The hippocampal formation has been shown to be particularly vulnerable to the neurotoxic effects of chronic ethanol consumption. It was hypothesized that this damage was due to the disruption of the expression of neurotrophic factors and certain other proteins within the hippocampus. By using real-time reverse transcription-polymerase chain reaction (RT-PCR) techniques, this study aimed to determine whether chronic ethanol consumption could alter the mRNA expression level of brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF), and oligodendrocyte myelin glycoprotein (OMgp) in the hippocampus. Wistar male rats received an unrestricted access to a liquid diet containing 5% (v/v) ethanol as the sole source of fluid from 10 to 29 weeks of age. Control rats had unlimited access to a liquid diet containing an isocaloric amount of sucrose. We found that chronic ethanol consumption did not cause significant changes in the levels of mRNA for BDNF and GDNF. However, OMgp mRNA showed a significant deficit in ethanol-treated animals. It is suggested that this deficit may be related to the demyelination that is commonly observed in human alcoholics and that this may contribute to the functional and cognitive deficits.

Rats of the Jcl: Wistar-TgN (ARGHGEN) 1Nts strain (Mini rats) are transgenic animals carrying an antisense RNA transgene for rat growth hormone (GH); they show poor somatic growth and a low blood GH level compared to age-matched wild-type Wistar (non-Mini) rats. The purpose of the present study was to investigate age-related changes in growth hormone-immunoreactive (GH-IR) cells in the anterior pituitary gland (AP) of Mini rats at four, six, and eight weeks of age. The body weight and size of the GH-IR cells of Mini rats was significantly lower than that of non-Mini rats at six and eight weeks of age; however, this difference was not observed at four weeks of age. The AP volume and the number of GH-IR cells in Mini rats were significantly smaller than those of the age-matched non-Mini rats at the three ages. These results suggest that the abnormal development of GH-IR cells in the AP induced by the GH antisense RNA transgene is responsible for the poor somatic growth and the low blood GH levels in Mini rats.

Chronic ethanol consumption has adverse effects on the central nervous system. Hippocampus is one of the target sites of ethanol neurotoxicity. Hippocampal damage is known to result in impairment of learning and memory. This study was aimed to determine whether chronic ethanol consumption could alter the expression levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) mRNAs in the hippocampus. Male Wistar rats were given unrestricted access to a liquid diet containing 5% (v/v) ethanol as the sole fluid source for 19 weeks beginning at 10 weeks of age. The expression levels of BDNF and GDNF mRNAs in the hippocampus were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. The present study revealed that chronic ethanol consumption did not result in significant changes in the expression levels of BDNF and GDNF mRNAs. Our present results showed no significant alteration in the expression of these neurotrophic factors; these results will lead to further studies to examine the possible alterations in the gene expression of various neurotrophins that are related to hippocampal functions including learning and memory.

Stereology is a group of mathematical and statistical methods that allows the extrapolation of three-dimensional structural information from two-dimensional sections (or slices). This allows researchers to derive important quantitative structural information, such as the volume, surface area or numbers of particular particles (e.g. cells) within defined regional boundaries. The need for such quantitative information in biology is of particular importance when evaluating the influence of various experimental treatments on specific organs, tissues and cells in the body. Knowledge of such changes has given important insights into the neural substrates that may be responsible for the functional and behavioral consequences of a disparate range of experimental treatments. Here, we describe some of these methods as applied to quantifying the total numbers of cells in defined regions of the hippocampal formation. The methods used for this evaluation were, first, the Cavalieri principle, which was used to determine the volumes of the various subdivisions of the rat hippocampus, and, second, the ‘physical disector’ method, which was used to estimate the numerical density of neurons within each subdivision. Once these values were derived, it was but a simple task to multiply them together to obtain estimates for the total numbers of cells in the given hippocampal region. We found that 16-and 30-day-old normal male rats had 176 800 and 152 700 pyramidal cells in the CA1 region, respectively. This decrease in the neuronal number was statistically significant. However, in the CA2+CA3 region, there were approximately 169 300 and 149 600 pyramidal cells in 16- and 30-day-old normal male rats, respectively, which was not significantly different. In the dentate gyrus, there were approximately 36 700 neurons in the hilus region and 483 000 granule cells in the granule cell layer, irrespective of the age of the rats. There were no significant differences between these estimates of hilus neurons and granule cells.

The effects of maternal deprivation (MD) during early postnatal life on the brain-derived neurotrophic factor (BDNF) level were investigated in the present study. Wistar rats were assigned to either maternal deprivation or mother-reared control (MRC) groups. MD manipulation was achieved by separating rat pups from their mothers for 3h a day during postnatal days (PND) 10–15. At 16, 20, 30, and 60 days of age, the level of BDNF mRNA in the hippocampal formation of each group was determined using real-time PCR analysis. Early postnatal maternal deprivation of rat pups resulted in a significant increase in body weight at 60 days of age. The expression of BDNF mRNA in the hippocampus was significantly decreased at 16 days of age, and increased at 30 and 60 days of age. These data indicate that even a brief period of maternal deprivation during early postnatal life can affect hippocampal BDNF expression.

Morphological features and functional implications of projections of the parabrachial nucleus to the central nucleus of the amygdala were investigated in the rat. The anatomical study was based on injections of the tracers horseradish peroxidase and biotinylated dextran amine. An extremely dense concentration of labeled fibers was found in the lateral and lateral capsular subdivisions of the central nucleus of the amygdala, originating mainly from the external lateral and ventral lateral subnuclei of the parabrachial nucleus. The parabrachial fibers exhibited the morphological characteristic of forming dense pericellular terminal arborizations. The functional implications of this pathway in cardiovascular functions were verified using Fos protein induction in response to hypotension induced by continuous intravenous administration of hydralazine-hydrochloride. In this paradigm, Fos immunoreactivity was found to be confined to the lateral and lateral capsular subdivisions of the central nucleus of the amygdala. Double immunostaining methods were used to visualize, at the electron microscopic level, terminals labeled by biotinylated dextran amine and Fos cell labeling. With this approach, we were able to confirm that Fos-immunoreactive neurons in the central nucleus of the amygdala receive axosomatic terminals from the parabrachial nucleus. The present findings point out that parabrachial inputs to the central nucleus of the amygdala play a relevant role in regulating cardiovascular function.

The suprachiasmatic nucleus (SCN) has long been recognized as the central mammalian circadian pacemaker that controls behavioral and physiological processes. The role of the SCN in circadian rhythms has been the subject of a wide range of physiological and behavioral studies, although the influence of homeostasis rhythms (such as fluctuating hormone levels) on the SCN of the hypothalamus is not entirely clear. The present study was undertaken to examine the morphological interactions between astroglial and neuronal elements in the SCN of mice after either a short-term excessive testosterone treatment (ETT) or castration, using glial fibrillary acidic protein (GFAP), and immediate early gene c-fos as well as calbindin-D28k (CB) immunohistochemistry. Both ETT and castration resulted in a significant increase in the accumulation of reactive astrocytes and Fos-imunoreactivity (IR), especially in the dorsomedial (DM) sub-region of the SCN. However, CB-IR neurons in the examined brain regions showed little change. These findings indicate that the DM sub-region of the SCN may be a possible center of hormonal regulation via a hypothalamic neuroendocrine circuit, and that a non-photic stimuli mechanism might play a role in circadian rhythm regulation.

OBJECTIVE:Toluene diisocyanate (TDI)-induced asthma is a common cause of occupational lung disease. In addition, a sore throat is one of the complaints of TDI-exposed workers. The aim of the present study was to determine whether TDI exposure induces laryngeal and/or tracheal lesions in experimental animals.MATERIAL AND METHODS:Guinea pigs underwent naris application of TDI three times, and their respiratory tracts were then examined using light and electron microscopy. Some animals simultaneously received vitamins C and E. which function as antioxidant agents.RESULTS:When TDI-treated animals showed the clinical sign of labored breathing, many eosinophils had appeared in the lamina propria and mucosa of both the larynx and trachea, which finally infiltrated the tract lumen through the ruptured epithelium. Laryngo-tracheal inflammation was more severe than that observed in the lungs. However, supplementation with antioxidant vitamins in TDI-treated animals ameliorated the respiratory eosinophilia.CONCLUSION:Naris application of TDI induced laryngotracheitis. which was significantly suppressed by the antioxidant vitamins, This implies a preventive effect of the vitamins on this occupational disease.

Exposure of the developing brain to X-irradiation in utero is known to cause various deleterious consequences. We have previously reported the effects of prenatal X-irradiation on the development of the cerebral cortex in rats. We have now extended this study to examine the effects of such X-irradiation on the development of the cerebellum. Wistar rats were exposed to 1.5 Gy X-irradiation either on days 14, 15 or 16 of gestation (E14, E15, E16). Sham-irradiated animals were used as controls. At seven postnatal weeks of age, male rats from each group were deeply anesthetized and killed by intracardiac perfusion with 2.5% glutaraldehyde in 0.1 M phosphate buffer. The unbiased stereological procedure known as the fractionator method was used to estimate the total number of Purkinje cells in the cerebellum of each animal. Body and cerebellar weights from E14 and E15, but not E16 irradiated rats showed significant deficits compared to control animals. Rats irradiated on E16 and control rats had about 285100-304800 Purkinje cells in the cerebellum. There was no significant difference between these values. However, E14 and E15 irradiated animals had about 117500 and 196300 Purkinje cells, respectively. These estimates were significantly different from those observed in both control and E16 irradiated rats. Given that the phase of division of Purkinje cell progenitors is mainly between E14-E15 and the phase of differentiation and migration is between E16-E20, it is concluded that the vulnerable period of the Purkinje cells to X-irradiation closely overlaps the phase of division of progenitors.