Passage of solutes through walls of Malpighian tubules of Rhodnius by paracellular and transcellular routes.

The walls of isolated upper Malpighian tubules of the insect, Rhodnius prolixus, are much more permeable to small relatively unchanged solutes (ethanol, xylose, and mannitol) than to larger or more charged solutes (acetate, glycine, tyrosine, and inulin). The more permeable solutes rapidly reach concentrations in the tubule cells equivalent to their concentrations in the bathing medium; the less permeable solutes do not penetrate into the cells. The time course of accumulation of permeable solute in the cells matches the time course of the appearance of solute in the lumen. Substances injected into the hemolymph of fed R. prolixus appear in the urine at concentrations predictable from the permeability of in vitro tubules, supporting the idea that the in vitro permeability of the tubules is representative of their properties in the intact insect. It is suggested that the rapid transcellular penetration of small solutes through the Malpighian tubules reflects the large areas of cell membrane. The area of cell membrane exceeds that of the paracellular clefts by a factor of 10(5).

Polymeric cryoprotectants in the preservation of biological ultrastructure. II. Physiological effects.

A study has been made of the physiological effects of three non-penetrating polymeric cryoprotective agents on sixteen different plant and animal cells and tissues. The cryoprotectants, when used at concentrations at which they are effective in preventing ice-crystal formation, generally have a lower toxicity to cells and tissue than similar concentrations of glycerol. The relatively low toxicity of these substances suggests that they would be more suitable as cryoprotectants for morphological and analytical studies than the commonly used low molecular weight compounds.

Excretion of alkaloids by malpighian tubules of insects.

Nicotine is transported at high rates by Malpighian tubules of larvae of Manduca sexta, Pieris brassicae and Rhodnius prolixus and the transport persists in the absence of alkaloid from the diet. In the fluid-secreting portion of Rhodnius tubules this transport is not coupled to ion transport, nor is it dependent on the physiological state of the animal. The transport, which can occur against a steep electrochemical gradient, shows saturation kinetics with a maximal rate of 700 pmol. min-1 per tubule and is half saturated at 2-3 mM. Nicotine transport independent of ion movements also occurs in the lower resorptive parts of Rhodnius tubules. Both portions of Rhodnius tubules can transport morphine and atropine. These alkaloids and nicotine compete with one naother and are presumed to be carried by the smae transport system. Nicotine transport in Rhodnius was unaffected by organic anions, such as amaranth and benzyl penicillin, or by the organic anion transport inhibitor, probenecid. Fluid secretion in 5-HT-stimulated tubules was reduced by atropine and nicotine, probably by blocking the 5-HT receptors. The Malpighian tubules of adult Calliphora erythrocephala and Musca domestica remove nicotine from bathing solutions, an unknown metabolic accumulating in the tubules. Adult P. brassicae and M. sexta do not exhibit transport of nicotine by their Malpighian tubules.

Induction of transport of organic anions in Malpighian tubules of Rhodnius.

1. The ability of Rhodnius Malpighian tubules to transport organic anions such as p-aminohippuric acid (PAH) and amaranth depends on the physiological state of the insect. Unfed insects progressively lose this ability, but the transport mechanisms rapidly become activated after a blood meal. 2. This induction occurs in adults as well as larval instars and is not prevented by decapitation. 3. No increase in rates of excretion of organic anions follows a meal of Ringer's solution or a single injection of 0-1 mumoles of PAH into the haemolymph. 4. Accelerated PAH excretion is induced in insects fed (a) blood plasma alone or a suspension of red blood cells in saline, (b) solutions of serum albumin or casein, and (c) whole or diluted milk. 5. These results show that the ingestion of a protein-rich meal is sufficient to induce an accelerated transport of organic anions such as PAH and they suggest that this induction is not controlled by a hormone released in response to abdominal distension but depends instead on the continued presence in the haemolymph of some product of digestion of the meal.