The influence of non-neuronal cells on catecholamine and acetylcholine synthesis and accumulation in cultures of dissociated sympathetic neurons

The effects of several non-neuronal cell types on neurotransmitter synthesis in cultures of dissociated sympathetic neurons from the new-born rat were studied. Acetylcholine synthesis from radioactive choline was increased 100- to 1000-fold in the presence of non-neuronal cells from sympathetic ganglia. This increase was roughly dependent on the number of ganglionic non-neuronal cells present. The effect did not appear to be due to an increased plating efficiency of neurons, since the non-neuronal cells were capable of increasing acetylcholine synthesis after only 48-hr contact with neurons that had been previously grown without non-neuronal cells for 2 weeks. C6 rat glioma cells were also able to stimulate acetylcholine synthesis, but 3T3 mouse fibroblast cells had little or no effect. None of the non-neuronal cell types synthesized detectable acetylcholine in the absence of the neurons. The ganglionic non-neuronal cells had no significant effect on catecholamine synthesis (which occurs in the absence of non-neuronal cells)

HEALTH, FOOD SAFETY AND MEAT DEMAND

For nearly two decades, the U.S. beef industry has faced a long term structural change, which has resulted in consumers shifting from beef to chicken. This shift has occurred due to consumer concerns on cholesterol consumption. More recently, this industry has confronted new challenges on the safety of beef, due to the potential presence of biological contaminants. This study incorporates a measure on food safety with a measure on health information in a meat demand system. Beef safety information is found to have a modest impact on beef demand, but is dominated by health information.meat demand, health, food safety, LA/AIDS, Food Consumption/Nutrition/Food Safety, Livestock Production/Industries,

Intrabody Gene Therapy Ameliorates Motor, Cognitive, and Neuropathological Symptoms in Multiple Mouse Models of Huntington's Disease

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease resulting from the expansion of a glutamine repeat in the huntingtin (Htt) protein. Current therapies are directed at managing symptoms such as chorea and psychiatric disturbances. In an effort to develop a therapy directed at disease prevention we investigated the utility of highly specific, anti-Htt intracellular antibodies (intrabodies). We previously showed that V_(L)12.3, an intrabody recognizing the N terminus of Htt, and Happ1, an intrabody recognizing the proline-rich domain of Htt, both reduce mHtt-induced toxicity and aggregation in cell culture and brain slice models of HD. Due to the different mechanisms of action of these two intrabodies, we then tested both in the brains of five mouse models of HD using a chimeric adeno-associated virus 2/1 (AAV2/1) vector with a modified CMV enhancer/chicken β-actin promoter. V_(L)12.3 treatment, while beneficial in a lentiviral model of HD, has no effect on the YAC128 HD model and actually increases severity of phenotype and mortality in the R6/2 HD model. In contrast, Happ1 treatment confers significant beneficial effects in a variety of assays of motor and cognitive deficits. Happ1 also strongly ameliorates the neuropathology found in the lentiviral, R6/2, N171-82Q, YAC128, and BACHD models of HD. Moreover, Happ1 significantly prolongs the life span of N171-82Q mice. These results indicate that increasing the turnover of mHtt using AAV-Happ1 gene therapy represents a highly specific and effective treatment in diverse mouse models of HD

Preliminary Studies on the Use of Monoclonal Antibodies as Probes for Sympathetic Development

The precise structural organization and proper functioning of the adult nervous system depend on the ability of neurones to make highly ordered synaptic connexions. To define molecules involved in the development of these connexions and to study their functional roles, we use primary cultures of dissociated rat sympathetic neurones grown in the virtual absence of non-neuronal cells. These neurones can develop adrenergic or cholinergic properties, depending on the environment in which they are grown. This ability to manipulate neuronal phenotype is being used in an attempt to identify cell surface macromolecules that are important in the development or function of adrenergic and cholinergic properties. We have produced monoclonal antibodies against the surface membranes of these neurones and are in the process of characterizing them. Results are presented on the binding specificity of one of these antibodies and on the effect of two other antibodies on neurotransmitter synthesis, uptake, and release

Antibody therapy in Neurodegenerative Disease

Advances in medical science have led to increased life expectancy and increased median age in the population. Because the symptoms of neurodegenerative diseases generally onset in mid- to late-life, a concomitant increase in the number of persons afflicted with these devastating diseases has occurred. Developing therapies for neurodegenerative diseases is of the highest priority due to the enormous cost of medical care required, as well as for the human suffering involved. Although caused by a variety of genetic and environmental insults, such diseases share commonalities. Many of these diseases are proteinopathies-diseases caused by misfolded, aggregating proteins. Antibodies that can recognize and remove misfolded proteins are ideally suited for proteinopathy therapeutics. The numerous intriguing advances in antibody-based therapies for neurodegenerative diseases are discussed in this review

Role of nerve growth factor in the development of rat sympathetic neurons in vitro. I. Survival, growth, and differentiation of catecholamine production

The effect of nerve growth factor (NGF) on the development of cholinergic sympathetic neurons was studied in cultures grown either on monolayers of dissociated rat heart cells or in medium conditioned by them. In the presence of rat heart cells the absolute requirement of neurons for exogenous NGF was partially spared. The ability of heart cells to support neuronal survival was due at least in part to production of a diffusable NGF-like substance into the medium. Although some neurons survived on the heart cell monolayer without added NGF, increased levels of exogenous NGF increased neuronal survival until saturation was achieved at 0.5 microgram/ml 7S NGF. The ability of neurons to produce acetylcholine (ACh) from choline was also dependent on the level of exogenous NGF. In mixed neuron-heart cell cultures, NGF increased both ACh and catecholamine (CA) production per neuron to the same extent; saturation occurred at 1 microgram/ml 7S NGF. As cholinergic neurons developed in culture, they became less dependent on NGF for survival and ACh production, but even in older cultures approximately 40% of the neurons died when NGF was withdrawn. Thus, NGF is as necessary for survival, growth, and differentiation of sympathetic neurons when the neurons express cholinergic functions as when the neurons express adrenergic functions

Intrabodies Binding the Proline-Rich Domains of Mutant Huntingtin Increase Its Turnover and Reduce Neurotoxicity

Although expanded polyglutamine (polyQ) repeats are inherently toxic, causing at least nine neurodegenerative diseases, the protein context determines which neurons are affected. The polyQ expansion that causes Huntington's disease (HD) is in the first exon (HDx-1) of huntingtin (Htt). However, other parts of the protein, including the 17 N-terminal amino acids and two proline (polyP) repeat domains, regulate the toxicity of mutant Htt. The role of the P-rich domain that is flanked by the polyP domains has not been explored. Using highly specific intracellular antibodies (intrabodies), we tested various epitopes for their roles in HDx-1 toxicity, aggregation, localization, and turnover. Three domains in the P-rich region (PRR) of HDx-1 are defined by intrabodies: MW7 binds the two polyP domains, and Happ1 and Happ3, two new intrabodies, bind the unique, P-rich epitope located between the two polyP epitopes. We find that the PRR-binding intrabodies, as well as VL12.3, which binds the N-terminal 17 aa, decrease the toxicity and aggregation of HDx-1, but they do so by different mechanisms. The PRR-binding intrabodies have no effect on Htt localization, but they cause a significant increase in the turnover rate of mutant Htt, which VL12.3 does not change. In contrast, expression of VL12.3 increases nuclear Htt. We propose that the PRR of mutant Htt regulates its stability, and that compromising this pathogenic epitope by intrabody binding represents a novel therapeutic strategy for treating HD. We also note that intrabody binding represents a powerful tool for determining the function of protein epitopes in living cells

Insulin Promotes Electrical Coupling between Cultured Sympathetic Neurons

Placing neurons in tissue culture is one way to study how environmental factors affect their differentiation. Replacement of serum- supplementation of the culture medium with defined ingredients extends the experimenter's control of the culture environment; however it also introduces additional potential influences. In this report, we confirm the observation of Higgins and Burton (Higgins, D., and H. Burton (1982) Neuroscience 7:2241–2253) of increased frequency of electrical coupling in serum-free compared to serum-supplemented cultures of rat sympathetic neurons. In addition, experiments were performed to determine whether this effect results from the removal of serum or from the addition of the defined medium components to the culture environment. The results of testing individual ingredients of the defined medium recipe adapted for use on sympathetic neurons (Bottenstein, J.E., and G. H. Sato (1979) Proc. Natl. Acad. Sci. U. S. A. 76:514–517) show that insulin is capable of inducing electrical coupling in serum-free cultures. Thus, the formation of electrical synapses by sympathetic neurons can be hormonally regulated

Big Sagebrush in Pinyon-juniper Woodlands: Using Forest Inventory and Analysis Data as a Management Tool for Quantifying and Monitoring Mule Deer Habitat

We used Interior West Forest Inventory and Analysis (IW-FIA) data to identify conditions where pinyon-juniper woodlands provide security cover, thermal cover, and suitable amounts of big sagebrush (Artemisia tridentata spp.) forage to mule deer in Utah. Roughly one quarter of Utah’s pinyon-juniper woodlands had a big sagebrush component in their understory. Security cover was the least abundant habitat component investigated and only an estimated three percent of Utah’s pinyon-juniper contained forage, security, and thermal cover concurrently. Area of potential mule deer habitat was generally distributed within ecoregion provinces in Utah in proportion to the spatial extent of each province. Quantile regression analysis suggests that when there is less than 22 percent crown cover, more than 20 percent of pinyon-juniper woodlands can be expected to have greater than 15 percent sagebrush cover. We demonstrate the utility of FIA data as an estimator of current habitat features and a long-term monitoring tool and show how quantile regression can be a useful tool for analyzing data with heterogeneous variation