Debra E. Gmerek

Studies on bombesin-induced grooming in rats

The gross behavior induced by centrally administered bombesin in rats was compared to that elicited by ACTH-(1-24) and the somatostatin analog, des AA1,2,4,5,12,13[D-Trp8]-somatostatin (ODT8-SS). Bombesin (0.001-1 microgram, ICV) caused dose-related excessive scratching which was qualitatively different from that associated with the other two groom-inducing agents. Bombesin-induced grooming was not markedly affected by behaviorally nondepressant doses of haloperidol, morphine, naloxone or neurotensin. Bombesin was active in genetically hypotrichotic (essentially furless) rats; and, again in such animals, even after numbing the area caudal to the shoulders with lidocaine. Tolerance and cross-tolerance studies with bombesin and ODT8-SS indicated that they produce scratching through different mechanisms. Bombesin caused scratching when injected directly into the periaqueductal gray, but not when administered intravenously. Neither hypophysectomy nor adrenalectomy markedly affected bombesin-induced grooming. This behavior appears to be initiated in the central nervous system and is produced independently of the pituitary-adrenal axis.

Effect of Inorganic Lead on Rat Brain Mitochondrial Respiration and Energy Production

Abstract: Toxicologically significant amounts of inorganic lead were added to rat brain mitochondrial preparations that did not contain EDTA or Pi. The binding of the lead to the mitochondria was measured by anodic stripping voltometry. In the presence of lead, the respiratory control ratios decreased, implying a decrease in the degree of dependence of respiration on a phosphate acceptor. Nucleotide contents were also measured, and in the presence of inorganic lead the actual amounts of ATP formed from ADP were found to be significantly decreased as well.

Intrathecal bombesin in rats: Effects on behaviour and gastrointestinal transit

When bombesin is given intracerebroventricularly to rats, it is known to cause excessive scratching and inhibit gastrointestinal transit. We have administered bombesin via a permanent indwelling cannula into the subarachnoid space of the lumbar spinal cord of rats. By this route, bombesin elicited immediate excessive scratching and rapidly inhibited passage of a charcoal meal along the gastrointestinal tract. The A50 values for these effects were 0.004 (0.001-0.018) micrograms/rat and 0.34 (0.22-0.55) micrograms/rat, respectively. Bombesin-induced scratching and inhibition of transit are therefore mediated at spinal, as well as supraspinal, levels.

A species difference in the slowing effect of intrathecal morphine on gastrointestinal transit.

Intrathecal administration of morphine, levorphanol, bremazocine, ethylketocyclazocine or [D-Ser2,Leu5,Thr6]enkephalin to rats, at doses 10-50 times greater than that necessary to elicit analgesia in the tail flick test, had no marked effect on gastrointestinal transit as determined by the charcoal meal test. In contrast, intrathecal administration of various doses of morphine to mice significantly antagonized transit (A50 (that dose which inhibited transit to 50% of controls) = 14.7 (0.71-2.89) micrograms/mouse). These results suggest (1) a lack of involvement of opioid sensitive spinal structures in the control of gastrointestinal transit in rats, and (2) a species difference in the slowing effect of intrathecal morphine on gastrointestinal transit.

Use of a programmable protocol timer and data logger in the monitoring of animal behavior

The quantitative assessment of animal behavior by continuous or intermittent observation often requires much time and intense concentration. We have developed an accurate and convenient system which allows one observer to monitor up to four animals and record two types of behavior simultaneously. The system uses an inexpensive portable microcomputer including keyboard, video monitor, and cassette recorder. A program written in BASIC generates timed visible and audible cues, and tabulated data entered through the keyboard. The total number of occurrences of a particular behavior (e.g., wet dog shakes) and the frequency of a second behavior (e.g., grooming) may be measured. Frequency data, bases on intermittent observations, is processed for histogram display. The results may be copied from the video display or recorded on tape for further statistical analysis. The system has provided a convenient way to time experiments and collect data on drug-induced behavior in over 800 rats.

Classification of opioids on the basis of their ability to antagonize bombesin-induced grooming in rats

Bombesin (0.001-0.125 microgram) elicits dose-related excessive grooming when administered i.c.v. to rats. In contrast to the grooming produced by various other endogenous and exogenous agents, bombesin-induced grooming is not markedly affected by behaviorally non-depressant doses (determined by a novel method) of morphine (3 and 10 mg/kg, s.c.). The benzomorphan, ethylketocyclazocine (0.25-0.5 mg/kg, s.c.), does attenuate bombesin-induced grooming in a dose-related and stereospecific manner. In this study, we have classified 27 opioids on the basis of their ability to antagonize bombesin-induced grooming. The results of the classification indicate that a benzomorphan nucleus is required for activity in this test. Our test provides evidence that postulated benzomorphan binding sites are functionally active as pharmacologic receptors.

ACTH-(1–24) and RX 336-M induce excessive grooming in rats through different mechanisms

ACTH-(1-24) (0.03-6 micrograms i.c.v.) and RX 336-M (7,8-dihydro-5,6-dimethylcyclohex-5-eno-1,2,8,14 codeinone) (1.5-6 mg/kg i.p.) induce dose-related excessive grooming and wet-dog shaking in rats. In the present study, the grooming associated with these compounds was compared and analyzed pharmacologically. Grooming caused by RX 336-M and by ACTH-(1-24) was antagonized when rats were pretreated with comparable doses of morphine (0.5-4 mg/kg s.c.), however, only ACTH-(1-24)-induced grooming was attenuated by naloxone (1 and 10 mg/kg s.c.). ICI 154,129 (N,N-bisallyl-Tyr-Gly-Gly-psi-(CH2S)-Phe-Leu-OH) (30 mg/kg s.c.), a selective delta-opiate receptor antagonist, was ineffective against both ACTH-(1-24) and RX 336-M. Although haloperidol is known to antagonize grooming elicited by ACTH-(1-24) (e.g., Wiegant et al., 1977, European J. Pharmacol. 41, 343), even a high dose of this neuroleptic agent (5 mg/kg s.c.) only partially attenuated grooming caused by RX 336-M. Tolerance developed to the grooming elicited by RX 336-M, and by ACTH-(1-24), but there was no cross-tolerance. Both agents were active in genetically hypotrichotic rats; and, again in such animals, even after numbing the area caudal to the shoulders with lidocaine. Given the divergent results with naloxone, and, possibly, with haloperidol, and the lack of cross-tolerance, we conclude that the excessive grooming induced in rats by ACTH-(1-24) and by RX 336-M is mediated by different mechanisms.

In vivo studies with ICI 154, 129, a putative delta receptor antagonist

We studied the in vivo pharmacology of ICI 154,129, a new antagonist that is claimed to show selectivity for delta opiate receptors. At s.c. doses of 30 and 100 mg/kg, ICI 154,129 had no marked effect on the gastrointestinal transit of a charcoal meal in mice. In this test, ICI 154,129 reversed the inhibitory action of metkephamid (a proposed delta receptor agonist) but not that of levorphanol. ICI 154,129 was proconvulsant in the mouse picrotoxin potentiation test; the dose-response curve had a low ceiling and was biphasic. Naloxone (1 mg/kg, s.c.) enhanced the proconvulsant action of ICI 154,129 (40 mg/kg, s.c.) by an unknown mechanism.

An animal model for preclinical screening of systemic antipruritic agents

Reliable antipruritic agents that can be given systemically are not available at present. This may be due to the lack of animal models for screening such compounds. Bombesin, a tetradecapeptide originally isolated from frog skin, induces dose-related excessive scratching when administered intracerebroventricularly (i.c.v.) to rats. With the help of a microcomputer, we monitored the scratching elicited by a standard, submaximal dose of bombesin (0.10 microgram, i.c.v.). This system provides 1) a sensitive and novel way of assessing drug-induced behavioral depression, and 2) a means of quantifying interactions between bombesin and possible antagonists. Thus, bombesin-induced grooming is antagonized by behaviorally nondepressant doses of methdilazine, trimeprazine, and chlorpromazine but not by morphine, haloperidol, diphenhydramine, hydroxyzine, mepyramine, cimetidine, or cyproheptadine. Methdilazine and trimeprazine are used clinically as antipruritic agents. The model therefore offers a means of evaluating new, systemic antipruritic agents, particularly those which may be active in treating histamine-independent pruritus.

A study of the shaking and grooming induced by RX 336-M in rats

Several endogenous peptides and experimental agents induce wet-dog shakes and excessive grooming after acute administration to rats, but quantitative information on a possible relationship between the two behaviors is lacking. RX 336-M (7,8-dihydro-5-6-dimethylcyclohex-5-eno-1,8,14 codeinone) is a novel compound which elicits dose-related shaking and grooming in the rat. We have measured and compared the shaking and grooming induced by several doses of RX 336-M (1.5-12 mg/kg, IP) in male Sprague Dawley rats at various stages of maturation. Analysis of the correlation between the number of wet-dog shakes and the frequency of grooming episodes indicates that a relationship may exist between the shaking and grooming. The excessive grooming induced by RX 336-M may be a mechanism by which the rats state of arousal (raised by the shaking) is lowered and homeostasis is maintained.