Identification of Marker Gene of Pars Tuberalis Morphogenesis in Chicken Embryo. The expression of Cytokine-like 1 and Gap junction protein alpha 5 in the pars tuberalis

Adenohypophysis delivered from oral ectoderm consists of pars distalis (PD), pars intermedia and pars tuberalis (PT). The mechanisms of development of PD has been well studied, and the cell differentiation of PD has been well understood. However, the morphogenesis and the differentiation of PT are still unclear, and the gene expression during the PT development remains largely unknown. In this study, we explored the specifically expressing genes in PT during development and analyzed its spatiotemporal expressions pattern. Microarray analysis on laser-captured PT and PD tissues obtained from chicken embryos on embryonic day 10 (E10.0) showed high expressing genes, Cytokine-like 1 (CYTL1) and Gap junction protein alpha 5 (GJA5) in PT. A detail analysis of spatiotemporal expressions pattern during chick embryo development by in situ hybridization revealed that CYTL1 mRNA was first detected in lateral head ectoderm and ventral head ectoderm in E1.5. The CYTL1 expressions moved into Rathke’s pouch at E2.5, then it was localized in PT primordium and continuously expressed in PT primordium until E12.0. On the other hand, GJA5 mRNA was transiently detected in PT primordium from E6 to E14.0, while the expression was not detected in PD during development. These results suggested that these genes may be involved in the regulation mechanism of PT development and could be a useful marker in the PT development

Competitive Binding Assay with an Umbelliferone-Based Fluorescent Rexinoid for Retinoid X Receptor Ligand Screening

Ligands for retinoid X receptors (RXRs), "rexinoids", are attracting interest as candidates for therapy of type 2 diabetes and Alzheimer's and Parkinson's diseases. However, current screening methods for rexinoids are slow and require special apparatus or facilities. Here, we created 7-hydroxy-2-oxo-6-(3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl)-2H-chromene-3-carboxylic acid (10, CU-6PMN) as a new fluorescent RXR agonist and developed a screening system of rexinoids using 10. Compound 10 was designed based on the fact that umbelliferone emits strong fluorescence in a hydrophilic environment, but the fluorescence intensity decreases in hydrophobic environments such as the interior of proteins. The developed assay using 10 enabled screening of rexinoids to be performed easily within a few hours by monitoring changes of fluorescence intensity with widely available fluorescence microplate readers, without the need for processes such as filtration

Runx3 regulates folliculogenesis and steroidogenesis in granulosa cells of immature mice

We previously demonstrated that female Runx3 knockout (Runx3-/-) mice were anovulatory and their uteri were atrophic and that Runx3 mRNA was expressed in granulosa cells. To clarify how Runx3 regulates folliculogenesis and ovulation, we examine the effects of Runx3 knockout on the gene expression of growth factors associated with folliculogenesis and enzymes associated with steroidogenesis. In Runx3-/- mouse ovaries, the numbers of primary and antral follicles were lower than those in wild-type (wt) mice at 3 weeks of age, indicating that the loss of Runx3 affects folliculogenesis. The expression of genes encoding activin and inhibin subunits (Inha, Inhba and Inhbb) was also decreased in ovaries from the Runx3-/- mice compared with that in wt mice. Moreover, the expression of the genes Cyp11a1 and Cyp19a1 encoding steroidogenic enzymes was also decreased. In cultured granulosa cells from 3-week-old mouse ovaries, Cyp19a1 mRNA levels were lower in Runx3-/- mice than those in wt mice. Follicle-stimulating hormone (FSH) treatment increased Cyp19a1 mRNA levels in both wt and Runx3-/- granulosa cells in culture but the mRNA level in Runx3-/- granulosa cells was lower than that in wt ones, indicating that granulosa cells could not fully function in the absence of Runx3. At 3 weeks of age, gonadotropin α subunit, FSHβ subunit and luteinizing hormone (LH) β subunit mRNA levels were decreased in Runx3-/- mice. These findings suggest that Runx3 plays a key role in female reproduction by regulating folliculogenesis and steroidogenesis in granulosa cells

Successful rituximab treatment of granulomatosis with polyangiitis with cranial neuropathies

Abstract Background In granulomatosis with polyangiitis (GPA), peripheral nerve involvement is common but central nervous system (CNS) involvement is extremely rare and treatment strategy has not been established. We report a case of intravenous cyclophosphamide (IVCY)-resistant GPA with associated cranial neuropathies that was successfully treated with rituximab (RTX). Case presentation A 37-year-old man with intractable sinusitis had several months of headache, hoarseness, and dysphagia; a month of right-sided deafness and nasal bleeding; and a week of dysarthria, steppage gait, and numbness in the right L5 distribution. A magnetic resonance imaging (MRI) examination of the head showed an infiltrative lesion in the right skull base encasing the carotid sheath. Computed tomography (CT) scan of the chest revealed a 23 mm nodule in the left upper lobe. Histology was inconclusive. Therefore, the patient was diagnosed as GPA. He was treated with glucocorticoids (GC) and IVCY. Three months later, he was readmitted for recurrence of headache and new left-sided hearing loss. He was treated with GC and RTX, and a 1-year remission followed. The molecular mechanism of RTX is not fully understood. In this case, RTX was more effective at rapidly and strongly suppressing B cells than CY. Since the B cell count was proportional to the patient’s clinical manifestations, B cells might represent a suitable target for the treatment of GPA with cranial neuropathies. Conclusions GPA with cranial neuropathies might be useful with RTX as induction therapy