Nerve Growth Factor Antiserum Research Articles

Regenerative repair in the severed optic nerve of the newt ( Triturus viridescens): Effect of nerve growth factor antiserum

At 14 days after transection those regenerating newt ( Triturus viridescens) optic nerves receiving anti-nerve growth factor treatment were easily distinguished from regenerating controls. Quantitative analysis revealed that antiserum treatment significantly reduced nerve diameter and cross-sectional areas compared to the control groups. Quantitation from electron microscope montages of nerve cross sections revealed similar results. In addition, antiserum treatment significantly reduced the area of regenerating axon fascicles per nerve cross section compared to the control groups. Most significantly, the mean number of regenerating axons per nerve decreased more than 50% in the antiserum-treated group. Regenerating axons appeared normal in all three groups. Axons were filled with clear cytoplasm containing neurofilaments, neurotubules, and an occasional mitochondrion. Axon density was not significantly different among the three groups and axon diameters were similar from 0.1 to 0.8 μm. Distention of glial cell processes surrounding fascicles of axons and increased intra- and extracellular debris may indicate an alteration of glial cell activity in the antiserum group. Many of the 14-day antiserum-treated nerves have the appearance of an untreated transected optic nerve 6 to 10 days after lesion.

Development of left ventricular hypertrophy in young spontaneously hypertensive rats after peripheral sympathectomy.

The effects of peripheral sympathectomy with nerve growth factor antiserum (NGFAS) on blood pressure, systemic hemodynamics, myocardial function, myocardial hypertrophy, and renin were studied in male spontaneously hypertensive (SH) rats of the Okamoto strain and normotensive control Kyoto-Wistar (WKY) rats. NGFAS prevented the developing of hypertension in the SH rats but did not alter blood pressure in the WKY rats. The NGFAS-treated SH rats developed the same hemodynamic abnormalities as the sham-treated rats, including increased peripheral vascular resistance and depressed cardiac output; Indices of left ventricular performance, including peak flow velocity, stroke power, stroke work, dP/dtmax, and flow acceleration (dF/dt), were diminished in the SH rats compared to the WKY rats. NGFAS treatment further depressed ventricular function in the SH rats, but had little effect on the WKY rats; Plasma renin activity in both the SH and WKY rats was unaffected by NGFAS treatment. Although NGFAS treatment effectively prevented the development of hypertension in the SH rats, it did not influence the development of left ventricular hypertrophy as reflected by increases in left ventricular mass, RNA, DNA, and hydroxyproline content. The data suggest that the development of myocardial hypertrophy and myocardial dysfunction in the SH rat is in part independent of hypertension and plasma renin activity.

Effects of nerve growth factor and nerve growth factor antiserum on tyrosine hydroxylase and dihydropterine reductase activities in rat superior cervical ganglia in vivo.

Effects of nerve growth factor and nerve growth factor antiserum on tyrosine hydroxylase and dihydropterine reductase activities in rat superior cervical ganglia in vivo.

Purification of nerve growth factor antibodies by affinity chromatography.

AbstractPure antibodies to nerve growth factor have been isolated from sheep nerve growth factorantiserum by affinity chromatography using 2.5 S nerve growth factor linked to Sepharose 4B by means of cyanogen bromide. The elution of the antibodies was accomplished either at low pH (pH 2) or by high salt concentration (4.5 wMgC12). The purity of the antibodies was established by SDS‐gel electrophoresis. Their immunological activity was tested by imrnunoprecipitation and their biological activity in a tissue culture assay using embryonic chick dorsal root ganglia.

Neonatal neuronal loss in rat superior cervical ganglia: retrograde effects on developing preganglionic axons and Schwann cells.

Beginning prenatally and during the first week after birth, there is normally a loss of axons in rat cervical sympathetic trunk. To test the hypothesis that this spontaneous axonal loss represents a natural process whereby an excessive number of immature preganglionic axons in the cervical sympathetic trunk adapts to the neuronal population in the superior cervical ganglion, the number of nerve cells in the superior cervical ganglion was reduced in newborn rats by administration of nerve growth factor antiserum, 6-hydroxy-dopamine or postganglionic anxotomy. Quantitative ultrastructural studies of these animals at later stages of development revealed that, with each method, the number of preganglionic axons and Schwann cells was reduced to nearly one-third of normal. These findings indicate that the superior cervical ganglion plays an important role in the development of the cervical sympathetic trunk. Removal of ganglionic cells causes a retrograde loss of preganglionic fibres. This process probably represents an exaggeration of the normal mechanism for elimination of redundant axons. Because the changes in axonal numbers are associated with similar reductions in the number of Schwann cells, it can also be concluded that postnatal Schwann cell proliferation is influenced by axonal populations.

Cardiac hemodynamics and hypertrophy in nerve growth factor antiserum treated spontaneously hypertensive rats

Cardiac hemodynamics and hypertrophy in nerve growth factor antiserum treated spontaneously hypertensive rats

Hypothalamic self-stimulation in rats following immunosympathectomy or central nerve growth factor antiserum injection.

In Experiment 1, immunosympathectomized rats self-stimulated at a much lower rate on high variable ratio schedules of reinforcement than injected controls. No differences were found for responding on continuous reinforcement or low variable ratio reinforcement schedules. In Experiment 2, a similar reduction in varialbe ratio response rate was found for subjects centrally injected with nerve growth factor-antiserum relative to controls. The results suggest a reduction in central catecholamine levels as a result of antiserum treatment.

Location of an isoproterenol-responsive cyclic AMP pool in adrenergic nerve cell bodies and its relationship to tyrosine 3-monooxygenase induction.

To decide whether adenosine 3':5'-cyclic monophosphate (cyclic AMP) plays a role as a second messenger in the trans-synaptic induction of tyrosine 3-monooxygenase (EC 1.14.16.2), it is desirable to discriminate between neuronal and extraneuronal changes in cyclic AMP concentration. Treatment of newborn rats with nerve growth factor antiserum or 6-hydroxydopamine, leading to destruction of 61-85% of the adrenergic nerve cell bodies in the superior cervical ganglion, led to a decrease in cyclic AMP of only 16-28%. This observation demonstrates that a relatively small portion of cyclic AMP is localized in the adrenergic neurons. However, administration of isoproterenol produced an increase (12-fold) in cyclic AMP only in this neuronal pool. Neither single nor repeated injections of isoproterenol led to induction of tyrosine monoxygenase. This, together with previous observations that experimental conditions leading to induction of the enzyme do not produce significant increases of cyclic AMP in the whole ganglion, is taken as an indication that cyclic AMP is not acting as a second messenger in the trans-synaptic induction of tyrosine monooxygenase in the rat superior cervical ganglion. In the rat adrenal medulla, treatment with reserpine led to both a shortlasting (60-90 min) increase in cyclic AMP and a subsequent induction of tyrosine monooxygenase. However, the increase in cyclic AMP was almost completely prevented (40 compared to 320%) by pretreatment of the rats with propranolol while the induction of tyrosine monooxygenase was not diminished. This observation also argues against an exclusive key-function of cyclic AMP in trans-synaptic induction of tyrosine monooxygenase in the adrenal medulla.

Role of sympathetic fibres and of adrenal medulla in the maintenance of cardio-vascular homeostasis in normotensive and hypertensive rats

The Concept of modulation of cardiovascular system by the autonomie nervous system has been accepted for a long time. However, the efferent sympatheticcomponent of the autonomie nervous system is more dynamically related to the peripheral cardiovascular system by a dense plexus of excitatory fibres distributed to the heart and to all vascular beds. Small arterioles which are crucial to the homeostasis of arterial blood pressure were found to be the most densely innervated segments of the vascular tree (N orberg, 1967). In addition to this network of excitatory fibres, the sympathetic nervous system is also responsible for the control of catecholamine secretion by the adrenal medulla which can also influence the heart and blood vessels by acting on specific receptors to catecholamines localized in these structures. Little is known about the relative contribution of each component of the sympathetic system in the maintenance of normal cardiovascular functions. In numerous previous studies, the effects of sympathectomy were studied after surgical denervation, after treatment with nerve growth factor antiserum or after administration of catecholamine depleting drugs such as reserpine. Most of these techniques however were either non specific to the peripheral fibres or, resulted in an incomplete sympathectomy so that the conclusions reached with these procedures may be questionable. The discovery that 6-hydroxydopamine (6-OH-DA) could specifically and selectively destroy adrenergic nerve fibres (T ranzer and Thoenen, 1967) has given a more specific and more useful tool for the study of the role of the sympathetic nervous fibres. In periphery, 6-OH-DA when given in sufficient amount, causes the degeneration of the great majority of sympathetic fibres without destroying the ganglion cell bodies or the adrenal medulla.

Trans-synaptic regulation of tyrosine hydroxylase activity in a developing mouse sympathetic ganglion: Effects of nerve growth factor (NGF), NGF-antiserum and pempidine

The response of the normal and surgically decentralized mouse superior cervical ganglion to nerve growth factor (NGF) and its antiserum have been studied in neonatal animals. NGF was able to increase the activity of the enzyme tyrosine hydroxylase, used as a marker for adrenergic neurones, in both normal and decentralized ganglia. However, it was unable to reverse the acute effects of surgical decentralization; a significant difference in tyrosine hydroxylase activity between the control and decentralized ganglia was observed even after the NGF treatment. In addition, 7 weeks after the NGF treatment had ceased, the enzyme activity returned to levels not significantly different from those in animals that had never received NGF. NGF antiserum was equally effective in causing a reduction in tyrosine hydroxylase activity in normal and decentralized ganglia. The ganglion blocking drug pempidine when administered to neonatal mice mimicked the effects of surgical decentralization; tyrosine hydroxylase activity of the superior cervical ganglia of animals treated with this drug failed to develop normally. It was also found that, as in surgically decentralized animals, NGF was unable to reverse the effects of pempidine. The effects of NGF were transient, while the effects of decentralization and pempidine were long lasting, suggesting that the most important regulation for ganglion development is the influence of trans-synaptic activity, most probably mediated by the depolarizing action of acetylcholine.

An assessment of sympathetic function in isolated tissues from mice given nerve-growth-factor antiserum and 6-hydroxydopamine.

1. Experiments were done on isolated tissues from mice injected with 0.9% w/v NaCl solution (saline), 6-hydroxydopamine (6-OHDA), nerve-growth-factor antiserum (NGF-As) or a combination of these agents (NGF-As+6-OHDA).2. Fluorescence histochemistry of vasa deferentia showed clear differences between each of the treatments but no such distinction was possible in cardiac ventricle or intestine.3. Compared with controls, the chronotropic responses of atria to field stimulation were reduced by all three treatments in the order NGF-As<6-OHDA<NGF-As+6-OHDA. Supersensitivity to noradrenaline occurred in atria from all treated groups.4. In contrast, vasa deferentia from NGF-As-treated mice responded almost identically with controls to both field stimulation and to exogenous noradrenaline. Considerable functional impairment and noradrenaline supersensitivity were obvious in both the 6-OHDA and the NGF-As+6-OHDA groups but no distinction could be made between them.5. The relaxation of superfused ileum following nerve stimulation was reduced in the NGF-As group, whilst preparations from the two groups given 6-OHDA usually gave a motor response which was blocked by atropine.6. None of the treatments employed in these experiments caused complete sympathectomy although 6-OHDA alone or in combination with NGF-As produced a more pronounced effect than NGF-As alone and the relative ineffectiveness of the latter is discussed. The implications of the motor response after 6-OHDA in ileum when the nerves were stimulated is considered in the light of the cholinergic link hypothesis.

Immunosympathectomy: Late effects on the composition of rat cervical sympathetic trunks and influence on axonal regeneration after crush

Uninjured and crushed cervical sympathetic trunks from immunosympathectomized and control rats 8 months old were compared by quantitative electron microscopic techniques. It was demonstrated that: 1. The unmyelinated axonal population, reduced by neonatal administration of nerve growth factor-antiserum, remains stable at approximately one-third of normal. 2. Median diameters of unmyelinated axons were significantly smaller for nerves from nerve grwoth factor-antiserum-treated animals than for control nerves. 3. The number of myelinated axons in cervical sympathetic trunks from nerve growth factor-antiserum-treated animals was reduced to approximately 1/20 of normal. 4. By 8 months of age, the configuration of Schwann cell complexes in nerves from immunosympathectomized animals had changed; the elongated processes and collagen pockets which were present at earlier stages were less prominent. 5. 2 weeks after crush injury, axonal sprouting was similar for immunosympathectomized and control cervical sympathetic trunks.

Comparative studies on the effect of the nerve growth factor on sympathetic ganglia and adrenal medulla in newborn rats

The effect of nerve factor (NGF) and nerve growth factor antiserum (NGF-AS) on sympathetic ganglia and adrenal chromaffin cells of newborn rats was investigated in a comparative study. In confirmation of previous experiments, NGF had a marked hypertrophic and hyperplastic effect in sympathetic ganglia. These morphological changes were accompanied by a selective induction of tyrosine hydroxylase and dopamine β-hydroxylase. The biochemical correlate with the degenerative changes elicited by NGF-AS in sympathetic ganglia was a reduction of all enzymes studied that are located in adrenergic neurones. In the adrenal medulla neither NGF nor NGF-AS produced any detectable morphological changes. However, both procedures resulted in a statistically significant increase in tyrosine hydroxylase and dopamine β-hydroxylase. The paradoxical finding that both NGF and its antiserum produce the same effect on the adrenal chromaffin cells is most plausibly explained by the assumption that both treatments elicit a stress-induced increase in the activity of splanchnic nerves supplying the adrenal medulla, and to a consequent neurally mediated induction of these key enzymes involved in the synsthesis of the adrenergic transmitter, as they do in adult animals under various experimental conditions that lead to an increased activity of the sympathetic nervous system.

Effects of NGF-antiserum in sympathetic neurons during early postnatal development

Mice at birth and on days 4, 7, and 14 were injected with nerve growth factor antiserum (A-NGF) to determine any changes in the nature of lesions during the early postnatal development of the superior cervical sympathetic ganglion. Correlative light and electron microscopic studies revealed both numerical and morphological changes between the first and second weeks of life. The alterations present in the two younger groups resembled those previously described in the newborn. The three zones were seen in many of the nuclei of affected neurons, resulting from nucleolar segregation. Zones A and B contained RNA and probably represent the pars granulosa and pars fibrosa, respectively, of the nucleolus, while zone C contained DNA, initially forming from the nucleolus-associated chromatin. Lesions were fewer in number and less severe in 7- and 14-day-old mice, but still included similar nucleolar alterations. Zone C differed significantly in structure, however, in that large numbers of vacuoles, many of which contained inclusions of varying form and density, were present throughout its substance. Evidence from 3H-uridine experiments, the similarity of nucleolar alterations to those produced by agents known to block DNA-dependent nucleolar RNA synthesis, and the absence of comparable lesions in the adult strongly suggest that A-NGF produces a block in rRNA synthesis in early neuronal development.

The effects of "immunosympathectomy" on blood pressure and vascular "reactivity" in normal and spontaneously hypertensive rats.

AbstractNewborn litters of spontaneously hypertensive rats (SHR) and normotensive control rats (NCR) were identically treated with sympathetic nerve growth factor antiserum (Wellcome) which markedly interferes with adrenergic cardiovascular control (Zaimis 1967). Blood pressure, measured intermittently during 8 months, was in treated SHR (SHRis) about 25 % higher than in NCRis, their respective pressures being about 40 % and 25 % lower than those of sham‐treated SHR and NCR.–The hindquarters of one SHRis, or NCRis, were then perfused at constant flow in parallel with those of ordinary NCR. Starting from maximal vasodilatation, resistance increases were induced by graded noradrenaline (NA) infusions, from “threshold” to maximal pressor responses. Compared to NCRis, SHRis showed an increased resistance at maximal dilatation, an increased slope of the NA dose‐response curve and an increased maximal pressor response, while their NA “thresholds” did not differ significantly. Thus, the structurally determined hemodynamic differences between ordinary SHR and NCR (Folkow et al. 1970 b) characterize also SHRis and NCRis, though to a reduced extent. Even when comparing SHRis with ordinary‐ NCR, which exhibited similar “resting” pressures, these differences partly remain, suggesting that the SHR resistance vessels might, for genetic reasons, be more prone to adapt structurally to pressure loads than those of NCR.

The role of post‐synaptic neurones in the biochemical maturation of presynaptic cholinergic nerve terminals in a mouse sympathetic ganglion

1. The role of post-synaptic adrenergic neurones in the biochemical maturation of presynaptic cholinergic nerve terminals has been investigated in mouse superior cervical ganglion in vivo.2. Selective destruction of ganglion adrenergic neurones chemically, with 6-hydroxydopamine (6-OH-DA), or immunologically, with nerve growth factor antiserum (NGF-antiserum) prevented the normal maturation of choline acetyl transferase (ChAc) activity in presynaptic endings during development. Enzyme activity remained depressed for at least 2 months.3. 6-OH-DA treatment failed to alter ChAc activity in the developing duodenum or diaphragm, organs in which cholinergic fibres do not synapse with adrenergic neurones, suggesting that destruction of post-synaptic neurones per se inhibited presynaptic maturation.4. Similarly, NGF-antiserum, which does not destroy adrenergic neurones in the adult did not alter ChAc activity in adult mouse ganglia.5. These observations suggest that post-synaptic adrenergic neurones regulate the biochemical development of presynaptic cholinergic nerve terminals.

Effects of nerve growth factor antiserum on peripheral unmyelinated nerve fibers.

Nerve Growth Factor (NGF) antiserum causes neuronal loss in the sympathetic ganglia of newborn rats. Pre and post ganglionic sympathetic fibers from eleven such animals and eight controls were studied by phase contrast and electron microscopy. Quantitative techniques were applied to the study of different parameters of the axons and Schwann cells. Treated animals showed only 24% of the axonal population of controls and 34% of Schwann cells. This fiber loss was non-selective, affecting all fiber sizes. Schwann cells from treated animals enclosed fewer axons than normal and some contained none. Contrary to what is seen following transection of an unmyelinated nerve, regenerative axonal sprouting was not observed during the 6 week period studied. Schwann cell processes were elongated and unfolded, frequently engulfing bundles of collagen. NGF antiserum experiments provide a tool for the study of axonal and Schwann cell behaviour after neuronal loss avoiding the disadvantages of the traumatic disruption of neural architecture that results from experimental surgical lesions.

The effects of "immunosympathectomy" on blood pressure and vascular "reactivity" in normal and spontaneously hypertensive rats.

Newborn litters of spontaneously hypertensive rats (SHR) and normotensive control rats (NCR) were identically treated with sympathetic nerve growth factor antiserum (Wellcome) which markedly interferes with adrenergic cardiovascular control (Zaimis 1967). Blood pressure, measured intermittently during 8 months, was in treated SHR (SHRis) about 25 % higher than in NCRis, their respective pressures being about 40 % and 25 % lower than those of sham-treated SHR and NCR.–The hindquarters of one SHRis, or NCRis, were then perfused at constant flow in parallel with those of ordinary NCR. Starting from maximal vasodilatation, resistance increases were induced by graded noradrenaline (NA) infusions, from “threshold” to maximal pressor responses. Compared to NCRis, SHRis showed an increased resistance at maximal dilatation, an increased slope of the NA dose-response curve and an increased maximal pressor response, while their NA “thresholds” did not differ significantly. Thus, the structurally determined hemodynamic differences between ordinary SHR and NCR (Folkow et al. 1970 b) characterize also SHRis and NCRis, though to a reduced extent. Even when comparing SHRis with ordinary- NCR, which exhibited similar “resting” pressures, these differences partly remain, suggesting that the SHR resistance vessels might, for genetic reasons, be more prone to adapt structurally to pressure loads than those of NCR.

Analysis of the effects of the antiserum to the nerve growth factor in adult mice

The effect of the nerve growth factor antiserum (NGF-AS) on adult mice has been studied at morphological and biochemical levels. A 5-day treatment of 3-month-old mice results in a marked volume reduction of sympathetic chain ganglia. This decrease in volume appears to be due in part to cell death, but to a major extent to atrophy of residual neurons. A severe impairment of the sympathetic function also obtains, as indicated by a marked decrease of catecholamine uptake and content in peripheral organs. A few weeks after the end of treatment, a gradual and progressive recovery of the sympathetic function ensues, and the nerve cells regain their normal appearance. These results are described in connection with the problem of the site of action of NGF antibodies in immature and in fully differentiated sympathetic neurons.

In utero sensitivity of rat pineal to nerve growth factor antiserum

In utero sensitivity of rat pineal to nerve growth factor antiserum