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C W Pool

Publications and source records attributed to C W Pool.

62 records · Page 4Linked to original sources
PubMed1978

The alpha-glucan-uridine diphosphate glucosyltransferase reation for the identification of glycogen-depleted muscle fibers.

This paper decribes the use of the alpha-glucan uridine di-phosphate glucosyl transferase reaction for enhancing the contrast between glycogen depleted and non-depleted muscle fibers in the periodic acid schiff (PAS) reaction. Muscle fiber glycogen was depleted by prolonged repetitive stimulation of single motor units of the extensor digitorum longus muscle from the rat.

Animals↗
PubMed1977

Detection of antigens and antibodies by an immuno-peroxidase method applied on thin longitudinal sections of SDS-polyacrylamide gels.

This paper describes a method for detection of antigens in thin sections of SDS-polyacrylamide gels. This method, called SGIP, involves longitudinal sectioning of SDS gels, fixation of the proteins in the gels, removal of the SDS, incubation of the sections with an antiserum and detection of antigens by the indirect immuno-peroxidase technique. The method is useful for assessing the affinity spectrum of a given antiserum against a heterogeneous mixture of proteins, and for the detection of proteins in tissue homogenates or other protein mixtures by means of well defined antisera. By applying the method to serial sections from a single SDS-polyacrylamide gel, a dilution dependent reactivity of antisera is demonstrated.

Acrylamides↗
PubMed1976

Muscle fibre typing with sera against myosin and actin. A comparison between enzyme- and immunohistochemical classification.

Sera raised against actin and myosin, extracted from white muscle of fish, were used for the immune-histochemical characterization of muscle fibers. It appeared that both, the actin- and the myosin serum are specific for white muscle fibres in fish. Further it was found that in both, the A- and the I-band of the sacromeres, fibre type specific proteins are present. The classification of muscle fibre types obtained with the antisera was compared with the classification obtained with some enzyme histochemical reactions. Muscle fibres that reacted positively with the two sera, also showed a high histochemical myofibrillar ATP-ase activity. The correlation with a low succinate dehydrogenase- and a high lactate dehydrogenase activity was not always found.

Actins↗
PubMed1975

Immunofluorescence of vasopressin and oxytocin in the rat hypothalamo-neurohypophypopseal system.

The present paper deals with the development of an immunofluorescence procedure that allows specific localization of vasopressin and oxytocin in the hypothalamo-neurohypophyseal system(hnx) of the rat. Antibodies against arginine vasopressin (AVP), lysine-vasopressin (LVP) and oxytocin were raised by injecting these hormones that were covalently bound to thyroglobulin into rabbits. The vasopressin-immunized rabbits showed periods of diabetes insipidus, while histoloty of the "hns revealed an intact neurosecretory system with signs of increased endogenous hormone synthesis in the supraoptic nucleus and increased release in the neuro-hypophysis of some rabbits. The daily water intake of the oxytocin-immunized rabbits was similar to that of control rabbits. The development of antibodies against vasopressin as measured in the immunofluorescence procedure showed a course that was quite different from the curve of the titer as determined by radioimmunoassay (RIA). Also the specificity of the antibodies used in the immunofluorescence procedure was found to be quite different from their specificity in a RIA system. Potency and specificity of the antibodies have to be studied therefore within the immunofluorescence procedure itself. Using freshly frozen acetone-postfixed hypotalami or pituitaries, no sharp localization of immunofluorescence could be obtained in the HNS. Therefore prefixation was performed. Both, the type and the duration of prefixation revealed quite different results regarding the immunofluorescence in the neurosecretory cell boides in the hypotalamus and of their endings in the neurohypophysis. The best immunofluorescence results were obtained using 6 hours glyoxal-prefixation for the hypothalamus and 24 hours formalin-prefixation for the pituitary. The cross-reaction of the antibodies for oxytocin or vasopressin was tested on synthetic hormones that were bound to CNBR-activated agarose beads and mounted on glass sides. All anti-plasmas showed cross-reaction on beads containing the heterologou- antigen. The plasmas were purified by incubation with beads containing the heterologous hormone until the cross-reacting component had been removed. Using purified antibodies, the distribution of oxytocin and vasopressin cells within the HNS was investigated. More oxytocin containing cells were localized in the rostral part and more vasopressin in the caudal part of both, the supraoptic (SON) and paraventricular nucleus (PVN). Comparable percentages of oxytocin and vasopressin containing cells were found in the SON and PVN. The absolute amount of oxytocin containing cells was 2.5 times more in the SON than in the PVN, which seems to contradict the "classical" view that the PVN predominantly or entirely synthetizes oxytocin. In addition, fluorescence was found using antobodies against vasopressin in the suprachiasmatic nucleus in Wistar rats and heterozygous Brattleboro rats, but not in this nucleus of homozygous Brattleboros.

Animals↗
PubMed1975

Specificity of oxytocin and vasopressin immunofluorescence.

A total inhibition of immunofluorescence could not be obtained using the technique of preincubating either anti-vasopressin or anti-oxytocin plasmas with their homologous antigens. An alternative test of specificity was therefore developed. Lysine-vasopressin (LVP), arginine-vasopressin (AVP) or oxytocin were covalently bound to CNB-activated agarose beads. These hormone-coupled beads were then either placed immediately on glass slides and treated in the same way as tissue sections for immunofluorescence, or pre-incubated in test-tubes with the antibodies and then prepared for immunofluorescence. Fluorescence was measured quantitatively using a Leitz orthoplan microscope with epi-illumination and a photometer attachment. Without pre-incubation the anti-oxytocin plasmas produced intensive fluorescence on those beads containing their homologous antigen (oxytocin) but only slight fluorescence with the heterologous antigens (AVP or LVP). Anti-vasopressin plasmas produced intensive fluorescence of AVP-, LVP- and oxytocin-containing beads. Because the neurohypophysial hormones were bound to agarose beads, antibodies binding to these hormones could be removed by simple centrifugation. Anti-oxytocin and anti-vasopressin lost their fluorescence capacity after pre-incubation with oxytocin- or vasopressin-containing beads respectively, showing that all the antibodies important for fluorescence were bound to homologous antigens. Pre-incubation of anti-oxytocin or anti-vasopressin with beads coupled to their heterologous hormones completely removed the components binding to the heterologous hormone, leaving antibodies that showed fluorescence only with oxytocin or vasopressin respectively. This purification showed that in contrast to the findings with homologous antigens, not all of the antibody population bound to its heterologous antigen. Using non-purified anti-vasopressin, immunofluorescence was observed in neurohypophyses of homozygous Brattleboro rats. This was evidently due to those antibodies which bind to oxytocin, since the fluorescence was abolished after pre-incubating the antibody-containing plasmas with oxytocin-coupled beads. Immunofluorescence was still observed, however, in the suprachiasmatic nucleus of both Wistar and heterozygous Brattleboro rats, if purified anti-vasopressin was used. This was probably due to either vasopressin storage or production in these hypothalamic cells.

Animals↗