Dissociation of opioid and nonopioid analgesic responses following adult monosodium glutamate pretreatment

Neonatal administration of monosodium glutamate (MSG: 2-4 mg/g, SC) selectively destroys circumventricular organs, especially the arcuate nucleus and median eminence of the hypothalamus, and also attenuates both nonopioid (continuous cold-water swim: CCWS) and opioid (morphine) analgesia when rats are tested as adults. The present study evaluated whether administration of MSG (1-6 g/kg, SC) or its equiosmotic control (2.37 M NaCl) to adult rats altered either basal nociception on the tail-flick and jump tests or analgesia following morphine (5 mg/kg, SC) or CCWS (2 degrees C for 3.5 min). MSG treatment dose-dependently produced small but significant increases in basal nociceptive thresholds in adult rats. Morphine analgesia was significantly reduced on both tests following pretreatment with MSG (30-32%) and hypertonic NaCl (17-25%). In contrast, MSG (55-247%), but not NaCl pretreatment potentiated both nonopioid CCWS analgesia on both tests and CCWS hypothermia. These data are discussed in terms of differential neonatal and adult MSG effects, dissociations between opioid and nonopioid pain-inhibition, and the role of MSG in altering adaptive mechanisms to environmental stressors.     

Role of the endogenous opioid system in the analgesic effect of α-tocopherol in dysmenorrhea

Laboratory of General Pathology of the Nervous System, Research Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR. No. 1 Department of Obstetrics and Gynecology, Department of Pathological Physiology, and Laboratory of Experimental Therapy, Institute of Experimental Cardiology, All-Union Cardiologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 105, No. 2, pp. 148–140, February, 1988.     

Environmentally induced analgesia: ontogeny of a nonopioid system

Previous research has demonstrated that exposure to 90 sec of hind paw shock activates an endogenous pain control system that involves cholinergic sites. The purpose of the present investigation was to examine the development of function of this nonopioid analgesic system. Research on the ontogeny of the cholinergic system suggests that this receptor system exhibits an extended period of postnatal development, with various neurochemical indexes reaching maturity between 30 to 50 days of age. Results revealed that exposure to hind paw shock produced very low levels of analgesia in 10- and 28-day-old rats. The analgesic response was more evident in 2-month-old rats, but the degree of hind paw shock-induced analgesia was not at its maximum until 3 months of age. Systemic injection of naltrexone had no effect on the degree of analgesia induced by hind paw shock, while a systemic injection of scopolamine significantly attenuated the analgesia displayed by the 28-day-old and 2- and 3-month-old rats. Thus, the neurochemical indexes of cholinergic development over-estimated the degree of functional maturity of the endogenous analgesia system activated by hind paw shock.     

The analgesic effects of automatically controlled rotating acupuncture in rats: mediation by endogenous opioid system

The technique of rotating acupuncture needles has long been used to enhance the effects of acupuncture in Oriental medicine. However, it is difficult to standardize and quantify this stimulation condition. Thus we developed an automatically controlled rotating acupuncture (ACRA) system. The present study was conducted to evaluate the analgesic effects of ACRA using 4 different stimulation conditions (i.e., angle and frequency of rotation: 90 degrees + 1 Hz, 90 degrees + 1/4 Hz, 360 degrees + 1 Hz, and 360 degrees + 1/4 Hz) in Sprague-Dawley rats. Tail-flick latency to a noxious radiant heat stimulus in lightly anesthetized rats was measured before and after 15 min of ACRA stimulation at the Zusanli (ST36) acupoint. ACRA stimulations under all of the conditions above produced more potent analgesic effects than plain acupuncture (PA, acupuncture needle insertion only), but only the 90 degrees + 1/4 Hz ACRA condition showed a statistically significant effect versus PA (P < 0.01). Further, the analgesic effect of 90 degrees + 1/4 Hz ACRA was reversed by pretreatment with naloxone (2 mg/kg, i.p.). These results indicate that the 90 degrees + 1/4 Hz ACRA stimulation has the most potent analgesic effect in rats and that this is mediated by the endogenous opioid system.     

Ontogeny and regulation of the immune system

Classic studies in embryology and contemporary research in immunology and molecular biology have disclosed the carefully orchestrated events leading to development of the immune system and immunoregulation that ultimately provide immunohomeostasis. During ontogeny, the pluripotential stem cell emerges and differentiates into all hematopoietic lineages, including three major immunologically relevant components: T-cell differentiation occurs within the thymus; B cells appear within fetal liver, adult bone marrow, and possibly other abdominal sites; and concurrently, the monocyte-macrophage system develops. Under the influence of an array of cytokines and cellular interactions, immune regulation is established. T and B lymphocytes elaborate genetically encoded messages that acquire specificity via transposable genetic elements. Receptors and cytokines provide immune recognition, communication, regulation, and memory for antigens. Inherited and acquired defects in ontogeny and immune regulation are the basis for immunodeficiency disorders.     

Ontogeny of REM sleep in rats: possible implications for endogenous depression

Nine neonatal Long-Evans rats had continuous (24 h/day) polysomnography for 2 weeks, from age 14 days through age 27 days. A new finding was that six more or less independent measures of REM sleep occurrence decreased in parallel from age 14 days to age 27 days. The measures included parallel decreases of four measures of 24-h REM duration (tonic REM sleep, phasic REM sleep, mean REM period duration, and number of REM periods) along with parallel increases of two measures of REM delay (REM latency and percent of nonsleep onset REM periods). A parsimonious interpretation of the correlated changes is that a common developmental REM sleep inhibitory process accounts for the six parallel changes over time. This hypothesis can be tested empirically by studying inhibitory processes that operate on the pedunculopontine tegmental/latero-dorsal tegmental nuclei, the generators of REM sleep. The study also noted that compared with (same species) normal adults, endogenous depressives had the same distinctive REM sleep characteristics as neonatal rats. The similarity suggests that an underdeveloped, relatively weak REM sleep inhibitory process may account for the REM sleep peculiarities of endogenous depression. This hypothesis can be tested in adult rats made "depressed" by neonatal treatment with antidepressant drugs. Thus, the ontogeny of REM sleep suggests a developmental process that may be altered in humans predisposed to endogenous depression, and may account for the (life-long) REM sleep abnormalities of the disorder.     

Ontogeny of the hypothalamic neuropeptide Y system

Early onset obesity and type II diabetes is rapidly becoming an epidemic, especially within the United States. This dramatic increase is likely due to many factors including both prenatal and postnatal environmental cues. The purpose of this review is to highlight some of the recent advances in our knowledge of the development of the hypothalamic circuits involved in the regulation of energy balance, with a focus on the neuropeptide Y (NPY) system. Unlike the adult rat, during the postnatal period NPY is transiently expressed in several hypothalamic regions, along with the expected expression within the arcuate nucleus (ARH). These transient populations of NPY neurons during the postnatal period may provide local NPY production to sustain the necessary energy intake during this critical growth phase. This may be physiologically important since ARH-NPY projections do not fully develop until the 3rd postnatal week. The significance of this ontogeny is that many peripheral metabolic signals have little effect of feeding prior to the development of the ARH projections. The essential questions now are whether prenatal and/or postnatal exposure to high levels of insulin or leptin during development can cause permanent changes in the function of hypothalamic circuits. It is vital to understand not only the natural development of the hypothalamic circuits that regulate energy homeostasis, but also their abnormal development caused by maternal and postnatal environmental cues. This will be pivotal for designing intervention and therapeutics to treat early onset obesity/type II diabetes, which may very well need to be different from those designed to prevent/treat adult onset obesity/type II diabetes.     

Ontogeny and genetics of the hemato/lymphopoietic system

During embryogenesis there is a sequential, temporal appearance of increasingly more-complex hematopoietic cells beginning with unipotential progenitors, proceeding to multipotential (myeloid, erythroid and lymphoid) progenitors and culminating with adult-repopulating hematopoietic stem cells. Current research has established an important role for the aorta-gonads-mesonephros region of the mouse embryo in the generation of multipotential progenitors and hematopoietic stem cells. Comparisons of normal and hematopoietic-cell-mutant mouse embryos have revealed several genes pivotal in hematopoietic stem cell generation/function. Other genes have been implicated in the critical generation of lymphoid lineage potential. Thus, an understanding of the cellular and molecular interactions within the midgestation aorta-gonads-mesonephros region offers insight into the mechanisms of hematopoietic lineage specification during ontogeny and perhaps will lead to a more complete knowledge of the adult hematopoietic system.     

Ontogeny of the immune system of the brushtail possum, Trichosurus vulpecula

The numbers and distribution of T and B cells in the thoracic thymus, spleen and intestinal tissue and the proliferation of T lymphocytes were examined during pouch life and in the adult to determine when the developing brushtail possum reaches immunological maturity. CD3-positive cells were observed in the thoracic thymus at day 2 post-partum indicating that the thymus produces T lymphocytes at or soon after birth. By day 25 the thymus was fully populated with CD3-positive T lymphocytes and they were observed in distinct regions of the cortex and medulla. By day 48 post-partum, B and T lymphocytes were identified in the follicles and parafollicular areas of the spleen. Although the numbers of T and B cells in the spleen increased significantly from day 25 to day 100 post-partum (P < 0.005), fewer cells were present at day 150 post-partum than in the adult (P < 0.05). Peyer's patches were not observed in the intestines up to day 73 post-partum. However, both T and B cells were observed in the intestinal lymph nodes. Although the T lymphocytes at weaning showed a proliferative response, the response was not as great as that observed in the adult possum. Thus, the immune system of the possum is not fully developed at weaning but continues its development after pouch life.     

Ontogeny of natural killer cell activity and antibody dependent cell mediated cytotoxicity in pigs

Cells from pigs of various ages were collected from peripheral blood, mesenteric lymph node, spleen and thymus and their ability to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) and natural killer (NK) cell activity was determined. ADCC against chicken red blood cell (CRBC) was present in cells from peripheral blood, lymph node and spleen, but was absent in thymic cells. There were no age-related differences in ADCC to CRBC and cells from fetal pigs had similar activities to cells from adult pigs. Maximal cytotoxicity against CRBC was found in the polymorphonuclear leukocyte (PMN) cell fraction. In contrast to the good response against CRBC, PMN cells were not lytic in the ADCC assay when PI3 virus-infected cells were used as target cells. Peripheral blood lymphocytes (PBL) had low but significant lytic activity against PI3 virus-infected cells in the presence of high concentrations of specific antiserum. NK cell activity against K562 target cells was readily detected in PBL of pigs older than 2 weeks but was not observed with cells from spleen, lymph node or thymus from pigs of any age. PBL of pigs younger than 2 weeks of age had low but detectable NK activity: however, fetal pigs had no NK activity against K562 target cells. In contrast, when PI3 virus infected Vero cells were used as target cells, NK cells were detected in spleen and PBL, but not lymph nodes or thymus, of pigs greater than one day of age. Similar to the absence of activity to K562, none of the lymphoid cells from fetal pigs had NK activity against PI3 virus-infected Vero cells. The present results suggest that the effector cells that mediate ADCC are distinct from those that mediate NK activity in that cells mediating ADCC develop earlier and are found in different organs than the NK cells. Additionally, the cells that mediate NK activity against viral infected cells may be different from those that mediate NK activity for K562 target cells; however, regardless of the target, NK cells are not present before birth in the pig.     

Ontogeny of the immune system in Acipenserid juveniles

Sturgeon aquaculture has increased considerably worldwide but little is known about their immunological development and competence in early life stages. Culture of larvae is one of the most critical stages in intensive sturgeon farming, often associated with high mortality rates. The objective of this study was to characterize the developmental morphology (light and transmission electron microscopy, LM and TEM) of the meningeal myeloid tissue, spleen and thymus in Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) from hatching until 5 months old (2895°C·day (dd)). The spleen was first visible on 541 dd larvae LM sections and the other two immune organs in 768 dd samples (approximately 400 and 600 dd after onset of feeding). Generally, younger fish had significantly higher percentages of undifferentiated cells (meningeal myeloid tissue and spleen) and effective adaptive immune competence would not be expected in these fish on the onset of feeding, but further functional immune assessment is needed.     

Ontogeny of the cranial system in Laonastes aenigmamus

Rodents, together with bats, are among the ecologically most diverse and most speciose groups of mammals. Moreover, rodents show elaborate specializations of the feeding apparatus in response to the predominantly fore-aft movements of the lower jaw. The Laotian rock rat Laonastes aenigmamus was recently discovered and originally thought to belong to a new family. The difficulties in classifying L. aenigmamus based on morphological characters stem from the fact that it presents a mixture of sciurognathous and hystricognathous characteristics, including the morphology of the jaw adductors. The origin of the unusual muscular organization in this species remains, however, unclear. Here, we investigate the development of the masticatory system in Laonastes to better understand the origin of its derived morphology relative to other rodents. Our analyses show that skull and mandible development is characterized by an overall elongation of the snout region. Muscle mass increases with positive allometry during development and growth, and so does the force-generating capacity of the jaw adductor muscles (i.e. physiological cross-sectional area). Whereas fetal crania and musculature are more similar to those of typical rodents, adults diverge in the elongation of the rostral part of the skull and the disproportionate development of the zygomaticomandibularis. Our data suggest a functional signal in the development of the unusual cranial morphology, possibly associated with the folivorous trophic ecology of the species.     

Are the analgesic effects of social defeat mediated by benzodiazepine receptors?

Social conflict in mice is associated with at least two forms of analgesia. A long-lasting opioid reaction is evident in intruder mice exposed to prolonged attack, whilst an acute non-opioid analgesia is seen in response to either defeat experience per se or the territorial scent-marking of an aggressive conspecific. Recent work from this laboratory has suggested that the non-opioid analgesic reaction to defeat experience may be mediated via benzodiazepine receptor mechanisms. The present studies were designed to further test this tentative hypothesis. Results confirmed that defeat analgesia is dose-dependently blocked by Ro15-1788 (20-40 mg/kg) and diazepam (2-4 mg/kg), and also indicated partial antagonism of the reaction by CGS8216 (2.5 mg/kg). The partial agonists CGS9896 (2.5-20 mg/kg) and ZK91296 (2.5-20 mg/kg) were ineffective in blocking the reaction, a finding also obtained with the full agonist ZK93423 (0.05-10 mg/kg). However, the antagonist/weak inverse agonist ZK93426 was found to possess significant intrinsic analgesic activity (10 mg/kg) and to enhance defeat analgesia (5-10 mg/kg). Although several interpretative frameworks for the current pharmacological profile are considered, it is concluded that full clarification of the substrates of defeat analgesia must await further investigations.     

Polyclonal activation of the murine immune system by an antibody to IgD. III. Ontogeny

It has recently been demonstrated that the injection of adult mice with an affinity-purified goat antibody to mouse IgD (GaM delta) stimulates activation of the humoral immune system that resembles, on a polyclonal level, specific B cell activation by a T cell-dependent antigen. One to 2 days after adult BALB/c mice are injected with 200 micrograms of GaM delta, their splenic B lymphocytes undergo a series of T-independent activation steps that include increases in surface (s) Ia expression, cell size and DNA synthesis. Seven days after GaM delta injection, these cells undergo T-dependent activation steps, that include further proliferation as well as differentiation into IgG1-secreting cells. We have now studied the ontogeny of the T-independent (day 2) and T-dependent (day 7) activation steps by injecting 100-200 micrograms of GaM delta into 3-day- to 10-week-old BALB/c mice. GaM delta failed to induce increases in B cell sIa expression or size 2 days after injection of mice 2 weeks old or younger and failed to stimulate increased DNA synthesis 2 days after injection of 4-week-old mice. In contrast, increases in spleen cell sIa expression, size and DNA synthesis were seen 7 days after injection of 6- to 8-day-old mice. Furthermore, increases in the numbers of spleen cells with large amounts of intracytoplasmic IgG1 were seen at the same time, although these increases were much less than were seen in GaM delta-treated adult mice. Thus, the ability of GaM delta to induce T help and to act in concert with such help to stimulate B cell proliferation and differentiation precedes in ontogeny the ability of GaM delta to directly induce B cell proliferation and early differentiative events. In addition, the early activating events that we have studied are not required for T-dependent B cell proliferation and antibody production to occur, although they appear to contribute to the magnitude of clonal expansion and antibody production.     

Opioid and nonopioid conditional analgesia: the role of spinal opioid, noradrenergic, and serotonergic systems.

The first aim of this study was to test whether opioid mediation of conditional analgesia is inversely related to the relative amount of training that rats received, a prediction derived from the severity hypothesis. Naltrexone attenuated the analgesia among subjects in the low-training group but had no effect in rats given an extended amount of training. The second aim of this study was to investigate whether noradrenergic, opiate, and serotonergic spinal systems play a role in conditional analgesia. Intrathecal administration of yohimbine, an alpha 2 antagonist, completely blocked both opioid and nonopioid forms of conditional analgesia. Spinal administration of either quaternary naltrexone or methysergide failed to have an effect. In addition to providing support for the severity hypothesis, these results indicate that both opioid and nonopioid forms of conditional analgesia are subserved by critical alpha 2-adrenoceptors at the spinal level.     

The sedative component of anesthesia is mediated by GABA(A) receptors in an endogenous sleep pathway

We investigated the role of regionally discrete GABA (gamma-aminobutyric acid) receptors in the sedative response to pharmacological agents that act on GABA(A) receptors (muscimol, propofol and pentobarbital; 'GABAergic agents') and to ketamine, a general anesthetic that does not affect GABA(A) receptors. Behavioral studies in rats showed that the sedative response to centrally administered GABAergic agents was attenuated by the GABA(A) receptor antagonist gabazine (systemically administered). The sedative response to ketamine, by contrast, was unaffected by gabazine. Using c-Fos as a marker of neuronal activation, we identified a possible role for the tuberomammillary nucleus (TMN): when gabazine was microinjected directly into the TMN, it attenuated the sedative response to GABAergic agents. Furthermore, the GABA(A) receptor agonist muscimol produced a dose-dependent sedation when it was administered into the TMN. We conclude that the TMN is a discrete neural locus that has a key role in the sedative response to GABAergic anesthetics.     

Reinvestigation of an endogenous meiotic drive system in the mosquito, Aedes aegypti (Diptera: Culicidae)

We have initiated efforts to determine the molecular basis for the M(D) meiotic drive system in the mosquito, Aedes aegypti. The effect of the M(D) gene is a highly male-biased sex ratio, but varies depending on the frequency and sensitivity of a susceptible responder m(s) allele. The M(D) system has potential as a mechanism for driving trangenes for pathogen resistance into natural Ae. aegypti populations. Because all previously existing laboratory strains carrying the M(D) gene have been lost, we have selected for a new strain, T37, that carries a strong driver. Matings between T37 males and drive-susceptible m(s) females result in progeny with highly biased sex ratios, wherein only approximately 14.7% females are produced. We discuss the potential for identifying M(D) candidate genes based on comparisons with the well-described Drosophila melanogaster segregation distorter (SD) meiotic drive system and considerations for release of transgenic Ae. aegypti into natural populations where M(D) and insensitive m3 alleles are likely segregating.     

Ontogeny of conditioned heart rate to an olfactory stimulus

When heart rate is used as the index of conditioning, rat pups younger than 15 days of age do not display an odor-shock association. This constitutes a marked delay relative to the development of a somatomotor conditioned response. The incapacity to display autonomic learning to an olfactory stimulus prior to day 15 is not due to the inability to perceive and to orient to the olfactory stimuli used, nor to the inability to make unconditioned phasic cardiac changes. Rather, the late development of the heart rate conditioned response may indicate that the central neural mechanisms mediating heart rate conditioning are distinct from, and mature later than those mediating 1) heart rate orienting and 2) somatomotor conditioning. Evidence from studies in adult species is used to support these speculations.