Román Vidaltamayo

Antidepressant-like effects of nicotine and transcranial magnetic stimulation in the olfactory bulbectomy rat model of depression

In this study, we compared the depression-like symptoms induced by olfactory bulbectomy (OBX) in the two inbred Wistar and Long Evans rat strains. We also analyzed the self-regulated oral intake of nicotine in these strains and the effect of nicotine on the depression-like symptoms of olfactory bulbectomy. Furthermore, we compared the antidepressant-like effects of nicotine on Wistar rats to those of transcranial magnetic stimulation (TMS), which has emerged as a therapeutic alternative for depression management. Our results show that Wistar rats develop depression-like symptoms, demonstrated by the forced swim test (FST), 4 weeks after OBX. However, in bulbectomized Long Evans rats these symptoms cannot be assessed due to a higher degree of variability of the swimming behavior of this strain. These results suggest that there are some innate differences in susceptibility to stress between these two rat strains. In Wistar rats, voluntary oral nicotine intake (1.2 mg/(kg day) for 14 days) as well as nicotine administered as a single daily i.p. injection (1.5 mg/(kg day) for 14 days) decrease the depression-like symptoms of OBX. Daily transcranial magnetic stimulation (60 Hz and 0.7 mT for 2h/day for 14 days) also decreases depression-like symptoms but is less effective than nicotine. In conclusion, our results support the idea that there are possible innate differences for depression susceptibility and that nicotine and TMS may be useful in the treatment of this syndrome.

Nerve growth factor increases sodium channel expression in pancreatic β cells: implications for insulin secretion1

The importance of nerve growth factor (NGF) modulation of pancreatic ß cells is demonstrated by the fact that these cells secrete and respond to this trophic factor. Among NGF effects on ß cells is an increase in Na+ and Ca2+ current densities. This study investigates the mechanisms involved in the NGF‐induced increase in Na+ current and the implications of this effect for insulin secretion. The following results were obtained in single ß cells cultured with NGF for 5–7 days: 1) A steady‐state level of mRNA coding for type III sodium channel α subunit increased twofold compared with that for control cells. 2) The increase in Na+ current density was blocked either by cycloheximide or by actinomycin D, indicating that it is mediated by protein synthesis and mRNA transcription. 3) NGF treatment strengthened, by nearly fourfold, the ß‐cell electrical activity; this effect is partially related to the increased Na+ current, because tetrodotoxin (TTX) decreased the duration of the depolarized plateau level. 4) Single ß cells secreted nearly two times more insulin in response to 5.6 or 15.6 mM glucose. This effect was inhibited by TTX, indicating that the enhanced Na+ current plays an important role. These data suggest that NGF could preserve an adequate expression of sodium channels and that impairment of NGF modulation could lead to deficient insulin secretion and diabetes mellitus.

Expression of Nerve Growth Factor in Human Pancreatic β Cells

Nerve growth factor (NGF) is an important modulator of rat pancreatic β-cell physiology in vitro. In this study, we analysed the expression of NGF, TrkA and insulin in human pancreatic islets from normal, ductal adenocarcinoma and insulinoma-afflicted samples, using double immunofluorescent labelling and confocal microscopy. We found that in normal human pancreas, insulin and NGF are co-expressed in β cells. Moreover, similar to previous observations in rat, the high affinity NGF receptor TrkA is also expressed in β cells. Pancreatic β cells in normal islets from adenocarcinoma and mucinous cystadenocarcinoma patients also expressed NGF. In 2 out of 15 exocrine tumour samples, NGF was detected also in the tissue surrounding the islets, while 2 out of 13 adenocarcinoma tumours expressed this growth factor. In five insulinoma samples, we observed weaker immunofluorescent labelling of insulin and NGF in the neoplastic tissue, compared to the islets not afflicted by the tumour, which may be a consequence of increased hormone secretion rate. We demonstrate that human β cells express TrkA and NGF. These findings are consistent with the hypothesis that NGF modulates insulin secretion through a paracrine/autocrine loop, similar to the one observed in cultured rat β cells.

Neuron-like phenotypic changes in pancreatic β-cells induced by NGF, FGF, and dbcAMP

We studied the effects of nerve growth factor (NGF), fibroblast growth factor (FGF), and dibutyryl-cAMP (dbcAMP) on rat pancreatic β-cell morphology and of NGF and dbcAMP on insulin secretion. After 2 wk in culture, nearly 3% of β-cells extended neurite-like processes spontaneously; when cells were treated with NGF, almost 30% of them extended processes. In the presence of dbcAMP, almost all β-cells flattened, and the extension of neurite-like processes was more pronounced in fetal than in adult cells. The most prominent effect, regardless of age, was observed in cells treated with NGF and dbcAMP together, since the percentage of neurite-like bearing β-cells increased to 50%. β-cells cultured under these conditions maintained their immunoreactivity to insulin and nearly all β-cells and their neurite-like processes were also positive to GABA, tubulin, tau protein, and N-CAM. FGF increased the percentage of adult β-cells bearing neurite-like processes to 13%, and FGF and dbcAMP applied together to 40%. β-cells treated with NGF and dbcAMP for 5 to 7 d preserved their capability to secrete the hormone in response to different extracellular glucose concentrations. Insulin secretion of dbcAMP-treated β-cells was 2.5-fold higher than in control cells. NGF-treated cells were able to discriminate between different glucose concentrations, a property lost in control cells with time in culture.

Supernumerary Formation of Olfactory Glomeruli Induced by Chronic Odorant Exposure: A Constructivist Expression of Neural Plasticity

It is accepted that sensory experience instructs the remodelling of neuronal circuits during postnatal development, after their specification has occurred. The story is less clear with regard to the role of experience during the initial formation of neuronal circuits, whether prenatal or postnatal, since this process is now supposed to be primarily influenced by genetic determinants and spontaneous neuronal firing. Here we evaluated this last issue by examining the effect that postnatal chronic exposure to cognate odorants has on the formation of I7 and M72 glomeruli, iterated olfactory circuits that are formed before and after birth, respectively. We took advantage of double knock-in mice whose I7 and M72 primary afferents express green fluorescent protein and β-galactosidase, correspondingly. Our results revealed that postnatal odorant chronic exposure led to the formation of permanent supernumerary I7 and M72 glomeruli in a dose and time dependent manner. Glomeruli in exposed mice were formed within the same regions of olfactory bulb and occupy small space volumes compared to the corresponding single circuits in non-exposed mice. We suggest that local reorganization of the primary afferents could participate in the process of formation of supernumerary glomeruli. Overall, our results support that sensory experience indeed instructs the permanent formation of specific glomeruli in the mouse olfactory bulb by means of constructivist processes.

Stem Cell Therapy for Parkinson’s Disease: A Road Map for a Successful Future

Cell transplant therapy for Parkinsons disease (PD) has been in use for over 2 decades as an experimental treatment. Different cell types have been proposed as better therapeutic alternatives. However, source availability and therapeutic value of the transplants as compared to current pharmacological options have precluded the use of this kind of surgery in the majority of PD patients. In this article, we discuss the suitability of different types of stem cells for PD therapy, the requirements that the donor cells should fulfill in order to improve upon current methods, and propose alternatives for evaluating the efficacy of PD cell therapy.

Odor memory stability after reinnervation of the olfactory bulb.

The olfactory system, particularly the olfactory epithelium, presents a unique opportunity to study the regenerative capabilities of the brain, because of its ability to recover after damage. In this study, we ablated olfactory sensory neurons with methimazole and followed the anatomical and functional recovery of circuits expressing genetic markers for I7 and M72 receptors (M72-IRES-tau-LacZ and I7-IRES-tau-GFP). Our results show that 45 days after methimazole-induced lesion, axonal projections to the bulb of M72 and I7 populations are largely reestablished. Furthermore, regenerated glomeruli are re-formed within the same areas as those of control, unexposed mice. This anatomical regeneration correlates with functional recovery of a previously learned odorant-discrimination task, dependent on the cognate ligands for M72 and I7. Following regeneration, mice also recover innate responsiveness to TMT and urine. Our findings show that regeneration of neuronal circuits in the olfactory system can be achieved with remarkable precision and underscore the importance of glomerular organization to evoke memory traces stored in the brain.

Aspects of the narcolepsy-cataplexy syndrome in O/E3-null mutant mice.

Orexins (hypocretins) are peptide neurotransmitters produced by a small group of neurons located exclusively in the lateral hypothalamus (LH). Orexins modulate arousal, and as a result, have profound effects on feeding behavior and the sleep-wake cycle. Loss of orexin producing neurons leads to a narcoleptic phenotype, characterized by sudden transitions from vigilance to rapid eye movement (REM) sleep (direct transition to REM, DREM) observed in electroencephalogram (EEG) and electromyogram (EMG) recordings. In this study, we demonstrate that mice lacking the basic helix-loop-helix transcription factor O/E3 (also known as ebf2) have a decrease in orexin-producing cells in the LH, in addition to a severely impaired orexinergic innervation of the pons. These changes in the orexinergic circuit of O/E3-null animals induce a narcoleptic phenotype, similar to the one arising in orexin-deficient and orexin-ataxin-3 mice. Taken together, our results suggest that O/E3 plays a central role during the establishment of a functional orexinergic circuit by controlling the expression of essential hypothalamic neurotransmitter and the correct development of the nerve fibers arising from the hypothalamus. This is the first report regarding the narcolepsy-cataplexy syndrome in O/E3-null mice, which adds the importance of transcription factors in the regulation of neural subpopulations that control the sleep-wake cycle.

Nerve Growth Factor Increases Sodium Current in Pancreatic β Cells

Abstract. Nerve growth factor (NGF) induces neuritelike process outgrowth in cultured adult pancreatic β cells. DbcAMP partially mimics this effect on cell morphology, and both compounds act synergistically to promote neuritelike process outgrowth. To determine if NGF- and dbcAMP-induced differentiation was accompanied by changes in β cell electrical activity, we studied the macroscopic Na current of adult rat β cells identified with the reverse hemolytic plaque assay and cultured for one week with these factors.After 5–7 days, β cells cultured in the presence of 2.5S NGF exhibited a 48% increase on the macroscopic Na current, which was due to an increase on Na current density. We did not observe changes on voltage dependence of current activation, nor on steady-state inactivation. Although dbcAMP also promotes changes on β cell morphology, it did not affect the Na current density.

Heterogeneity in glutamic acid decarboxylase expression among single rat pancreatic beta cells

Aims/hypothesis. An isoform of glutamic acid decarboxylase, (GAD)65 has been identified as a pancreatic beta-cell autoantigen in Type I (insulin dependent) diabetes mellitus. We investigated the expression of GAD isoforms among single rat beta cells in culture, under different conditions and the correlation between GAD65 expression and insulin secretion-rate. Results. Independent of culture conditions, 100 % of fresh and cultured beta cells express GAD67. In contrast, considerable heterogeneity in GAD65 expression among single beta cells was observed. After 2 days in culture in 2.6 mmol/l glucose, only 24 % of the beta cells express GAD65. This percentage increases to 39 % in 5.6 mmol/l glucose and to 54 % and 56 % in 11.6 mmol/l and 20.6 mmol/l glucose, respectively. Moreover, reducing glucose concentration from 11.6 to 2.5 mmol/l for 2 days, reduces GAD65 expression by nearly 30 %. After 11 days in culture with 11.6 mmol/l glucose, 50 % of beta cells continue expressing GAD65, this percentage is further increased to nearly 75 % by including either nerve growth factor or dibutyryl cyclic AMP or both in the culture medium. When beta cells are challenged for 1 h with 20.6 mmol/l glucose, 67 % respond forming insulin-immunoplaques. More than two-thirds of insulin-secretors are GAD65-positive, in contrast to only 11 % of the non-secreting cells. Moreover, 87 % of beta cells that have a high insulin secretory rate express GAD65. Conclusion/interpretation. These results show that the most active beta cells, which secrete more insulin, also express GAD65 and that manipulating extracellular glucose may modify the expression of the enzyme and possibly the autoimmune attack in Type I diabetes. [Diabetologia (1999) 42: 1086–1092]