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N Volkmer

Publications and source records attributed to N Volkmer.

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Hypothalamic proline endopeptidase activity is not changed by various behavioral procedures.

Proline endopeptidase (E.C.3.4.21.26) is an enzyme which cleaves several neuropeptides at the carboxyl-side of proline residues. Some peptide substrates of this enzyme may be found in the rat hypothalamus (thyrotropin releasing hormone, neurotensin, substance P, oxytocin, vasopressin, beta-endorphin). Recent research has shown that the hypothalamic levels of some of these substances (e.g., vasopressin, beta-endorphin) change by a variety of training procedures. We studied the effect of various forms of training on the activity of proline endopeptidase of rat hypothalamus. The present results show that the activity of this enzyme is not altered by electroconvulsive shock or inhibitory avoidance training when measured, 0, 1, or 3 hr after these procedures. Other behavioral procedures (habituation to an open field, two-way active avoidance conditioning, or 1 min of inescapable footshock) also had no effect on hypothalamic proline endopeptidase activity measured immediately after training or test sessions. We conclude that proline endopeptidase probably does not play a regulatory role in the effect of synaptically released hypothalamic neuropeptides on behavior.

Animals

Distribution of proline endopeptidase activity in sub-synaptosomal fractions of rat hypothalamus.

1. Proline endopeptidase (E.C.3.4.21.26) is an enzyme which cleaves several peptides at the carboxyl side of proline residues. Because brain contains relatively large amounts of this enzyme and because of its specificity it has been suggested that it plays a role in the metabolism of neuropeptides, acting both on their processing and their degradation. 2. Since the final steps of neuropeptide processing occur in the synaptic vesicles and the degradation of most of these peptides is believed to occur in the synaptic cleft, we studied the distribution of proline endopeptidase activity in sub-fractions of rat hypothalamus. 3. Proline endopeptidase activity is present in synaptosomal fractions and is released by hypo-osmotic shock. Its specific activity is higher in the synaptoplasma than in synaptic membranes or vesicles (7.98 vs 0.18 and 0.24 nmol min-1 mg protein-1 carbobenzoxy-glycyl-prolyl-sulfamethoxazole hydrolysis). 4. Inhibitory avoidance training, a situation which releases hypothalamic vasopressin and beta-endorphin, both in vitro substrates, did not affect the specific or total activity of proline endopeptidase in synaptosomal plasma membranes.

Animals

Response of the rat brain beta-endorphin system to novelty: importance of the fornix connection.

In control rats, a step-down inhibitory avoidance training trial using a 0.8 mA footshock, or simple exposure to the training apparatus without footshock, was followed by a decrease of beta-endorphin-like immunoreactivity measured in the hypothalamus and ventral thalamus. The effect of inhibitory avoidance training was also measured in rats submitted to a brain sham operation, to bilateral transection of the dorsal fornix, to anterior or to posterior hypothalamic deafferentation, to adrenal medullectomy, to an adrenal sham operation, to 16 daily ip injections of 0.2 mg/kg dexamethasone, or to 16 daily ip injections of 1 ml/kg saline. The diencephalic beta-endorphin-like immunoreactivity response to training was abolished by fornix transection and was unaffected by all other treatments. This suggests that the response is not mediated by anterior or posterior neural afferents to the hypothalamus, or by a hypersecretion of epinephrine by the adrenal medullae, or of ACTH by the pituitary gland. The response, instead, appears to require the integrity of the pathway that sends projections from the septo-hippocampal system to the hypothalamus. Previous evidence had suggested that the diencephalic beta-endorphin-like immunoreactivity response to training is a result of novelty, and the septo-hippocampal system has been postulated to play a role in the registration of novelty.

Adrenal Medulla

The course of the decrease of hypothalamic beta-endorphin induced by training, and the development of the effect of beta-endorphin on the retrieval of inhibitory avoidance in rats.

Step-down inhibitory avoidance training or the simple exposure of rats to the training apparatus is followed by a decrease of hypothalamic beta-endorphin immunoreactivity at 0.1, 1.0 or 2.0 h after training. Immunoreactivity returns to normal at 6.0 h. The ip administration of 1.0 microgram/kg of human beta-endorphin 6 min prior to training produces an inhibition of the retrieval of the step-down task at 6.0 h, but not at 0, 1.0 or 2.0 h after training. This effect is reversed by a second injection of the substance immediately before testing. The possible physiological significance of this parallel development of the effect of beta-endorphin on retrieval and the depletion of the substance caused by training is discussed. The data indicate that retrieval is insensitive to the peptide when its hypothalamic stores are depleted.

Animals

Interferon induction in mouse fibroblast L-A9 cells.

Mouse L-A9 cell interferon was induced by infection with Newcastle disease virus. Interferon production was 1.5 X 10(5) IU/10(7) cells. Interferon was partially purified by precipitation with ammonium sulphate, chromatography on CM-Sephadex and hydrophobic chromatography on octyl-agarose. The specific activity of the final preparation was 1.7 X 10(7) IU/mg protein. Treatment of L-A9 cells with 20 IU/ml interferon prior to viral infection inhibited the intracellular accumulation of reovirus-specific double-stranded RNA. Dose-response studies of the cells to interferon indicated that L-A9 cells require 10, 13 and 15 IU/ml to obtain 50% viral plaque reduction for Marituba virus, vesicular stomatitis virus and reovirus, respectively. The present results demonstrate the potential of mouse L-A9 cells as an interferon-producing system and also as a model for the study of the effect of cellular response to exogenous interferon treatment on the replication of RNA viruses.

Animals