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PubMed1990

Type VI collagen. In situ hybridizations and immunohistochemistry reveal abundant mRNA and protein levels in human neurofibroma, schwannoma and normal peripheral nerve tissues.

Cutaneous neurofibromas contain an extensive extracellular matrix composed of collagenous and non-collagenous macromolecules. In this study, the expression of type VI collagen genes in cutaneous neurofibromas was examined by a combination of in situ hybridizations and immunohistochemistry. In situ hybridizations with a 32P-labeled human type VI collagen-specific cDNA revealed that the majority of cells within neurofibromas expressed the gene for alpha 2(VI) collagen chain. The number of cells expressing clearly detectable levels of alpha 2(VI) collagen mRNA was considerably higher than that of cells actively expressing the pro alpha 1(I) or pro alpha 1(III) collagen genes. The presence of type VI collagen epitopes within the neurofibromas was also demonstrated by immunostaining with specific polyclonal antibodies. The expression of type VI collagen genes in neural tissues was further examined by immunostaining of a benign schwannoma tissue specimen consisting of Schwann cells. The results indicated close association of type VI collagen epitopes with the neoplastic Schwann cells. Immunolocalization of type VI collagen epitopes within normal human peripheral nerve revealed pericellular staining of perineurial cells and Schwann cells, suggesting synthesis of type VI collagen by these cell types. These results suggest that the expression of type VI collagen gene is active in nerve-derived tissues, and that type VI collagen may be a major component of the extracellular matrix in neural connective tissues.

Collagen↗
PubMedDate not supplied

[Activity of PMSF-inhibited carboxypeptidase in brain tissues and regions of the European hedgehog (Erinaceus europaeus)].

The activity of the PMSF-inhibited carboxypeptidase in different tissues and brain regions of Erinaceus europaeus has been studied. The highest PMSF-inhibited carboxypeptidase activity, has been discovered in the pituitary gland and in some peripheral tissues: tests, lung, kidney, adrenal gland, spleen. The PMSF-inhibited carboxypeptidase activity in the brain regions is approximately two folds as low in the peripheral tissues. The activity of PMSF-inhibited carboxypeptidase in the brain regions also is 3-4 fold as low as that in the pituitary gland. The PMSF-inhibited carboxypeptidase activity in brain regions rich for the neuron bodies is higher than that in the conduction paths. In skeletal muscle and in blood serum the PMSF-inhibited carboxypeptidase activity has not been found.

Animals↗
PubMed1986

Cold exposure reverses the diabetogenic effects of high-fat feeding.

Long-term cafeteria feeding, cold exposure, and the combination of treatments increased energy intake in female Wistar rats by 25%, 113%, and 150%, respectively, in comparison with controls (P less than 0.01). Although cafeteria feeding at room temperature markedly increased the insulin response to an intravenous glucose tolerance test (IVGTT), glucose tolerance was deteriorated (P less than 0.01). In contrast, cold exposure significantly improved glucose tolerance in the presence of a reduced insulin response in Purina- and cafeteria-fed animals. Moreover, cold exposure also decreased body weight gain and increased brown adipose tissue mass, total cytochrome-oxidase activity, and cellularity by approximately 600-800%. The results suggest that cold exposure enhances insulin sensitivity of peripheral tissues, whereas hyperphagia on a high-fat, low-protein diet leads to insulin resistance. In addition, the results demonstrate that prolonged stimulation of energy expenditure by cold exposure not only reverses the diabetogenic effects of cafeteria feeding but also improves glucose tolerance. This phenomenon could result from a combination of two factors: (1) a cold-induced prevention of obesity; and (2) an enhanced disposal of circulating glucose into peripheral tissues, including brown adipose tissue.

Adipose Tissue↗
PubMed1989

Receptor sites for atrial natriuretic factors in brain and associated structures: an overview.

1. Recent data have clearly shown the existence of specific receptor binding sites for atrial natriuretic factors (ANF) or polypeptides in mammalian brain tissues. 2. Ligand selectivity pattern and coupling to cGMP production suggest that brain ANF sites are similar to high-affinity/low-capacity sites found in various peripheral tissues (kidney, adrenal gland, blood vessels). These brain ANF sites possibly are of the B-ANP subtype. 3. High densities of ANF binding sites are found especially in areas of the central nervous system associated with the control of various cardiovascular parameters (such as the subfornical organ and area postrema). However, high densities of sites are also present in other regions such as the hippocampus, cerebellum, and thalamus in the brain of certain mammalian species, suggesting that brain ANF could act as a neuromodulator of noncardiovascular functions. 4. The density of brain ANF binding sites is modified in certain animal models of cardiovascular disorders and during postnatal ontogeny, demonstrating the plasticity of these sites in the central nervous system (CNS). 5. Specific ANF binding sites are also found in various other CNS-associated tissues such as the eye, pituitary gland, and adrenal medulla. In these tissues ANF appears to act as a modulator of fluid production and hormone release. 6. Thus, ANF-like peptides and ANF receptor sites are present in brain and various peripheral tissues, demonstrating the existence of a family of brain/heart peptides.

Animals↗
PubMed1994

Localization of messenger RNA for three distinct alpha 2-adrenergic receptor subtypes in human tissues. Evidence for species heterogeneity and implications for human pharmacology.

BACKGROUND: alpha 2-Adrenergic receptor (alpha 2AR) agonists have become important adjuncts as anesthetic agents. They act by binding to alpha 2ARs on the surface of cell membranes and cause centrally mediated sedation and analgesia. alpha 2ARs also contribute to other aspects of physiologic regulation. Three subtypes of alpha 2ARs (alpha 2-C2, alpha 2-C4, and alpha 2-C10) have been described using molecular and pharmacologic techniques. We recently demonstrated species heterogeneity in the distribution of alpha 1-adrenergic receptor subtypes, therefore making it imperative to analyze the distribution of alpha 2AR subtypes in human tissues. This information may have importance in the understanding of potential side effects of administration of alpha 2AR subtype-selective agonists for anesthesia in humans. METHODS: RNA extracted from human tissues was analyzed by using quantitative ribonuclease protection assays to determine alpha 2AR subtype messenger RNA (mRNA) expression in cardiovascular, central nervous system, and peripheral tissues. RESULTS: alpha 2AR mRNA is present in greatest concentrations in human kidney, followed by aorta > spleen > heart = lung. alpha 2-C4 mRNA predominates in heart, lung, aorta, cerebral cortex, cerebellum, spleen, kidney, and adrenal gland; alpha 2-C2 mRNA in liver; and alpha 2-C10 mRNA in pancreas and small intestine. Hence alpha 2AR subtype mRNA distribution is tissue-selective and differs from that reported for rat. CONCLUSIONS: (1) On comparison with previous research we find possible species heterogeneity in alpha 2AR subtype mRNA distribution (rat vs. human) for all three alpha 2AR subtypes. (2) We demonstrate the presence and subtype heterogeneity of alpha 2AR subtype mRNA in both brain and peripheral tissues. (3) Significant concentrations of alpha 2AR mRNA are present in adult human heart. These findings have important implications for our understanding of human adrenergic physiology, provide a possible explanation for the existence of pharmacologically similar yet distinct alpha 2AR subtypes, and may be important for the rational development of alpha 2AR subtype-selective anesthetics and other therapeutic agents for use in treating human diseases.

Adult↗
PubMed2003

Neurokinin 1 receptor and relative abundance of the short and long isoforms in the human brain.

Substance P exerts its various biochemical effects mainly via interactions through neurokinin-1 receptors (NK1). Recently, the NK1 receptor has attracted considerable interest for its possible role in a variety of psychiatric disorders including depression and anxiety. However, little is known regarding the anatomical distribution of NK1 in the human central nervous system (CNS). Riboprobe in situ hybridization, quantitative PCR and in vitro autoradiography were performed. Highest NK1 mRNA levels were localized in the locus coeruleus and ventral striatum, while moderate hybridization signals were observed in the cerebral cortex (most abundant in the visual cortex), hippocampus and different amygdaloid nuclei. Very low levels of the NK1 mRNA were detected in the cerebellum and thalamus. In view of the existence of a long and short isoform of the NK1 receptor, it was of interest to assess whether there was a differential distribution of the two splice variants in the human CNS and peripheral tissues. A quantitative TaqMan PCR analysis showed that the long NK1 isoform was the most prevalent throughout the human brain, while in peripheral tissues the truncated form was the most represented. 3H-Substance P autoradiography revealed a good correlation between receptor binding sites and NK1 mRNA expression throughout the brain, with the highest levels of binding in the locus coeruleus. These results provide the anatomical evidence that the NK1 receptors have a strong association with neuronal systems relevant to mood regulation and stress in the human brain, but do not suggest a region-specific role of the two isoforms in the CNS.

Adult↗
PubMed1997

Pubertal changes in insulin secretion and peripheral insulin sensitivity.

Using a simple and standardized method we estimated both insulin secretion and insulin sensitivity in peripheral tissues in relation to Tanner pubertal stages. Early insulin response, mean blood glucose (MBG), mean serum insulin (MSI), glucose uptake rate in peripheral tissues and insulin sensitivity index (SI) in response to the standard oral glucose tolerance test were evaluated in 73 normal girls. Study subjects were divided into 4 groups: group 1 (Tanner stage I, n = 20); group 2 (Tanner stage II, n = 14); group 3 (Tanner stages III and IV, n = 15), and group 4 (Tanner stage V, n = 24). Steroid levels and insulin-like growth factors were determined to characterize clinical pubertal development. MBG was similar in all groups but MSI increased at stage II and retained similar values throughout puberty, with those of group I being statistically lower than in the other groups (p < 0.001). When MSI values were adjusted per kilogram of body weight, a significant increase was observed in group II (p < 0.05). The MSI adjusted values were: group 1, 1.0 +/- 0.4; group 2, 1.4 +/- 0.4; group 3, 1.0 +/- 0.3, and group 4, 1.0 +/- 0.4 mU/l/kg. SI values were similar in groups 1 and 2 and significantly lower than in groups 3 and 4 (p < 0.001). Our results confirm both that insulin secretion is related to age and that an insulin-resistant state occurs during puberty. Thus, the insulin-resistant state coincides with Tanner stage II. In conclusion, this mathematical approach is considered to be a simple and reliable method for analyzing the possible alterations in insulin secretion and action in children and adolescents in whom more sophisticated procedures must be limited in this early period of life for ethical reasons.

Adolescent↗
PubMed1996

NTP summary report on the metabolism, disposition, and toxicity of 1,4-butanediol (CAS No. 110-63-4).

1,4-Butanediol is an industrial chemical used in the manufacture of other organic chemicals. It was nominated by the National Cancer Institute and selected for evaluation by the NTP because of high production volume, the potential for worker exposure, the lack of adequate toxicological characterization, and the lack of evaluation for carcinogenic potential. As documented in the scientific literature, 1,4-butanediol is rapidly absorbed and metabolized to gamma-hydroxybutyric acid in animals and humans. A metabolism and disposition study conducted in F344/N rats by the NTP confirmed the rapid and extensive conversion of 1-[14C]-1,4-butanediol to 14CO2. Because of this rapid and extensive conversion, the toxicological profile of 1,4-butanediol reflects that of gamma-hydroxybutyric acid. gamma-Hydroxybutyric acid is a naturally occurring chemical found in the brain and peripheral tissues which is converted to succinate and processed through the tricarboxylic acid cycle. Although the function of gamma-hydroxybutyric acid in peripheral tissues is unknown, in the brain and neuronal tissue it is thought to function as a neuromodulator. gamma-Hydroxybutyric acid readily crosses the blood-brain barrier, and oral, intraperitoneal, or intravenous administration elicits characteristic neuropharmacologic responses. These same responses are observed after administration of 1,4-butanediol. The lactone of gamma-hydroxybutyric acid, gamma-butyrolactone, is also rapidly converted to gamma-hydroxybutyric acid by enzymes in the blood and liver of animals and humans. gamma-Butyrolactone was previously evaluated by the NTP in 14-day and 13-week toxicology studies and 2-year toxicology and carcinogenesis studies in F344/N rats and B6C3F1 mice. No organ-specific toxicity occurred in the toxicology studies. In the carcinogenesis studies, an equivocal response occurred in male mice, based on a marginal increase in the incidence of pheochromocytomas of the renal medulla. Because of the rapid and extensive conversion of gamma-butyrolactone to gamma-hydroxybutyric acid, the evaluation of gamma-butyrolactone was in fact an evaluation of gamma-hydroxybutyric acid. This summary report presents a review of the current literature which documents that both 1,4-butanediol and gamma-butyrolactone are rapidly metabolized to gamma-hydroxybutyric acid, and the pharmacologic and toxicologic responses to these chemicals are due to their metabolic conversion to gamma-hydroxybutyric acid. Because the toxicity and carcinogenicity of gamma-hydroxybutyric acid was fully evaluated in the NTP studies of gamma-butyrolactone, and a lack of organ-specific toxicity or carcinogenic potential was demonstrated, it is concluded that there is a high likelihood that 1,4-butanediol would be negative in a similar set of studies. For these reasons, it is the opinion of the NTP that 1,4-butanediol should be considered not carcinogenic in animals and no further evaluation of 1,4-butanediol is needed at this time.

Animals↗
PubMed2006

Lysozyme gene expression by hemocytes of Pacific white shrimp, Litopenaeus vannamei, after injection with Vibrio.

The purpose of this study was to quantify the gene expression of lysozyme, an important antibacterial protein produced by shrimp hemocytes, within tissues of Litopenaeus vannamei Boone in response to a pathogen challenge. We quantified lysozyme transcripts with a real-time PCR method and used these data, along with total hemocyte counts, to infer patterns of hemocyte trafficking during the immune response. Transcript expression was detected by in situ hybridization of mRNA in circulating hemocytes, and within tissues with high hemocyte concentrations. Lysozyme gene expression was monitored in 5 tissues and in circulating hemocytes for 48 h following challenge with the shrimp pathogen Vibrio campbellii Baumann. The results suggest that lysozyme is expressed in most if not all hemocytes in circulation and in peripheral tissues. Injection with V. campbellii produced a significant decrease in transcriptional signal in circulating hemocytes and peripheral tissues 4 h after injection. Over the same early time period lysozyme signal increased significantly in the muscle at the site of injection and remained high for the duration of the time-course, suggesting that hemocytes are recruited to the site of injection early during the course of the immune response. After 4 h, lysozyme signal increased in circulating hemocytes and tissues, with a return to control levels noted for all tissues except the muscle at the site of injection.

Animals↗
PubMed1997

Nervous system-specific expression of a novel serine protease: regulation in the adult rat spinal cord by excitotoxic injury.

A full-length cDNA clone of a previously unidentified serine protease, myelencephalon-specific protease (MSP), has been isolated by using a PCR cloning strategy and has been shown to be expressed in a nervous system and spinal cord-specific pattern. Sequence analysis demonstrated that MSP is most similar in sequence to neuropsin, trypsin, and tissue kallikrein and is predicted to have trypsin-like substrate specificity. MSP mRNA was found to be approximately 10-fold greater in the CNS of the rat and human, as compared with most peripheral tissues, and within the CNS was found to be highest by a factor of four in the medulla oblongata and spinal cord. Levels of mRNA encoding tissue plasminogen activator (tPA) also were elevated in the spinal cord but were more widespread in peripheral tissues as compared with MSP. In the adult rat lumbosacral spinal cord, in situ localization of MSP mRNA demonstrated 2-fold higher levels in the white, as compared with the gray, matter. MSP mRNA expression was shown to increase 3-fold in the white matter and 1.5-fold in the gray laminae at 72 hr after intraperitoneal injection of the AMPA/kainate glutamate receptor-specific agonist, kainic acid (KA). MSP mRNA remained elevated in the ventral gray matter, including expression associated with the motor neurons of lamina IX, at 7 d after the initial excitotoxic insult. Together, these observations indicate that MSP is in a position to play a fundamental role in normal homeostasis and in the response of the spinal cord to injury.

Amino Acid Sequence↗
PubMedDate not supplied

Isolation and determination of tissue spermidine using Bio-Rex 70.

Bio-Rex 70, a weak cation exchange resin, has been used to extract spermidine from rat tissues. This method compares favourably to other extraction procedures with respect to selectivity, reproducibility and time needed to perform the procedure and, because it combines the optimum of these procedures, it appears more adaptable for routine laboratory use. Following extraction, spermidine is quantified fluorometrically after condensation with o-phthaldialdehyde (OPT). The sensitivity of the present procedure allows for the chromatographic isolation of 20 ng spermidine to give fluorescence twice that of blank. The procedure is selective for spermidine because the fluorescence of other OPT-reactive substances has been eliminated by either the extraction or the assay procedures. The stability and reproducibility of the adsorption and elution characteristics of Bio-Rex 70 enables the determination of 30 tissue samples in a working day. This procedure has been used to determine the levels of spermidine in whole and regional brain areas. In addition, the concentration of spermidine in peripheral tissues was also determined. Of the brain regions, the medulla oblongata/pons had the highest level of spermidine (68.24 +/- 7.67 microgram/g), and in peripheral tissues the highest level of this polyamine was found in the heart (48.59 +/- 4.32 microgram/g).

Animals↗
PubMed1993

Expression of NGF in sympathetic neurons leads to excessive axon outgrowth from ganglia but decreased terminal innervation within tissues.

The effects of nerve growth factor (NGF) on sympathetic axon growth were investigated by generating transgenic mice in which the beta subunit of NGF was expressed in sympathetic neurons using the human dopamine beta-hydroxylase (DBH) promoter. In DBH-NGF mice, the sympathetic trunk and nerves growing to peripheral tissues were enlarged and contained an increased number of sympathetic fibers. Although sympathetic axons reached peripheral tissues, terminal sympathetic innervation within tissues was decreased in DBH-NGF mice. This effect could be reversed in the pancreas by overexpression of NGF in pancreatic islets. The observations are consistent with a model in which NGF gradients are not required to guide sympathetic axons to their targets, but are required for the establishment of the normal density and pattern of sympathetic innervation within target tissues.

Adrenal Glands↗
PubMed1982

Treponema pallidum in peripheral nerve tissue of syphilitic chancres.

Biopsies for electron microscopy were obtained from syphilitic chancres of 7 patients, 6 males and one female. In 4 of the patients, Treponemata pallida were seen gathering around peripheral nerves and invading the spaces between Schwann cells and their basal lamina. No definite degenerative changes were seen in the axons. In all patients, Treponemata pallida were gathering around the small blood vessels. The fine structure of Treponema pallidum in the lesions included a twined cytoplasmic cylinder with an inner and an outer lamina, an axial fibril consisting of three to five parallel filaments, each consisting of six microtubules, and a covering, outpouching periplastic membrane.

Adult↗
PubMed1992

[Glucose transporters. Physiology and physiopathology].

Glucose transport is an important step in the regulation of glucose homeostasis. Two types of transport systems are described: active transport accumulates glucose in specific cells, whereas facilitative transport equilibrates blood glucose and intracellular glucose inside all mammalian cells. At the present time, different levels of facilitative transport regulation are known. Facilitative transport is achieved by 5 different isoforms; each isoform has its own characteristics and is subjected to tissue-specific regulation. Alteration of glucose transporters expression may be involved in a physiopathological situation such as diabetes which is characterized by insulin resistance of peripheral tissues and impaired insulin secretion by beta pancreatic cells. Thus, Glut 2 expression is reduced in the beta cells of diabetic rats. The reduction of Glut 2 expression correlates with, and may contribute to the loss of glucose-stimulated insulin secretion. However, Glut 2 expression in liver remains unaltered. The insulin resistance of peripheral tissues may be explained in adipose tissue by a decrease in Glut 4 expression. In skeletal muscle, Glut 4 expression remains constant whatever the physiological or physiopathological situation.

Biological Transport, Active↗
PubMed2002

Effect of treatment with different doses of 17-beta-estradiol on the insulin receptor.

The mechanism for the development of insulin resistance in normal pregnancy is complex and is associated with serum levels of sex hormones. However, the influence of these hormones on the early steps of insulin action has not been extensively studied, although the potentially beneficial effect of estradiol on glucose homeostasis has been reported. In this paper, we attempted to determine the effect of 17-beta-estradiol on the insulin receptor of ovariectomized rats treated with different doses of hormones. Our results showed a tissue-dependent response to estradiol. We found that low doses of estradiol increased the amount of insulin receptors in liver and muscle on days 6 and 11 of treatment but not in adipose tissue, and after 16 days only the muscle responsed in this way. On the other hand, high doses of estradiol significantly decreased the amount of insulin receptors, at least in muscle and adipose tissue. We believe that the low concentrations of 17-beta-estradiol (similar to early pregnancy) could be responsible for the increase in insulin sensitivity by increasing the amount of insulin receptors in peripheral tissues. When the hormone levels were high (similar to late pregnancy) the amount of insulin receptors decreased in peripheral tissues, and insulin sensitivity is diminished just as in late pregnancy. The specific molecular mechanism for this action is as yet unknown.

Adipose Tissue↗
PubMed2002

[Adipose tissue within peripheral nerves. Study of the human sciatic nerve].

AIM: To describe the distribution of intraneural adipose cells in relation to nerve fascicles in a portion of peripheral nerve usually involved in accomplishing an anesthetic blockade of a lower extremity. METHOD: Using a scanning electron microscope, we studied sciatic nerve samples from the point of amputation of a lower limb of three patients. The samples were obtained at the upper angle of the popliteal fossa, 10-15 cm cephalad to the knee joint line. RESULTS: During dissection of the sciatic nerve samples, we observed a solitary trunk, but examination of the cross-sections under the microscope revealed the components of two clearly separated branches joined by supporting tissue. The sciatic nerve had an oval form in the portion under study, measuring 6.5 to 7.5 mm by 3.6 to 3.9 mm. Between the fascicles, the adipose tissue varied in thickness from 0.5 mm in the central zones to 0.2 mm in the peripheral zones. The adipocytes, which were all similar in the size with diameters of 40 microns, were empty, as a result of elimination of the lipid vacuoles during fixation. The adipose tissue was distributed inside the epineurium to surround isolated fascicles or groups of fascicles. CONCLUSIONS: The adipose tissue inside a nerve surrounded the fascicles to form adipose sheaths that separated the fascicles from one another. The thicknesses of these adipose sheaths varied from one fascicle to another. Cells join to make it possible to create a compact adipose sheet that can delay the diffusion of local anesthetic injected near a nerve and that can therefore interfere with the characteristics of an anesthetic blockade.

Adipose Tissue↗