Mukaila A. Akinbami

Peroxisome Proliferator-activated Receptor δ Is Up-regulated during Vascular Lesion Formation and Promotes Post-confluent Cell Proliferation in Vascular Smooth Muscle Cells

Although peroxisome proliferator-activated receptor (PPAR) δ is widely expressed in many tissues, the role of PPARδ is poorly understood. In this study, we report that PPARδ was up-regulated in vascular smooth muscle cells (VSMC) during vascular lesion formation. By using Northern blot analysis, we demonstrated that PPARδ was increased by 3–4-fold in VSMC treated with platelet-derived growth factor (PDGF) (20 ng/ml). In addition, PDGF-induced PPARδ mRNA expression neither needs de novo protein synthesis nor affects the stability of PPARδ mRNA in VSMC. Preincubation of VSMC with phosphatidylinositol 3-kinase inhibitor (LY294002, 50 μmol/liter) or infection of VSMC with an adenovirus carrying the gene for a dominant negative form of Akt abrogated PDGF-induced PPARδ mRNA expression, suggesting that phosphatidylinositol 3-kinase/Akt signaling pathway is involved in the regulation of PDGF-induced PPARδ mRNA expression in VSMC. To explore the role of PPARδ in VSMC, we generated rat vascular smooth muscle cells (A7r5) stably overexpressing PPARδ and the control green fluorescent protein. Overexpression of PPARδ in VSMC increased post-confluent cell proliferation by increasing the cyclin A and CDK2 as well as decreasing p57 kip2 . Taken together, the results suggest that PPARδ plays an important role in the pathology of diseases associated with VSMC proliferation, such as primary atherosclerosis and restenosis.

A Longitudinal Study of Leptin During Development in the Male Rhesus Monkey: The Effect of Body Composition and Season on Circulating Leptin Levels

Abstract The objective of this study was to examine longitudinal changes in serum leptin concentrations during development and to correlate those changes with sexual development in male rhesus monkeys housed under natural environmental conditions. Blood samples were drawn from 8 control animals approximately every other month from 10 to 30 mo of age and thereafter monthly through 80 mo of age. Leptin levels declined through the juvenile period until the onset of puberty and were negatively correlated with body weight. Seven of the eight animals became sexually mature during the breeding season of their fourth year of life. Puberty was delayed in the other animal until the subsequent breeding season. There were no significant fluctuations in leptin levels prior to or in association with the pubertal rise in LH and testosterone (T) secretion. During the peripubertal period, levels of leptin varied between 2 and 3 ng/ml. The animal that exhibited delayed puberty had the lowest body weight and highest leptin levels during this period. With the achievement of sexual maturity, leptin levels varied seasonally, with peak levels in the late winter (Jan–Mar) and a nadir in the late summer (Aug–Sept). A late winter rise in leptin was also evident in most of the animals during Years 2 and 3, but not during Year 4. In the fall of Years 5 and 6, the seasonal rise in leptin concentrations lagged 3–4 mo behind the seasonal increase in LH and T. In the fall of Year 5, but not thereafter, leptin levels were positively related to percent body fat and negatively correlated with lean body mass. The data do not support the hypothesis that increasing leptin concentrations trigger the onset of puberty in the male rhesus monkey. During the juvenile period and after sexual maturation, but not during the peripubertal period, leptin secretion varied with season in the animals; but the environmental factors that cue or drive this rhythm remain to be determined.

Peroxisome proliferator-activated receptors and the cardiovascular system

Insulin resistance syndrome (also called syndrome X) includes obesity, diabetes, hypertension, and dyslipidemia and is a complex phenotype of metabolic abnormalities. The disorder poses a major public health problem by predisposing individuals to coronary heart disease and stroke, the leading causes of mortality in Western countries. Given that hypertension, diabetes, dyslipidemia, and obesity exhibit a substantial heritable component, it is postulated that certain genes may predispose some individuals to this cluster of cardiovascular risk factors. Emerging data suggest that peroxisome proliferator-activated receptors (PPARs), including alpha, gamma, and delta, are important determinants that may provide a functional link between obesity, hypertension, and diabetes. It has been well documented that hypolipidemic fibrates and antidiabetic thiazolidinediones are synthetic ligands for PPAR alpha and PPAR gamma, respectively. In addition, PPAR natural ligands, such as leukotriene B4 for PPAR alpha, 15-deoxy-delta 12,14-prostaglandin J2 for PPAR gamma, and prostacyclin for PPAR delta, are known to be eicosanoids and fatty acids. Studies have documented that PPARs are present in all critical vascular cells: endothelial cells, vascular smooth muscle cells, and monocyte-macrophages. These observations suggest that PPARs not only control lipid metabolism but also regulate vascular diseases such as atherosclerosis and hypertension. In this review, we present structure and tissue distribution of PPAR nuclear receptors, discuss the mechanisms of action and regulation, and summarize the rapid progress made in this area of study and its impact on the cardiovascular system.

Effect of a Peripheral and a Central Acting Opioid Antagonist on the Testicular Response to Stress in Rats

The possible involvement of opioid receptors in mediating the inhibitory effects of immobilization stress on testicular steroidogenesis was determined in adult male rats. Unstressed controls and animals exposed to 3 h of immobilization stress were injected subcutaneously with either vehicle or 1 or 10 mg/kg body weight (BW) of naloxone or naltrexone methobromide (NMB; an opioid receptor antagonist that does not cross the blood-brain barrier) at the beginning of and at 1.5 h of the stress period. Animals were sacrificed at 2 h (30 min after the second injection of antagonist) or 3 h (90 min after the second injection of antagonist) of stress. Plasma LH was not affected by stress, but 30 min after naloxone (1 or 10 mg/kg BW) injection, LH was elevated in both control and stressed rats above levels in vehicle-injected animals. By 90 min after naloxone injection, plasma LH had declined to levels comparable to those in vehicle-injected animals. NMB had no effect on plasma LH concentrations in either control or stressed rats. Three hours of stress reduced plasma testosterone (T) levels by 60% in vehicle-injected animals. This effect of stress on plasma T levels was antagonized by the 10 mg/kg BW dose of naloxone and 1 or 10 mg/kg BW of NMB. The ability of naloxone to reverse the effect of stress on plasma T levels was likely related to its ability to stimulate LH secretion, but NMB normalized plasma T values in stressed animals without altering plasma LH concentrations. Only the highest dose of NMB increased plasma T levels in unstressed control animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Endocrine–Immune Interaction: Alterations in Immune Function Resulting From Neonatal Treatment with a GnRH Antagonist and Seasonality in Male Primates

PROBLEM: The effect of neonatal gonadotropin releasing hormone (GnRH) antagonist (Ant) treatment and seasonality on immune system development and function was investigated in male primates.
 METHOD OF STUDY: Neonatal male rhesus monkeys and marmosets were treated with Ant, and its effect on immune system morphology, circulating lymphocyte subsets, and cell‐ and humorally‐mediated immune responses was assessed during development. In adult rhesus monkeys, we correlated seasonal changes in immune function with circannual fluctuations in immunoactive hormones.
 RESULTS: In neonatal marmosets, Ant reduced the number of B cells and T cells in the thymic medulla and T cells in the periarterial lymphatic sheaths (PALS) of the spleen. Ant also altered the development of, but did not permanently impair, the proliferative index (PI) of blood lymphocytes to mitogens. In vitro treatment of control lymphocytes with GnRH analogues altered their response to these proliferative agents. In neonatal rhesus monkeys, Ant treatment increased the frequency of clinical problems, lowered circulating levels of lymphocytes, total T cells, CD8+ T cells and B cells, and altered the PI of lymphocytes to mitogens. As adults, the cell‐ and humorally‐mediated immune responses remained impaired. We also documented seasonal fluctuations in the prevalence of diseases, circulating immune cells and immune function in rhesus monkeys. The number of cases of campylobacteriosis and shigellosis was lowest in the winter and highest in the spring. Circulating numbers of white blood cells (WBC) and neutrophils and the PI of lymphocytes to mitogens were higher in the winter than in the summer. Natural killer cell activity also varied with season. Cortisol and leptin secretion exhibited circannual rhythms, rising in concert with decreasing photoperiod and increasing testicular activity in the fall. Conversely, prolactin levels declined with decreasing photoperiod and then rose in the spring.
 CONCLUSION: Neonatal exposure of male primates to Ant appears to alter early postnatal programming of immune function. In the rhesus monkey, immune function shows seasonal fluctuations that may be driven by circannual changes in the secretion of immunoactive hormones.

Effect of neonatal treatment with a gonadotropin releasing hormone antagonist on developmental changes in circulating lymphocyte subsets: a longitudinal study in male rhesus monkeys

We have examined changes in circulating lymphocyte subsets from the neonatal period until adulthood (4 months until 5.5 years of age) in male rhesus monkeys, and the impact of neonatal treatment with a GnRH antagonist (Ant) or Ant and androgen (Ant/And) on these parameters. Absolute numbers of lymphocytes, B cells, total T lymphocytes, and CD4+ T cells decreased, neutrophils increased, and CD8+ T cells did not change with age. WBC counts increased between 4 mo and 2 years of age and then fell to neonatal levels over the next two years. The decline of CD4 + T cells in association with stable CD8+ T cell levels resulted in an age-related decrease in the CD4+/CD8+ T cell ratio. At 4 months of age, WBCs, lymphocytes, total T cells, CD8+ T cells and B cells were lower in Ant- and Ant/And-treated animals compared to controls. With the exception of WBC counts, these values had normalized by 2 years of age. Reduced WBC levels in treated animals persisted through adulthood. CD4+ T cell levels tended to be lower in Ant-treated and higher in Ant/And-treated animals than in controls at 4 months of age. CD4+ T cells remained lower in Ant- than in Ant/And-treated animals at most ages. The higher CD4 + T cell counts in Ant/And-treated animals resulted in an elevated CD4 + /CD8 + T cell ratio that persisted until the onset of year 5. During years 5 and 6, seasonal fluctuations in WBCs and neutrophils were observed with counts being higher in the breeding (fall) than in the nonbreeding (summer) season. The data document that developmental changes in circulating immune cells in the rhesus monkey are qualitatively similar to those reported in humans, and provide further evidence that neonatal treatment of male rhesus monkeys with Ant or Ant/And may alter early programming of the immune system.

Neonatal testosterone and handedness in yearling rhesus monkeys (Macaca mulatta)

This study investigated the relationship between neonatal testosterone (T) and hand bias in young rhesus monkeys (Macaca mulatta). Subjects (n = 8 per group) included: neonatally androgen-suppressed males, using a Nal-Lys gonadotropin releasing hormone (GnRH) antagonist (Antide); androgen-suppressed males receiving T replacement by a long-acting T preparation (CDB); control males; and control females. Antide suppressed T to the female range, whereas CDB replacement produced supranormal levels. Visually guided reaching, in a social context, showed a population-level left-hand bias. Males with elevated T did not show a stronger left-hand bias than males with normal T, but did show a stronger bias for the preferred hand whether left or right. Males with Antide-suppressed T showed an intermediate degree of hand bias. Results suggest that high neonatal T levels affect laterality and raise the possibility that GnRH analogues influence brain development. These data suggest a broad influence of the CNS-pituitary-testicular axis on brain asymmetries and provide support for an early neonatal period of T-influenced brain differentiation.

Expression of mRNA and proteins for testicular steroidogenic enzymes and brain and pituitary mRNA for glutamate receptors in rats exposed to immobilization stress

The objectives of this study were to determine whether stress attenuates the pituitary LH response to excitatory amino acids by altering expression of glutamate receptor 1 (GluR1) and N-methyl-D-aspartic acid (NMDA) receptor mRNA levels in the hypothalamus or pituitary, and assess whether stress influences testicular levels of mRNA or protein for steroidogenic enzymes. Three hours (h) of immobilization stress was associated with a greater than 7-fold increase in serum corticosterone, and a marked reduction in serum testosterone (T) concentrations. Stress did not significantly alter hypothalamic or pituitary GluR1 and NMDA receptor mRNA levels. Although transcript levels for P450SCC and P45017alpha mRNA in the testis were unchanged in stressed rats, western blotting of testicular fractions revealed reduced amounts of P450SCC and 3beta-HSD, but not P45017alpha. The data suggest that immobilization stress reduces T production by suppressing the translation of transcripts for P450SCC and 3beta-HSD, but the attenuated LH response of stressed animals to NMDA is not mediated by altered hypothalamic or pituitary expression of GluR1 and NMDA receptor levels.

Changes in lymphoid tissue after treatment with a gonadotropin releasing hormone antagonist in the neonatal marmoset (Callithrix jacchus).

PROBLEM: The effect of neonatal treatment with a gonadotropin releasing hormone (GnRH) antagonist on the morphology and distribution of lymphocytes in lymphoid tissue of the infant marmoset was examined.

Effect of neonatal treatment with a GnRH antagonist on development of the cell-mediated immune response in marmosets

PROBLEM: We examined the effect of neonatal treatment with a gonadotropin‐releasing hormone (GnRH) antagonist (antide) on the development of cell‐mediated immunity in male marmosets.