GCH1 deficiency activates brain innate immune response and impairs tyrosine hydroxylase homeostasis

The Parkinson’s disease (PD) risk gene GTP cyclohydrolase 1 (GCH1) catalyzes the rate-limiting step in tetrahydrobiopterin (BH4) synthesis, an essential cofactor in the synthesis of monoaminergic neurotransmitters. To investigate the mechanisms by which GCH1 deficiency may contribute to PD, we generated a loss of function zebrafish gch1 mutant (gch1-/-), using CRISPR/Cas technology. gch1-/- zebrafish develop marked monoaminergic neurotransmitter deficiencies by 5 dpf, movement deficits by 8 dpf and lethality by 12 dpf. Tyrosine hydroxylase protein levels were markedly reduced without loss of ascending dopaminergic (DAergic) neurons. L-Dopa treatment of gch1-/- larvae improved survival without ameliorating the motor phenotype. RNAseq of gch1-/- larval brain tissue identified highly upregulated transcripts involved in innate immune response. Subsequent experiments provided morphological and functional evidence of microglial activation in gch1-/-. The results of our study suggest that GCH1 deficiency may unmask early, subclinical parkinsonism and only indirectly contribute to neuronal cell death via immune-mediated mechanisms. Our work highlights the importance of functional validation for GWAS risk factors and further emphasises the important role of inflammation in the pathogenesis of PD

GCH1 deficiency activates brain innate immune response and impairs tyrosine hydroxylase homeostasis

The Parkinson's disease (PD) risk gene GTP cyclohydrolase 1 (GCH1) catalyzes the rate-limiting step in tetrahydrobiopterin (BH4) synthesis, an essential cofactor in the synthesis of monoaminergic neurotransmitters. To investigate the mechanisms by which GCH1 deficiency may contribute to PD, we generated a loss of function zebrafish gch1 mutant (gch1–/–), using CRISPR/Cas technology. gch1–/– zebrafish develop marked monoaminergic neurotransmitter deficiencies by 5 d postfertilization (dpf), movement deficits by 8 dpf and lethality by 12 dpf. Tyrosine hydroxylase (Th) protein levels were markedly reduced without loss of ascending dopaminergic (DAergic) neurons. L-DOPA treatment of gch1–/– larvae improved survival without ameliorating the motor phenotype. RNAseq of gch1–/– larval brain tissue identified highly upregulated transcripts involved in innate immune response. Subsequent experiments provided morphologic and functional evidence of microglial activation in gch1–/–. The results of our study suggest that GCH1 deficiency may unmask early, subclinical parkinsonism and only indirectly contribute to neuronal cell death via immune-mediated mechanisms. Our work highlights the importance of functional validation for genome-wide association studies (GWAS) risk factors and further emphasizes the important role of inflammation in the pathogenesis of PD

Phylogenetic Distinctiveness of Middle Eastern and Southeast Asian Village Dog Y Chromosomes Illuminates Dog Origins

Modern genetic samples are commonly used to trace dog origins, which entails untested assumptions that village dogs reflect indigenous ancestry or that breed origins can be reliably traced to particular regions. We used high-resolution Y chromosome markers (SNP and STR) and mitochondrial DNA to analyze 495 village dogs/dingoes from the Middle East and Southeast Asia, along with 138 dogs from >35 modern breeds to 1) assess genetic divergence between Middle Eastern and Southeast Asian village dogs and their phylogenetic affinities to Australian dingoes and gray wolves (Canis lupus) and 2) compare the genetic affinities of modern breeds to regional indigenous village dog populations. The Y chromosome markers indicated that village dogs in the two regions corresponded to reciprocally monophyletic clades, reflecting several to many thousand years divergence, predating the Neolithic ages, and indicating long-indigenous roots to those regions. As expected, breeds of the Middle East and East Asia clustered within the respective regional village dog clade. Australian dingoes also clustered in the Southeast Asian clade. However, the European and American breeds clustered almost entirely within the Southeast Asian clade, even sharing many haplotypes, suggesting a substantial and recent influence of East Asian dogs in the creation of European breeds. Comparison to 818 published breed dog Y STR haplotypes confirmed this conclusion and indicated that some African breeds reflect another distinct patrilineal origin. The lower-resolution mtDNA marker consistently supported Y-chromosome results. Both marker types confirmed previous findings of higher genetic diversity in dogs from Southeast Asia than the Middle East. Our findings demonstrate the importance of village dogs as windows into the past and provide a reference against which ancient DNA can be used to further elucidate origins and spread of the domestic dog

Atomic Energy Commission Reports

This report summarizes only that portion of the injection-casting experiments in which the castings were made with air pressure

The new fundamental gravimetric network of Slovenia

The paper presents all stages of the development and processing of the fundamental gravimetric network of Slovenia, which consists of a zero order network, which has six absolute gravity stations, and twenty nine first order gravimetric stations. Descriptions are given of the design of the network, the geological assessment of the gravimetric stations, the gravity survey of the first order network, and the post-processing and adjustment of the gravimetric observations, which was performed in two stages. First the observations in the zero order network were adjusted as a free network, and then a standard adjustment of the first order network was performed. Finally, the adjusted gravity values at the stations were analysed against the Potsdam system, which was the basis of all previous gravimetric calculations in Slovenia. In the analyses an equation for the transformation of gravity values between the Potsdam system and the IGSN71 system (International Gravity Standardization Network 1971) has been derived

Ab initio calculation of NMR shielding in phosphaalkenes X-P = CY2

Rozhenko AB, Schoeller W, Povolotskii MI. Ab initio calculation of NMR shielding in phosphaalkenes X-P = CY2. MAGNETIC RESONANCE IN CHEMISTRY. 1999;37(8):551-563.P-31 and C-13 chemical shielding values, sigma(iso), were calculated for a series of phosphaalkenes X-P=CY2 with a variety of substituents such as X=H, CH3, F, Cl, Br, OH, NH2, PH2, SiH3, CN and Y=H, CH3, NH2, SiH3, using the GIAO procedure at the RHF/6-311 + G(d,p) and RHF/6-311 + G(2d,2p) levels. The trends governing the Variation of the geometric parameters and the natural charges and Wiberg indices are discussed. The contributions of the various molecular orbitals in P-31 and C-13 chemical shielding in the phosphaalkene H-P=CH2 and ethylene, respectively, were compared. The P-31 chemical shielding variation was determined by mixing of the ground and excited states. Good correlations were found between the experimentally measured SC and SP values in the series X'-P=C(SiMe3)(2) and those calculated for the series X-P=C(SiH3)(2), but on average the calculated P-31 chemical shifts seem to be overestimated. A comparison with the calculated data (at the same level) for X-P=C(SiMe3)(2) indicates only a partial improvement. The C-13 chemical shifts in phophaalkenes correlate within the subseries X-P=CH2, X-P=C(CH3)(2) and X-P=C(SiH3)(2) with the charge variation at the corresponding carbon atom, reflecting the X-(P=C) pi-mesomeric interactions. The absence of the corresponding correlation with all phophaalkenes investigated manifestates limited validity of this generally used approach. Copyright (C) 1999 John Wiley & Sons, Ltd