A simplified method for raising antibodies to rat dopamine‐β‐hydroxylase

Helle, Karen B.; Fillenz, Marianne; Stanford, S Clare; Pihl, K.E.; Srebro, Bolek

doi:10.1111/j.1471-4159.1979.tb11068.x

Cited by 23 publications

(3 citation statements)

References 16 publications

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“…By CIE no identity has been detected between the bovine and sheep enzymes, on the one side, and DPH of man, the dog, cat, rabbit, rat, mouse, hamster, guinea pig, and hedgehog, on the other, with any of our antisera against bovine DPH. Partial identity has, on the other hand, been demonstrated between the sheep and bovine enzymes and that of man and the smaller mammals by radio-immunoassays [28, 68, 82, 861, enzyme inhibition [20,43,701, and immunohistochemical studies [41,42,47,58,62,921. An example of the latter is given in Fig.…”

Section: (B) Dph Of the Sheep And Pigmentioning

confidence: 99%

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Immunochemical Characterization of Dopamine β‐Hydroxylase

1982

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Section: (B) Dph Of the Sheep And Pigmentioning

confidence: 99%

“…The anti-rat DPH raised to the total granule DPH without further purification [58] has been used for comparison of immunofluorescence in the rat brain with heterologous ( Fig. 3C) and homologous (Fig.…”

Section: (B) Anti-rat Dphmentioning

confidence: 99%

Immunochemical Characterization of Dopamine β‐Hydroxylase

1982

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“…In synaptic vesicles (Klein et al, 1977;Kirksey et al, 1978) and in the chromaffin granules of many species [e.g. the rat (Ciaranello et al, 1975;Helle et al, 1979)] dopamine fl-hydroxylase is recovered largely as a component of the vesicle membrane. The bestcharacterized granules, however, are those of bovine origin, and in this case about 50% of the enzyme activity is recovered as a soluble component of the granule matrix (Hortnagl et al, 1972).…”

mentioning

confidence: 99%

Reduction of membrane-bound dopamine β-hydroxylase from the cytoplasmic surface of the chromaffin-granule membrane

Grouselle¹,

Phillips²

1982

Biochemical Journal

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Resealed bovine chromaffin-granule 'ghosts' were used for assaying the membrane-bound form of dopamine beta-hydroxylase. Hydroxylation of the substrate tyramine is dependent on its accumulation within the 'ghosts', where the active site of the enzyme is located. Free tyramine in the medium is at a low concentration, low ionic strength and a relatively high pH (7.0), so that even in the presence of a reducing agent (co-reductant) the unaccumulated amine is hydroxylated at a negligible rate. 'Ghosts' contain an endogenous co-reductant, which is shown to be catecholamine remaining in the membrane itself after granule lysis. Catecholamine that is free in solution in the medium or in the interior of the 'ghosts' is not effective as co-reductant, nor is ascorbate, in contrast with the situation with soluble dopamine beta-hydroxylase. Ferrocyanide is an active co-reductant, however, giving a hydroxylation rate approximately equal to the tyramine accumulation rate: it does not enter the 'ghosts', nor does the enzyme appear to utilize ferrocyanide sealed inside the 'ghosts'. A mechanism must therefore exist for transferring electrons across the membrane from the cytoplasmic surface to the matrix surface. NADH is not an electron donor for the enzyme, nor is cytochrome b-561 involved.

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An immunohistochemical study of the organization of catecholaminergic cells and terminal fields in the paraventricular and supraoptic nuclei of the hypothalamus

Swanson

Sawchenko

Bérod

et al. 1981

J of Comparative Neurology

387

112

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The distribution of catecholaminergic fibers and cell bodies in the paraventricular and supraoptic nuclei of the hypothalamus was investigated with immunohistochemical methods in the adult albino rat. Sections through the nuclei were stained with antisera to the catecholamine synthesizing enzymes tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT). The results suggest that adrenergic (PNMT-stained) fibers innervate the entire parvocellular division of the paraventricular nucleus, although the highest density of fibers was found in the medial part of the division. Only widely scattered adrenergic fibers are found in the magnocellular division of the nucleus and in the supraoptic nucleus. Noradrenergic fibers appear to innervate the periventricular zone of the paraventricular nucleus and those parts of the paraventricular and supraoptic nuclei that contain predominantly vasopressinergic neurons in both the normal and in the homozygous Brattleboro rat. Significant numbers--somewhat more than 500--of dopaminergic (TH-stained) neurons are found in the paraventricular nucleus; the cells are distributed throughout the nucleus but are concentrated in the medial and periventricular parts of the parvocellular division. Double-labeling experiments with the retrogradely transported tracer true blue indicate that between 4% and 8% of the dopaminergic neurons in the paraventricular nucleus project to the region of the dorsal vagal complex and/or thoracic levels of the spinal cord. It is concluded that adrenergic inputs to the paraventricular nucleus may influence cells that project to the median eminence and to preganglionic autonomic cell groups in the medulla and spinal cord. Noradrenergic inputs to the supraoptic and paraventricular nuclei may influence primarily vasopressinergic cells that project to the posterior lobe of the pituitary, as well as cells in the periventricular part of the paraventricular nucleus that project to the median eminence.

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A simplified method for raising antibodies to rat dopamine‐β‐hydroxylase

Cited by 23 publications

References 16 publications

Immunochemical Characterization of Dopamine β‐Hydroxylase

Immunochemical Characterization of Dopamine β‐Hydroxylase

Reduction of membrane-bound dopamine β-hydroxylase from the cytoplasmic surface of the chromaffin-granule membrane

An immunohistochemical study of the organization of catecholaminergic cells and terminal fields in the paraventricular and supraoptic nuclei of the hypothalamus

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