Glial cell activity is maintained during prolonged inflammatory challenge in rats

We evaluated the expression of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), ionized calcium binding adaptor protein-1 (Iba-1), and ferritin in rats after single or repeated lipopolysaccharide (LPS) treatment, which is known to induce endotoxin tolerance and glial activation. Male Wistar rats (200-250 g) received ip injections of LPS (100 µg/kg) or saline for 6 days: 6 saline (N = 5), 5 saline + 1 LPS (N = 6) and 6 LPS (N = 6). After the sixth injection, the rats were perfused and the brains were collected for immunohistochemistry. After a single LPS dose, the number of GFAP-positive cells increased in the hypothalamic arcuate nucleus (ARC; 1 LPS: 35.6 ± 1.4 vs control: 23.1 ± 2.5) and hippocampus (1 LPS: 165.0 ± 3.0 vs control: 137.5 ± 2.5), and interestingly, 6 LPS injections further increased GFAP expression in these regions (ARC = 52.5 ± 4.3; hippocampus = 182.2 ± 4.1). We found a higher GS expression only in the hippocampus of the 6 LPS injections group (56.6 ± 0.8 vs 46.7 ± 1.9). Ferritin-positive cells increased similarly in the hippocampus of rats treated with a single (49.2 ± 1.7 vs 28.1 ± 1.9) or repeated (47.6 ± 1.1 vs 28.1 ± 1.9) LPS dose. Single LPS enhanced Iba-1 in the paraventricular nucleus (PVN: 92.8 ± 4.1 vs 65.2 ± 2.2) and hippocampus (99.4 ± 4.4 vs 73.8 ± 2.1), but had no effect in the retrochiasmatic nucleus (RCA) and ARC. Interestingly, 6 LPS increased the Iba-1 expression in these hypothalamic and hippocampal regions (RCA: 57.8 ± 4.6 vs 36.6 ± 2.2; ARC: 62.4 ± 6.0 vs 37.0 ± 2.2; PVN: 100.7 ± 4.4 vs 65.2 ± 2.2; hippocampus: 123.0 ± 3.8 vs 73.8 ± 2.1). The results suggest that repeated LPS treatment stimulates the expression of glial activation markers, protecting neuronal activity during prolonged inflammatory challenges.

GnRH neurones population in the diencephalon of the coypu myocastor coypus

El objetivo del presente estudio fue investigar la distribución y morfología de las neuronas que sintetizan y almacenan el factor de liberación de gonadotrofinas (GnRH), en el diencéfalo del coipo (Myocastor coypus), roedor sudmericano del suborden histricomorpha. Para tales fines los encéfalos de tres coipos machos, adultos, fueron fijados por perfusión intra-arterial, utilizando una solución de paraformaldehído y ácido pícrico. Los bloques del hipotálamo fueron separados del resto del encéfalo y seccionados en micrótomo de congelación, obteniendo láminas coronales de 40 µm de espesor. Las secciones fueron sometidas a un proceso de recuperación antigénica por ultrasonido, para desenmascarar los antígenos ocultos, luego las secciones fueron procesadas utilizando una técnica de inmunohistoquímica para evidenciar las neuronas GnRH, usando un anticuerpo monoclonal (LRH 13). Un grupo de secciones seriadas fue coloreada con violeta de cresilo (técnica de NissI), mientras que las otras secciones fueron coloreadas usando el método de Kuver Barrera (azul luxol rápido y violeta de cresilo), para observar los núcleos y tractos nerviosos del hipotálamo, con el microscopio óptico. Los análisis morfométrico y cuantitativo de los cuerpos neuronales fue realizado usando un analizador de imágenes. Las neuronas GnRH inmunoreactivas observadas eran bipolares y alargadas. El número total de cuerpos neuronales para esta especie fue estimado en 1072 ñ 27. Todas las células fueron localizadas en el hipotálamo rostral, principalmente en el área preóptica. Pocas neuronas fueron observadas en el núcleo de la estría terminal y en el núcleo medial preóptico hipotalámico. Las fibras inmunorreactivas fueron visibles en la capa externa de la eminencia media. De acuerdo a los datos obtenidos, se concluye que la distribución de las neuronas GnRH en el coipo coinciden con la de otros roedores, tales como la rata

Regional specificity seen within hypothalamus in neuroimmunomodulation.

Wistar strain male albino rats lesioned and sham lesioned at Ventromedial hypothalamus (VMH) were used to study the neuroimmunomodulation by this brain region. Except the decrease in thymus weight/body weight ratio (P < 0.01), and its cellularity (P < 0.02) in immunized VMH lesion animals, the rest of the parameters like plaque forming cells (PFC), antibody titre, leukocyte migration inhibition index, foot pad thickness in the lesioned as well as the lesion immunized animals never deviated from their respective sham and immunized sham animals. Sham operated belongs to VMH when compared to control rats, showed marked decrease in spleen weight (P < 0.001), thymus weight (P < 0.02) and decrease in popliteal lymph node weight (P < 0.001) ratios. After immunization, the immunized sham animals showed a marked decrease in antibody titre (P < 0.05), PFC (P < 0.05), spleen (P < 0.05) and thymus (P < 0.001) weight ratios with the significant increase in splenic cell count (P < 0.01) compared to immunized control rats. VMH may be one of the information receiving center. However, from these results, it is inferred that VMH could not be a modulating center for the many of the parameters studied as far as neuroimmunomodulation is concerned.

Cellular localization of atrial natriuretic peptide and tissue kallikrein in the human hypothalamus

In the kidney, renal atrial natriuretic peptide (ANP) is considered to play an important role wter and salt homeostasis. Immunoreactive ANP in the brain of lower invertebrates, such as the rat, has been shown to be localizaed in the hypothalamus and septum. Several studies have investigated the possibility of a regulatory system in the brain similar to that of the kidney. Since neuronal function is acutely sensitive to disturbances of the intracranial water and salt balance we have attempted to immunolocalize ANP-containing cells in the normal human hypothalamus, using a polyclonal antiserum specific to ANP. Also, we have observed tissue kallikrein (TK), using a polylonal antiserum specific to TK, in the same areas as ANP. A regulatory role for TK on prolactin has been suggested as the rationale for the co-localization of these two hormones in human prolactinomas. Therefore, it could be suggested that TK plays a similar role in the processing of precursor ANP in the brain. It is contemplated to examine the status of these peptides in patients with cerebral oeodema