyroid hormone levels in spite of total recovery in thyroid iodine contents, while plasma TSH levels exceeded control values. A significant amelioration in thyroid histological aspects was observed in pups and their dams.     

Oxidative stress and thyroid impairment after gibberellic acid treatment in pregnant and lactating rats and their offspring

Gibberellic acid (GA?) has been worldwide used in agriculture as a plant growth regulator. The purpose of this study is to assess the effects of GA? on the morphology and the thyroid hormone levels in adult rats and their suckling pups. Animals were given daily 200 ppm GA? in drinking water from the 14th day of pregnancy until day 14 after delivery. Compared with a control group, GA?-treated mothers and pups showed an increase in body and thyroid weights, a decrease in plasma FT? and FT? levels, which were more pronounced in pups than in their mothers. Thyroid iodine content was also decreased in pups. These biochemical modifications corresponded histologically; the majority of follicles had cubical epithelial cells, which surrounded empty vesicular cavities. Toxicity was objectified by a significant increase in plasma malondialdehyde, protein carbonyls, and advanced oxidation protein products levels in GA?-treated dams and their suckling pups; while, the activities of superoxide dismutase, catalase, and glutathione peroxidase were decreased in plasma of both dams and their pups. Moreover, a significant decline was observed in plasma glutathione, nonprotein thiols, and vitamin C levels. We conclude that GA? treatment affects thyroid function and plasma antioxidant status in adult rats and their progeny.     

Effect of weaning time on the growth rate and food intake of the spiny mouse pup

The spiny mouse Acomys cahirinus produces well-developed pups although it is a relatively small mammal (45 g). We envisioned two opposing hypotheses on the effect of early weaning on the growth rate of pups. The first predicts little effect since the increase in energy intake of dams above non-reproducing females is relatively low, suggesting that pups consume a large portion of their energy as solid food, and the pups are very well developed at birth. The second predicts a substantial effect since the 'index of precociality', that is the energy intake for maintenance of a pup as a proportion of that predicted for a rodent of its body mass, falls within values for altricial rodents, suggesting an extended maternal dependence of the young. To test these hypotheses, we measured the growth rate and food intake of pups weaned after either 7, 14, 21 or 28 days. Only three of 12 pups weaned after 7 days survived and, consequently, the latter hypothesis was supported. All pups weaned at 14–28 days survived. There was a significant decrease in growth rate during the first day after weaning in pups weaned at 7, 14 and 21 days but not after 28 days, suggesting that pups did not require parental care by day 28. Peak growth rate in pups weaned at 14 days occurred in the second week but occurred in the third week in pups weaned at 21 and 28 days. In spite of these differences, pups in all treatments had similar body mass at 64 days, indicating compensatory growth. We concluded that pups of A. cahirinus are precocial from a morphological aspect in that they are well developed at birth but altricial from a nutritional aspect in that they require extended maternal support.     

Olfactory bulb serotonin level modulates olfactory recognition in the neonate rat

Role of serotonin in olfactory recognition was tested by depleting the olfactory bulb serotonin during postnatal day (PND) 1 - 4 following administration of 5,7-dihydroxytryptamine. Significant difference in the olfactory recognition test was observed during PND5-7; control pups successfully recognized and oriented towards their mother; whereas treated pups failed to recognize their mother odour. Later on, during PND12-14, both group of pups responded equally in the recognition test. Levels of olfactory bulb serotonin were depleted (53.3%) in the treated pups on PND-8, which was restored on PND-14 with only 15% variation. Further analysis demonstrated that depletion of serotonin in olfactory bulb did not affect the normal suckling and weight gain, it only modulates olfactory recognition.     

棕色田鼠幼仔对父鼠的早期识别及条件位置偏爱

群居性动物的幼仔对亲本的识别有利于获得最大化的亲本抚育。雄性棕色田鼠对幼仔具有较高水平的父本投资,那么幼仔在断乳前是否能够识别父鼠并形成条件位置偏爱(Conditioned place preference,CPP)呢?本研究通过双向选择测试、社会互作测试以及CPP 测试探讨了这一问题。双向选择结果显示,与陌生雄鼠相比,出生后14 d 和21 d 的幼仔对父鼠访问的持续时间及潜伏期有显著性差异(P < 0.05);社会互作结果表明,14 d幼仔对父鼠的跟随行为较陌生雄鼠多,但探究行为及亲密行为没有显著性差异(P > 0.05),21 d 幼仔对父鼠的探究行为较少、跟随行为和亲密行为较多(P < 0.05)。CPP 结果表明,在连续4 d (出生后13 ～ 16 d;19 ～ 22 d)的父鼠强化后,第17 d 和第23 d 的幼仔不能形成对父鼠的位置偏爱(P > 0.05)。这些结果说明断乳前的棕色田鼠幼仔已能够识别父鼠和陌生雄鼠,但不能形成对父鼠的位置偏爱。     

Effects of excess methionine on collagen metabolism: a study on newborn rat skin

We studied the biochemical effects of excess methionine intake on the skin of newborn rats. Group 1 pups were intubated with methionine dissolved in 0.1 ml physiological saline solution in the amount of 0.1 g/100 body wt as a control using a gastric needle. Group 2 pups were given 0.2 g/100 g in the same manner as group 1 as an experimental group. They were intubated every other day from day 3 to 13. On Day 15, [14C]proline was injected intraperitoneally into pups and their skin was removed. 14C total and hydroxyproline uptake was examined in the tissue as well as in the sequential extracts. Although excess methionine intake by the pups did not alter the collagen content of their skin, it caused an increase in the content of type III collagen and a decrease in crosslinked collagen. In addition, newly synthesized collagen in the neutral salt extract increased in the excess methionine group, indicating that crosslinked collagen decreases as excess methionine was intubated. The present study demonstrated that excess methionine in the early lactational period altered the nature of the skin collagen of suckling newborn rat pups.     

Evidence that suckling pups, through an exteroceptive mechanism, inhibit the milk stimulatory effects of prolactin in the rat during late lactation

Lactating rats were removed from the animal room on postpartum Day 20 and placed in a room where there were no other rats. On postpartum Day 21, these rats released prolactin either in response to exteroceptive signals from their own pups which were placed underneath or in response to those from 25 to 30 other lactating rats which along with their litters were placed in a rack 3 ft in front of the mothers' cage. Milk secretion was stimulated in the isolated rats on Day 21 by the prolactin released in response to exposure to the rack of lactators, but paradoxically was not stimulated by that released in response to exposure of the mother to her own pups. In fact, the stimulatory effects upon milk secretion, resulting from exposure to the rack of lactators was totally prevented if the mother was exposed to her pups 3–4 min before exposure to the rack of lactators. The blocking effect of the pups, however, did not occur when the pups were placed in cages alongside the mothers' cage. From subsequent experiments, it was concluded that sensory cues, from the pups, appeared to activate the sympathetico-adrenal system of the mother to release catecholamines which then blocked the milk stimulatory effects of prolactin which already had been discharged into the circulation. The pups did not inhibit milk secretion on Day 14 which indicates that the inhibiting mechanism becomes established sometime between Days 14 and 21 of lactation. These data suggest that a peripheral mechanism may operate to reduce milk secretion in the rat during late lactation, and thus, may be involved in the normal weaning process.     

Effects of acute and repeated exposure to stress on the hypothalamo-pituitary-adrenocortical activity in mice during postnatal development

Hypothalamo-pituitary-adrenocortical (HPA) response to a mild stressful procedure was investigated in mice at Days 8, 10, 12, and 14 of postnatal development. Pups that were removed from the dam and exposed to a novel odor (clean bedding) for 15 min showed higher plasma corticosterone levels than pups whose mother was removed from the cage for 15 min or unhandled pups at all ages, although statistically significant differences were only evident at Days 12 and 14. Lower HPA axis responding in younger mice was not due to immaturity since 8-day-old mice showed a significant and larger increase of plasma corticosterone levels when separated from the mother and isolated from littermates in the absence of bedding. Mice daily exposed to clean bedding (15 min) for the first 13 days of life did not show reduced plasma corticosterone response when reexposed to the stressor at 14 days of age. Conversely, increased plasma corticosterone levels in dams in response to removal of pups was not detectable after repeated exposure to this manipulation (14 days) regardless of the procedure their pups were submitted to, thus ruling out a role of maternal corticosterone passing through the milk on which the pups were fed. These results demonstrate that 15 min exposure to clean bedding is a noninvasive procedure able to elicit HPA axis response in developing mice over a wide age range without producing habituation.     

Maternal Separation in Guinea-Pigs: A Study in Behavioural Endocrinology

The aim of this study was to elucidate the modulation of behaviour and endocrine stress responses of guinea‐pig pups by social and spatial factors in a maternal separation paradigm. The animals were kept in two colonies (each colony: nine males, 13 females and their offspring; enclosure size: 6 m²). Blood samples were taken from the ear vessels of eight male and eight female pups (aged 13–14 d) immediately before and 2 h after they were removed from their colony and were placed singly into a novel environment. Furthermore, eight male and eight female pups were tested in their home colony before and after 2 h of sepration from their mothers. Blood samples were taken from control animals (eight male and eight female pups) which remained in the home colonies together with their mothers. Additionally, the behaviour of 16 male and 16 female pups was recorded in their home colonies when their mothers were either present or absent. Male and female pups separated from their mothers showed a significantly higher locomotor activity, with females showing higher frequencies of distress calls, than pups whose mothers were present. The mother's absence did not cause a significant increase in serum cortisol concentrations in the male or the female offspring. This result is entirely different from the endocrine stress response of pups that were placed singly in an unknown environment. These animals showed a significant increase in their serum cortisol concentrations after 2 h of separation from their mothers. The absence of the mother led to distinct changes in the behaviour of the guinea‐pig pups. However, staying in the familiar social and spatial environment buffered the endocrine stress response that normally occurs because of maternal separation.     

Malnutrition in utero and lactation: relation between the weight gained by the mothers and the development of their offspring

Physiological weight changes in rat dams and their offspring as sequelae of malnutrition during pregnancy and lactation have been studied. Daily monitoring of the body weight as well as the consumption of food (malnutrition dams 14 g during pregnancy and 20 g during lactation) and drink in control and malnutrition dams was conducted. The number of pregnant dams that completed their pregnant period successfully was registered as well as the number and weight of the pups at birth and their evolution over a period of a month. The percentage of mortality of the pups during that period has been studied. The present results indicate a highly significant decrease in body weight in experimental dams, which determines a retardation in the physiological development of the pups, and yields a higher percentage of mortality in the experimental animals than in controls. It can thus be concluded that malnutrition in utero and during lactation affects the ratio between weight gain for dams and the physical development of their pups.     

A chemically derived milk substitute that is compatible with mouse milk for artificial rearing of mouse pups.

The object of this study was to prepare a chemically derived milk substitute that is compatible with mouse-milk. Milk was independently collected from ICR, BALB/c, and FVB/N mice, and analyzed for the protein, fat, and mineral contents to formulate a milk substitute. Thereafter, ICR mouse pups were artificially reared on the milk substitute to evaluate the rate of increase of their body weights. A gastric cannula tube was placed through the esophageal way into 8-day-old ICR pups, and the mice were fed with the milk substitute by computer-regulated infusion pumping by the pup-in-a-cup method. The analytical mean values of total protein and total fat in milk from ICR, BALB/c, and FVB/N mice were 10.23 +/- 0.49% and 21.34 +/- 1.31%, respectively. The milk substitute was constituted from purified bovine casein and whey proteins, five edible oils, including MCT oil, minerals, and vitamins. After 8 days of artificial rearing with the new milk substitute, 36 of the 42 pups had survived, and the growth rate of these mice was not significantly different from that of maternally reared littermate pups. In conclusion, we have succeeded in the preparation of a chemically derived milk substitute for mice pups which is available for clarifying the roles of dietary components such as milk-bone substance during the suckling period in mice pups including those of knockout and transgenic mice.     

Hormonal correlates of parental behavior in male rats

Groups of seven intact male Long-Evans hooded rats were housed with pups for 1, 7, 14, or 22 days and their parental behavior was recorded each day. All but one male showed parental behavior. Twenty-eight control males were housed without pups. At the end of each observation period the males were decapitated and their trunk blood was collected for hormone analysis by radioimmunoassay. Males with pups had lower prolactin levels than control males on all 4 days but did not differ in leutinizing hormone, follicle-stimulating hormone, or testosterone levels from males without pups. Both prolactin and testosterone levels were correlated positively with the frequency of parental behavior, suggesting that the males which are behaviorally the most responsive to infants are also hormonally the most responsive.     

THE RELIABILITY OF SKINFOLD-CALIPERS FOR MEASURING BLUBBER THICKNESS OF STELLER SEA LION PUPS (EUMETOPIAS JUBATUS)

Twelve dead Steller sea lion pups ( Eumetopias jubatus ) aged 3-14 d were recovered from rookeries in Southeast Alaska. They had a wide range of body sizes and conditions (small to large and fat to no fat). The ability of calipers to estimate the thickness of their blubber layer was assessed with a set of skinfold calipers. Average error of measurement for skin and blubber thickness was an acceptable 5.4%, but the skin and blubber of the pups were highly compressible. Skinfold thickness increased with body mass but did not necessarily reflect the development of blubber, given that pups with no blubber also showed an increase in skinfold thickness with increases in body mass. Skinfold thickness of sea lion pups appears to predict body size better than it predicts blubber thickness, making it difficult if not impossible to develop a simple index of body condition or a calculation of percent body fat for Steller sea lion pups from skinfold caliper measurements.     

Tissue-specific effects of starvation and refeeding on the disposal of oral [1-14C]triolein in the rat during lactation and on removal of litter.

1. The effects of starvation and refeeding on the disposal of oral [14C]triolein between 14CO2 production and 14C-lipid accumulation in tissues of virgin rats, lactating rats and lactating rats with pups removed were studied. 2. Starvation (24 h) increased 14CO2 production in lactating rats and lactating rats with pups removed to values found in virgin rats. This increase was accompanied by decreases in 14C-lipid accumulation in mammary gland and pups of lactating rats and in white and brown adipose tissue of lactating rats with pups removed. 3. Short-term (2 h) refeeding ad libitum decreased 14CO2 production in lactating rats and lactating rats with pups removed, and restored the 14C-lipid accumulation in mammary glands plus pups and in white and brown adipose tissue respectively 4. Insulin deficiency induced with mannoheptulose inhibited the restoration of 14C-lipid accumulation in white adipose tissue on refeeding of lactating rats with pups removed, but did not prevent the restoration of 14C-lipid accumulation in mammary gland. 5. Changes in the activity of lipoprotein lipase in mammary gland and white adipose tissue paralleled the changes in 14C-lipid accumulation in these tissues. 6. It is concluded that 14C-lipid accumulation in mammary gland may not be affected by changes in plasma insulin concentration and that it is less sensitive to starvation than is lipogenesis or lactose synthesis. This has the advantage that the milk lipid content can still be maintained from hepatic very-low-density lipoprotein for a period after withdrawal of food. The major determinant of the disposal of oral 14C-triolein appears to be the total tissue activity of lipoprotein lipase. When this is high in mammary gland (fed lactating rats) or white adipose tissue (fed lactating rats with pups removed), less triacylglycerol is available for the muscle mass and consequently less is oxidized.     

Effects of acute and chronic administration of exorphin C on behavior and learning in white rat pups

We studied the behavior of 21- and 35-day-old white rat pups in the “open field” and the learning of 36- to 41-day-old pups in a maze with food reinforcement. An opioid fragment of wheat gluten exorphin C (YPISL) was injected to pups chronically from day 1 to day 14 of their life or immediately prior to testing. We found that an acute peptide injection did not change animal behavior. The chronic intraperitoneal administration of the peptide at the same dose of 5 mg/kg significantly increased exploratory activity, decreased anxiety, and improved learning. Delayed exorphin C effects were more expressed in female rats.     

哺乳期棕色田鼠对非亲缘幼仔的行为反应

通过观察产后2 日(L2)、6 日(L6)、10 日(L10)、14 日(L14)和20 日(L20)的棕色田鼠母鼠对2 日龄、6 日龄、10 日龄和14 日龄非亲缘幼仔的攻击行为和衔回行为,探讨了哺乳期棕色田鼠的母性行为与非亲缘幼仔的发育(日龄或个体大小)及母鼠产后时间的关系。结果表明,在哺乳前期(L2,L6),非亲缘幼仔的日龄大小显著影响母鼠的攻击行为(P < 0.05);在哺乳中期(L10,L14)及后期(L20),非亲缘幼仔的日龄对母鼠的攻击行为没有显著影响(P > 0. 05)。在整个哺乳期,母鼠对14 日龄非亲缘幼仔的攻击频次最高(P < 0.05)。产后不同时间的母鼠对同一日龄非亲缘幼仔的攻击频次没有显著差异(P > 0.05)。这些结果表明,哺乳期内的棕色田鼠对非亲缘幼仔的攻击与幼仔的发育有关,与母鼠的产后时间无关。     

Demonstration of a quiescent period for feeding controls in the developing rat

Feeding behaviour of rats during development was assessed by weighing pups before and after a 4 h feeding session. During the first postnatal week, fasted pups gained significantly more weight than fed pups. This difference disappeared during the second week, but reappeared during the third week and persisted through the fourth week. In another series, pups were weighed at 2 and 4 h after beginning feeding. This showed that fasted pups aged 6 days compensate by suckling longer than fed pups. At 10 and 14 days of age there were no differences between fed and fasted pups at either 2 or 4 h, but from 16 days onward, fasted pups had eaten significantly more than fed pups at both times. A control experiment showed that the lack of compensation by fasted pups aged 10-14 days did not reflect lack of availability of milk. Video-analysis of suckling behaviour at ages 6, 10 and 15 days provided further evidence for lack of feeding controls during the second postnatal week: at 6 and 15 days fasted pups spent more time actively sucking than did fed pups. Whereas at 10 days, there were no differences between fed and fasted pups. It is concluded that feeding controls are present during the first postnatal week, become quiescent during the second week and reappear during the third week.     

Effect of maternal iron deficiency on GABA shunt pathway of developing rat brain

Iron deficiency during pregnancy and lactation (35 mg iron/kg diet) produced a significant reduction in liver nonheme iron in dams as well as in fetus and young ones. The body, liver and brain weights of fetus, new-born, and developing pups remained unaffected. However, the body weight and PCV were reduced only in 21-day-old pups. The enzyme activities of GDH, GAD, GABA-transaminase, and NAD(+)-linked ICDH were reduced in 14 and 21-day-old pups. The enzyme activities of NADP(+)-linked ICDH activities remained unaffected in the fetus and developing pups brain. Maternal rehabilitation on iron sufficient diet for 1 week from day 14 to 21 of lactation period did not reverse these changes. The maternal iron deficiency during lactation period alone did not cause any alteration in all parameters assayed, however, there was a reduction in liver non-heme iron of pups on days 14 and 21.     

Canine parvovirus enteritis, canine distemper, and major histocompatibility complex genetic variation in Mexican wolves

The endangered Mexican wolf (Canis lupus baileyi) was recently reintroduced into Arizona and New Mexico (USA). In 1999 and 2000, pups from three litters that were part of the reintroduction program died of either canine parvovirus or canine distemper. Overall, half (seven of 14) of the pups died of either canine parvovirus or canine distemper. The parents and their litters were analyzed for variation at the class II major histocompatibility complex (MHC) gene DRB1. Similar MHC genes are related to disease resistance in other species. All six of the surviving pups genotyped for the MHC gene were heterozygous while five of the pups that died were heterozygous and one was homozygous. Resistance to pathogens is an important aspect of the management and long-term survival of endangered taxa, such as the Mexican wolf.     

Energy intake and utilisation by nursing bearded seal (Erignathus barbatus) pups from Svalbard,Norway

In this study we measure energy intake via milk in nursing bearded seal (Erignathus barbatus) pups and determine how this energy is allocated into metabolism and storage of new tissues. This was accomplished using longitudinal mass gain records and the doubly labelled water technique on nursing pups in combination with cross-sectional data on changes in milk composition from bearded seal mothers. The pups (n=3) were all less than a week old at the start of the experiments. Pups gained 3.3±0.4 kg·day^-1 of which 50% was fat, 14% protein and 36% water. Average daily water influx for the pups was 69.5±9.0 ml · kg^-1· day^-1. Average CO₂ production during the study period was 0.99±0.10 ml·g^-1·h^-1, which corresponds to a field metabolic rate of 642±67 kJ·kg^-1· day^-1, or 6.0±0.5 times the predicted basal metabolic rate according to Kleiber (1975). The pups drank an average of 7.6±0.5 kg of milk daily. This corresponds to a daily energy intake of 154±8 MJ, 47±14% of which was stored as new body tissue. Despite this high energy intake bearded seal pups do not get as fat as do other nursing phocids. This is in part due to their larger body size but also due to their very active aquatic lifestyle and the lower and more consistent fat content of the milk compared to other phocid species. Bearded seal mothers forage during lactation and may also be involved in teaching their pups to feed independently. All these data suggest that the lactation strategy of bearded seals differs from the phocid norm.