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I C Robinson

Publications and source records attributed to I C Robinson.

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Extraction of small amounts of oxytocin from biological fluids by means of agarose-bound neurophysin.

The use of agarose-bound neurophysin for the extraction of oxytocin from biological fluids is described. Oxytocin can be extracted from plasma, urine and cerebrospinal fluid with a high rate of recovery and samples varying widely in volume and oxytocin concentration can be tested by the method. Columns can be used to extract and concentrate dilute samples, or to help identify small amounts of neurohypophysial hormones by affinity chromatography. The oxytocin can be eluted from the column directly into the buffer used for subsequent bioassay. The composition of the final extract is constant and independent of the composition of the sample. The specificity of the binding is high. It is suggested that the method has many advantages over others in current use.

Animals

Calcium and stimulus secretion coupling in the neurohypophysis. V. The effects of the Ca2+ ionophores A23187 and X537 A on vasopressin release and 45Ca2+ efflux; interactions with sodium and a verapamil analogue (D600).

Slices from ox neurohypophyses were incubated in a calcium-free medium with the ionophores A23187 or X537A. X537A (5 X 10(-5) mol/l) caused a marked release of vasopressin, neurophysin and protein to the medium. A23187 (2 X 10(-5) mol/l) did not cause any release by itself, but when Ca2+ was added to the medium in the presence of the ionophore, an increase in the release of vasopressin, neuorphysin and protein occurred. Release of lactate dehydrogenase and peptidase were not affected by the ionophores. The secretion caused by A23187 was abolished by D600 (a verapamil analogue) (2 X 10(-5) mol/l) whereas the effect of X537A was unchanged. The effects of X537A were strongly inhibited by removal of sodium from the medium. Re-addition of sodium to the medium caused a marked release. Gramicidin (10(-6) or 5 X 10(-5) mol/l) had no effect on secretion. Efflux of 45Ca2+ from pre-loaded slices was drastically reduced in a sodium-free medium. X537A caused an increase in the efflux rate of 45Ca2+ both in medium with a normal concentration of sodium and when slices had been incubated in a sodium-free medium. A23187 and X537A both released 45Ca2+ from a neurohypophyseal mitochondrial fraction. When sodium in a concentration of 20 mmol/l was added to this fraction, the Ca2+ accumulation was inhibited. This effect was reduced by inorganic phosphate up to a concentration of 2 mmol/l.

Animals