Postnatal Protein Deprivation Research Articles

Effects of pre- and postnatal protein deprivation and postnatal refeeding on atrial natriuretic peptide (ANP) granules of auricular cardiocytes

Background & aims: Atrial natriuretic peptide (ANP-granules) of auricular cardiocytes was examined by morphometry in undernourished and refed rats. Methods: The study was conducted on 21 and 42-day-old rats from mothers exposed to 73% protein restriction during the gestation and lactation with and without refeeding and on age-matched control animals. The body and the heart were weighted and the ANP-granules are located by transmission electron microscopy. Results: Protein deprivation in the 42 postnatal days reduced the body weights and the heart weights of the pups. Refeeding did not restore body weight or heart weight to control. In the undernourished group, the number of ANP-granules/field (27.3 ± 6.22) was significantly (p < 0.05) greater at 42 days than in the control (10.5 ± 1.89). In the refed group, the number of ANP-granules was also significantly greater, than that in the group on undernutrition. The diameter of ANP-granules in the undernourished animals was similar to the refed rats at 42 days and significantly greater than in the control (p < 0.05). Conclusion: Refeeding between 21 and 42 days increased the number of ANP-granules and maintained their diameter even when there is a reduction in heart weight caused by protein deprivation. © 2010 European Society for Clinical Nutrition and Metabolism.

Effects of perinatal protein deprivation and recovery on esophageal myenteric plexus

To evaluate effects of pre- and postnatal protein deprivation and postnatal recovery on the myenteric plexus of the rat esophagus. Three groups of young Wistar rats (aged 42 d) were studied: normal-fed (N42), protein-deprived (D42), and protein-recovered (R42). The myenteric neurons of their esophagi were evaluated by histochemical reactions for nicotinamide adenine dinucleotide (NADH), nitrergic neurons (NADPH)-diaphorase and acetylcholinesterase (AChE), immunohistochemical reaction for vasoactive intestinal polypeptide (VIP), and ultrastructural analysis by transmission electron microscopy. The cytoplasms of large and medium neurons from the N42 and R42 groups were intensely reactive for NADH. Only a few large neurons from the D42 group exhibited this aspect. NADPH detected in the D42 group exhibited low reactivity. The AChE reactivity was diffuse in neurons from the D42 and R42 groups. The density of large and small varicosities detected by immunohistochemical staining of VIP was low in ganglia from the D42 group. In many neurons from the D42 group, the double membrane of the nuclear envelope and the perinuclear cisterna were not detectable. NADH and NADPH histochemistry revealed no group differences in the profile of nerve cell perikarya (ranging from 200 to 400 microm(2)). Protein deprivation causes a delay in neuronal maturation but postnatal recovery can almost completely restore the normal morphology of myenteric neurons.

The relative effects of severe burn injury and pre- and post-natal protein deprivation on mandibular condyle morphology

The mandible has a mixed embryological origin, and its growth is associated with the secondary cartilage of the condyle process (CP). In this area, growth depends on an array of intrinsic and extrinsic factors that influence protein metabolism. In the present study, we used an adolescent rat model to evaluate the growth and development of the CP under conditions of pre- and postnatal protein deficiency, combined with or without the stress of severe burn injury (BI). We found that protein deficiency severely undermined the growth of the CP, by altering the thickness of its constituent layers. BI is also capable of affecting CP growth, although the effect is less severe than protein deficiency. Interestingly, the summed effect of protein deficiency and BI on the CP is less severe than protein deficiency alone. A possible explanation is that the increased carbohydrates in a hypoproteic diet stimulate the production of endogenous insulin and protein synthesis, which partially compensates for the loss of lean body mass caused by BI.

Immunoreactive vasoactive intestinal polypeptide and vasopressin cells after a protein malnutrition diet in the suprachiasmatic nucleus of the rat

The aim of the present study was to evaluate the effects of prenatal and postnatal protein deprivation on the morphology and density of vasopressin (VP) and vasoactive intestinal polypeptide (VIP) immunoreactive neurons in the suprachiasmatic nucleus (SCN) of young rats. Female Wistar rats were fed either 6% (malnourished group) or 25% (control group) casein diet five weeks before conception, during gestation and lactation. After weaning, the pups were maintained on the same diet until sacrificed at 30 days of age. The major and minor axes, somatic area and the density of VP- and VIP-immunoreactive neurons were evaluated in the middle sections of the SCN. The present study shows that chronic protein malnutrition (ChPM) in VP neurons induces a significant decrease in number of cells (-31%,) and a significant increase in major and minor axes and somatic area (+12.2%, +21.1% and +15.0%, respectively). The VIP cells showed a significant decrease in cellular density (-41.5%) and a significant increase in minor axis (+13.5%) and somatic area (+10.1%). Our findings suggest that ChPM induces abnormalities in the density and morphology of the soma of VP and VIP neurons. These alterations may be a morphological substrate underlying circadian alterations previously observed in malnourished rats.

Effects of pre- and postnatal protein deprivation on atrial natriuretic peptide- (ANP-) granules of the right auricular cardiocytes

The atrial natriuretic peptide (ANP) is a peptide hormone that is mainly produced in the cardiocytes of the atria and auricles, where it is stored within secretory granules. The aim of the present work was to analyze the effects of pre- and postnatal protein malnutrition on weight gain, the size of the heart and the number and sizes of the ANP-granules in the cardiocytes of the rat's right auricle. This study was conducted on 21-day-old rats from mothers exposed to 73% protein restriction during the gestation and lactation and on age-matched control animals. At this stage, both control and protein-deprived animals were killed by a lethal intraperitonial injection of sodic pentobarbital. The weight and the size of the heart were determined and ANP-granules of auricular cardiocytes were examined by transmission electron microscopy and ultrastructural morphometry. Protein deprivation of the mother throughout pregnancy, and the mother and unweaned rat pups in the first 21 postnatal days reduced the weights of pups to about 60% of the normally fed group. The weight and size of the heart were also reduced, by about 50%. Despite this, perinatal malnutrition did not significantly affect the numbers of ANP-granules/field in the cardiocytes. However, there was a small but significant reduction in the sizes of granules. It is concluded that ANP-granules are protected from loss even when there is a substantial reduction in body weight and organ size caused by protein deprivation.

Effects of combined pre- and post-natal protein deprivation on the myenteric plexus of the esophagus of weanling rats: A histochemical, quantitative and ultrastructural study

To evaluate the effects of protein deprivation on the myenteric plexus of the esophagus of weanling rats. Pregnant female Wistar rats were divided into 2 groups: nourished (N), receiving normal diet, and undernourished (D), receiving a protein-deprived diet, which continued after birth. At twenty-one days of age, 13 esophagi from each group were submitted to light microscopy and morphometrical analysis employing the NADH diaphorase, NADPH diaphorase and acetylcholinesterase techniques. Three other esophagi from each group were evaluated with transmission electron microscopy (TEM). In both the NADH- and the NADPH-reactive mounts, the neurons of the N mounts were more intensely stained, while in the D esophagi only the larger neurons were reactive. Many myenteric neurons of N were intensely reactive for AChE activity but only a few neurons of D exhibited these aspects. Ultrastructural analysis revealed that the granular reticulum of N showed large numbers of ribosomes aligned on the outer surface of its regularly arranged membrane while the ribosomes of D were disposed in clusters. The chromatin was more homogeneously scattered inside the neuron nucleus of N as well as the granular component of the nucleolus was evidently more developed in this group. Statistically significant differences between N and D groups were detected in the total estimated number of neurons stained by the NADPH technique. The morphological and quantitative data shows that feeding with protein-deprived diet in 21-d old rats induces a delay in the development of the myenteric neurons of the esophagus.

Effects of pre- and postnatal protein deprivation and postnatal refeeding on myenteric neurons of the rat small intestine: A quantitative morphological study

We investigated weight gain, the size of the small intestine and numbers and sizes of enteric neurons in rats whose mothers had been deprived of protein during pregnancy and who themselves were deprived postpartum. Postnatally, protein deprivation was for 42 days, or for 21 days with refeeding for a further 21 days. Control animals received normal nourishment. Neurons were located by nicotinamide adenine dinucleotide (NADH) diaphorase staining, by acetylcholinesterase (AChE) activity and immunoreactivity for choline acetyltransferase (ChAT). The collagen and elastic fibers in the myenteric ganglia were evaluated histologically. The myenteric ganglia were regular and uniform in the nourished and refed groups. In the undernourished group, the myenteric ganglia were irregularly arranged and the cytoplasm of most of the neurons showed less intense staining for NADH diaphorase, AChE and ChAT. AChE activity and ChAT immunoreactivity showed that most ganglionic neurons were stained in nourished and refed groups, but the neurons of undernourished rats were unstained or moderately stained. The distribution of the connective tissue of the ganglionic capsule was similar in the three groups. There was a decrease in weight of undernourished rats, which was restored in refed rats. The size of the small intestine of the undernourished group was smaller than in the normally fed group, by about 45%, but it was similar in nourished and refed rats. After 42 days of protein deprivation the numbers of neurons that were revealed by NADH diaphorase were fewer than in well nourished rats, but numbers were not different between nourished and refed rats. These observations indicate that protein deprivation alters histological features and acetylcholinesterase activity of neurons and also reduces body weight but these were restored by refeeding.

Effects of Undernutrition on Glycine Metabolism in the Cerebellum of Rats

Undernutrition is a worldwide problem affecting millions of unborn and young children during the most vulnerable stages of brain development. Total restriction of protein during the perinatal period of life can alter the development of the mammalian fetus and have marked repercussions on development of the central nervous system (CNS). The brain is vulnerable to undernutrition with altered morphologic and biochemical maturation, leading to impaired functions. The focus of this study is to investigate [U-¹⁴C]glycine metabolism in undernourished rats submitted to pre- and postnatal protein deprivation (diet: 8% protein with and without addition of L-methionine; control group: 25% protein). Although undernutrition produced a reduction in cerebellar weight and alterations in the DNA concentration, the present study shows that glycine metabolism in this structure is partially protected because the undernourished group with L-methionine did not show modifications in glycine metabolism at all ages studied. However, L-methionine deficiency alters glycine metabolism at 7 and 21 days, but in the adult age both undernourished groups presented no differences in oxidation to CO₂, conversion to lipids and incorporation into protein from glycine, compared to the control group.

Effects of pre- and postnatal protein deprivation and postnatal refeeding on myenteric neurons of the rat large intestine: a quantitative morphological study.

We investigated the effect of protein deprivation and refeeding on weight gain, the size of the colon, and the numbers and sizes of enteric neurons. Neurons were located by reduced nicotinamide adenine dinucleotide (NADH) diaphorase staining. Protein deprivation of the mother throughout pregnancy, and the mother and unweaned rat pups in the first 21 postnatal days, reduced the weights of pups to about 50% of control. The size of the colon was also reduced, by about 40%. Despite this, total numbers of neurons in the colon were not reduced. However, there was a small, but significant, 15% reduction in the areas of neuron profiles. After 21 days the remaining pups were removed from the mothers, and either maintained on the control diet, maintained on the protein-deprived diet, or changed from the protein-deprived diet to a normal diet (refed group). These rats were examined after a further 21 days. Refeeding restored body weight to 20% below control, restored colon size, and restored nerve cell size. After a total of 42 days of protein deprivation, nerve cell numbers were not significantly different from control. In undernourished rats at 21 and 42 days, neurons were less well stained than control for NADH diaphorase. Refeeding between 21 and 42 days restored the normal appearance of the neurons. It is concluded that enteric neurons are protected from loss even when there is a substantial reduction in body weight and organ size caused by protein deprivation. The neurons become smaller, but recover size after refeeding.

Effect of undernutrition on GMP-PNP binding and adenylate cyclase activity from rat brain.

1. Undernutrition is an insult that affects brain development and functioning. Considering that signaling through metabotropic receptors/G proteins is critical for normal synaptic transmission and contributes to CNS development and synaptic plasticity, the present study investigated the effects of pre- and postnatal protein deprivation (diet: 8% protein; normonourished group: 25% protein) on brain signal transduction by G proteins. 2. Undernutrition decreased the [3H] GMP-PNP binding to G proteins and AC activity, in neural plasma synaptic membranes of 21- and 75-day-old rats. This effect was less pronounced or even absent in old rats. 3. Ontogenetically, the dietary treatment effect might be interpreted as a retarded development associated with protein malnutrition.

Effects of pre- and postnatal protein deprivation on rat's hard palatine mucosae: a scanning electron microscopic study of the connective tissue papillae

The primary (connective tissue papillae) and secondary projections of the subepithelial connective tissue of the lamina propria were studied in the hard palate of normal (control group), and in protein-deficient pups (experimental group) obtained through maternal protein deprivation during pregnancy and lactation. At birth, the primary projections in the control group are more developed in width whereas in the experimental group they are thin with a crest-like shape. On day 7 of lactation, the primary projections are abundant, well developed and regularly arranged in the control group. They are few in number and are irregular in shape and disposition in the experimental group. The secondary projections are located posterior to the primary ones in the control group and are intercalated with them in the experimental group. On day 21 of lactation, the characteristic foliate aspect of the primary projections are noted in the control group. In the experimental group they remain thin with a crest-like aspect.

Effect of protein malnutrition on CA3 hippocampal pyramidal cells in rats of three ages

Prenatal and postnatal protein deprivation effect on CA3 hippocamapal pyramidal pyramidal cells were investigated in 30-, and 90- and 220-day old rats Female rats were fed either a 6% or a 25% casein diet 5 wk before conception and the litters were maintained on their respective diet until sacrificed. In 216 rapid Golgi-impregnatd cells, we measured somal size, length and diameter of apical dendrite, number of apical dendrites intersecting 10 concentric rings 38 μm apart, thorny excrescence area and length, head diameter and density of synaptic spines on 50-μm segments of apical dendrite. The present experiments showed that malnutrition produced significant reductions of somal size in animals at 220 days of age. There were significant reductions of apical dendrite diameters in animals of 30 and 90 days, and of density and head diameter of synaptic spines at the three ages studied, and significant decrease of the thorny excrescence area at 220 days of age. At this latter age, dendritic branching was significantly decreased in the last four rings representing the area into which the perforant pathway projects. In 30-day malnourished rats, dendritic branching showed a significant increase in rings 4–6 representing the area in which the Schaffer collaterals synapse. The location of the deficit spines corresponds to the sites where mossy fibers synapse on the apical dendrites of CA3 neurons. Age-related changes normally observed in control rats (e.g., the 30-day-old control group showed the smallest somal size and 220-day-old controls the largest size) failed to occur in the malnourished rats. The deficits in spine density and dendritic branching (in animals of 220 days old) were similar to those found in our previous studies on fascia dentata.

Postnatal protein deprivation in rhesus monkeys

We studied the growth of 97 rhesus monkeys (M. mulatta) that from birth until 120 days of age were fed a diet containing 13.4%, 6.7%, 3.35% protein or a commercial simulated human-milk formula (SMA) affording 9% protein. After 120 days, all animals were fed SMA. Females fed the diet lowest in protein, but not the other diets, were moderately affected, gaining less weight than their well-fed counterparts did, but they recovered their deficit during the repletion period, so that at 240 days no group differences remained. Males fed the two lowest-protein diets were severely and profoundly affected, in keeping with the depth of their protein deficiency. Moreover, unlike the females, they recovered none of their weight deficit during the repletion period. Food intake during deprivation was lower in the animals eating the low-protein diets. During SMA repletion, intakes followed no simple rule, but they did not converge. Except for the males fed the 3.35% protein diet, relative food intake (ml formula/kg body weight) did not differ substantially between diets or sexes at any time during the deprivation and diminished as the animals got older. Those males' relative intakes did not decline. During repletion, the SMA females ate the most in proportion to weight and the 13.4% group the least. The lowest-protein males, in contrast, ate the most in proportion to their weight during both deprivation and repletion. Males fed the lowest-protein diet gained little for what they ate; those fed the highest-protein diet gained much. Females were more efficient than males were when eating the low-protein diet and less efficient when eating the high-protein diet. When fed SMA during repletion, males' food efficiencies (grams gained/liter of diet) were nearly equal; females still differed: the SMA group was the least efficient and the low-protein group most efficient. Its counterpart, protein efficiency, was greatest (during deprivation) for females eating the low-protein diet and least (among females) for those eating the high-protein diet. Males were least protein efficient if eating the low-protein diet. Evidently, a 4 month bout of protein deprivation had prolonged effects on the amount of food the animals needed to produce a given gain in weight.

Cellular enzymatic differentiation in brain vessels of normal and protein-deprived rats. Biochemical studies.

Brain capillary development was studied in normal and protein-deprived rats using the specific activity of alkaline phosphatase (EC 3.1.3.1) and gamma-glutamyl transpeptidase (EC 2.3.2.1) in whole brain homogenates and microvessels obtained by gradient centrifugation according to Orlowski et al. (1974). Pre- and postnatal protein deprivation was induced by a 50% reduction in the dietary protein content. The density of microvessel fragments changed during development. Most of the early developmental increase in the specific activity of both enzymes in whole brain homogenates of normal rats can probably be explained by a rapid formation of new capillary segments. The increase in specific activity of gamma-glutamyl transpeptidase in microvessels was interpreted as a sign of cellular differentiation. Protein deprivation resulted in reduced specific activity of both enzymes in whole brain homogenates of 30-day-old rats, probably as a result of the decreased length per volume of the cerebral capillary network at this age (Conradi et al. 1979a). Signs of impaired endothelial growth were also present in the protein-deprived rats since the distribution of microvessel fragments in the 30-day-old protein-deprived rats was similar to that in 3-week-old normal rats. The specific activity of alkaline phosphatase was decreased in the microvessel fractions of 30- and 96-day-old protein-deprived rats, apparently signifying an effect of the protein deprivation on the endothelial cells. These effects of protein deprivation on the brain capillary endothelial transport system may have negative consequences for growth and function in the brain.