https://ejh.it/index.php/ejh/issue/feed European Journal of Histochemistry 2018-08-15T15:19:09+02:00 Nadia Moscato nadia.moscato@pagepress.org Open Journal Systems <h3>The&nbsp;Proceedings of the <a href="/index.php/ejh/issue/view/172">64th Congress of the Italian Embryological Group (GEI)</a> are now available.</h3> https://ejh.it/index.php/ejh/article/view/2932 Distribution of choline acetyltransferase (ChAT) immunoreactivity in the brain of the teleost Cyprinus carpio 2018-08-15T15:19:08+02:00 Arianna Casini arianna.casini@uniroma1.it Rosa Vaccaro rosa.vaccaro@uniroma1.it Mattia Toni mattia.toni@uniroma1.it Carla Cioni carla.cioni@uniroma1.it <p>Cholinergic systems play a role in basic cerebral functions and its dysfunction is associated with deficit in neurodegenerative disease. Mechanisms involved in human brain diseases, are often approached by using fish models, especially cyprinids, given basic similarities of the fish brain to that of mammals. In the present paper, the organization of central cholinergic systems have been described in the cyprinid <em>Cyprinus carpio</em>, the common carp, by using specific polyclonal antibodies against ChAT, the synthetic enzyme of acetylcholine, that is currently used as a specific marker for cholinergic neurons in all vertebrates. &nbsp;In this work, serial transverse sections of the brain and the spinal cord were immunostained for ChAT. Results showed that positive neurons are present in several nuclei of the forebrain, the midbrain, the hindbrain and the spinal cord. Moreover, ChAT-positive neurons were detected in the synencephalon and in the cerebellum. In addition to neuronal bodies, afferent varicose fibers were stained for ChAT in the ventral telencephalon, the preoptic area, the hypothalamus and the posterior tuberculum. No neuronal cell bodies were present in the telencephalon. The comparison of cholinergic distribution pattern in the <em>Cyprinus carpio</em> central nervous system has revealed similarities but also some interesting differences with other cyprinids. Our results provide additional information on the cholinergic system from a phylogenetic point of view and may add new perspectives to physiological roles of cholinergic system during evolution and the neuroanatomical basis of neurological diseases.</p> 2018-07-24T08:39:13+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2930 Activation of the renin-angiotensin system in mice aggravates mechanical loading-induced knee osteoarthritis 2018-08-15T15:19:08+02:00 Kotaro Yamagishi k-yamagishi@med.kindai.ac.jp Ichiro Tsukamoto tsukamoto@med.kindai.ac.jp Fumihisa Nakamura fumihisa@med.kindai.ac.jp Kazuhiko Hashimoto hazzhiko@med.kindai.ac.jp Kazuhiro Ohtani kch-03@kishiwada-hospital.com Masao Akagi makagi@med.kindai.ac.jp <p>Epidemiological studies have shown an association between hypertension and knee osteoarthritis (OA). The purpose of this study was to investigate whether activation of the renin–angiotensin system (RAS) can aggravate mechanical loading-induced knee OA in mice. Eight-week-old male Tsukuba hypertensive mice (THM) and C57BL/6 mice were divided into running and non-running groups. Mice in the running group were forced to run (25 m/min, 30 min/day, 5 days/week) on a treadmill. All mice in the four groups (n=10 in each group) were euthanized after 0, 2, 4, 6, or 8 weeks of running or natural breeding. Cartilage degeneration in the left knees was histologically evaluated using the modified Mankin score. Expression of Col X, MMP-13, angiotensin type 1 receptor (AT1R), and AT2R was examined immunohistochemically. To study the effects of stimulation of the AT1R in chondrocytes by mechanical loading and/or Angiotensin II (AngII) on transduction of intracellular signals, phosphorylation levels of JNK and Src were measured in bovine articular chondrocytes cultured in three-dimensional agarose scaffolds. After 4 weeks, the mean Mankin score for the lateral femoral condylar cartilage was significantly higher in the THM running group than in the C57BL/6 running group and non-running groups. AT1R and AT2R expression was not detected at 0 weeks in any group but was noted after 4 weeks in the THM running group. AT1R expression was also noted at 8 weeks in the C57BL/6 running group. The expression levels of AT1R, COL X, and MMP-13 in chondrocytes were significantly higher in the THM running group than in the control groups. Positive significant correlations were noted between the Mankin score and the rate of AT1R-immunopositive cells, between the rates of AT1R- and Col X-positive cells, and between the rates of AT1R- and AT2R-positive cells. The phosphorylation level of JNK was increased by cyclic compression loading or addition of AngII to the cultured chondrocytes and was reversed by pretreatment with an AT1R blocker. A synergistic effect on JNK phosphorylation was observed between compression loading and AngII addition. Transgene activation of renin and angiotensinogen aggravated mechanical load-induced knee OA in mice. These findings suggest that AT1R expression in chondrocytes is associated with early knee OA and plays a role in the progression of cartilage degeneration. The RAS may be a common molecular mechanism involved in the pathogenesis of hypertension and knee OA.</p> 2018-07-24T00:00:00+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2945 Effect of high-fat mixed lipid diet and swimming on fibre types in skeletal muscles of rats with colon tumours 2018-08-15T15:19:07+02:00 Vika Smerdu vika.smerdu@mf.uni-lj.si Martina Perše martina.perse@mf.uni-lj.si <p>Skeletal muscle fibre types, whose characteristics are determined by myosin heavy chain (MyHC) isoforms, can adapt to changed physiological demands with changed MyHC isoform expression resulting in the fibre type transitions. The endurance training is known to induce fast-to-slow transitions and has beneficial effect in carcinogenesis, whereas the effect of an excessive fat intake and its interaction with the effect of swimming are less conclusive<em>.</em> Therefore, we studied the effect of high-fat mixed lipid (HFML) diet and long-term (21-week) swimming on fibre type transitions and their average diameters by immunohistochemical demonstration of MyHC isoforms in slow soleus (SOL), fast extensor digitorum longus (EDL), and mixed gastrocnemius medialis and lateralis (GM, GL) muscles, divided to deep and superficial portions (GMd, GMs, GLd, GLs), of sedentary and swimming Wistar rats with experimentally (dimethylhydrazine) induced colon tumours and fed either with HFML or low-fat corn oil (LFCO) diet. HFML diet induced only a trend for fast-to-slow transitions in SOL and in the opposite direction in GMd. Swimming triggered significant transitions in unexpected slow-to-fast direction in SOL, whereas in GMs the transitions had tendency to proceed in the expected fast-to-slow direction. The average diameters of fibre types were mostly unaffected. Hence, it can be concluded that if present, the effects of HFML diet and swimming on fibre type transitions were counteractive and muscle-specific implying that each muscle possesses its own adaptive range of response to changed physiological conditions.</p> 2018-07-25T08:12:48+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2956 Interaction between sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in experimental intestinal fibrosis. An in vivo immunohistochemical study 2018-08-15T15:19:06+02:00 Roberta Sferra roberta.sferra@univaq.it Simona Pompili pompili.simona@virgilio.it Luca Ventura luca.ventura@tin.it Caroline Dubuquoy carolinedubuquoy@hotmail.com Silvia Speca silviaspeca@icloud.com Eugenio Gaudio eugenio.gaudio@uniroma.it Giovanni Latella giolatel@tin.it Antonella Vetuschi antonella.vetuschi@univaq.it <p>A concomitant action of multiple profibrotic mediators appears crucial in the development and progression of fibrosis. Sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways are both involved in pathogenesis of fibrosis in several organs by controlling differentiation of fibroblasts to myofibroblasts and the epithelial to-mesenchymal transition. However, their direct involvement in chronic colitis-associated fibrosis it is not yet known. In this study we evaluated the immunohistochemical expression of some proteins implicated in sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in Dextrane Sodium Sulphate (DSS)-induced colorectal fibrosis in mice. Compared to control mice, DSS-induced chronic colitis mice developed a marked intestinal fibrosis associated with a concomitant overexpression of TGF-β, p-Smad3, α-SMA, collagen I-III, SPHK1, RhoA, PI3K, Akt, p-Akt, p-mTOR. This study highlights the relationship between the two pathways and the possible role of SPHK1 in the intestinal fibrosis.&nbsp; These results, if confirmed by <em>in vitro</em> studies, may have important clinical implications in the development of new therapeutical approaches in inflammatory bowel disease.</p> 2018-07-31T12:29:13+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2942 Expression of acetylated tubulin in the postnatal developing mouse cochlea 2018-08-15T15:19:05+02:00 Wenjing Liu Liuwj38@163.com Chuanxi Wang 15212239815@163.com Hao Yu yuhao1990510@163.com Shaofeng Liu liusf_cn@163.com Jun Yang yangjun@xinhuamed.com <p>Acetylation tubulin is one of the major post-translational modifications of microtubules. Stable microtubules are well known to contain acetylated tubulin. Here, we examined the spatiotemporal expression of acetylated tubulin in the mouse cochlea during postnatal development. At postnatal day 1 (P1), acetylated tubulin was localized primarily to the auditory nerve inside the cochlea and their synaptic contacts with the inner and outer hair cells (IHCs and OHCs). In the organ of Corti, acetylated tubulin occurred first at the apex of pillar cells. At P5, acetylated tubulin first appeared in the phalangeal processes of Deiters’ cells. At P8, staining was maintained in the phalangeal processes of Deiters’ cells. At P10, labeling in Deiters’ cells extended from the apices of OHCs to the basilar membrane. Labeling was expressed throughout the cytoplasm of pillar cells. At P12, acetylated tubulin displayed prominent and homogeneous labeling along the full length of the pillar cells. Linear labeling was present mainly in the Deiters’ cell bodies underlying OHCs. Between P14 and P17, acetylated tubulin was strongly expressed in inner and outer pillar cells and Deiters’ cells in a similar pattern as observed in the adult, and labeling in these cells were arranged in bundles. In addition, acetylated tubulin was intensely expressed in stria vascularis, root cell bodies, and a small number of fibrocytes of the spiral ligament until the adult. In the adult mouse cochlea, immunostaining continued to predominate in Deiters’ cells and pillar cells. Immunolabeling formed cups securing OHCs basal portions, and continued presence of acetylated tubulin-labeled nerve terminals below IHCs was shown. Our results presented here underscored the essential role played by acetylated tubulin in postnatal cochlear development, auditory neurotransmission and cochlear mechanics.</p> 2018-08-08T10:36:23+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2947 Alveolar bone regeneration during three weeks following transfer of BMP-2/7 gene via in vivo electroporation 2018-08-15T15:19:04+02:00 Mariko Kawai kawai-m@cc.osaka-dent.ac.jp Yohei Kataoka pqxh5pcp@s.okayama-u.ac.jp Junya Sonobe sonobe@kuhp.kyoto-u.ac.jp Hiromitsu Yamamoto hyamamotokuhp@mac.com Toshio Yamamoto ytoshio@md.okayama-u.ac.jp Kazuhisa Bessho bes@kuhp.kyoto-u.ac.jp Kiyoshi Ohura ohura@cc.osaka-dent.ac.jp <p>Alveolar bone is not spontaneously regenerated following trauma or periodontitis. We previously proposed an animal model for new alveolar bone regeneration therapy based on the non-viral BMP-2/7 gene expression vector and <em>in vivo</em> electroporation, which induced the formation of new alveolar bone over the course of a week. Here, we analysed alveolar bone during a period of three weeks following gene transfer to periodontal tissue. Non-viral plasmid vector pCAGGS-BMP-2/7 or pCAGGS control was injected into palatal periodontal tissue of the first molar of the rat maxilla and immediately electroporated with 32 pulses of 50 V for 50 msec. Over the following three weeks, rats were double bone-stained by calcein and tetracycline every three days and mineral apposition rates (MAR) were measured. Double bone-staining revealed that MAR of alveolar bone was as similar level three days before BMP-2/7 gene transfer as three days after gene transfer. However, from 3 to 6 days, 6 to 9 days, 9 to 12 days, 12 to 15 days, 15 to 18 days, and 18 to 20 days after, MARs were significantly higher than prior to gene transfer. Our proposed gene therapy for alveolar bone regeneration combining non-viral BMP-2/7 gene expression vector and <em>in vivo</em> electroporation could increase alveolar bone regeneration potential in the targeted area for up to three weeks.</p> 2018-08-09T08:54:28+02:00 ##submission.copyrightStatement## https://ejh.it/index.php/ejh/article/view/2896 Damaged muscle fibers might masquerade as hybrid fibers – a cautionary note on immunophenotyping mouse muscle with mouse monoclonal antibodies 2018-08-15T15:19:09+02:00 Morium Begam morium.begam@wayne.edu Joseph A. Roche joseph.roche@wayne.edu <p>We report that, labeling mouse muscle tissue, with mouse monoclonal antibodies specific to slow or fast myosin heavy chain (sMyHC and fMyHC, respectively), can lead to artefactual labeling of damaged muscle fibers, as hybrid fibers (sMyHC+ and fMyHC+).&nbsp; We demonstrate that such erroneous immunophenotyping of muscle may be avoided, by performing colabeling or serial-section-labeling, to identify damaged fibers. The quadriceps femoris muscle group (QF) in 7-month-old, male, C57BL/6J mice had: 1.21 ± 0.21%, 98.34 ± 1.06%, 0.07 ± 0.01%, and 0.53 ± 0.85% fibers, that were, sMyHC+, fMyHC+, hybrid, and damaged, respectively.&nbsp; All fibers in the tibialis anterior muscle (TA) of 3-month-old, male, C57BL/6J mice were fMyHC+; and at 3 days after injurious eccentric contractions, there was no fiber-type shift, but ~ 18% fibers were damaged.</p> 2018-07-24T00:00:00+02:00 ##submission.copyrightStatement##