Activation of locus coeruleus neurons by peripheral stimuli: modulation by a collateral inhibitory mechanism.

Noradrenergic neurons of the rat locus coeruleus (LC) respond to noxious stimuli or peripheral nerve stimulation with a burst of spikes followed by a period of suppressed activity. During this period of post-activation suppression, responses to additional stimuli were attenuated. After antidromic activation of the LC there was also a period of reduced responsivity, presumably mediated by inhibitory recurrent LC collaterals. The suppression of LC unit firing which follows nerve stimulation was reduced by piperoxane, an α-adrenergic antagonist which is known to block the norepinephrine-mediated autoinhibitory action of recurrent LC axon collaterals. The specificity of piperoxane in blocking norepinephrine was shown by the fact that it did not antagonize several other putative transmitters in the LC (i.e., GABA, glycine, and met-enkephalin). It is concluded that the post-activation reduction of LC neuronal responsivity may be mediated in part through noradrenergic autoinhibitory mechanisms within the LC.     

Specificity of coenzyme analogues and fragments in promoting or impeding the refolding of clostridial glutamate dehydrogenase

NAD+ facilitates high-yield reactivation of clostridial glutamate dehydrogenase (GDH) after unfolding in urea. The specificity of this effect has been explored by using analogues and fragments of NAD+. The adenine portion, unlike the nicotinamide portion, is important for reactivation. Alteration in the nicotinamide portion, in acetylpyridine adenine dinucleotide, has little effect, whereas loss of the 6-NH2 substitution on the adenine ring, in 6-deamino NAD, diminishes the effectiveness of the nucleotide in promoting refolding. Also ADP-ribose, lacking nicotinamide, promotes reactivation whereas NMN-phosphoribose, lacking the adenine, does not. Of the smaller fragments, those containing an adenosine moiety, and especially those with one or more phosphate groups, impede the refolding ability of NAD+, and are able to bind to the folding intermediate though unable to facilitate refolding. These results are interpreted in terms of the known 3D structure for clostridial glutamate dehydrogenase. It is assumed that the refolding intermediate has a more or less fully formed NAD+-binding domain but a partially disordered substrate-binding domain and linking region. Binding of NAD+ or ADP-ribose appears to impose new structural constraints that result in completion of the correct folding of the second domain, allowing association of enzyme molecules to form the native hexamer.     

Inhibition of myostatin in adult mice increases skeletal muscle mass and strength

A human therapeutic that specifically modulates skeletal muscle growth would potentially provide a benefit for a variety of conditions including sarcopenia, cachexia, and muscular dystrophy. Myostatin, a member of the TGF-beta family of growth factors, is a known negative regulator of muscle mass, as mice lacking the myostatin gene have increased muscle mass. Thus, an inhibitor of myostatin may be useful therapeutically as an anabolic agent for muscle. However, since myostatin is expressed in both developing and adult muscles, it is not clear whether it regulates muscle mass during development or in adults. In order to test the hypothesis that myostatin regulates muscle mass in adults, we generated an inhibitory antibody to myostatin and administered it to adult mice. Here we show that mice treated pharmacologically with an antibody to myostatin have increased skeletal muscle mass and increased grip strength. These data show for the first time that myostatin acts postnatally as a negative regulator of skeletal muscle growth and suggest that myostatin inhibitors could provide a therapeutic benefit in diseases for which muscle mass is limiting.     

New observations on the fine structure of the liver in goldfish (Carassius auratus)

Summary A re-examination of goldfish liver was made through the use of SEM of fractured samples and TEM of ultrathin-sections and freeze-etch replicas. Several new hepatic fine structures described in the present study are morphologically similar to those reported previously in many higher vertebrates including mammals. Hepatic sinusoids of goldfish contain fenestrations which are arranged into sieve plates. Although the hepatic plates are made up of two layers of hepatocytes, the parenchymal cells of goldfish liver are morphologically similar to mammalian hepatocytes, particularly with respect to the sinusoidal surfaces which are studded with numerous microvilli. The intercellular surfaces of hepatocytes have both nexus and desmosomal junctions, similar to those found in various epithelial cells of higher vertebrates, as cell attachments and communication foci. Tight junctions are found mainly between the openings of the intracellular bile canaliculi and the intralobular bile ductules which are situated in the center of the bicellular hepatic plate.Supported in part by Grants # GM92 and ES07017     

Iatrogenic molar borings in 18th and early 19th century Native American dentitions

Six iatrogenic dental borings were identified in four individuals of a Native American skeletal collection from an 18th and early 19th century Middle Columbia River burial site. The borings, all in maxillary first molars with severe dental attrition and secondary dentin, demonstrate striated walls and associated periapical alveolar lesions. An ethnographic review of the subsistence pattern during the burial period indicates a diet that is consistent in dental attrition with other riverine fisher-hunter-gathers. Histological changes of dental pulp tissue during the process of attrition may result in dental necrosis. Access into the pulp chamber is a technique used to drain necrotic fluid. A common Euro-American therapeutic dental practice of the 18th and 19th centuries for diseases of the pulp was dental extraction. Multiple dental borings indicate that the practice of molar drilling into the pulp chamber was an effective and independent technique used by the Wishram and Wasco people.     

The fine structure of the pit connections ofBatrachospermum sirodotii skuja

Summary A new type of pit connection is described in the fresh water red algaBatrachospermum sirodotii and reported inTuomeya sp. consisting of a doughnut-shaped pit ring with a central pore that is occluded by a rivet-shaped pit plug. This structure occurs in pit connections between cells of the indeterminant axes and also cells of whorled assimilatory branches of limited growth, the pleuridia, but not between axial cells and the basal cells of the pleuridia, the periaxial cells. The pit plug and pit ring ultimately break down in pit connections of the main axis reopening the septal pore to intercellular cytoplasmic connection between vegetative axial cells. A similar breakdown of pit connections may also occur in the pleuridia. The functioning of a truncated cone-shaped network of endoplasmic reticulum in the development of the pit connection is described.This report represents a portion of a dissertation submitted in partial fulfillment of the requirements of a Doctor of Philosophy degree, University of North Carolina, Chapel Hill, NC.     

Severe deficits in 5-HT2A -mediated neurotransmission in BDNF conditional mutant mice

BDNF is thought to provide critical trophic support for serotonin neurons. In order to determine postnatal effects of BDNF on the serotonin system, we examined a line of conditional mutant mice that have normal brain content of BDNF during prenatal development but later depletion of this neurotrophin in the postnatal period. These mice show a behavioral phenotype that suggests serotonin dysregulation. However, as shown here, the presynaptic serotonin system in the adult conditional mutant mice appeared surprisingly normal from histological, biochemical, and electrophysiological perspectives. By contrast, a dramatic and unexpected postsynaptic 5-HT2A deficit in the mutant mice was found. Electrophysiologically, serotonin neurons appeared near normal except, most notably, for an almost complete absence of expected 5-HT2A -mediated glutamate and GABA postsynaptic potentials normally displayed by these neurons. Further analysis showed that BDNF mutants had much reduced 5-HT2A receptor protein in dorsal raphe nucleus and a similar deficit in prefrontal cortex, a region that normally shows a high level of 5-HT2A receptor expression. Recordings in prefrontal slice showed a marked deficit in 5-HT2A -mediated excitatory postsynaptic currents, similar to that seen in the dorsal raphe. These findings suggest that postnatal levels of BDNF play a relatively limited role in maintaining presynaptic aspects of the serotonin system and a much greater role in maintaining postsynaptic 5-HT2A and possibly other receptors than previously suspected.     

Antagonistic Roles of Neurofilament Subunits NF-H and NF-M Against NF-L in Shaping Dendritic Arborization in Spinal Motor Neurons

Dendrites play important roles in neuronal function. However, the cellular mechanism for the growth and maintenance of dendritic arborization is unclear. Neurofilaments (NFs), a major component of the neuronal cytoskeleton, are composed of three polypeptide subunits, NF-H, NF-M, and NF-L, and are abundant in large dendritic trees. By overexpressing each of the three NF subunits in transgenic mice, we altered subunit composition and found that increasing NF-H and/or NF-M inhibited dendritic arborization, whereas increasing NF-L alleviated this inhibition. Examination of cytoskeletal organization revealed that increasing NF-H and/or NF-M caused NF aggregation and dissociation of the NF network from the microtubule (MT) network. Increasing NF-H or NF-H together with NF-M further reduced NFs from dendrites. However, these changes were reversed by elevating the level of NF-L with either NF-H or NF-M. Thus, NF-L antagonizes NF-H and NF-M in organizing the NF network and maintaining a lower ratio of NF-H and NF-M to NF-L is critical for the growth of complex dendritic trees in motor neurons.     

MELATONIN: DEACETYLATION TO 5-METHOXYTRYPTAMINE BY LIVER BUT NOT BRAIN ARYL ACYLAMIDASE

Abstract— Rat liver and brain slices were incubated in vitro with [³H]melatonin. Liver slices synthesized small amounts of [³H]5-methoxyindoleacetic acid ([³H]5-MIAA) along with other melatonin metabolites including 6-hydroxymelatonin. Pretreatment of animals prior to killing with the irreversible monoamine oxidase inhibitor pargyline allowed [³H]5-methoxytryptamine ([³H]5-MT) to be recovered from the incubation. No [³H]5-MIAA or [³H]5-MT could be detected in incubations with hypothalamic slices or following intraventrieular injection of [³H]melatonin. The possibility that the deacetylase aryl acylamidase was in part responsible for the deacetylation occurring in liver slices was examined. Liver aryl acylamidase was able to utilize [³H]melatonin as substrate to produce [³H]5-MT. Furthermore, the liver enzyme was inhibited by melatonin ( K_i. 1 m m ) when tested with the alternate substrate o -nitroacetanalide. Brain aryl acylamidase did not generate any detectable [³H]5-MT nor was it inhibited by melatonin. These results suggest that 5-MT is not formed in brain from melatonin although trace amounts of 5-MT in the periphery could be derived from this precursor.