Susan K McCune

Expression of multiple alpha adrenergic receptor subtype messenger RNAs in the adult rat brain

Multiple subtypes of alpha adrenergic receptors with CNS expression (alpha 1A, alpha 1B, alpha 2A and alpha 2C) have been identified through pharmacological and molecular biological means. To characterize the localization of these subtypes and attempt to correlate subtype expression with physiological significance, the expression of the mRNAs encoding the alpha 1A, alpha 1B, alpha 2A and alpha 2C adrenergic receptor subtypes was examined in the adult rat brain by in situ hybridization histochemistry. Each subtype demonstrated a unique pattern of distribution, with the alpha 1 adrenergic receptors more restricted in their distribution and the alpha 2 receptors more widespread. The alpha 1A was primarily localized in the olfactory bulb, intermediate layers of the cortex, the hippocampus and the reticular nucleus of the thalamus. The alpha 1B was expressed in intermediate and deep layers of the cortex, thalamus, hippocampus, dorsal raphe and cerebellum. Although the alpha 2A message was relatively low in abundance, it was identified in the olfactory bulb, cortex, hippocampus, locus coeruleus, pons and cerebellum. The alpha 2C messenger RNA was localized in the cortex (particularly cingulate), hippocampus, caudoputamen, pons and cerebellum. Multiple alpha adrenergic receptor subtypes have significant sequence homology and similar pharmacologic properties; however, they each possess a unique pattern of messenger RNA distribution throughout the brain. The multiplicity of subtypes of alpha adrenergic receptors in specific brain regions may dictate the physiological and pharmacological responses to catecholamines.

Maternal vasoactive intestinal peptide and the regulation of embryonic growth in the rodent.

Vasoactive intestinal peptide (VIP) has been shown to regulate early postimplantation growth in rodents through central nervous system receptors. However, the source of VIP mediating these effects is unknown. Although VIP binding sites are present prenatally, VIP mRNA was not detected in the rat central nervous system before birth and was detected in the periphery only during the last third of pregnancy. In the present study, the embryonic day (E11) rat embryo/trophoblast was shown to have four times the VIP concentration of the E17 fetus and to have VIP receptors in the central nervous system. However, no VIP mRNA was detected in the E11 rat embryo or embryonic membranes by in situ hybridization or reverse transcriptase-PCR. RIA of rat maternal serum revealed a peak in VIP concentration at days E10-E12 of pregnancy, with VIP rising to levels 6-10-fold higher than during the final third of pregnancy. After intravenous administration of radiolabeled VIP to pregnant female mice, undegraded VIP was found in the E10 embryo. These results suggest that maternal tissues may provide neuroendocrine support for embryonic growth through a surge of VIP during early postimplantation development in the rodent.

The rat α2-C4 adrenergic receptor gene encodes a novel pharmacological subtype

A rat gene and brain cDNA (pA2d) encoding the homologue of the human α‐C4 adrenergic receptor subtype were isolated and characterized. RNA blots indicate that this gene is expressed in brain, heart and kidney but not in lung, liver or pancreas. Yohimbine, WB‐4101 and prasozin all exhibited high affinity for this receptor in binding studies. Clonidine was more potent and efficacious than norepinephrine in inhibiting forskolin‐stimulated cAMP production in CHO cells expressing pA2d. Together, these data suggest that the α2‐C4 gene product defines a previously undescribed pharmacological subtype α2‐adrenergic receptor.

Learning and Sexual Deficiencies in Transgenic Mice Carrying a Chimeric Vasoactive Intestinal Peptide Gene

The molecular mechanisms responsible for behavior are largely unknown. A state of the art model, paving the path from genes to behavior, is offered by transgenic animals. Candidate molecules are classic neuropeptides, such as vasoactive intestinal peptide (VIP). Transgenic mice harboring a chimeric VIP gene driven by the polyoma promoter were produced. Behavioral studies revealed learning impairment and prolonged retardation in memory acquisition in the genetically altered animals. Furthermore, reduced performance was observed when the male transgenic mice were tested for sexual activity in the presence of receptive females. Surprisingly, radioimmunoassays showed an approx 20% decrease in the VIP content of the transgenic mice brains. To directly assess genetically reduced VIP content as a cause for learning impairment, transgenic mice carrying diphtheria toxia-encoding sequences driven by the rat VIP promoter were created. These animals had reduced brain VIP and exhibited deficiencies in learning abilities, strongly supporting an important neurobiological function for VIP in vivo.

gp120 as an etiologic agent for NeuroAIDS: Neurotoxicity and model systems

The search for an agent that can mediate the symptoms of NeuroAIDS has been directed at gp120, the major envelope protein from HIV. The toxicity associated with gp120 was examined as a model and predictor of the neuropathological and neuropsychiatric manifestations of AIDS. Studies of the neurotoxic effects of purified gp120 on neurons from the rodent CNS cell cultures indicated the following: potent and selective killing of subpopulations of hippocampal neurons; varying potency of gp120s obtained from various HIV isolates; complete and potent protection from gp120 killing action after treatment with peptides related to vasoactive intestinal peptide; and obligatory presence of glia for gp120-related toxicity. Investigations of gp120 treatment of rodents revealed: cortical neurodystrophy with reduced arborizations and swollen processes; delays in developmental behaviors involving motor skills; peptide T prevention or attenuation of the morphological and behavioral deficits/delays produced by administration of gp120; and impairment of learning in the Morris swim maze. In addition, studies of subcutaneously administered, radiolabeled gp120 in neonatal animals demonstrated the presence of toxic fragments of gp120 in the developing brain. With the use of model test systems of non-human derived cell cultures and neonatal rats, we have captured and predicted a number of the morphological and behavioral deficits associated with AIDS. These multi-disciplinary studies of the actions of gp120 and associated fragments in rodents and rodent cells predict that the loss of cognitive and neurological function in patients with AIDS are attributed in part to interference of critical brain functions by the envelope protein, gp120.

Extracorporeal membrane oxygenation therapy in neonates with septic shock

Neonatal septic shock has significant morbidity and mortality with current therapeutic measures. At Childrens National Medical Center, from June 1984 to October 1986, 10 of 100 patients treated with venoarterial extracorporeal membrane oxygenation (ECMO) had a documented diagnosis of septic shock. All of these infants fulfilled criteria consistent with 80% mortality using conventional intensive medical management. However, the survival rate for the septic neonates in this study was 100%. Compared with other groups of infants treated with ECMO, these septic neonates required significantly more ventilatory support after ECMO and had a higher incidence of chronic lung disease (30% v 12%). The septic neonates were also at higher risk for intracranial hemorrhage than the other infants treated with ECMO (40% v 26%). The necessity for prolonged intubation after ECMO for patients with septic shock suggests that this condition may be associated with additional structural damage not seen with meconium aspiration syndrome or respiratory distress syndrome. Nevertheless, for neonatal patients with septic shock unresponsive to conventional medical management, ECMO must be considered a viable alternative treatment.

Blockade of VIP during Neonatal Development Induces Neuronal Damage and Increases VIP and VIP Receptors in Brain

Vasoactive intestinal peptide (VIP) is a 28-amino-acid peptide that is involved in diverse regulatory functions, including vasodilation, gastric secretion, and glycogenolysis.’ In the central nervous system (CNS), VIP exhibits neurotransmitter and neuromodulator functions, and recent work has highlighted an important role for VIP in the regulation of CNS development. In CNS primary culture experiments, subnanomolar concentrations of VIP were shown to stimulate neuronal survival and astrocyte mitogenesis and to induce the secretion of trophic factors by astrocytes.z-s In the micromolar concentration range, VIP treatment was shown to stimulate neuronal mitosis, neurite extension, and neuronal survival in sympathetic and neuroblastoma cultures.kx In addition, cultured whole embryo studies have demonstrated that a four-hour exposure to VIP resulted in a dramatic increase in growth.

Vasoactive intestinal peptide in the brain of a mouse model for Down syndrome

The most common genetic cause of mental retardation is Down syndrome, trisomy of chromosome 21, which is accompanied by small stature, developmental delays, and mental retardation. In the Ts65Dn segmental trisomy mouse model of Down syndrome, the section of mouse chromosome 16 most homologous to human chromosome 21 is trisomic. This model exhibits aspects of Down syndrome including growth restriction, delay in achieving developmental milestones, and cognitive dysfunction. Recent data link vasoactive intestinal peptide malfunction with developmental delays and cognitive deficits. Blockage of vasoactive intestinal peptide during rodent development results in growth and developmental delays, neuronal dystrophy, and, in adults, cognitive dysfunction. Also, vasoactive intestinal peptide is elevated in the blood of newborn children with autism and Down syndrome. In the current experiments, vasoactive intestinal peptide binding sites were significantly increased in several brain areas of the segmental trisomy mouse, including the olfactory bulb, hippocampus, cortex, caudate/putamen, and cerebellum, compared with wild-type littermates. In situ hybridization for VIP mRNA revealed significantly more dense vasoactive intestinal peptide mRNA in the hippocampus, cortex, raphe nuclei, and vestibular nuclei in the segmental trisomy mouse compared with wild-type littermates. In the segmental trisomy mouse cortex and hippocampus, over three times as many vasoactive intestinal peptide-immunopositive cells were visible than in wild-type mouse cortex. These abnormalities in vasoactive intestinal peptide parameters in the segmental trisomy model of Down syndrome suggest that vasoactive intestinal peptide may have a role in the neuropathology of Down-like cognitive dysfunction.

Vasoactive intestinal peptide regulation of nerve growth factor in the embryonic mouse

Vasoactive intestinal peptide (VIP), a regulator of embryonic growth, increased the concentration of nerve growth factor (NGF)-like immunoreactivity in the conditioned medium of cultured explanted embryonic day (E) 9.5 neural tube preparations compared to control preparations. VIP treatment also induced an increase of NGF-like immunoreactivity (NGF-IR) within the neural tube preparation tissue. A 60 kDa isoform was the primary form of NGF detected. VIP is shown to be a regulator of NGF in the E9.5 embryonic mouse and stimulates the release of a high molecular weight isoform of NGF.

Acquired Immune Deficiency Syndrome and the Developing Nervous System

Publisher Summary The study of AIDS has contributed substantially to a growing awareness of the relationship between the immune and nervous systems. This chapter discusses a comparison of the impairments in the nervous and immune systems that arise from HIV infection. The chapter also emphasizes the examination of the evidence for the role of the major envelope glycoprotein of HIV—gp120—as a causative agent for the pathology of AIDS, which may occur as the result of an interference with neuroimmune effectors. The latter idea serves to highlight the thesis that the nervous and immune systems share many common chemical mediators. Although the identity and action of these substances are uncertain, interference in their action may contribute to, if not define, the development and course of AIDS. The chapter focuses on the identification of such substances as well as possible roles for peptides as protective agents against the deleterious effects of HIV in the nervous system.