Project description:While the thyroid has the capacity to grow in response to a stimulus that perturbs the pituitary-thyroid axis, cells involved in this process have not been well characterized. In this study, partial thyroidectomy was used to produce regeneration. Histological and gene profiling analyses were carried out using normal mouse thyroid as well as those after partial thyroidectomy. In the central area of the right or left normal thyroid lobe, a proliferative center was found with the occasional presence of micro-follicles, where BrdU-positive and/or C cells were mainly localized. Two weeks after partial thyroidectomy, the number of BrdU-positive cells and cells with clear or faintly eosionphilic cytoplasm, markedly increased in the central area of the thyroid lobe and the overlapping area close to the cutting edge. Clear-looking cells had scant cytoplasmic components as determined by electron microscopy; some retained characteristic features of C cells such as the presence of neuroendocrine granules while others retained follicular cell specific features such as those directly facing a lumen with microvilli at the apical side of the cell. Some of the clear-looking cells were BrdU-positive and expressed FOXA2, the definitive endoderm lineage marker. Microarray analysis further revealed that partial thyroidectomy affected the expression pattern of genes involved in cellular and/or metabolic process and their regulation categorized as Endocrine System Disorders and Embryonic Development, by Pathway analysis. These results suggest that cells previously C cells and/or follicular cells may be altered after partial thyroidectomy to become immature cells that participate in the repair and/or regeneration of the thyroid, a process similar to embryonic thyroid development. Total RNAs used for microarray analysis were prepared from laser capture microdissected mouse thyroids of the central (C) and peripheral (P) areas before (N: normal) and after (D: dissected) partial thyroidectomy. The RNAs obtained were first amplified, which were then subjected to hybridization using 5 NC (central area of the right or left lobe of normal thyroid), 3 NP (peripheral area of the right or left lobe of normal thyroid), 5DC (central area of the right or left lobe of dissected thyroid), 5 DP (peripheral area of the right or left lobe of dissected thyroid), and universal mouse RNAs as a reference. The 4-way analysis was carried out: NC vs NP vs DC vs DP.

Project description:The epilepsies represent one of the most common neurological disorders. Mesial temporal lobe epilepsies (MTLE) are the most frequent form of partial epilepsies and display frequent resistance to anti-epileptic drugs thus representing a major health care problem. In TLE, the origin of seizure activity typically involves the hippocampal formation, which displays major neuropathological features, described with the term hippocampal sclerosis (HS). HS is the most frequent pathological substrate of refractory mesial temporal lobe epilepsy. Complex partial seizures (CPS) are the predominant seizure type associated with medial temporal lobe epilepsy. MTLE is commonly due to mesial temporal sclerosis (MTS). The biology underlying the epilepstic seizures and the transcriptome associated to the seizure in intractable medial temporal lobe epilepsy is ill understood. The aim of the study was to identify potential biomarkers that could identify epileptic seizure. Thus we performed transcriptome profiling of ten medial temporal lobe epilepsy cases which are resistant to the drug and underwent temporal lobectomy. The cases constitutes of patients with intractable complex partial seizure, treated medically and have undergone detailed presurgical evaluation and subjected to surgery for standard temporal lobectomy and amygdalo-hippocampectomy. The spiking areas identified after the electrocorticography will form the test tissues, which compared with the nonspiking areas removed during the surgery, from the same patient. This could probably form one of the appropriate controls, as test and control are from same patient, which eliminates the genome variations that could incur due to the comparison with the tissues from the another patient. Also this could get rid of expression changes due to the treatments undergone by the patient. We performed two color microarray wherein we labled seizure focus (spiking area) with Cy5 and non-seizure region tissues (non-spiking) with Cy3. As a strategy to test the possibility of potential diagnostic biomarkers we are intended to test the differentially regulated molecules in an independent set of epilepsy samples. Two color experiment

Project description:The epilepsies represent one of the most common neurological disorders. Mesial temporal lobe epilepsies (MTLE) are the most frequent form of partial epilepsies and display frequent resistance to anti-epileptic drugs thus representing a major health care problem. In TLE, the origin of seizure activity typically involves the hippocampal formation, which displays major neuropathological features, described with the term hippocampal sclerosis (HS). HS is the most frequent pathological substrate of refractory mesial temporal lobe epilepsy. Complex partial seizures (CPS) are the predominant seizure type associated with medial temporal lobe epilepsy. MTLE is commonly due to mesial temporal sclerosis (MTS). The biology underlying the epilepstic seizures and the transcriptome associated to the seizure in intractable medial temporal lobe epilepsy is ill understood. The aim of the study was to identify potential biomarkers that could identify epileptic seizure. Thus we performed transcriptome profiling of ten medial temporal lobe epilepsy cases which are resistant to the drug and underwent temporal lobectomy. The cases constitutes of patients with intractable complex partial seizure, treated medically and have undergone detailed presurgical evaluation and subjected to surgery for standard temporal lobectomy and amygdalo-hippocampectomy. The spiking areas identified after the electrocorticography will form the test tissues, which compared with the nonspiking areas removed during the surgery, from the same patient. This could probably form one of the appropriate controls, as test and control are from same patient, which eliminates the genome variations that could incur due to the comparison with the tissues from the another patient. Also this could get rid of expression changes due to the treatments undergone by the patient. We performed two color microarray wherein we labled seizure focus (spiking area) with Cy5 and non-seizure region tissues (non-spiking) with Cy3. As a strategy to test the possibility of potential diagnostic biomarkers we are intended to test the differentially regulated molecules in an independent set of epilepsy samples.

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Proliferation of mouse thyroid cells after partial thyroidectomy

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