Using genetically-defined rodent strains for the identification of hippocampal traits relevant for two-way avoidance behavior: a non-invasive approach

Genetically-defined rodent strains permit the identification of hippocampal traits which are of functional relevance for the performance of two-way avoidance behavior. This is exemplified here by analyzing the relationship between infrapyramidal mossy fibers (a tiny projection terminating upon the basal dendrites of hippocampal pyramidal neurons) and two-way avoidance learning in about 800 animals. The necessary steps include 1) identification of structural traits sensitive to selective breeding for extremes in two-way avoidance, 2) testing the robustness of the associations found by studying individual and genetical correlations between hippocampal traits and behavior, 3) establishing causal relationships by Mendelian crossing of strains with extreme structural traits and studying the behavioral consequences of such structural 'randomization', 4) confirming causal relationships by manipulating the structural variable in inbred (isogenic) strains, thereby eliminating the possibility of genetic linkage, and 5) ruling out the possibility of spurious associations by studying the correlations between the hippocampal trait and other behaviors known to depend on hippocampal functioning. In comparison with the classical lesion approach for identifying relationships between brain and behavior, the present procedure appears to be superior in two aspects: it is non-invasive, and it focuses automatically on those brain traits which are used by natural selection to shape behaviorally-defined animal populations, i.e., it reveals the natural regulators of behavior.     

Kinetics of lymphocyte division in blood cultures studied with the BrdU-giemsa technique

Summary Normal human lymphocytes were cultured for 72 h with different doses of BrdU. The analysis of metaphases processed with the BrdU-Giemsa method shows that in leukocyte cultures 3 different lymphocyte populations coexist which are able to perform 1, 2 or 3 rounds of replication in vitro. Moreover, it was concluded that 5 g/ml is the minimal dose of BrdU inducing good differentiation in the areas of sister chromatid exchanges.This work was supported by grants from the International Atomic Energy Agency, the Consejo Nacional de Investigaciones Cientificas and the Comisión de Investigaciones Cientificas.     

ACTH response induced by interleukin-1 is mediated by CRF secretion stimulated by hypothalamic PGE

Summary We investigated whether hypothalamic prostaglandin E₂ (PGE₂) and corticotropin releasing factor (CRF) are responsible for the development of the adrenocorticotropic hormone (ACTH) response induced by interleukin-1 (IL-1). The present results show that ACTH responses induced by intravenous injection of IL-1 were suppressed by systemic pretreatment with indomethacin and that intrahypothalamic injection of PGE₂ stimulates the secretion of ACTH. Furthermore, systemic pretreatment with anti-CRF antibody significantly suppressed the ACTH response induced by intrahypothalamic injection of PGE₂. These data suggest that the ACTH response induced by IL-1 is mediated by CRF secretion stimulated by hypothalamic PGE₂.     

Genetic and pharmacological models of cholinergic supersensitivity and affective disorders

Summary Increased muscarinic sensitivity has been associated with altered hormonal states (hypothyroidism and hyperadrenocorticism), chronic administration of muscarinic antagonists or antidepressants with muscarinic actions, selective breeding for anticholinesterase sensitivity, and certain inbred strains of rats and mice. Thus, both genetic and environmental factors may influence muscarinic receptor sensitivity. The reasonably detailed studies on the selectively-bred rats have revealed that the Flinders Sensitive Line (FSL) rats weigh less, are less active, are more sensitive to muscarinic agonists and to stressors, and have higher concentrations of hippocampal and striatal muscarinic receptors than normal, or the selectively-bred, Flinders Resistant Line (FRL) rats. Thus, there are a number of parallels between FSL rats and depressed humans. The FSL rats may be the first animal model of depression to mimic the actual trait of depression, and not just the state.     

Mitochondrial DNA mutators

In this article we review our current knowledge of the mechanisms by which point mutations arise in the mitochondrial DNA (mtDNA) of Saccharomyces cerevisiae and discuss to what extent these mechanisms operate in human mtDNA mutagenesis. The 3–5 exonuclease proofreading activity of Pol ensures accuracy of mtDNA replication in both yeast and humans, while the role of base excision repair in mtDNA error avoidance remains debated. The mitochondrial mismatch repair Msh1 protein, which removes transitions in yeast, is absent in humans, a particularity that might cause accumulation of transitions, while the most frequent substitution in yeast mtDNA is A:T to T:A transversion. Proofreading-deficient mutator human cell lines and knockin mice have been created. They will be useful for studying the mechanisms by which mtDNA mutations accumulate in diseases, ageing, malignancy and drug therapy.Received 25 May 2004; received after revision 21 June 2004; accepted 7 July 2004     

Effects of angiotensin II and of an angiotensin II receptor antagonist on Simian virus 40-induced tumor growth in vivo

Summary The effects of angiotensin II and of the competitive angiotensin II receptor antagonist saralasin on in vivo tumor growth were investigated in hamsters. Angiotensin II strongly inhibited tumor growth while saralasin stimulated it, though the high dose used had partial agonistic angiotensin II-like actions. Lower doses of saralasin were without significant effect on tumor weights.Acknowledgments. These studies were supported by the Deutsche Forschungsgemeinschaft (DFG) within the SFB 90 Cardiovaskuläres System. Saralasin was a gift from Dr A.W. Castellion, Norwich Pharmacal Company, USA. We thank S. Thiele, P. Baumgärtl and D. Kurz for excellent technical assistance.     

Genetic control of hippocampal cholinergic and dynorphinergic mechanisms regulating novelty-induced exploratory behavior in house mice

Summary Neurobehavioral genetics endeavors to trace the pathways from genetic and eenvironmental determinants to neuroanatomical and neurophysiological systems and, thence, to behavior. Exploiting genetic variation as a tool, the behavioral sequelae of manipulating these neuronal systems by drugs and antisera are analyzed. Apart from research in rats, this paper deals mainly with the genetically-influenced regulation in mice of exploratory behaviors that are adaptive in novel surroundings and are hippocampally-mediated. Special attention is paid to neuropeptidergic, GABAergic, and cholinergic synaptic functions in the mouse hippocampus.The behaviorally different inbred mouse strains C57BL/6 and DBA/2 show opposite reactions (reductions and increases, respectively, in exploration rates) to peripheral and intrahippocampal injections with agents that interfere with peptidergic, cholinergic, and GABAergic neurotransmission. These findings can be explained by an interdependent over-release of opioids, arrested GABA release, and excess acetylcholine in the hippocampal neuronal network of DBA/2 mice, as compared to C57BL/6 mice where these systems are functionally well balanced. Very similar results have been obtained with the lines SRH and SRL, derived from C57BL/6 and DBA/2, and genetically selected for rearing behavior. Most probably, the opioids act to disinhibit exploratory responses. An additional genetic approach is mentioned, in which four inbred mouse strains and one derived heterogeneous stock are used for estimating genetic correlations between structural properties of the hippocampal mossy fibers and levels of hippocampal dynorphin B, on the one hand, and frequencies of exploratory responses to environmental novelty, on the other.     

Genetic control of hippocampal cholinergic and dynorphinergic mechanisms regulating novelty-induced exploratory behavior in house mice

Neurobehavioral genetics endeavors to trace the pathways from genetic and environmental determinants to neuroanatomical and neurophysiological systems and, thence, to behavior. Exploiting genetic variation as a tool, the behavioral sequelae of manipulating these neuronal systems by drugs and antisera are analyzed. Apart from research in rats, this paper deals mainly with the genetically-influenced regulation in mice of exploratory behaviors that are adaptive in novel surroundings and are hippocampally-mediated. Special attention is paid to neuropeptidergic, GABAergic, and cholinergic synaptic functions in the mouse hippocampus. The behaviorally different inbred mouse strains C57BL/6 and DBA/2 show opposite reactions (reductions and increases, respectively, in exploration rates) to peripheral and intrahippocampal injections with agents that interfere with peptidergic, cholinergic, and GABAergic neurotransmission. These findings can be explained by an interdependent over-release of opioids, arrested GABA release, and excess acetylcholine in the hippocampal neuronal network of DBA/2 mice, as compared to C57BL/6 mice where these systems are functionally well balanced. Very similar results have been obtained with the lines SRH and SRL, derived from C57BL/6 and DBA/2, and genetically selected for rearing behavior. Most probably, the opioids act to disinhibit exploratory responses. An additional genetic approach is mentioned, in which four inbred mouse strains and one derived heterogeneous stock are used for estimating genetic correlations between structural properties of the hippocampal mossy fibers and levels of hippocampal dynorphin B, on the one hand, and frequencies of exploratory responses to environmental novelty, on the other.     

Correlations between glucose-inhibition and control parameters of α-glucosidase kinetics inApis mellifica haemolymph (Hymenoptera: Insecta)

Summary Kinetics studies of haemolymph -glucosidase inhibition by D. glucose led to general correlations between inhibition and control parameters, all over the honeybee development. Maximum velocities are not affected by the inhibition, while affinity constants are always significantly increased, and Hill coefficients tend to decrease, especially in foraging adults and in prenymphs. In this later case, the n-type effect tends to break the manifestation of the K-type mechanism.     

An approach for the in vitro screening of drugs for activity against leprosy

Summary Slow growing strains of mycobacteria isolated from leprous tissues present a characteristic resistance pattern to antibacterial agents that is comparable to drug sensitivity ofM. leprae in man.The Hansen Chair of Research is a foundation of Le Secours aux Lépreux (Canada) Inc..     

Phasic hippocampal activity during paradoxical sleep in the rat: selective suppression after diazepam administration

Summary The effects of diazepam on tonic (T) and phasic (P) components of the paradoxical-sleep hippocampal theta rhythm were studied in the rat. Results show that diazepam, a benzodiazepine analogue known to interfere with the putative neurotransmitter -aminobutyric acid (GABA) in the mammalian central nervous system, selectively abolishes P. They confirm previous data according to which sedative/anaesthetic drugs cause, an apparent dissociation of T and P of the paradoxical-sleep hippocampal theta rhythm. Moreover, they suggest a possible involvement of GABA in the neurochemical mechanisms underlying P in the rat.     

The locus ceruleus: a possible neural focus for genetic differences in emotionality

Summary The Maudsley Reactive and Non-Reactive strains have been developed as a model for the study of individual variations in stress-reactivity, and many differences in biobehavioral systems have been found between them. This review discusses limitations of the emotionality construct in accounting for differences between the Maudsley strains and offers an alternative, theoretical approach. Amaral and Sinnamon have proposed that the locus ceruleus (LC) plays a stress-attenuating role in mediating behavioral, physiological and neuroendocrine response to prepotent, emergency-provoking stimuli and, building upon this formulation, it is proposed that the LC has been an important focus for gene action in the Maudsley model. It is suggested that the LC of the Non-Reactive strain is more strongly activated by stressful stimuli than the LC of Reactive rats, and is the basis of many of the behavioral and physiological differences between them. Behavioral and biochemical evidence consistent with this proposition is reviewed. Identification of the LC as a target for gene-action in the Maudsley model has an important advantage. It substitutes variations at a specific anatomic location in the brain for a loosely defined construct like emotionality, and the hypothesis is amenable to empirical tests by a variety of experimental approaches.Supported by MH-39210 from the National Institute of Mental Health to DAB     

The inability of nuclear dehydrogenating clostridia to oxidize bile salt hydroxyl groups

Summary In a survey of the intracellular bile salt oxidoreductase activity in fecal bacteria, 16 strains of nuclear dehydrogenating clostridia and 2 strains of non-nuclear dehydrogenatingC. paraputrificum were demonstrated unable to oxidize cholate at any of the 3 OH groups. Since nuclear dehydrogenation at the -1 and -4 position requires a 3-oxo precursor steroid, it appears that these organisms require the presence of a 3-hydroxysteroid dehydrogenating organism for nuclear dehydrogenation.This work is supported by the National Cancer Institute of Canada (IAM) and by the Cancer Research Campaign (MJH).     

Sensitivity of the developing chick myocardium to the positive inotropic effects of calcium and isoproterenol

Summary The present results show that the sensitivity of the chick myocardium to the positive motropic effect of Ca⁺⁺ decreases during development and that the Ca⁺⁺ concentration of the physiological solution used must be lowered below normal to study the effects of positive inotropic agents in preparations from younger embryos. Isoproterenol elicits positive inotropic responses in 7–9-day embryonic ventricle and in newborn chick atria; however, the 4-day embryonic myocardium is unresponsive to isoproterenol.This work was supported by Grant No. HL-15995 from the National Heart Institute (USPHS).The authors would like to thank Dr.F. E. Shideman for the isolated tissue baths used in these experiments.     

GMP-stimulation of the cyanide-insensitive mitochondrial respiration in heat-shocked conidia ofNeurospora crassa

Summary In mitochondria of heat-shocked conidia ofNeurospora exogenous NADH and succinate were oxidized mainly via the alternative, hydroxamate-sensitive pathway (70%) and only 30% via the cytochromic, cyanide-sensitive pathway which was predominant in untreated conidia; the alternative oxidase pathway was markedly stimulated by guanosine 5-monophosphate (GMP).We are grateful to Mrs A. Cattaneo for her skillful technical assistance.     

Changes in mouse liver superoxide dismutase activity and lipid peroxidation during embryonic and postpartum development

Summary In inbred mice possessing high and low tissue superoxide dismutase (SOD) activity, it was observed that the difference in the SOD activities of the liver homogenates during development attains the maximum characteristic of the strain by about the 150th day. Subsequently, the SOD activity change displays a tendency in contrast with the age and the basic state. In the course of the development, a difference was also observed between the 2 mouse strains in the lipid peroxidation variation.Part XVIII of the series Properties of enzymes.     

Individual recognition and incest avoidance in eusocial common mole-rats rather than reproductive suppression by parents

Non-reproductive females in families of eusocial common mole-rats (Cryptomys sp., Rodentia) are not suppressed by their mother, (either behaviourally or pheromonally) as is generally assumed. They do not mate with their father and brothers simply because they are not sexually attractive for them (and vice versa). The incest avoidance is based on the capability to recognize (and keep in memory for up to three weeks) each family member individually. A sterile daughter may conceive and deliver young in her parental family if given the opportunity to mate with an unfamiliar mate in a separate cage. In this way, two females may breed side by side in one family.     

The role of repair in radiobiology

Summary Apart from cancer and mutation induction, radiobiological effects on mammals are mostly attributable to cell death, defined as loss of proliferative capacity. Survival curves relate retention of that capacity to radiation dose, and often manifest a quasi-threshold (shoulder). The shoulder is attributable to an initial mechanism of repair (Q-repair) which is gradually depleted as dose increases. Another form of repair, which is not depleted (P-repair), increases the dose required to deliver an average of one lethal event per cell (dose D₀). Neither form of repair can unambiguously be linked with repair of defects in isolated DNA. An important initial lesion may well be disruption of the complex structural relationship between the DNA, nuclear membrane and associated proteins. One form of P-repair may be restoration of that structural relationship.     

Correlation between car ownership and leukaemia: Is non-occupational exposure to benzene from petrol and motor vehicle exhaust a causative factor in leukaemia and lymphoma?

Although there is widespread agreement that many cancers have environmental causes we are often unable to see associations between specific cancers and exposure to environmental chemicals. One might also speulate that the more widespread, common-place and normal a chemical exposure is perceived to be then the less likely it will be that the exposure is recognised, let alone be considered to cause cancer. Widespread contamination of air by chemicals associated with internal combustion may be an example of one such invisible carcinogenic exposure. Yet evidence is available which suggests that many leukaemia and lymphoma cases, as well as other cancers, may be caused by this mundane and ubiquitous environmental contamination. The hypothesis is developed that leukaemia clustering as well as national leukaemia incidence may be related to non-occupational exposure to benzene formed by petrol combustion and resulting from petrol evaporation¹. The possible association between exposure to fuel vapours, internal combustion products and cancer merits much closer examination than it receives at present.     

Transfilter associations of nonleguminous plants with rhizobia: Channelling of N2 fixed by rhizobia into the nitrogen-metabolism ofPortulaca grandiflora callus tissues

Summary In transfilter associations ofPortulaca grandiflora callus tissues withRhizobium sp. cowpea 32H1 the nitrogenium fixed by the bacteria was channelled into the normal pathway of nitrogen metabolism of higher plants. In associations kept in an¹⁵N₂ containing atmosphere 10% of the¹⁵N taken up into the plant cells was incorporated intoPortulaca proteins. The results demonstrate that cells of nonleguminous plants are profiting from the quasi-symbiotic situation.