Cyclic AMP Phosphodiesterase in Leydig Cell Preparations of the Rat Testis

Enzymatic digestion of the interstitial tissue of early juvenile and adult rat testes resulted in an enrichment of the Leydig cell population. The cells of the intertubular preparation from adult testes were separated by centrifugal elutriation, according to differences in sedimentation velocity, a counter-flow centrifugation technique leading to 70% Leydig cell purity. Using this approach, it was possible to demonstrate that Leydig cells from adult testes contain only low affinity isoenzymes of cyclic AMP phosphodiesterase (PDE; E.C.: 3.1.4.17), an intracellular regulator of cAMP. Starch gel electrophoresis showed that the isozyme of cAMP PDE of Leydig cells is masked in crude testis homogenates due to the relatively low level of these cells in the total population. In Leydig cells, there are two different electrophoretic forms expressed which resemble two of eleven different molecular forms of cAMP PDE demonstrated for comparison in 21 different organs of the adult rat.
An interstitial cell preparation from early juvenile testes, with a Leydig cell content of up to 20%, was also investigated electrophoretically with regard to molecular forms of cAMP PDE, the properties of which were characterized by kinetic analysis of cAMP hydrolysis. The results presented are discussed in relation to the onset of testosterone synthesis in Leydig cells of prepubertal rats leading to the initiation of male puberty.     

Discrimination of conspecific-built chimneys from human-built ones by the burrowing crayfish, Fallicambarus fodiens (Decapoda, Cambaridae)

Abstract. The vast majority of tested juvenile crayfish of Fallicambarus fodiens consistently preferred conspecific-built mud chimneys over similar-looking human-built chimneys. When the chimneys were surrounded by transparent acetate sheets, the crayfish no longer discriminated between conspecific-built and human-built chimneys. This suggests that visual cues were not important in allowing the crayfish to discriminate between the two chimney types. In the absence of chimneys, significantly more crayfish showed an initial preference for mud saturated with water that had contained conspecifics, over similar mud saturated with distilled water. This indicates that chemical cues play a role in allowing these crayfish to discriminate between the conspecific-conditioned mud and the control wet mud. The preference of F. fodiens individuals for conspecific-built chimneys appears to be based, at least in part, on chemical cues, possibly in addition to tactile cues.     

Recombination Between Guanidine-resistant and Dextran Sulfate-resistant Mutants of Type 1 Poliovirus

Mixed infection of monkey kidney cells with two mutants of the LSc2ab strain of poliovirus, one resistant to guanidine and the other resistant to both dextran sulfate and 2-(alpha-hydroxybenzyl)-benzimidazole (HBB), yielded progeny in which the number of gua(r)dex(r) particles exceeded by a factor of 7 to 10 the expected number of similar particles occurring through spontaneous mutation; recombination would explain the fairly high excess of doubly mutant particles that was obtained. Scoring of HBB resistance in 50 gua(r)dex(r) clones suggested that, during recombination, resistance to dextran sulfate is not associated with HBB resistance.     

A simple polymerase chain reaction apparatus based on a computer printer.

We describe a very simple laboratory-made polymerase chain reaction (PCR) apparatus. The reaction tubes are placed in a holder fixed through a mechanical arm to the tape cartridge of a computer printer. A computer controls the horizontal movement of the tube carrier by sending the proper printing commands. The holder is raised or lowered by a frame fixed to the paper-advancing roller of the printer. The system allows the programmed movement of the test tubes within the holder, successively through three thermal baths placed in front of the printer. DNA from single lambda gtll lysis plaques was successfully amplified with this system in our laboratory.