Changes in Activities of Glucose Metabolising Enzymes in Germ Cells during Spermatogenesis in Rats

The rate of ¹⁴CO₂, liberation from [¹⁴C-1]glucose was identical to that from [¹⁴C-6]glucose in spermatids, but more than the latter in spermatogonia. Rotenone (1 μM) completely inhibited ¹⁴CO₂ release from [¹⁴C-1]glucose in spermatids, but decreased it only 30% in spermatogonia. The activity of glucose-6-phosphate dehydrogenase, but not 6-phosphogluconate dehydrogenase, was markedly lower in spermatocytes and spermatids than in spermatogonia. The activities of the glycolytic enzymes, glucosephosphate isomerase, fructose diphosphatase, glyceraldehyde-3-phosphate dehydrogenase and enolase, differed only slightly in spermatids and spermatogonia. It is concluded that the low glucose-6-phosphate dehydrogenase activity may contribute to the low activity of the pentose cycle in spermatocytes and spermatids.     

Studies on the metabolism of a sulfur-oxidizing bacterium VI1. Fractionation and reconstitution of the elementary sulfur-oxidizing system of Thiobacillus thiooxidans

The sulfur-oxidizing system of a strain of Thiobacillus thiooxidanswas obtained in cell-free state. The system is resolved intothree fractions and can be reconstituted from these fractions.Both the soluble and particulate fractions are required forthe oxidation of elementary sulfur. The soluble fraction wasfurther separated into two fractions, the collodion membrane-permeable(S-P)and the impermeable(S-IP). S-P contains a low molecular weight,relatively heat stable substance(s) which is indispensable forthe reconstitution of the sulfur-oxidizing system and was identifiedas a pyridine nucleotide. The function of S-P can be replacedby NAD or NADP, but not by cysteine nor GSH. Oxidation of NADH₂ and NADPH₂ is catalyzed by the particulatefraction. Oxidation of the latter is much more rapid than thatof the former. Oxidation of NADPH₂ as well as sulfur oxidationis inhibited by cyanide, pCMB and CO, the CO-inhibition beingphoto-irreversible. However, strong inhibitors of sulfur oxidationsuch as DDC, 8-hydroxyquinoline and salicylaldoxime have noeffect on the oxidation of NADPH₂. The optimum pH values for sulfur and sulfite oxidations by thecell-free extract are shifted to the neutral side in comparisonwith pH values by intact cells. ¹V = References(I). ²Partly supported by a grant from the Ministry of Education. (Received April 3, 1969; )     

Continuous Cell Lines from Larval Hemocytes of the Cabbage Armyworm, Mamestra brassiae

Two continuous cell lines, NIAS-MaBr-92 and NIAS-MaBr-93, were obtained from larval hemocytes of the cabbage armyworm, Mamestra brassicae . The cells grew in suspended state. Spherical cells were predominant, although there were spindle shaped and irregular shaped cells. The chromosome numbers of the cell lines varied very much with the mode of 100 of 120. The population doubling times of these cell lines were about 36hr. The cells could grow in media free of serum and lacking sterols, fatty acids and protein. The cell lines consumed ammonia and produced glutamine when cultured in MM medium. The cells could be stored at -100°C for more than two year, and at 5°C for two months. The cells were sensitive to Autographa californica nuclear polyhedrosis virus and Chilo iridescent virus.     

MULTIPLE FORMS OF β -GALACTOSIDASE AND THE CHANGES IN ITS PATTERN DURING DEVELOPMENT OF DICTYOSTELIUM DISCOIDEUM

Four forms of β-galactosidase (EC 3.2.1.23) were separated from cell extracts of Dictyostelium discoideum by polyacrylamide gel electrophoresis. Changes in the pattern of multiple forms of this enzyme during differentiation and dedifferentiation were studied. The band closest to the anode (Band 1) existed throughout development, but the other three bands appeared and disappeared at certain stages. One of those bands was specific for the vegetative stage (Band 3), and another for the morphogenetic stages (Band 2). The last one predominantly appeared during dedifferentiation of disaggregated slug cells (Band 4), although it was slightly detected during culmination. During the process of dedifferentiation, the increase in activity of Band 4 coincided with the decrease in Band 2. After the completion of dedifferentiation, Band 4 also disappeared, only Band 1 remaining. These multiple forms were not only electrophoretically separable but different in their sensitivities to various inhibitors. All forms of the enzyme were localized in subcellular particles, probably in lysosomes and phagosomes, and the relative activities in these two fractions varied during development.