Three-dimensional organisation of the vasculature of the rat spermatic cord and testis

Summary The vascular architecture of the rat testis and spermatic cord was studied by a corrosion cast technique combined with scanning and transmission electron microscopy, and light microscopy. The casts preserve the endothelial impressions of the vessels and enable them to be differentiated into the various vascular components. Frequent arterio-arterial anastomotic arcades and occasional arterio-venous anastomotic channels are seen. A well defined hexagonal pattern of intertubular and peritubular vessels surround the seminiferous tubules. Prominent large endothelial nuclei protrude into the arterial lumina at branching sites, but their functional significance is not known. The outermost vascular layer of the testis consists of large veins, venules, and capillaries, but lacks any arterial branches; it also contains loosely arranged veno-venous anastomotic networks. We have named this vascular layer the sub-albugineal venous plexus. The testicular artery increases in luminal diameter as it approaches the testis. The periarterial capillary plexus, which lies between the pampiniform plexus and the testicular artery, is drained by two types of venules.     

Action of delta 9-tetrahydrocannabinol on the pool of acid soluble nucleotides

The effects of delta 9-tetrahydrocannabinol (THC) treatment on acid soluble pools of uridine nucleoside and nucleotides were investigated in Tetrahymena pyriformis and in isolated mouse lymphocytes and spermatogenic cells. In THC treated Tetrahymena and mouse lymphocytes the uptake of labelled precursor into acid soluble pools of uridine nucleoside and nucleotides fluctuated, whereas in pachytene spermatocytes and round spermatid cells the labelled pool was reduced. The reduction in the labelled pool measured in mouse spermatogenic cells was attributed primarily to a reduction in radioactively labelled uridine nucleoside. Treatment of Tetrahymena in high concentrations of THC (960 and 3,200 microM) resulted in an increase of labelled uridine nucleoside and a reduction in the amount of labelled uridine nucleotides. Expansion of the acid soluble pool with radioactive uridine resulted in small differences in labelled nucleoside and nucleotides in control and THC treated Tetrahymena and mouse lymphocytes. The results are discussed in terms of the effects of THC on macromolecular synthesis in various cellular systems.     

OrthoMaM: A database of orthologous genomic markers for placental mammal phylogenetics

Background  

Molecular sequence data have become the standard in modern day phylogenetics. In particular, several long-standing questions of mammalian evolutionary history have been recently resolved thanks to the use of molecular characters. Yet, most studies have focused on only a handful of standard markers. The availability of an ever increasing number of whole genome sequences is a golden mine for modern systematics. Genomic data now provide the opportunity to select new markers that are potentially relevant for further resolving branches of the mammalian phylogenetic tree at various taxonomic levels.     

Structural basis for VIPP1 oligomerization and maintenance of thylakoid membrane integrity

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Genotyping and development of single-nucleotide polymorphism (SNP) markers associated with blast resistance genes in rice using GoldenGate assay

Development and large-scale genotyping of single-nucleotide polymorphism (SNP) is required to use identified sequence variation in the alleles of different genes to determine their functional relevance to the candidate gene(s). In the present study, Illumina GoldenGate assay was used to validate and genotype SNPs in a set of six major rice blast resistance genes, viz. Pi-ta, Piz(t), Pi54, Pi9, Pi5(1) and Pib, distributed over five chromosomes, to understand their functional relevance and study the population structure in rice. All the selected SNPs loci (96) of six blast (Magnaporthe oryzae) resistance genes were genotyped successfully in 92 rice lines with an overall genotype call rate of 92.0 % and minimum GenTrain cutoff score of ≥0.448. The highest genotyped SNPs were found in japonica type (97.1 %) rice lines, followed by indica (92.12 %), indica basmati (91.84 %) and minimum in case of wild species (82.0 %). Among the genotyped loci, the highest score (98.68 %) was observed in case of Piz(t), followed by Pi-ta, Pi5(1), Pib, Pi54 and Pi9. Polymorphism was obtained in 87.5 % SNPs loci producing 7,728 genotype calls. Minor allele frequency ranged from 0.01 to 0.49 and has good differentiating power for distinguishing different rice accessions. Population structure analysis revealed that a set of genotypes from four rice subpopulations had “admix” ancestry (>26 %) with more than one genetic background of indica, japonica and wild types. SNPs markers were validated in a set of 92 rice lines and converted into CAPS markers which can be used in blast resistance breeding programme.     

Highly variable SSR markers suitable for rice genotyping using agarose gels

Simple sequence repeats (SSR) are the DNA markers of choice for genetic analysis in rice (Oryza sativa L.) due to their abundance, high polymorphism and simple assays using agarose gel electrophoresis. In an attempt to find most variable SSR loci for the agarose gel system, the relationship between SSR length and level of polymorphism was evaluated in a set of eight diverse rice genotypes using 201 random SSR loci of different repeat motifs and lengths, representing both genic and intergenic sequences from the 12 rice chromosomes. There was a positive correlation between SSR length and average number of alleles per locus but linearity of this relationship was limited to the SSR length range of 10–70 bp. The highest level of polymorphism was in the SSR length range of 51–70 bp, beyond which there was stabilization and then decline of polymorphism in SSRs longer than 70 bp. Proportion of polymorphic loci in the different SSR length groups also followed similar pattern with even sharper decline of polymorphism in the highest size range. Here we describe a genome wide set of 436 validated highly variable SSR (HvSSR) markers with repeat lengths of 51–70 bp for their consistent amplification and high polymorphism. In the parental lines of three different mapping populations, the HvSSR loci showed more than twice the level of polymorphism than random SSR markers with average repeat length of 34 bp, and therefore are suitable for QTL mapping and fingerprinting studies in rice employing agarose gels.     

SNP haplotypes of the <Emphasis Type="Italic">BADH1</Emphasis> gene and their association with aroma in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Betaine aldehyde dehydrogenase (BADH) is a key enzyme involved in the synthesis of glycinebetaine—a powerful osmoprotectant against salt and drought stress in a large number of species. Rice is not known to accumulate glycinebetaine but it has two functional genes coding for the BADH enzyme. A non-functional allele of the BADH2 gene located on chromosome 8 is a major factor associated with rice aroma. However, similar information is not available regarding the BADH1 gene located on chromosome 4 despite the similar biochemical function of the two genes. Here we report on the discovery and validation of SNPs in the BADH1 gene by re-sequencing of diverse rice varieties differing in aroma and salt tolerance. There were 17 SNPs in introns with an average density of one per 171 bp, but only three SNPs in exons at a density of one per 505 bp. Each of the three exonic SNPs led to changes in amino acids with functional significance. Multiplex SNP assays were used for genotyping of 127 diverse rice varieties and landraces. In total 15 SNP haplotypes were identified but only four of these, corresponding to two protein haplotypes, were common, representing more than 85% of the cultivars. Determination of population structure using 54 random SNPs classified the varieties into two groups broadly corresponding to indica and japonica cultivar groups, aromatic varieties clustering with the japonica group. There was no association between salt tolerance and the common BADH1 haplotypes, but aromatic varieties showed specific association with a BADH1 protein haplotype (PH2) having lysine₁₄₄ to asparagine₁₄₄ and lysine₃₄₅ to glutamine₃₄₅ substitutions. Protein modeling and ligand docking studies show that these two substitutions lead to reduction in the substrate binding capacity of the BADH1 enzyme towards gamma-aminobutyraldehyde (GABald), which is a precursor of the major aroma compound 2-acetyl-1-pyrroline (2-AP). This association requires further validation in segregating populations for potential utilization in the rice breeding programs.