Dual signal transduction pathways activated by TSH receptors in rat primary tanycyte cultures

Tanycytes play multiple roles in hypothalamic functions, including sensing peripheral nutrients and metabolic hormones, regulating neurosecretion and mediating seasonal cycles of reproduction and metabolic physiology. This last function reflects the expression of TSH receptors in tanycytes, which detect photoperiod-regulated changes in TSH secretion from the neighbouring pars tuberalis. The present overall aim was to determine the signal transduction pathway by which TSH signals in tanycytes. Expression of the TSH receptor in tanycytes of 10-day-old Sprague Dawley rats was observed by in situ hybridisation. Primary ependymal cell cultures prepared from 10-day-old rats were found by immunohistochemistry to express vimentin but not GFAP and by PCR to express mRNA for Dio2, Gpr50, Darpp-32 and Tsh receptors that are characteristic of tanycytes. Treatment of primary tanycyte/ependymal cultures with TSH (100 IU/l) increased cAMP as assessed by ELISA and induced a cAMP-independent increase in the phosphorylation of ERK1/2 as assessed by western blot analysis. Furthermore, TSH (100 IU/l) stimulated a 2.17-fold increase in Dio2 mRNA expression. We conclude that TSH signal transduction in cultured tanycytes signals via Gαs to increase cAMP and via an alternative G protein to increase phosphorylation of ERK1/2

Circadian clock mechanism driving mammalian photoperiodism.

The annual photoperiod cycle provides the critical environmental cue synchronizing rhythms of life in seasonal habitats. In 1936, Bünning proposed a circadian-basis for photoperiodic synchronization. Here, light-dark cycles entrain a circadian rhythm of photosensitivity, and the expression of summer or winter biology depends on whether light coincides with the phase of high photosensitivity. Formal studies support the universality of this so-called coincidence timer, but we lack understanding of the mechanisms involved. Here we show in mammals that coincidence timing takes place in the pars tuberalis of the pituitary, through a melatonin-dependent flip-flop switch between circadian transcriptional activation and repression. Long photoperiods produce short night-time melatonin signals, leading to induction of the circadian transcription factor BMAL2, in turn triggering summer biology through the eyes absent / thyrotrophin (EYA3 / TSH) pathway. Conversely, short photoperiods produce long melatonin signals, inducing circadian repressors including DEC1, in turn suppressing BMAL2 and the EYA3/TSH pathway, triggering winter biology. These actions are associated with progressive genome-wide changes in chromatin state, elaborating the effect of the circadian coincidence timer. Hence, circadian clock interactions with pituitary epigenetic pathways form the basis of the mammalian coincidence timer mechanism. Our results constitute a blueprint for circadian-based seasonal timekeeping in vertebrates

Differential response patterns of kisspeptin and RFamide-related peptide to photoperiod and sex steroid feedback in the Djungarian hamster ( Phodopus sungorus )

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Spontaneous perikaryal neurofilament phosphorylation in the septofimbrial nucleus of the rat.

Phosphorylation of the 200 kDa neurofilament peptide NF-H usually only occurs in axons. We describe the spontaneous presence of phosphorylated NF-H in a population of small spindle-shaped neurons of the rat septofimbrial nucleus. A similar phenomenon has been observed in axotomized neurons and in human neurodegenerative diseases. Our observations, as well as previous studies by other authors, indicate that perikaryal neurofilament phosphorylation is not necessarily linked to pathological conditions

Paired helical filament-like inclusions and Hirano bodies in the mesencephalic nucleus of the trigeminal nerve in the aged rat.

The alterations appearing in trigeminal mesencephalic primary sensory neurons during ageing have been analyzed by electron microscopy in the Wistar-Louvain rat. Two phases have been distinguished, similar to those observed in dorsal root ganglion neurons. Up to 24 months, the mesencephalic trigeminal neurons progressively accumulate lipofuscins, while filamentous inclusions start to appear around 24 months of age. Hirano bodies and paired helical filament-like structures have been identified in animals aged 24 months or more. This time-course parallels the one observed previously in dorsal root ganglion neurons, indicating that the blood-brain barrier does not seem to influence the ageing of mesencephalic trigeminal neurons. The relationship between the paired helical filament-like inclusions and Hirano bodies, as well as similar structures already described by other authors, is discussed

[Adriamycin Effects On Axotomized Neurons in the Regeneration Chamber Model]

Peripheral nerve tubulization is mainly used to study regeneration. We used this model to study the effects of adriamycin on axotomized dorsal root ganglion neurons after local administration of this drug in silicone chambers placed on the proximal stump of a sectioned nerve. No massive neuronal degeneration has been detected in the dorsal root ganglia. However, adriamycin induced a significant atrophy in axotomized neurons. Our observations indicate that this effect is due to blood borne transport of adriamycin, rather than retrograde axonal transport, and that it affects also axotomized neurons that are not directly exposed to adriamycin at the sectioned stump of their axon

Regulation of MT melatonin receptor expression in the foetal rat pituitary.

During development, melatonin receptors are transiently expressed in multiple neuroendocrine tissues, suggesting a novel role for melatonin in developmental physiology. The best characterised model of melatonin signalling during development is the pars distalis of the rat pituitary. However, although many studies have characterised the postnatal decline of melatonin receptors in the rat pars distalis, the mechanism(s) that time the developmental onset of receptor expression during embryogenesis are unknown. Analysis of these mechanisms may yield important information regarding the putative role of melatonin in neuroendocrine development. Here, we report the expression of MT(1) melatonin receptor mRNA in the rat pituitary from embryonic day 15.5 (e15.5). Prior to e15.5, the homeodomain transcription factor Msx-1, an inhibitor of cellular differentiation, is widely expressed throughout the pituitary. In transient transfection experiments, Msx-1 potently inhibited pituitary homeobox-1 (Pitx-1)-induced MT(1) promoter activity and therefore may represent a key inhibitor of MT(1) expression in early pituitary development. During late embryogenesis, MT(1) mRNA was expressed in both the ventral and dorsal pituitary. Analysis of a 1.5-kb fragment of the rat MT(1) promoter revealed four putative cis-elements for the POU domain factor Pit-1, which is associated with mid-dorsal cell lineages. Although Pit-1 induced a strong, dose-dependent stimulation of MT(1) promoter activity in vitro, dual-labelled in situ hybridisation revealed no colocalisation of MT(1) and Pit-1 mRNAs in vivo at e19.5. By contrast, all MT(1) positive cells colocalised with alphaGSU and most with betaTSH mRNA. Our data therefore implicate the decline of Msx-1 expression as a key event that times the onset of melatonin receptor expression to the differentiation of endocrine cells types in the developing pituitary gland, and suggest that the melatonin-sensitive cells in the embryonic pituitary are primarily Pit-1-independent thyrotrophs in the rostral pituitary, with a secondary population of pars distalis gonadotrophs

Phosphorylated neurofilament epitopes in neuronal perikarya in the septum, mesencephalon and dorsal root ganglia of mammals and birds.

We and other researchers have previously described the presence of axon-specific phosphorylated neurofilament epitopes in the cell bodies of three neuronal types in the rat: bipolar septofimbrial neurons and the large light A-type cells in the dorsal root ganglia and the mesencephalic nucleus of the Vth nerve. This spontaneous presence of phosphorylated neurofilaments at the level of the perikaryon contrasts with the induced appearance of these epitopes in axotomized neurons. We have undertaken a study of this phenomenon in rat, mouse, gerbil, rabbit, pig and chicken to analyse its species distribution. Phosphorylated neurofilament positive perikarya could be detected in the dorsal root ganglia and mesencephalic nucleus of the Vth nerve in all analysed species. Although this labelling has been shown to be specific for A-type cells in rat, in pig small cells were preferentially labelled, whereas the largest cells were mostly completely devoid of label. In the septofimbrial nucleus, phosphorylated neurofilament positive perikarya were seen in rat, mouse, gerbil and rabbit. In the pig, only a phosphatase-insensitive neurofilament antibody labelled these neurons. In the chicken, the labelling was completely absent. These observations establish the widespread species distribution of perikaryal phosphorylated neurofilament epitopes in the dorsal root ganglia and mesencephalic nucleus of the Vth nerve. In the septofimbrial nucleus however, this phenomenon seems to be restricted to rodents and lagomorphs. We discuss possible explanations for these cytoskeletal singularities in dorsal root ganglia, the mesencephalic nucleus of the Vth nerve and septofimbrial neurons