The human neuroblastoma clonal cell line SH-SY5Y expresses both mu- and delta-opioid receptors (ratio approximately 4.5:1). Differentiation with retinoic acid (RA) was previously shown to enhance the inhibition of adenylyl cyclase (AC) by mu-opioid agonists. We tested here the inhibition of cyclic AMP (cAMP) accumulation by morphine under a variety of conditions: after stimulation with prostaglandin E1 (PGE1), forskolin, and vasoactive intestinal peptide (VIP), both in the presence and in the absence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). Morphine inhibition of the forskolin cAMP response (approximately 65%) was largely unaffected by the presence of IBMX. In contrast, deletion of IBMX enhanced morphine's inhibition of the PGE1 and VIP cAMP response from approximately 50 to approximately 80%. The use of highly mu- and delta-selective agents confirmed previous results that inhibition of cAMP accumulation by opioids is mostly mu, and not delta, receptor mediated in SH-SY5Y cells, regardless of the presence or absence of IBMX. Because of the large morphine inhibition and the high cAMP levels even in the absence of IBMX, PGE1-stimulated, RA-differentiated SH-SY5Y cells were subsequently used to study narcotic analgesic tolerance and dependence in vitro. Upon pretreatment with morphine over greater than or equal to 12 h, a fourfold shift of the PGE1-morphine dose-response curve was observed, whether or not IBMX was added. However, mu-opioid receptor number and affinity to the mu-selective [D-Ala2, N-Me-Phe4, Gly5-ol]enkephalin were largely unaffected, and Na(+)- and guanyl nucleotide-induced shifts of morphine-[3H]naloxone competition curves were unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Hereditary sensory and autonomic neuropathies (HSANs) are a genetically and clinically diverse group of disorders defined by peripheral nervous system (PNS) dysfunction. HSAN type III, known as familial dysautonomia (FD), results from a single base mutation in the gene IKBKAP that encodes a scaffolding unit (ELP1) for a multi-subunit complex known as Elongator. Since mutations in other Elongator subunits (ELP2 to ELP4) are associated with central nervous system (CNS) disorders, the goal of this study was to investigate a potential requirement for Ikbkap in the CNS of mice. The sensory and autonomic pathophysiology of FD is fatal, with the majority of patients dying by age 40. While signs and pathology of FD have been noted in the CNS, the clinical and research focus has been on the sensory and autonomic dysfunction, and no genetic model studies have investigated the requirement for Ikbkap in the CNS. Here, we report, using a novel mouse line in which Ikbkap is deleted solely in the nervous system, that not only is Ikbkap widely expressed in the embryonic and adult CNS, but its deletion perturbs both the development of cortical neurons and their survival in adulthood. Primary cilia in embryonic cortical apical progenitors and motile cilia in adult ependymal cells are reduced in number and disorganized. Furthermore, we report that, in the adult CNS, both autonomic and non-autonomic neuronal populations require Ikbkap for survival, including spinal motor and cortical neurons. In addition, the mice developed kyphoscoliosis, an FD hallmark, indicating its neuropathic etiology. Ultimately, these perturbations manifest in a developmental and progressive neurodegenerative condition that includes impairments in learning and memory. Collectively, these data reveal an essential function for Ikbkap that extends beyond the peripheral nervous system to CNS development and function. With the identification of discrete CNS cell types and structures that depend on Ikbkap, novel strategies to thwart the progressive demise of CNS neurons in FD can be developed.

A coorelation has been observed between increased blood ketones and the tolerance of mice to hypoxia (4-5% oxygen). In previous studies fasted mice, alloxan diabetic mice and mice given 1,3-butanediol were found to be ketotic and to have increased tolerance to hypoxia. We attempted to induce a similar increased hypoxic tolerance by direct elevation of blood ketones with IV and IP beta-hydroxybutyrate (BHB). No increase in hypoxic tolerance was observed with BHB alone. Inasmuch as fasting and alloxan diabetes are both associated with elevated blood glucagon (G), hypoxic tolerance tests were made 30 min after G alone or a combination of G plus BHB. The mice given G alone or BHB alone had hypoxic survival times not different from saline controls. The mice given G plus BHB had increased survival times that could not be explained on the basis of a G mediated alteration in blood BHB.

The loss of behaviors that organisms use to avoid predation may serve as a sensitive indicator of pollution. We tested the hypothesis that a correlation exists in the field between heavy metal levels and antipredator behaviors. We examined the antipredator behavior of aquatic caddisfly larvae and snails at sites in the Coeur d'Alene basin of Northern Idaho which varied in their levels of heavy metals. We tested the antipredator response of Physella columbiana snails at 10 polluted lakes downstream from the Bunker Hill Environmental Protection Agency (EPA) Superfund cleanup site. We then compared their behavior to snails at 14 reference lakes. We placed the snails in a plastic testing apparatus, exposed them to an extract of crushed snail, and then monitored their movements to a normally preferred shaded area. We also tested the behavior of caddisfly larvae from 36 sites from a total of 6 streams/rivers adjacent to the Superfund site. Sites were located upstream and downstream of abandoned mines. We located active larvae of four genera, simulated predation by grasping the animals between thumb and forefinger (the larvae respond to being grasped by withdrawing into their case), lifted them from the water for 3 s, and then placed them in an adjacent, slower section of the stream. We then recorded how long it took each larvae to partially emerge from its case and attempt to move away. Unlike reference site snails, snails from heavy metal-polluted environments failed to exhibit antipredator behaviors in response to crushed conspecifics. These results are consistent with previous laboratory studies. We found no effect of heavy metals on the antipredatory behavior of caddisfly larvae.

In this paper we propose the hypothesis that pathogen-induced host defense responses result in altered host behaviors and enhanced predation. In particular we examine the effects of the acute phase response (whose effects include fever, reduced activity and malaise) on antipredatory behavior in bullfrog (Rana catesbeiana) tadpoles. This host response is associated with the preliminary stages of infection with many pathogens yet its behavioral effects have received little attention. Bullfrog tadpoles were injected with alcohol-killed bacteria to induce a response to infection and their ability to detect and avoid capture by predatory salamanders (Taricha granulosa) was explored. We predicted that acute phase responses increase tadpole vulnerability to predation by influencing thermoregulatory behavior and their ability to detect, and avoid capture by, salamanders. We found that the sterotypical effects of the acute phase response can lead to increased predation. Malaise affected the refuge seeking behavior of the tadpoles in the presence of salamanders. We suggest that for tadpoles provided with refuges, altered behaviors are a liability. This endogenous response may afford some parasites a potential pathway to their next host.