A real time PCR based approach for the quantitative detection of FUS-CHOP fusion transcripts in human liposarcoma.

PURPOSE: Histology still forms the backbone for the diagnosis of the myxoid and round cell subtypes of liposarcoma but the molecular identification of the different transcript variants remains a challenge, due, in part, to the complexity of multiple overlapping exons that they share between them. This study was conducted to develop and evaluate a more sensitive platform than existing semi-quantitative approaches for detecting FUS-CHOP transcripts. MATERIALS AND METHODS: In the present investigation we describe a novel approach using real-time PCR to identify and differentiate the fusion transcripts formed in the t(12; 16)(q13; p11) chromosomal translocation. This method is founded on the basis of transcript individualized primers and probes, which were designed to detect specifically the different variants in both frozen and FFPE tissues. RESULTS: Our results show that the method is highly specific, sensitive, and superior to the widely used nested PCR approach, and is accurately able to differentiate the most common variants, as well as quantify copy numbers. Primer amplification and probe detection of FUS-CHOP from genomic DNA of human, mouse, cocker spaniel and chicken sources all resulted in completely negative results indicating this technique is specific for human RNA derived transcripts. CONCLUSION: This new method offers an additional tool in the investigation of liposarcoma that may impact considerably on missed diagnosis and it's accompanying clinical ramifications.

Induction of c-fos gene expression by the selective sigma receptor ligand EMD 57445 in rat brain.

Based on animal studies it has been reasoned that ligands to sigma binding sites might be effective in the treatment of schizophrenic disorders and may also be used to investigate this largely elusive disorder on a molecular level. Expression patterns of c-fos in rat brain were studied following treatment with single doses of the sigma ligand EMD 57445 (0.3, 1, 3, 30 mg/kg s.c.). Specific c-fos gene expression was detected at all concentrations tested in various cortical areas. The signals observed were dose-dependent with the highest intensities in the piriform cortex. Strong signals were also detected in hippocampal areas CA 1,2,3 and the gyrus dentatus, as well as in the medial habenula nuclei. In the caudate putamen, nucleus accumbens and lateral septal nucleus signals were detectable after administration of doses > or = 1 mg/kg. Furthermore, c-fos hybridization was visible in the amygdala, in the mammillary bodies, the islands of Calleja and in the olfactory tubercle. In the hypothalamus, c-fos expression was seen in the median eminence area after 30 mg/kg EMD 57445. No hybridization signals were obtained in brainstem or cerebellum. Since c-fos expression induced by EMD 57445 resembled the pattern obtained with atypical neuroleptics and studies on animal behavior point to antipsychotic activity, it is concluded that the drug might be suitable in the treatment of schizophrenia.

Anterograde neuronal tracing of retinohypothalamic projections in the hamster--possible innervation of substance P-containing neurons in the suprachiasmatic nucleus.

The retinohypothalamic tract (RHT) in the Djungarian hamster Phodopus sungorus was studied using anterograde neuronal transport of cholera toxin subunit B (CTB) following unilateral intraocular injection. A major projection terminates bilaterally in the hypothalamic suprachiasmatic nuclei (SCN). In the anterior ventral SCN, a light ipsilateral predominance was evident. In the medial SCN, labelling was concentrated laterally where it was seen over the dorso-ventral extension of the nuclei, pronounced contralaterally to the site of CTB injection, which was even more characteristic in the posterior aspects of the nuclei. Labelled fibers and terminals were observed in the supraoptic nuclei, but not in lateral and paraventricular hypothalamic regions. Additional experiments utilizing double immunofluorescence of CTB and of substance P (SP) in the SCN showed that SP-containing perikarya were particularly observed in a central portion of the nucleus, where CTB-stained terminals were accumulated in the vicinity of immunoreactive cell bodies, fibers and terminals. Our data provide preliminary morphological evidence for the regulation of SCN function by retinal afferents and may explain the circadian and photoperiodic fluctuations in the amount of SP in the SCN.