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Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans
Published on: March 30, 2013
Quantitative analysis of RNA, glycogen and nucleotides from different developmental stages of Ascaris lumbricoides
Abstract:
Cold acid phenol extractions (Kirby, K.S. (1968) in Methods in Enzymology, Vol. XII B, pp. 87--99, Academic Press, New York) of homogenates from oocytes, zygotes and larval stages of Ascaris lumbricoides yield predominantly glycogen, some glycogen . phosphate complexes and guanine nucleotides, but only small amounts of RNA. Oocytes contain 10.0 pg of extractable 19 S and 26 S rRNA and 1.1 pg 4 S and 5 S RNA per cell. During the maturation of the oocyte to the zygote stage, the amount of 19 S and 26 S rRNA increases by 57%, and that of 4 S and 5 S RNA by 130%. Larval stages contain 62% more 19 S and 26 S rRNA, and 47% more 4 S and 5 S RNA than zygotes. The amount of the extractable glycogen decreases by about 80% and nucleotides by about 70% from oogenesis to the larval stage. The Kirby extract from homogenates of spermatids contains only traces of glycogen, glycogen . phosphate complexes and nucleotides, and 115 times less RNA per spermatid than oocytes. The nucleotide pool sizes of oocytes, zygotes and larval stages were determined; the pools consist of 71--88% guanine nucleotides. Spermatids contain only adenine nucleotides and an unidentified, nucleotide-like compound.

