Glutamine Synthetase is Expressed by Primary Sensory Neurons from Chick Embryos In Vitro but not In Vivo: Influence of Skeletal Muscle Extract.

Glutamine synthetase (GS) catalyses the ATP-dependent formation of glutamine from glutamate and ammonia. To determine whether dorsal root ganglion (DRG) cells from chick embryos express the enzyme in vivo or in vitro, GS was detected by immunocytochemical reaction either in vibratome sections of DRG or in dissociated DRG cell cultures. The immunocytochemical detection of GS showed that in vivo the DRG taken from chick embryos at day 10 (E10), E14, E18 or from chickens after hatching were free of any GS-positive ganglion cells; in contrast, in neuron-enriched cultures of DRG cells grown in vitro at E10, virtually all the neuronal cells (98.6 +/- 1.0%) express GS at 3, 5 or 7 days of culture. In mixed DRG cell cultures, only 83.6+/-4.6% of the neurons displayed a GS-immunoreactivity. In both culture conditions, neither the presence of horse serum nor the age of the culture appeared to affect the percentage of neurons which displayed a GS-immunoreactivity. After [3H]glutamine uptake, radioautographs revealed that only 80% of the neurons were labelled in neuron-enriched DRG cell cultures while 96% of the neurons were radioactive in mixed DRG cell cultures. Furthermore the most heavily [3H]glutamine-labelled neurons were exclusively found in mixed DRG cell cultures. Combination of both immunocytochemical detection of GS and radioautography after [3H]glutamine uptake showed that strongly GS-immunostained neurons corresponded to poorly radioactive ones and vice versa. When skeletal muscle extract (ME) was added to DRG cell cultures, the number of GS-positive neurons was reduced to 77.5 +/- 2.5% in neuron-enriched cultures or to 43.6 +/- 3.8% in mixed DRG cell cultures; in both types of culture, the intensity of the neuronal immunostaining was depressed. Furthermore, combined action of ME and non-neuronal cells potentiates the enzyme repression exerted separately by ME or non-neuronal cells. Since GS-immunoreactivity is expressed in DRG cells grown in vitro, but not in vivo, it is suggested that microenvironmental factors influence the expression of GS. More specifically, the repression of GS by primary sensory neurons grown in vitro may be strongly induced by soluble factors present in skeletal muscle, and to a lesser extent in brain, and potentiated by non-neuronal cells.

A role for altered TLR gene expression in association with increased expression of CD200R in the induction of mucosal tissue CD4(+) Treg in aged mice following gavage with a liver extract along with intramuscular monophosphoryl lipid A (MPLA) injection.

Previous studies showed a fetal sheep liver extract (FSLE), in association with LPS, injected into aged (>20 months) mice reversed the altered polarization (increased IL-4 and IL-10 with decreased IL-2 and IFN-gamma) in cytokine production seen from ConA stimulated lymphoid cells of those mice. Aged mice show a >60% decline in numbers and suppressive function of both CD4(+)CD25(+)Foxp3(+)Treg and so-called Tr3 (CD4(+)TGFbeta(+)). Their number/function is restored to levels seen in control (8-week-old) mice by FSLE. We have reported at length on the ability of a novel pair of immunoregulatory molecules, members of the TREM family, namely CD200:CD200R, to control development of dendritic cells (DCs) which themselves regulate production of Foxp3(+) Treg. The latter express a distinct subset of TLRs which control their function. We report that a feature of the altered Treg expression following combined treatment with FSLE and monophosphoryl lipid A, MPLA (a bioactive component of lipid A of LPS) is the altered gene expression both of distinct subsets of TLRs and of CD200Rs. We speculate that this may represent one of the mechanisms by which FSLE and MPLA alter immunity in aged mice.

Fenología del anillo de crecimiento de Pinus uncinata Ramond y Pinus sylvestris L. en un gradiente altitudinal en los Pirineos Centrales

En el presente trabajo, describimos el desarrollo y la estructura del anillo anual de crecimiento de Pinus uncinata Ramondy Pinus sylvestris L. en un gradiente altitudinal en los Pirineos centrales. Medimos mensualmente el aumento de la anchura del anillo y el número de traqueidas añadidas a lo largo de 1993. En ambas especies, la mayor parte del anillo se forma desde mediados de jimio hasta mediados de julio de 1993. El desarrollo de la madera tardía -formación, coloración y engrasamiento y lignificación de las paredes celulares- comienza en julio y acaba en octubre. El anillo comienza antes su formación en los bosques estudiados de P. sylvestris que en P. uncinata, el cual vive a mayor altitud. P. uncinata forma anillos más estrechos, con menor múmero de células y con mayor heterogeneidad de color de la madera tardía que P. sylvestris. Existe una gran variabilidad entre árboles en la posición de las células de las maderas temprana y tardía a lo largo del anillo de crecimiento. Los distintos patrones fenológicos en el crecimiento del anillo anual, a lo largo del gradiente altitudinal seleccionado, pueden explicarse en parte, por la influencia de distintas condiciones climáticas, principalmente térmicas.

Crecimiento estacional y caída de acículas en Pinus uncinata Ram. y Pinus sylvestris L.

En este trabajo se estudia la formación mensual del anillo del tronco, la caída de acículas y la elongación de las ramas en poblaciones de Pinus sylvestris L. y Pinus uncinata Ram. en el Pirineo Central Español durante 1993. La formación del anillo se produce de principios de junio hasta final de septiembre, si bien la mayoría de su crecimiento se lleva cabo de mediados de junio hasta mediados de julio. P. sylvestris comienza el desarrollo de su anillo antes que P. uncinata. El inicio del desarrollo de los brotes de P. uncinata se adelanta ligeramente al del crecimiento en grosor del tronco. La caída de acículas es máxima en el otoño y se produce más tarde en P. uncinata que en P. sylvestris. Este retraso, junto al mayor peso de las acículas y el menor diámetro de las traqueidas de P. uncinata, denotan su adaptación a un mayor estrés ambiental que P. sylvestris.