Morphological studies of the neuromuscular mechanism shifting from sucking to biting of mice

In order to give a neuroanatomical evidence to the mechanism of shifting from sucking to biting, we investigated in prenatal, newborn and postnatal mice whether there is a time difference in the neurogenesis of the neurons relative to sucking and biting or in the histogenesis of their peripheral effector organs by the HRP labeling technique and electron microscopy. The results obtained are as follows. (1) At birth the facial motoneurons exceed the trigeminal motoneurons in cell area and development. (2) After birth, the trigeminal motoneurons grow rapidly and outstrip the growth of the facial motoneurons at the age of 6 days. (3) Thereafter, the cell area of both neuron types continues to increase gradually. (4) The initial sign of the alpha motor end plates is found in the orbicularis oris muscle innervated by the facial nerve in 17-day-old fetuses, while that of the trigeminal nerve is delayed in the masseter muscle of 18-day-old fetuses. (5) The initial sign of the muscle spindle appears with the sensory terminals in the masseter muscle of 17-day-old fetuses and the fundamental structure of the muscle spindle is formed in 4-day-old youngs. (6) Myelination of the facial nerve begins in 3-day-old youngs, while that of the trigeminal nerve becomes apparent in 4- or 5-day-old youngs. From these bases, it is obvious that the facial nerve elements related to sucking are firstly developed at birth and that the differentiation of the trigeminal nerve elements related to biting is rapidly accelerated after birth.     

Postnatal development of the anulospiral endings of Ia fibers in muscle spindles of mice

The formation of the anulospiral ending of Ia fibers in muscle spindles was investigated in the masseter muscle of developing mice. Before 15 days after birth, the complete anulospiral ending was not observed in almost all of the muscle spindles examined. With the growth of mice, the Ia fiber began to construct the spiral ending, and by the 40th postnatal day after weaning, almost all of the Ia fibers of the muscle spindles had complete coiled endings, though the formation still continued in some spindles. The continuous formation of anulospiral endings for a long period after weaning indicates that muscle spindle morphogenesis may be affected by muscle tension in the masseter muscle due to the movement activated after weaning.     

Ontogeny of flight in the little brown bat,Myotis lucifugus: behavior,morphology, and muscle histochemistry

Summary Postnatal changes in wing morphology, flight ability, muscle morphology, and histochemistry were investigated in the little brown bat, Myotis lucifugus. The pectoralis major, acromiodeltoideus, and quadriceps femoris muscles were examined using stains for myofibrillar ATPase, succinate dehydrogenase (SDH), and mitochondrial -glycerophosphate dehydrogenase (-GPDH) enzyme reactions. Bats first exhibited spontaneous, drop-evoked flapping behavior at 10 days, short horizontal flight at 17 days, and sustained flight at 24 days of age. Wing loading decreased and aspect ratio increased during postnatal development, each reaching adult range before the onset of sustained flight. Histochemically, fibers from the three muscles were undifferentiated at birth and had lower oxidative and glycolytic capacities compared to other age groups. Cross-sectional areas of fibers from the pectoralis and acromiodeltoideus muscles increased significantly at an age when dropevoked flapping behavior was first observed, suggesting that the neuromuscular mechanism controlling flapping did not develop until this time. Throughout the postnatal growth period, pectoralis and acromiodeltoideus muscle mass and fiber cross-sectional area increased significantly. By day 17 the pectoralis muscle had become differentiated in glycolytic capacity, as indicated by the mosaic staining pattern for -GPDH. By contrast, the quadriceps fibers were relatively large at birth and slowly increased in size during the postnatal period. Fiber differentiation was evident at the time young bats began to fly, as indicated by a mosaic pattern of staining for myosin ATPase. These results indicate that flight muscles (pectoralis and acromiodeltoideus) are less well developed at birth and undergo rapid development just before the onset of flight. By contrast the quadriceps femoris muscle, which is required for postural control, is more developed at birth than the flight muscles and grows more slowly during subsequent development.     

ACTH/MSH(4-10) peptide increases postnatal metabolic activity of EDL muscle following early prenatal administration

ACTH/MSH(4-10) (10 micrograms/kg/b.i.d.; IP), administered to pregnant Sprague-Dawley rats during gestational days (GD) 3 to 12, significantly increased the metabolic activity of extensor digitorum longus (EDL) muscle at postnatal day 14. ACTH/MSH peptide, administered from day of birth to postnatal day 13, had no effect on EDL muscle metabolic activity using the 2,3,5-triphenyltetrazolium chloride indicator. By postnatal day 30, no differences were seen between the early prenatally treated group and saline controls. These results confirm our previous electrophysiological studies that showed that early prenatal ACTH/MSH(4-10) administration accelerates EDL muscle maturation.     

IGF and myostatin pathways are respectively induced during the earlier and the later stages of skeletal muscle hypertrophy induced by clenbuterol,a β2‐adrenergic agonist

Clenbuterol, a β₂‐adrenergic agonist, increases the hypertrophy of skeletal muscle. Insulin‐like growth factor (IGF) is reported to work as a potent positive regulator in the clenbuterol‐induced hypertrophy of skeletal muscles. However, the precise regulatory mechanism for the hypertrophy of skeletal muscle induced by clenbuterol is unknown. Myostatin, a member of the TGFβ super family, is a negative regulator of muscle growth. The aim of the present study is to elucidate the function of myostatin and IGF in the hypertrophy of rat masseter muscle induced by clenbuterol. To investigate the function of myostatin and IGF in regulatory mechanism for the clenbuterol‐induced hypertrophy of skeletal muscles, we analysed the expression of myostatin and phosphorylation levels of myostatin and IGF signaling components in the masseter muscle of rat to which clenbuterol was orally administered for 21 days. Hypertrophy of the rat masseter muscle was induced between 3 and 14 days of oral administration of clenbuterol and was terminated at 21 days. The expression of myostatin and the phosphorylation of smad2/3 were elevated at 21 days. The phosphorylation of IGF receptor 1 (IGFR1) and akt1 was elevated at 3 and 7 days. These results suggest that myostatin functions as a negative regulator in the later stages in the hypertrophy of rat masseter muscle induced by clenbuterol, whereas IGF works as a positive regulator in the earlier stages. Copyright © 2012 John Wiley & Sons, Ltd.     

Contractile properties of the rat external abdominal oblique and diaphragm muscles during development.

We studied the in vitro contractile and fatigue properties of the rat external abdominal oblique (EAO) and costal diaphragm (DIA) muscles during postnatal development. Isometric twitch contraction (CT) and half-relaxation (RT1/2) times were longer in both the EAO and DIA muscles during the early postnatal period and decreased with age. In the first postnatal week, the CT and RT1/2 were longer in the EAO than the DIA muscle. At 14 days of age and thereafter, the CT and RT1/2 were shorter in the EAO than in the DIA muscle. Force-frequency relationships of the EAO and DIA muscles changed during postnatal development such that the relative force (percent maximum) generated at lower frequencies (less than 15 pulses/s) decreased with age. Moreover the relative force generated by the EAO muscle at lower frequencies was greater than that of the DIA muscle during the early postnatal period but less than that of the DIA muscle in adults. The specific force of both the EAO and DIA muscles increased progressively with age. There were no differences in specific force between the EAO and DIA muscles at any age. The fatigability of the EAO and DIA muscles was comparable during the early postnatal period and increased in both muscles with postnatal development. In adults the EAO muscle was more fatigable than the DIA muscle. We conclude that the contractile and fatigue properties of the EAO and DIA muscles undergo significantly different postnatal transitions, which may reflect their functional involvement in sustaining ventilation.     

Postnatal thyroxine modifies effects of early androgen on lordosis

The sensitivity of female rats to the organizational effects of postnatal androgen was examined after néonatal manipulations known to affect the rate of brain development: thyroxine (T₄) administration and handling at birth. Testosterone propionate (TP) was injected subcutaneously in oil on postnatal day 6 to littermates that (i) had been undisturbed at birth; (ii) had received saline injections (S) and associated handling (H) on the day of birth (postnatal day 1) and the following day; and (iii) had received T₄ in saline (1 μg/g body wt) and H on postnatal days 1 and 2. Estrous cycles at 45 and 90 days of age, ovulation at Day 100 and sexual receptivity (lordosis score) at Days 115 and 125 were used to evaluate changes in TP effects. The majority of animals treated with 100 μg TP on postnatal day 6 exhibited persistent estrus (PE) at 45 and 90 days of age as expected. Neither T₄ nor S pretreatment on postnatal days 1 and 2 changed the incidence of PE. A reduction in ovarian weights and incidence of ovulation at 100 days of age supported cycle data in that only one out of 25 androgenized rats showed ovulation, compared with 16 of 22 controls. At approximately 115 days of age 2 μg of estradiol benzoate were administered for 3 days and 0.5 mg progesterone given on the 4th day 4 hr prior to placing females with sexually vigorous males. T₄-TP females exhibited higher (< .01) median lordosis scores than their S-TP littermates. The latter results were replicated in a second test conducted 10 days later (p = .002). In addition, the second test indicated that the S-TP group had lower (p = .005) lordosis scores than littermates given only TP neonatally. The results of these studies demonstrate that pretreatment with T₄ and handling, which are known, respectively, to hasten and retard the chronology of brain maturation, can exert differential effects on behavioral manifestations of postnatal TP without modifying androgen-induced sterility.     

Changes in protein kinase activity in rat testes during development.

Protein kinase activity in rat testes remained fairly constant from day 16 1/2 of embryonic life up to 10 days after birth. At the 21st postnatal day a nadir of activity was observed, and after an increase at 35 days of age a decrease in activity at 60 days was seen. The enzyme reached maximal specific activity in the testes of 90-day-old rats.     

Masticatory muscles of domestic sheep and swine in ontogenesis

Age changes of morphometrical parameters of the masticatory muscles have been analyzed in domestic sheep and pigs of white large breed in the following age groups: 2-, 3-, 4-month-old fetuses, newborns, 4-month-old lambs, 10-month-old pigs, 18-month-old lambs, mature she-sheep and brood-sows. Uneven weight growth of the masticatory muscles in the sheep and pigs during the prenatal ontogenesis should be considered as a consequence of recapitulation of their phylogenesis, and in the postnatal ontogenesis it depends on changes in life conditions, type of nutrition, character of food and type of life. In newborn sheep the digastric, lateral, pterygoid and temporal muscles grow intensively, and in pigs--medial pterygoid and temporal ones. When they pass to roughage, in the former the mass of the musculus masseter major and medial pterygoid muscle increases, and in the latter--that of the musculus masseter major and temporal one. The masticatory muscles of the species studied increase in their mass especially intensively during the middle of the prenatal ontogenesis and during suckling period of their development. This should be taken into consideration in stock-breeding practice. In domestic pigs there is only one muscular belly in the digastric muscle. In sheep there are two bellies, separated one from another by means of a tendinous intersection, owing to crossing of the latter by the stylohyoid muscle.     

棕色田鼠脑和行为不同发育阶段副嗅球和主嗅球的细胞活动

本文运用免疫组化显示Fos蛋白的方法首次研究了棕色田鼠脑和行为不同发育阶段副嗅球和主嗅球的细胞活动。当不同年龄阶段的幼鼠同时暴露于自己家庭的熟悉底物和另一家庭的陌生底物时 ,嗅闻和呆在自己熟悉底物上的时间较多 ,直到产后 15d、 2 0d和 2 5d时 ,幼鼠探究不同底物的行为显示出显著性差异。脑的大小随着日龄增加而增加 ,但从产后 1到 15d ,脑重、脑宽和嗅球大小随着日龄增加特别显著。当不同日龄幼鼠暴露于陌生底物或者暴露于自己的熟悉底物时 ,从产后 5到 15日龄 ,主嗅球僧帽细胞层、颗粒细胞层、副嗅球僧帽细胞层和颗粒细胞层Fos免疫阳性细胞随着日龄明显增加 ,但直到 15和 30日龄时 ,和对照组相比 ,陌生底物可引起幼鼠主嗅球Fos免疫阳性细胞明显增加 ,从 2 0日龄起 ,陌生底物可引起副嗅球Fos免疫阳性细胞明显增加。主嗅球颗粒细胞层Fos免疫阳性细胞随着日龄的增加从边缘到中心逐渐出现 ,而副嗅球Fos免疫阳性细胞随着日龄的增加从顶部到底部逐渐出现。以上结果说明产后第 1d到 15d左右可能是棕色田鼠脑结构发育的重要阶段 ,而从此以后棕色田鼠主嗅球和副嗅球就具有区别熟悉气味和陌生气味的能力 ,表明棕色田鼠行为、脑发育和细胞活动间有紧密关系     

Resting tension characteristics in differentiating intact rat fast- and slow-twitch muscle fibers.

The postnatal changes in resting muscle tension were investigated at 20 degrees C by using small muscle fiber bundles isolated from either the extensor digitorum longus or the soleus of both neonatal (7-21 days old) and adult rats. The results show that the tension-extension characteristics of the bundles depended on the age of the rats. For example, both the extensor digitorum longus and soleus bundles of rats older than 14 days showed characteristic differences that were absent in bundles from younger rats. Furthermore, the tension-extension relation of the adult slow muscle fiber bundles were similar to those of the two neonatal muscles and were shifted to longer sarcomere lengths relative to those of the adult fast-fiber bundles. Thus, at the extended sarcomere length of 2.9 microm, the adult fast muscle fiber bundles developed higher resting tensions (5.6 +/- 0.5 kN/m2) than either the two neonatal ( approximately 3 kN/m2) or the adult slow (3.1 +/- 0.4 kN/m2) muscle fiber bundles. At all ages examined, the resting tension responses to a ramp stretch were qualitatively similar and consisted of three components: a viscous, a viscoelastic, and an elastic tension. However, in rats older than 14 days, all three tension components showed clear fast- and slow-fiber type differences that were absent in younger rats. Bundles from 7-day-old rats also developed significantly lower resting tensions than the corresponding adult ones. Additionally, the resting tension characteristics of the adult muscles were not affected by chemical skinning. From these results, we conclude that in rats resting muscle tension, like active tension, differentiates within the first 3 wk after birth.     

A Halothane-Related Effect on Rat Brain Myelination: A Comparison of Chronic Prenatal or Postnatal Exposure

Rats were exposed to 0.5% halothane in air for 8 h per day during the intervals (1) 5 days postconception to birth, (2) birth to 5 days postnatal age, or (3) birth to 10 days postnatal age. Controls were exposed to an equivalent flow of air. Prenatal exposure had no significant effect on body or brain weight and no subsequent effect on the relative synthesis of brain subcellular membranes. Five days of postnatal exposure caused a 10% reduction in body and brain weight and a 10% relative reduction in the synthesis of brain myelin. The effect persisted throughout the period of rapid postnatal brain myelination. Ten days of postnatal exposure produced equivalent, more severe effects on body and brain weights and a more severe effect on myelin synthesis. Postnatal exposure had no apparent effect on the relative synthesis of non-myelin particulate proteins.     

Prenatal and postnatal development of the small intestine in the insectivore Suncus murinus

The development of the small intestine in the insectivore Suncus murinus was noted during the period from 21 days' gestation to 20 days after birth. At 21 days of gestation, the proximal small intestine exhibited the beginning of villus formation, whereas the distal small intestine preserved the stratified epithelium. Stratified epithelium in the distal small intestine changed into a single layer by 24 days' gestation. At 26 days' gestation, each epithelial cell was immature; but by 28 days mature-looking epithelial cells were found. The shape of the villi changed from cuboid to columnar during the same period. The connective-tissue cores of the villi began to develop at 7 days after birth in the proximal small intestine and at 15 days after birth in the distal small intestine. Crypts appeared at 15 days after birth. Endocytosis of epithelial cells took place at 28 days of gestation. In the proximal small intestine, supranuclear vesicle clusters were observed first at birth; they began to decrease both in number and size at 10 days' gestation and then disappeared completely by 20 days after birth. In the distal small intestine, large supranuclear vacuoles were observed first at 28 days of gestation. Although these vacuoles invariably were found up to 15 days after birth, they also disappeared completely by 20 days. Epithelial cells showed a structure similar to those of the adult after weaning.     

Quantitative analysis of development of mitochondrial ultrastructure in differentiating mouse hepatocytes during postnatal period

Between birth and 10 days of age, the volume density (volume/unit cytoplasmic volume) of the matrix, and the surface density (area/unit cytoplasmic volume) of the inner membrane and cristae increased in both periportal and perihepatic hepatocytes, and did not differ significantly between the cells of the two zones. After 10 days of age, however, the volume density of the matrix decreased in perihepatic cells and remained unchanged in periportal cells, and, therefore, it became greater in periportal cells than in perihepatic cells in 20-day-old and adult animals. The surface density of the inner membrane and cristae decreased in the cells of both zones. Further, the hepatocyte volume increased markedly, especially in perihepatic zones between 20 days of age and the adult. The results show that, in postnatally differentiating hepatocytes, mitochondria are likely to develop during early postnatal period, then the structural heterogeneity of mitochondria arises, and hepatocyte volume increases markedly during late postnatal period after weaning. Thus, the process of postnatal hepatocyte differentiation includes such several phases of development.     

Severity and timing of early thyroid deficiency as factors in the induction of learning disorders in rats

In Experiment 1 (N = 277 rats), more extreme deficits in maze learning than heretofore shown appeared in the adult offspring of mothers exposed for 16 pre- and 16 postnatal days to thiouracil-treated mash diets in doses up to 0.3%; the same offspring also displayed deficits in single-alternation pattern learning and a modified operant discrimination task. Surprisingly small maze learning deficits, however, were found in the offspring of mothers which received thiouracil during the 16 postnatal days only, despite previous findings indicating that the postnatal half of the total 32-day perinatal period was the more critical in determining later learning impairments. Reconciliation was provided by Experiment 2 (N = 54), in which the manipulation of a 0.2% thiouracil diet starting at birth or 3, 6, 10, or 15 days before birth indicated a lower age boundary of the critical period for the induction of maze learning deficit by thyroid deficiency at approximately the fetal age at which thyroid tissue becomes functional-around the 18th day of gestation.     

Temporal and spatial dimensions of postnatal growth of the mouse urinary bladder urothelium

Postnatal growth and renewal of mouse urothelium start on the day of birth. In the present study, temporal and spatial dimensions of urothelial growth were studied during the first two postnatal weeks. Quantitative analysis showed that the rate of urothelial cell proliferation is significantly higher during all 14 postnatal days than in adult mice. Three peaks of proliferative and mitotic activity were revealed: on the day of birth and postnatal day 1, on days 6 and 7, and on day 14. The high proliferation rate around the day of birth and at postnatal days 6 and 7 coincides with cell death in the urothelium. Semiquantitative analysis showed that during all 14 postnatal days, the urothelial proliferative response is mostly confined to the basal cell layer. Urothelial cells divide predominantly in parallel to the plain of the urothelium on all chosen postnatal days. Increased portions of urothelial cells, dividing perpendicularly to the urothelium were observed only on the day of birth and on postnatal day 7. Our results suggest that postnatal growth of mouse urothelium is particularly the result of an increasing number of cells in individual cell layers and not the result of an increasing number of cell layers.     

Lack of the bone remodeling in osteopetrotic (op/op) mice associated with microdontia.

Osteopetrosis is an inherited metabolic disease characterized by an excessive accumulation of bone. This is associated with an osteoclast deficiency. Osteopetrosis is always accompanied by the failure and/or delay of tooth eruption. The present study was conducted to examine in detail the morphological and histological changes of growth of the third molars in the osteopetrosis (op/op) mouse. At the age of 10 days, normal and op/op mice showed no detectable difference in the shape of the third molar follicles. However, the third molars in the op/op mouse became obscured by the proliferation of neighboring bone trabeculae. Moreover, no tartrate-resistant acid phosphatase-positive cells were detected on the bone surfaces of 10-day-old op/op mice. Ankylosis between the root dentin and proliferating bone trabeculae was a common feature in the 20- and 30-day-old op/op mice. The third molars erupted into the oral cavity before the age of 30 days in normal mice, and the crowns, roots, and periodontal ligaments appeared well developed. Throughout the experiment, it seemed that the primary cause of the microdontia and ankylosis of the developing root in the mutant mouse was a deficiency of osteoclasts, with attendant lack of bone remodeling.     

Sedentary behavior during postnatal life is determined by the prenatal environment and exacerbated by postnatal hypercaloric nutrition

The discovery of a link between in utero experience and later metabolic and cardiovascular disease is one of the most important advances in epidemiology research of recent years. There is now increasing evidence that alterations in the fetal environment have long-term consequences on metabolic and endocrine pathophysiology in adult life. This process has been termed "fetal programming," and we have shown that undernutrition of the mother during gestation leads to obesity, hypertension, hyperphagia, hyperinsulinemia, and hyperleptinemia in offspring. Using this model of maternal undernutrition throughout pregnancy, we investigated whether prenatal influences may lead to alterations in postnatal locomotor behavior, independent of postnatal nutrition. Virgin Wistar rats were time mated and randomly assigned to receive food either ad libitum (ad libitum group) or at 30% of ad libitum intake (undernourished group). Offspring from UN mothers were significantly smaller at birth than AD offspring. At weaning, offspring were assigned to one of two diets [control or hypercaloric (30% fat)]. At ages of 35 days, 145 days, and 420 days, voluntary locomotor activity was assessed. At all ages studied, offspring from undernourished mothers were significantly less active than offspring born of normal birth weight for all parameters measured, independent of postnatal nutrition. Sedentary behavior in programmed offspring was exacerbated by postnatal hypercaloric nutrition. This work is the first to clearly separate prenatal from postnatal effects and shows that lifestyle choices themselves may have a prenatal origin. We have shown that predispositions to obesity, altered eating behavior, and sedentary activity are linked and occur independently of postnatal hypercaloric nutrition. Moreover, the prenatal influence may be permanent as offspring of undernourished mothers were still significantly less active compared with normal offspring at an advanced adult age, even in the presence of a healthy diet throughout postnatal life.     

Diaphragm muscle fatigue resistance during postnatal development.

Changes in the contractile and fatigue properties of the cat diaphragm muscle were examined during the first 6 wk of postnatal development. Both twitch contraction time and half-relaxation time decreased progressively with age. Correspondingly, the force-frequency curve was shifted to the left early in development compared with adults. The ratio of peak twitch force to maximum tetanic force decreased with age. Fatigue resistance of the diaphragm was highest at birth and then progressively decreased with age. At birth, most diaphragm muscle fibers stained darkly for myofibrillar adenosinetriphosphatase after alkaline preincubation and thus would be classified histochemically as type II. During subsequent postnatal development, the proportion of type I fibers (lightly stained for adenosinetriphosphatase) increased while the number of type II fibers declined. At birth, type I fibers were larger than type II fibers. The size of both fiber types increased with age, but the increase in cross-sectional area was greater for type II fibers. On the basis of fiber type proportions and mean cross-sectional areas, type I fibers contributed 15% of total muscle mass at birth and 25% in adults. Thus postnatal changes in diaphragm contractile and fatigue properties cannot be attributed to changes in the relative contribution of histochemically classified type I and II fibers. However, the possibility that these developmental changes in diaphragm contractile and fatigue properties correlated with the varying contractile protein composition of muscle fibers was discussed.