Enhancement of proliferation in cultures of Chinook salmon embryo cells by interactions between inosine and bovine sera

Abstract

The influence of inosine on DNA synthesis by Chinook salmon embryo cells (CHSE-214) was investigated because previously cell number was shown to increase from six- to thirtyfold if inosine was added to the basal medium (L-15) supplemented with either dialyzed fetal bovine serum (dFBS), calf serum (CS), or dCS. Relative to L-15, 3H-thymidine incorporation was inhibited by these sera alone but elevated in nondialyzed (intact) FBS. Inosine at 10 microM stimulated 3H-thymidine incorporation from ten- to seventyfold in dFBS, CS, and dCS but was only slightly stimulatory in FBS and in L-15 alone. As well as inosine, hypoxanthine, cIMP, IMP, IDP, and ITP were just as stimulatory, but the nonsalvageable purines (xanthine, xanthosine, and XMP) were not. The stimulatory action of inosine was highest in low density cultures. Dipyridamole and S-(p-nitrobenzyl)-6-thioinosine (NBTI), inhibitors of facilitated nonconcentrative nucleoside transport, did not completely block the enhancement of cell number by inosine and by themselves increased proliferation in CS and dCS. Overall, these results suggest that exogenous inosine promoted CHSE-214 proliferation by overcoming factors in the nondialyzable fraction of sera that led to purine loss and by raising intracellular purine nucleotides to levels necessary for cells to respond to growth factors in the nondialyzable fraction of sera.

Electrochemical oxidation of inosine-5′-triphosphate at pyrolytic graphite electrode

“…The diol (6) then undergoes series of follow-up chemical steps causing pyrimidine ring opening to give allantoin (7), CO 2 and ribose-triphosphate as shown in Scheme 1. Allantoin possesses five silylable sites and GC-MS data clearly indicate that all the reactive hydrogen atoms are replaced with trimethylsilyl groups on silylation with BSTFA/acetonitrile to give pentasilylated allantoin having m/z 518 (9). The formation of ribose as one of the products appears to be from the hydrolysis of ribosetriphosphate moiety.…”

“…The mass spectrum of the silylated material exhibited a clear peak at R t ∼ 8.51 min having m/z 518 (M + , 3.12%) along with a large peak at m/z 503 (M + -CH 3 , 31.10%), which corresponds to pentasilylated allantoin (9). The dried material also gave important bands in the FT-IR spectrum at 3436 (NH 2 ), 3341, 3220 and 3058 (NH), 2759 (CH), 1781, 1711 and 1602 (CO), 1528, 1433, 1390, 1322, 1282 (CN) and 1182 (C C) cm −1 .…”

“…The molar mass of 518 corresponds to pentasilylated allantoin (9), which was formed due to the rupture of the pyrimidine ring. The species responsible for molar masses 766, 414, 558 and 438 were due to hexasilylated C O O C (24), disilylated N N (19) and tetrasilylated C C linked (16) dimers and tetrasilylated d-ribose (11).…”

“…5 -ITP, an inosyl nucleotide causes chlorea toxin and prostaglandin E 1 to enhance the activity of thyroid adenylate cyclase [7]. It is also claimed to cause an increased yield of nucleic acid sequence-base amplification (NASBA) products of base-paired viroid RNA [8] and promote enhanced cell proliferation in Chinook Salmon embryo cells [9]. 5 -ITP causes the synthesis of mRNAs by initiation and elongation of reovirus * Corresponding author.…”

In Vitro and in Vivo Characterization of Inosine Monophosphate Delivered by a Nanoparticle in Rainbow Trout

“…While nucleotide mixtures, such as Optimûn (Chemoforma, Augst, Switzerland), that contain cytokine monophosphate, uridine monophosphate, adenine monophosphate, IMP, and guanine monophosphate, are relatively well studied, the individual components are not. At the in vitro level, IMP stimulated the proliferation of a Rainbow Trout cell line as measured by a significant increase in 3H-thymidine incorporation into DNA (Ganassin et al 1994). An in vivo study by Song et al (2012) found that IMP-supplemented diets significantly increased myeloperoxidase and lysozyme activity in Olive Flounder Paralichthys olivaceus (Song et al 2012).…”