CaMKII controls neuromodulation via neuropeptide gene expression and axonal targeting of neuropeptide vesicles

Ca2+/calmodulin-dependent kinase II (CaMKII) regulates synaptic plasticity in multiple ways, supposedly including the secretion of neuromodulators like brain-derived neurotrophic factor (BDNF). Here, we show that neuromodulator secretion is indeed reduced in mouse α- and βCaMKII-deficient (αβCaMKII double-knockout [DKO]) hippocampal neurons. However, this was not due to reduced secretion efficiency or neuromodulator vesicle transport but to 40% reduced neuromodulator levels at synapses and 50% reduced delivery of new neuromodulator vesicles to axons. αβCaMKII depletion drastically reduced neuromodulator expression. Blocking BDNF secretion or BDNF scavenging in wild-type neurons produced a similar reduction. Reduced neuromodulator expression in αβCaMKII DKO neurons was restored by active βCaMKII but not inactive βCaMKII or αCaMKII, and by CaMKII downstream effectors that promote cAMP-response element binding protein (CREB) phosphorylation. These data indicate that CaMKII regulates neuromodulation in a feedback loop coupling neuromodulator secretion to βCaMKII- and CREB-dependent neuromodulator expression an

Improved generation of rat gene knockouts by target-selected mutagenesis in mismatch repair-deficient animals

BACKGROUND: The laboratory rat (Rattus norvegicus) is one of the preferred model organisms in physiological and pharmacological research, although the availability of specific genetic models, especially gene knockouts, is limited. N-ethyl-N-nitrosourea (ENU)-driven target-selected mutagenesis is currently the most successful method in rats, although it is still very laborious and expensive. RESULTS: As ENU-induced DNA damage is normally recognized by the mismatch repair (MMR) system, we hypothesized that the effectiveness of the target-selected mutagenesis approach could be improved by using a MMR-deficient genetic background. Indeed, Msh6 knockout rats were found to be more sensitive to ENU treatment and the germ line mutation rate was boosted more than two-fold to 1 mutation per 585 kb. In addition, the molecular mutation spectrum was found to be changed in favor of generating knockout-type alleles by approximately 20%, resulting in an overall increase in efficiency of approximately 2.5 fold. The improved effectiveness was demonstrated by high throughput mutation discovery in 70 Mb of sequence in a set of only 310 mutant F1 rats. This resulted in the identification of 89 mutations of which four introduced a premature stopcodon and 64 resulted in amino acid changes. CONCLUSION: Taken together, we show that the use of a MMR-deficient background considerably improves ENU-driven target-selected mutagenesis in the rat, thereby reducing animal use as well as screening costs. The use of a mismatch repair-deficient genetic background for improving mutagenesis and target-selected knockout efficiency is in principle applicable to any organism of interest

Chronic Loss of Melanin-Concentrating Hormone Affects Motivational Aspects of Feeding in the Rat

Current epidemic obesity levels apply great medical and financial pressure to the strenuous economy of obesity-prone cultures, and neuropeptides involved in body weight regulation are regarded as attractive targets for a possible treatment of obesity in humans. The lateral hypothalamus and the nucleus accumbens shell (AcbSh) form a hypothalamic-limbic neuropeptide feeding circuit mediated by Melanin-Concentrating Hormone (MCH). MCH promotes feeding behavior via MCH receptor-1 (MCH1R) in the AcbSh, although this relationship has not been fully characterized. Given the AcbSh mediates reinforcing properties of food, we hypothesized that MCH modulates motivational aspects of feeding

Systematic biases in DNA copy number originate from isolation procedures

BACKGROUND: The ability to accurately detect DNA copy number variation in both a sensitive and quantitative manner is important in many research areas. However, genome-wide DNA copy number analyses are complicated by variations in detection signal. RESULTS: While GC content has been used to correct for this, here we show that coverage biases are tissue-specific and independent of the detection method as demonstrated by next-generation sequencing and array CGH. Moreover, we show that DNA isolation stringency affects the degree of equimolar coverage and that the observed biases coincide with chromatin characteristics like gene expression, genomic isochores, and replication timing. CONCLUSION: These results indicate that chromatin organization is a main determinant for differential DNA retrieval. These findings are highly relevant for germline and somatic DNA copy number variation analyses

Melanocortin Receptor 4 Deficiency Affects Body Weight Regulation, Grooming Behavior, and Substrate Preference in the Rat

Obesity is caused by an imbalance between energy intake and expenditure and has become a major health-care problem in western society. The central melanocortin system plays a crucial role in the regulation of feeding and energy expenditure, and functional loss of melanocortin receptor 4 (MC4R) is the most common genetic cause of human obesity. In this study, we present the first functional Mc4r knockout model in the rat, resulting from an N-ethyl-N-nitrosourea mutagenesis–induced point mutation. In vitro observations revealed impaired membrane-binding and subsequent nonfunctionality of the receptor, whereas in vivo observations showed that functional loss of MC4R increased body weight, food intake, white adipose mass, and changed substrate preference. In addition, intracerebroventricular (ICV) administration of Agouti-Related Protein79–129 (AgRP79–129), an MC4R inverse agonist, or Melanotan-II (MTII), an MC4R agonist, did affect feeding behavior in wild-type rats but not in homozygous mutant rats, confirming complete loss of MC4R function in vivo. Finally, ICV administration of MTII induced excessive grooming behavior in wild-type rats, whereas this effect was absent in homozygous mutant rats, indicating that MTII-induced grooming behavior is exclusively regulated via MC4R pathways. Taken together, we expect that the MC4R rat model described here will be a valuable tool for studying monogenic obesity in humans. More specifically, the relative big size and increased cognitive capacity of rats as compared to mice will facilitate complex behavioral studies and detailed mechanistic studies regarding central function of MC4R, both of which ultimately may help to further understand the specific mechanisms that induce obesity during loss of MC4R function

White dwarfs in the building blocks of the Galactic spheroid

Aims. The Galactic halo likely grew over time in part by assembling smaller galaxies, the so-called building blocks (BBs). We investigate if the properties of these BBs are reflected in the halo white dwarf (WD) population in the solar neighbourhood. Furthermore, we compute the halo WD luminosity functions (WDLFs for four major BBs of five cosmologically motivated stellar haloes). We compare the sum of these to the observed WDLF of the Galactic halo, derived from selected halo WDs in the SuperCOSMOS Sky Survey, aiming to investigate if they match better than the WDLFs predicted by simpler models. Methods. We couple the SeBa binary population synthesis model to the Munich-Groningen semi-analytic galaxy formation model applied to the high-resolution Aquarius dark matter simulations. Although the semi-analytic model assumes an instantaneous recycling approximation, we model the evolution of zero-age main sequence stars to WDs, taking age and metallicity variations of the population into account. To be consistent with the observed stellar halo mass density in the solar neighbourhood (ρ0), we simulate the mass in WDs corresponding to this density, assuming a Chabrier initial mass function (IMF) and a binary fraction of 50%. We also normalize our WDLFs to ρ0. Results. Although the majority of halo stars are old and metal-poor and therefore the WDs in the different BBs have similar properties (including present-day luminosity), we find in our models that the WDs originating from BBs that have young and/or metal-rich stars can be distinguished from WDs that were born in other BBs. In practice, however, it will be hard to prove that these WDs really originate from different BBs, as the variations in the halo WD population due to binary WD mergers result in similar effects. The five joined stellar halo WD populations that we modelled result in WDLFs that are very similar to each other. We find that simple models with a Kroupa or Salpeter IMF fit the observed luminosity function slightly better, since the Chabrier IMF is more top-heavy, although this result is dependent on our choice of ρ0

Espécies novas de Blaptica do Rio Grande do Sul, Brasil (Blattaria, Blaberidae, Blaberinae) New species of Blaptica from Rio Grande do Sul, Brazil (Blattaria, Blaberidae, Blaberinae)

Cinco novas espécies de Blaptica Stål, 1875 do Estado do Rio Grande do Sul, Brasil são descritas e apresentadas ilustrações de Blaptica dubia.<br>Five new species of Blaptica Stål, 1875 from Rio Grande do Sul State, Brazil are described and illustrations of Blaptica dubia are presented

Binary White Dwarfs in the Galactic Halo

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