Immune functions in beluga whales (Delphinapterus leucas): evaluation of mitogen-induced blastic transformation of lymphocytes from peripheral blood, spleen and thymus.

A quantitative assay was developed to evaluate mitogen-induced lymphoblastic transformation in beluga whales (Delphinapterus leucas) using peripheral blood mononuclear cells, splenocytes and thymocytes. Optimal concentrations of four different mitogens (Con-A, PHA, LPS and PWM) were determined with the use of standard curves. Addition of human recombinant IL-2 (rhIL-2) after 48 h in culture with the different mitogens suggests that Con-A, PHA and PWM, but not LPS, stimulate T cells in belugas, as they do in other animal species. The addition of 2-mercaptoethanol did not enhance significantly the proliferation of cells stimulated by Con-A, PHA and LPS, while it did with the cells stimulated by PWM and those cultured without mitogen. The proliferative response of cells was suppressed when the culture medium was supplemented by beluga serum instead of fetal calf serum. This assay will be useful to assess the status of the immune functions in different populations of beluga whales as well for further in vitro immunotoxicological experiments.

Immune function of bovine leukocytes after in vitro exposure to selected heavy metals.

OBJECTIVE: To study effects of in vitro exposure of bovine leukocytes to mercury, cadmium, and lead on phagocytosis, natural killer cell activity, and lymphocyte proliferation. SAMPLE POPULATION: Leukocytes from 6 nonpregnant Holstein heifers. PROCEDURE: Leukocytes were exposed in vitro to the aforementioned metals, and leukocyte functions were assessed. RESULTS: Phagocytosis was suppressed by 10(-5) to 10(-7) M CdCl2 and by 10(-5) and 10(-6) M HgCl2, but not 10(-7) M HgCl2 nor 10(-4) to 10(-6) M PbCl2. Spontaneous and concanavalin A- or phytohemagglutinin-stimulated proliferation of metal-treated bovine blood mononuclear cells was not significantly different from that of nontreated control cells, except for enhanced spontaneous proliferation in response to 10(-5) M HgCl2. When proliferation was expressed as a stimulation index, a dose-dependent increase of spontaneous proliferation was observed in response to exposure to HgCl2 and PbCl2. Compared with response to 10(-6) or 10(-7) M CdCl2, reduction of mitogen-induced and spontaneous proliferation was observed on exposure to 10(-5) M CdCl2. Natural killer cell activity against YAC-1 target cells, evaluated by flow cytometry, was decreased only in cells exposed to 10 M HgCl2. CONCLUSION AND CLINICAL RELEVANCE: Bovine leukocytes are susceptible to the immunomodulatory effects of in vitro exposure to heavy metals at concentrations equal to or higher than those at which similar effects are seen for leukocytes from most other animal species for which data are available for comparison. Exception is phagocytosis, which is severely affected by low concentrations of CdCl2 and HgCl2 in cattle. Reduction of defense mechanisms on exposure to metals could lead to increased susceptibility to potential pathogens.

Mesocortical dopamine depletion and anxiety-related behavior in the rat: sex and hemisphere differences.

The mesocortical dopamine (DA) system of the rat plays an important role in prefrontal cortex (PFC) regulation of stress and emotion and exhibits functional hemispheric asymmetry for such processing. Since few studies examine sex differences in this context, we compared the effects of left vs. right unilateral PFC DA depletion in males and females in several behavioral situations associated with anxiety or aversion. Adult rats received unilateral injections of 6-hydroxydopamine (6-OHDA) or vehicle in the ventromedial (vm) PFC. Behavioral tests included a predator odor burying test, elevated plus maze and sucrose consumption with simple taste aversion. Tissue analysis confirmed that vmPFCs injected with 6-OHDA were depleted of DA (75-85%) compared to controls. Burying behavior and sucrose consumption were affected only by left lesions, similarly in both sexes. However, risk assessment behaviors were affected by right lesions in opposite directions in males and females. Behaviors modified preferentially by the left cortex thus showed less evidence of sex differences than those modulated by the right. While mesocortical DA depletion effects are lateralized, the nature of these effects can vary with sex and specific behavior. Such findings may be clinically significant, given the large gender differences in the incidence of mood and anxiety disorders, which also show many lateralized prefrontal abnormalities.

Role of sex in the neurochemical and neuroendocrine correlates of paw preference in the rat.

Handedness in humans and paw preference (PP) in rodents have been associated with neural and physiological correlates, which frequently appear to be sex-specific. The present study examines sex differences in the effects of differential PP on post mortem measures of regional monoamine activity in adult Long-Evans rats. The effects of PP on neuroendocrine function were also assessed by measuring plasma adrenocorticotropic hormone (ACTH) in response to a 30-min restraint stress. Most rats showed strong individual PP. Males (n=27) and females (n=26) did not differ in the direction or strength of their PP with nearly equal numbers of left and right-pawed rats. However, many Sex×PP interactions were noted in regional neurochemical measures, and most effects of PP were bilateral in nature and sex-specific. In males, Left PP (relative to Right PP) was associated with reduced striatal dopamine (DA) levels and amygdala DA metabolism bilaterally. In females, Left PP was associated with a bilateral upregulation of DAergic metabolism in both prefrontal cortex and nucleus accumbens, increased amygdala serotonin metabolism, and a (right) unilateral increase in amygdala DA. As in previous studies, some correlations were also noted between PP and asymmetrical or unilateral monoamine measures. As well, rats with strong PP, independent of direction or sex, had lower basal ACTH and more robust stress responses than rats with weak PP, suggesting a possible adaptive advantage to strong lateralization. Overall, sex differences were found only regarding effects of the direction of PP, not its magnitude. The findings suggest that the direction of PP affects distinct bilateral networks of structures sex-dependently. Such fundamental influences of PP on functional brain organization have implications for a wide array of processes under monoaminergic modulation in these brain regions, and may further our understanding of the numerous human examples of gender and handedness interactions across several modalities.

Comparative sensitivity of harbour and grey seals to several environmental contaminants using in vitro exposure.

In this study, we investigated the effects of cadmium chloride (CdCl(2)), mercury chloride (HgCl(2)), methylmercury chloride (CH(3)HgCl), and PCBs on lymphocyte proliferation in phocids. PBMCs isolated from harbour and grey seals were exposed in vitro to varying concentrations of contaminants. A reduction of viability occurred when cells were exposed to 10(-4)M HgCl(2) or CH(3)HgCl or to 50ppm of Aroclor 1254. In both grey and harbour seals, T-lymphocyte proliferation was suppressed when their cells were incubated with 5 x 10(-5)M CdCl(2) or 10(-4)M HgCl(2). An inhibition of proliferation occurred with CH(3)HgCl from 10(-6)M in grey seals and from 10(-5)M in harbour seals. In grey seals, Aroclor 1254 reduced lymphocyte proliferation at 15ppm. In both harbour and grey seals, CH(3)HgCl was ten times more immunotoxic that HgCl(2). From IC(50), chemicals were ranked in terms of toxicity as followed: CH(3)HgCl>CdCl(2)>HgCl(2)>Aroclor 1254.