Histology and histopathology Vol.35, nº2 (2020)

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Comparative study of calcium and calcium-related enzymes with differentiation markers in different ages and muscle types in mdx mice.
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Gaglianone, Rhayanna B.; Bloise, Flavia Fonseca; Carvalho, Tania Maria Ortiga; Santos, Thereza Quirico; Costa, Manoel Luis; Mermelstein, Claudia
Sarcolemma instability and increased calcium influx in muscle fibers are characteristics of the Duchenne muscular dystrophy. Excessive calcium activates calcium-dependent enzymes, such as calpains (CAPN) and matrix metalloproteases (MMP). Here, we analyzed calcium deposits, the activity of CAPN and MMP and the expression of Myh, SERCA and myogenic regulatory factors in different skeletal muscles during myonecrosis (4-weeks) and regeneration (12-weeks) phases of the mdx muscular pathology. Alizarin red staining was used to assess calcium deposits, casein and gelatin zymography were performed to evaluate CAPN and MMP activity, and qPCR was used to evaluate the expression of Myh, Capn, Atp2a1 and Atp2a2, Myod1 and Myog. We observed the following characteristics in mdx muscles: (i) calcium deposits almost exclusively in mdx muscles, (ii) lower CAPN1 activity in mdx muscles, (iii) higher CAPN2 activity in mdx muscles (only at 12 wks), (iv) autolyzed CAPN activity exclusively in mdx muscles, (v) lower expression of Capn1 and higher expression of Capn2 in mdx muscles; (vi) lower expression of Atp2a1 and Atp2a2 in mdx muscles, (vii) higher MMP (pre pro MMP2, pro MMP2, MMP2 and MMP9) activity in mdx muscles, (viii) MMP2 activity exclusively in mdx muscles at 12 wks, (ix) MMP9 activity exclusively in mdx muscles, (x) higher expression of Myog in mdx muscles at 12 wks, and (xi) lower expression of Myh (Myh7, Myh2, Myh1, Myh4) in mdx muscles, particularly Myh7 and Myh2. The collection of our results provides valuable information for a better characterization of mdx pathology phenotype
Open Access
Long noncoding RNA small nucleolar RNA host gene 12 promotes papillary thyroid carcinoma cell growth and invasion by targeting miR16-5p
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Feng, Xiaocheng; Dong, Xuehong; Wu, Dingting; Zhao, Hanxin; Xu, Changqin; Li, Hong
Emerging evidence has shown that long noncoding RNA (lncRNA) plays an important role in various types of malignant cancer. Small nucleolar RNA host gene 12 (SNHG12) was found to be upregulated and to act as an oncogene in several cancers. However, the function and regulatory mechanism of SNHG12 remain unclear in papillary thyroid carcinoma (PTC). In this study, SNHG12 was found to be increased in PTC tissues and cell lines using quantitative real-time PCR. Knockdown of SNHG12 significantly inhibited PTC cell proliferation, migration and invasion and induced apoptosis in vitro. Mechanistic investigations revealed that SNHG12 functions as a competing endogenous RNA (ceRNA) to sponge miR-16-5p, which was downregulated in PTC tissues. In addition, rescue assays further confirmed that SNHG12 contributed to the progression of PTC through regulating miR-16-5p expression. These results indicated that SNHG12 might contribute to tumor progression in PTC by acting as a ceRNA to sponge miR-16-5p.
Open Access
Roles of microRNAs as non-invasive biomarker and therapeutic target in colorectal cancer.
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Wan, Timothy Ming-Hun; Iyer, Deepak Narayanan; Ng, Lui
MicroRNAs are endogenous, short non- coding RNA molecules that function as critical regulators of various biological processes. There is a strong functional evidence linking the involvement of dysregulated miRNAs to the occurrence, development and progression of colorectal cancer. Studies indicate that while overexpression of oncomiRs, and repression of tumor suppressor miRNAs tends to drive the overall tumorigenic process, the global picture of aberrant miRNA expression in colorectal cancer can classify the disease into multiple molecular phenotypes. Moreover, the expression pattern of miRNAs in colorectal cancer makes them viable disease determinants as well as potential therapeutic targets. Through this review, we will summarize the importance of miRNAs in the etiology and progression of colorectal cancer. Specifically, we will explore the key role played by these RNA molecules as likely therapeutic avenues and the strategies presently available to target them. Finally, we will investigate the role of miRNAs as potential non- invasive diagnostic and prognostic biomarkers in colorectal cancer.
Open Access
Fiber type diversity in skeletal muscle explored by mass spectrometry-based single fiber proteomics
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Schiaffino, Stefano; Reggiani, Carlo; Murgia, Marta
Mammalian skeletal muscles are composed of a variety of muscle fibers with specialized functional properties. Slow fibers are suited for long lasting and low intensity contractile activity, while various subtypes of fast fibers are optimized to produce high force and power even with a significant fatigue. The functional specialization of muscle fibers is based on selective gene expression regulation, which provides each fiber with a specific protein complement. The recent refinement of small-scale sample preparation, combined with the development of mass spectrometers characterized by high sensitivity, sequencing speed and mass accuracy, has allowed the characterization of the proteome of single muscle fibers with an unprecedented resolution. In the last few years, the first studies on the global proteomics of individual fibers of different types have been published. In this short review we discuss the methodological advancements which have opened the way to single fiber proteomics and the discovery power of this approach. We provide examples of how specific features of single fibers can be overlooked when whole muscle or multi- fiber samples are analyzed and can only be detected when a single fiber proteome is analyzed. Thus, novel subtype-specific metabolic features, most prominently mitochondrial specialization of fiber types, have been revealed by single fiber proteomics. In the same way, specific adaptive responses of single fibers to aging or loss of neural input have been detected when single fibers were individually analyzed. We conclude that the fiber type-resolved proteomes represent a powerful tool which can be applied to a variety of physiological and pathological conditions.
Open Access
Comparative study of the eyelids and orbital glands morphology in the okapi (Okapia johnstoni, Giraffidae), Père David's deer (Elaphurus davidianus, Cervidae) and the Philippine mouse-deer (Tragulus nigricans, Tragulidae).
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Nawrot, Joanna Elżbieta Klećkowska; Harłajczuk, Karolina Goździewska; Barszcz, Karolina
The accessory organs of the eye represent part of the protective system of the eyeball. In the present study, an examination of the accessory organs of the eye of three species of captive ruminants was performed using light microscopy. In the okapi, the superficial gland of the third eyelid and lacrimal gland were complex branched multilobar tubular glands formed by mucous units with tubular secretory portions and no plasma cells. The deep gland of the third eyelid was absent in the okapi and present in both the Père David’s deer and the Philippine mouse-deer. In the Philippine mouse-deer, the deep gland had a very thick connective capsule and thick interlobar septae. It contained fewer lobes forming the gland parenchyma compared to Père David’s deer and other ruminants. Organized lymphoid follicles were present within the upper and lower eyelids only in the okapi and Père David’s deer, while diffuse lymphocytes were observed in the Philippine mouse-deer. The orbital glands in the Père David’s deer had a multilobar tubuloacinar structure with numerous plasma cells and a mucoserous character. In contrast to the Philippine mouse-deer, these glands had a serous character. The presence of several macroscopic and microscopic structural differences of the examined accessory organs of the eye in the three captive ruminant species may be understood within an ecological context and may be associated with different habitat-specific environmental conditions.

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