DigitalUM :: Browsing by Subject "Mouse"

Browsing by Subject "Mouse"

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Open Access
A metabolic signature predicts biological age in mice
(Wiley Open Access, 2013-02) Tomas-Loba, Antonia; Bernardes de Jesus, Bruno; Mato, Jose M; Blasco, Maria Antonia; Fisiología
Our understanding of the mechanisms by which aging is produced is still very limited. Here, we have determined the sera metabolite profile of 117 wild-type mice of different genetic backgrounds ranging from 8 to 129 weeks of age. This has allowed us to define a robust metabolomic signature and a derived metabolomic score that reliably/accurately predicts the age of wild-type mice. In the case of telomerase-deficient mice, which have a shortened lifespan, their metabolomic score predicts older ages than expected. Conversely, in the case of mice that overexpress telomerase, their metabolic score corresponded to younger ages than expected. Importantly, telomerase reactivation late in life by using a TERT-based gene therapy recently described by us significantly reverted the metabolic profile of old mice to that of younger mice, further confirming an anti-aging role for telomerase. Thus, the metabolomic signature associated with natural mouse aging accurately predicts aging produced by telomere shortening, suggesting that natural mouse aging is in part produced by presence of short telomeres. These results indicate that the metabolomic signature is associated with the biological age rather than with the chronological age. This constitutes one of the first aging-associated metabolomic signatures in a mammalian organism.
Open Access
A method to collect to high volumes of milk from mice (Mus musculus)
(Murcia: Servicio de Publicaciones de la Universidad de Murcia, 2013) Gómez Gallego, Carlos; Ilo, T.; Ulla-Marjut, Jaakkola; Salminen, Seppo; Periago Castón, María Jesús; Ros Berruezo, Gaspar Francisco; Frías Beneyto, Rafael; Facultad de Veterinaria
ABSTRACT Collecting milk samples from mice (Mus musculus) may be interesting for a variety of preclinical research. References in the literature for protocols describing how to milk a dam are scarce, and a major limitation of such protocols is the small sample volume that is generally collected. The aim of our study was to develop a practical protocol to collect substantial amounts of milk from mice. Adult female outbred NMRI and inbred BALB/c mice with nursing litters were used in this study. The milking was carried out on days 7–12 after parturition. The pups were separated from their mothers for 6–12 h before milking to allow accumulation of milk in the glands. Dams were anesthetized using either an injectable mixture of midazolam and ketamine, or by use of the inhalational agent isoflurane. To induce milk flow, the mice were given 2-8 IU of oxytocin intraperitoneally. The milk was collected using an electric human breast pump that was modified to accommodate mouse nipples and to handle small liquid volumes. With this protocol, the total amount of milk collected from each dam per each milking ranged between 0.2 and 1.5 mL. We concluded that this milking method provides an excellent means for acquiring substantial amounts of mouse milk.
Open Access
Aberrant expressions of delta-protocadherins in the brain of Npc1 mutant mice
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Yan, Xin; Lukas, Jan; Lin, Juntang; Ernst, Mathias; Koczan, Dirk; Witt, Martin; Fuellen, Georg; Wree, Andreas; Rolfs, Arndt; Luo, Jiankai
Niemann-Pick type C1 (NPC1) disease is an autosomal recessive disorder characterized by dysmyelination and neurodegeneration, which can result in the death of patients in early childhood in some cases. Members of the delta-protocadherins (Pcdhs) play important roles in neurogenesis and brain development. In this study, we compared expression profiles of Pcdhs in the brain between wild-type and Npc1 mutant mice from postnatal day (P) 9 onwards by in situ hybridization. Our data show that laminar distribution of some Pcdhs in the cerebral cortex of Npc1 mutated mice is different from that of wild-type mice. Furthermore, expressions of Pcdhs by oligodendrocytes in the corpus callosum and by Purkinje cells and granular cells in the cerebellum are strongly decreased in Npc1 mutated mice at later stages. Taken together, our data suggest that aberrant expression of Pcdhs is a pathological process accompanied by neurodegeneration in Npc1 mutant mice.
Open Access
Age-related morphometric changes in the pineal gland. A comparative study between C57BLI6J and CBA mice
(Murcia : F. Hernández, 2000) Cernuda-Cernuda, R.; Huerta, J.J.; Muñoz Llamosas, M.; Alvarez-Uría, M.; García-Fernández, J.M.
Relatively little is known about the effects of melatonin on the aging of the pineal, the organ which is the main place for synthesis of this hormone. Using simple morphometric methods, some parameters of the pineal gland, such as total volume, number of pinealocytes and pinealocyte volume were estimated in two mice strains: normal CBA and melatonin-deficient C57BLl6J. Two age groups, 6 weeks and 10 months, were studied in order to evaluate possible differential age-related changes between both strains. Pineals of both strains have similar morphometric and morphological features at 6 weeks of age. This suggests that pineal development, which has already concluded at 6 weeks of age, is not affected by the absence of melatonin synthesis in the pinealocytes. Later on, CBA pineal showed an increase in size caused by cellular hypertrophy. In contrast, the C57BLl6J pineal volume decreased by loss of pinealocytes in the same period of time. Semithin sections analysed by light microscopy did not show that this cell death was evident in the C57BL16J strain at any of the ages studied. Thus, a gradual loss of pinealocytes could be hypothesised in these pineals. These results suggest that pineal melatonin could have a role in the maintenance of pinealocyte viability and the increase of pineal size which takes place after development. The abnormal pattern observed in the C57BL16J pineal should be taken into account in future studies on this gland.
Open Access
Cellular compartmentation of lysozyme and a-amylase in the mouse salivary glands. lmmunogold approaches at light and electron microscopy level
(Murcia : F. Hernández, 2000) Marchetti, L.; Gabrielli, M.G.; Materazzi, G.; Menghi, Giovanna
The research was planned to study the subcellular distribution of enzymatic secretory products within the secretory structures of the mouse major salivary glands at light and electron microscopy level by immunogold silver stain (IGSS) technique and doublesided post-embedding immunogold binding and silver amplification in order to speculate about their compartmentation. In particular, we experimented the above immunogold labeling approaches to localize the lysozyme and to verify its distribution patterns in relation to another secretion enzyme, a-amylase. Co-presence of lysozyme and a-amylase was observed in the convoluted granular tubule cells of the submandibular gland and in the demilunar cells of the sublingual gland as well as in the electron-dense regions of the mottled secretory granules in the parotid gland. Exclusive binding patterns of lysozyme were observed in the acinar cells of the submandibular and sublingual glands where a-amylase did not occur.
Open Access
Changes in intestinal endocrine cells in the mouse after unilateral cervical vagotomy
(Murcia : F. Hernández, 1999) Qian, B.F.; El-Salhy, M.; Danielsson, A.; Shalaby, A.; Axelsson, H.
The effect of right or left unilateral cervical vagotomy on the intestinal endocrine cells was studied in 23 mice at 2 and 8 weeks after operation, respectively. The results were compared with that from 10 sham operated mice. Various types of endocrine cells in duodenum and proximal colon were detected by immunohistochemistry and quantified by computerized image analysis. In mouse duodenum, chromogranin-, CCKIgastrin-, GIP- and somatostatin-cells were significantly decreased at 2 weeks after right vagotomy, but returned to the control levels at 8 weeks. Serotonincells were reduced at both 2 and 8 weeks after right vagotomy. The amount of the duodenal endocrine cells did not change after left vagotomy with the exception of secretin-cells, which were diminished at 8 weeks after both right and left vagotomy. In the proximal colon, chromogranin-cells were also decreased at 2 weeks after right vagotomy. Serotonin-cells were reduced at 8 weeks after left vagotomy but not right vagotomy. There was no significant difference between the unilaterally vagotomized and the sham operated mice with regard to PYY- and glucagon-cells. It was concluded that vagotomy affected the intestinal endocrine cells in mouse. The influence was more pronounced in the small intestine than the proximal colon. The right vagus nerves seemed to exert more effect on the intestinal endocrine cells than the left ones.
Open Access
Effects of four polyphenols on mouse wound healing and the gene expression profile of resveratrol action
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2024) Hu, Xiao; Zhang, Hanbin; Chen, Zhenguo
Studies have demonstrated the potent effects of polyphenols on cutaneous wound healing. However, the molecular mechanisms underlying polyphenol activity are incompletely understood. Herein, mice were experimentally wounded, intragastrically treated with four polyphenols, resveratrol, tea polyphenols, genistein, and quercetin; and monitored for 14 days. Resveratrol was the most effective compound, promoting wound healing starting at day 7 after wounding, by enhancing cell proliferation and reducing apoptosis and subsequently promoting epidermal and dermal repair, collagen synthesis and scar maturation. RNA sequencing was performed in control and resveratrol-treated tissues on day 7 after wounding. Resveratrol treatment upregulated 362 genes and downregulated 334 genes. Gene Ontology enrichment analysis showed that differentially expressed genes (DEGs) were associated with different biological processes (keratinization, immunity, and inflammation), molecular functions (cytokine and chemokine activities), and cellular components (extracellular region and matrix). Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that DEGs were predominantly enriched in inflammatory and immunological pathways, including cytokine-cytokine receptor interaction, chemokine signaling, and tumor necrosis factor (TNF) signaling. These results show that resveratrol accelerates wound healing by promoting keratinization and dermal repair and attenuating immune and inflammatory responses.
Open Access
Further characterization of spontaneous arthritic changes in murine squamo-mandibular joint: histopathological aspects
(Murcia : F. Hernández, 1986) Livne, E.; Silbermann, M.
The appearance of age-related ulcerative changes in the mouse mandibular condyle were evaluated by light and electron microscopy examinations. Fibrillations appeared along the articular surface and in deeper tissue regions, as early as at six months of age. Such changes were characterized by a marked loss of the tissue's cellularity and by a marked reduction in matrix metachromasia and safranin-0 staining. These microscopical changes were accompanied by a reduced reactivity for both ruthenium red and colloidal iron binding, as noted ultrastructurally. At the same time, increasing numbers of erythrocytes appeared to be adhered to the surface irregularities and were also found in deeper regions within the articular lesions. Using morphological criteria, it became apparent that the degenerative changes of aging articular cartilage started at the more superficial regions of the tissue and only thereafter proceeded toward the chondro-osseous junction. Also, with the advancement of age, the degenerative changes became more severe
Open Access
Golgi apparatus localization of ZNT7 in the mouse cerebellum
(Murcia : F. Hernández, 2009) Gao, Hui-Ling; Feng, Wan-Yu; Li, Xiao-Ling; Xu, He; Huang, Liping; Wang, Zhan-You
We have recently reported that four members of the zinc transporter (ZNT) family, ZNT1, ZNT3, ZNT4, and ZNT6, are abundantly expressed in the mouse cerebellum. In the present study, we reported that ZNT7 was present throughout the cerebellar cortex. ZNT7 immunoreactivity was predominately present in the somas and primary dendrites of the Purkinje cells. ZNT7 was also present in the Bergmann glial cell bodies as well as their radial processes, which extended into the molecular cell layer. Confocal immunofluorescence results demonstrated that the expression of ZNT7 overlapped with that of TGN38 in the somas of the Purkinje cells and granule cells. Immuno-electron microscopic study showed that ZNT7 was localized to the membrane of the Golgi apparatus in the somas of the Purkinje cells, Bergmann glial cells, and granule cells. Western blot analysis demonstrated that a considerable amount of ZNT7 was expressed in the cerebellum. These findings suggest a significant role of ZNT7 in zinc homeostasis in the mouse cerebellum.
Open Access
Morphological changes in mouse ovary due to hormonal hypersecretion and matrix metalloproteinase-2 activity
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Kim, Sang-Hwan; Yoon, Jong-Taek
We analyzed whether aberrant gonadotropin secretion affects the morphological remodeling of murine ovarian tissues facilitated by activated matrix metalloproteinase (MMP) enzymes. Six mice were intraperitoneally injected with 5 IU of pregnant mare serum gonadotropin (PMSG) or human chorionic gonadotropin (HCG) every two days after estrus synchronization. Morphology and expression of various MMPs were assessed following the successful induction of hormonal secretion in these tissues. HCG treatment, but not PMSG treatment, resulted in the expanded production of granulose second follicular cells. In addition, the number of developing follicular cells in the HCG group increased compared with that in the PMSG group. Ovarian diameters were also very small in the PMSG group. Immunohistochemistry revealed decreased MMP-2 protein activity in the HCG group and increased MMP-2 activity in the PMSG group. Activity was particularly high in theca and granulose cells of the PMSG group, but only partial activity was observed in the theca cells of the HCG group. Vascular endothelial growth factor activity was increased in both the external and internal theca cell walls in the PMSG group while the HCG group showed high overall expression of this protein in the internal theca cells. These data indicate that follicular cell activity and remodeling of the ovaries differ based on the type of secretory hormone signals they receive. Inappropriate gonadotropin secretion may induce functional changes in the ovaries, and follicular remodeling may be facilitated by the activity of various MMPs.
Open Access
Neurotrophin-4 dependency of intraepithelial vagal sensory nerve terminals that selectively contact pulmonary NEBs in mice
(Murcia : F. Hernández, 2010) Oztay, Fusun; Brouns, Inge; Pintelon, Isabel; Raab, Marion; Neuhuber, Winfried; Timmermans, Jean Pierre; Adriaensen, Dirk
Important physiological functions of neurotrophins (NTs) in airways and lungs are the early development, differentiation and maintenance of peripheral sensory neurons. The main pulmonary sensory innervation is of vagal origin, with several nerve fibre populations that selectively contact complex morphologically well-characterized receptor end-organs, called neuroepithelial bodies (NEBs). NEBs in mouse lungs are innervated by at least two separate myelinated vagal sensory nerve fibre populations, of which the neurochemical coding is suggestive of a mechanosensory function. Since neurotrophin-4 (NT-4) has been especially described to be important for the maintenance of mechanosensory nerve terminals, the present study aimed at investigating the NT-4 dependency of the two myelinated vagal sensory nerve fibre populations innervating mouse pulmonary NEBs. Multiple immunostaining in 21-day-old and adult mouse lungs revealed the expression of the NT-4 receptor TrkB on the two different myelinated vagal sensory nerve fibre populations, i.e., the vesicular glutamate transporter/calbindin-positive and the P2X2/3- positive fibres, which selectively contact pulmonary NEBs. Examination of the effect of the lack of NT-4 on these NEB-related nerve fibre populations, by comparing adult NT-4-/- and wild-type mice, revealed that in NT-4-/- mice the percentage of NEBs contacted by P2X2/3+ is reduced by 75%, while the VGLUT+/CB+ population seemed to be unaffected. This study demonstrated that although mouse pulmonary NEBs are contacted by two distinct TrkB expressing populations of vagal myelinated afferents, only one is distinctly reduced in NT-4 deficient mice, suggesting the involvement of NTs. In view of the growing evidence for the involvement of NTs in neuronal plasticity associated with airway diseases, pulmonary NEBs innervated by NT-sensitive vagal afferents may play a significant role.
Open Access
Primordial germ cell specification, the importance of being blimped
(Murcia : F. Hernández, 2008) Chuva de Sousa Lopes, Susana M.; Roelen, Bernard A.J.
In mouse embryos, the expression of Blimp1 has recently revealed a population of allocated primordial germ cell precursors 24 hours earlier than previously thought. Those ‘blimped’ precursors have been shown to give rise, by mitotic division, to germ cells only and no other cell lineages. Here, we try to understand the events that lead to Blimp1 expression in the primordial germ cell precursors and speculate on what can be the role of Blimp1 during primordial germ cell specification and gastrulation in the mouse. Finally, we discuss the possible involvement of Blimp1 in the two know modes of germ line segregation (epigenesis and preformation).
Open Access
Role of HTRA1, a serine protease, in the progression of articular cartilage degeneration
(Murcia : F. Hernández, 2010) Polur, Llona; Lee, Peter L.; Servais, Jacqueline M.; Xu, Lin; Li, Yefu
This study is to investigate the possible role of high temperature requirement A 1 (HtrA1) in the articular cartilage degeneration. Paraffin sections were prepared from the knee and temporomandibular (TM) joints of four mouse OA models; two of the models had a genetic mutation (type IX collagen-deficient and type XI collagen-haploinsufficient) and two were surgically induced (destabilization of the medial meniscus of knee joint and discectomy of TM joint). The HtrA1 protein expression profiles of the prepared sections were examined by immunohistostaining. The level of HtrA1 mRNA in the articular cartilage taken from the knee joints of one of the genetically mutated OA models was determined by real-time PCR. Double immunohistostaining was used to examine the expression of co-localization of HtrA1 with type VI collagen and HtrA1 with discoidin domain receptor 2 (Ddr2) in the articular cartilage of knee joints from the genetically mutated OA model. The expression of HtrA1 was found to be increased in the knee and TM joints of these four models at early stages of the disease. An examination of the knee joint of a mutant mouse indicated an 8-fold increase in the level of HtrA1 mRNA, when compared to the levels observed in the knee joints of its wild-type littermates. Pericellular type VI collagen was not present in chondrocytes expressing HtrA1. Meanwhile, the expression of HtrA1 was associated with the expression of Ddr2 in the chondrocytes. Results indicate that HtrA1 may disrupt the pericellular matrix network, resulting in alteration of chondrocyte metabolisms. This eventually leads to OA
Open Access
Role of skeletal muscle in lung development
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Baguma-Nibasheka, Mark; Gugic, Dijana; Saraga-Babic, Mirna; Kablar, Boris
Skeletal (striated) muscle is one of the four basic tissue types, together with the epithelium, connective and nervous tissues. Lungs, on the other hand, develop from the foregut and among various cell types contain smooth, but not skeletal muscle. Therefore, during earlier stages of development, it is unlikely that skeletal muscle and lung depend on each other. However, during the later stages of development, respiratory muscle, primarily the diaphragm and the intercostal muscles, execute so called fetal breathing-like movements (FBMs), that are essential for lung growth and cell differentiation. In fact, the absence of FBMs results in pulmonary hypoplasia, the most common cause of death in the first week of human neonatal life. Most knowledge on this topic arises from in vivo experiments on larger animals and from various in vitro experiments. In the current era of mouse mutagenesis and functional genomics, it was our goal to develop a mouse model for pulmonary hypoplasia. We employed various genetically engineered mice lacking different groups of respiratory muscles or lacking all the skeletal muscle and established the criteria for pulmonary hypoplasia in mice, and therefore established a mouse model for this disease. We followed up this discovery with systematic subtractive microarray analysis approach and revealed novel functions in lung development and disease for several molecules. We believe that our approach combines elements of both in vivo and in vitro approaches and allows us to study the function of a series of molecules in the context of lung development and disease and, simultaneously, in the context of lung’s dependence on skeletal muscle-executed FBMs.
Open Access
Role of skeletal muscle in mandible development
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Rot, Irena; Mardesic-Brakus, Snjezana; Costain, Willard J.; Saraga-Babic, Mirna; Kablar, Boris
As a continuation of the previous study on palate development (Rot and Kablar, 2013), here we explore the relationship between the secondary cartilage mandibular condyles (parts of the temporomandibular joint) and the contributions (mechanical and secretory) from the adjacent skeletal musculature. Previous analysis of Myf5-/- :MyoD-/- mouse fetuses lacking skeletal muscle demonstrated the importance of muscle contraction and static loading in mouse skeletogenesis. Among abnormal skeletal features, micrognathia (mandibular hypoplasia) was detected: small, bent and posteriorly displaced mandible. As an example of Waddingtonian epigenetics, we suggest that muscle, in addition to acting via mechanochemical signal transduction pathways, networks and promoters, also exerts secretory stimuli on skeleton. Our goal is to identify candidate molecules at that muscle-mandible interface. By employing Systematic Subtractive Microarray Analysis approach, we compared gene expression between mandibles of amyogenic and wild type mouse fetuses and we identified up- and downregulated genes. This step was followed by a bioinformatics approach and consultation of webaccessible mouse databases. We searched for individual tissue-specific gene expression and distribution, and for the functional effects of mutations in a particular gene. The database search tools allowed us to generate a set of candidate genes with involvement in mandibular development: Cacna1s, Ckm, Des, Mir300, Myog and Tnnc1. We also performed mouse-to-human translational experiments and found analogies. In the light of our findings we discuss various players in mandibular morphogenesis and make an argument for the need to consider mandibular development as a consequence of reciprocal epigenetic interactions of both skeletal and non-skeletal compartments.
Open Access
Role of skeletal muscle in palate development
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2013) Rot, Irena; Kablar, Boris
The involvement of skeletal muscle in the process of palatal development in mammals is an example of Waddingtonian epigenetics. Our earlier study showed that the cleft palate develops in the complete absence of skeletal musculature during embryonic development in mice. This contrasts with previous beliefs that tongue obstruction prevents the elevation and fusion of the palatal shelves. We argue that the complete absence of mechanical stimuli from the adjacent muscle, i.e., the lack of both static and dynamic loading, results in disordered palatogenesis. We further suggest that proper fusion of the palatal shelves depends not only on mechanical but also on paracrine contributions from the muscle. The muscle’s paracrine role in the process of palatal fusion is achieved through its being a source of certain secreted and/or circulatory proteins. A cDNA microarray analysis revealed differentially expressed genes in the cleft palate of amyogenic mouse fetuses and suggested candidate molecules with a novel function in palatogenesis (e.g., Tgfbr2, Bmp7, Trim71, E2f5, Ddx5, Gfap, Sema3f). In particular, we report on Gdf11 mutant mouse that has cleft palate, and on several genes whose distribution is normally restricted to the muscle (completely absent in our amyogenic mouse model), but which are found down-regulated in amyogenic mouse cleft palate. These molecules probably present a subset of paracrine cues that influence palatogenesis from the adjacent muscle. Future studies will elucidate the role of these genes in muscle-palate crosstalk, connecting the cues produced by the muscle with the cartilage and bone tissue’s responses to these cues, through various degrees of cell p
Open Access
Survivin and Cyclooxygenase-2 are co-expressed in human and mouse colon carcinoma and in terminally differentiated colonocytes
(Murcia : F. Hernández, 2007) Mori, F.; Piro, F.R.; Della Rocca, C.; Mesiti, G.; Giampaoli, S.; Silvestre, G.; Lazzaro, D.
In the evolution of colon rectal cancer (CRC) the imbalance between cell proliferation and apoptosis is considered one of the prominent causes of tumor induction and/or progression. In order to establish the role of anti apoptotic proteins in colon cancer development, we studied with immunohistochemical techniques the expression of Survivin in a mouse model of colon carcinogenesis induced by 1,2-dimethylhydrazine treatment. In this mouse model Survivin was over-expressed during tumor development, showing a distribution mimicking that described in the correspondent human malignancies. We also correlated Survivin distribution with COX-2 and ß-Catenin expression patterns. The co-localization of COX-2/ß-Catenin/Survivin in the same epithelial cells in tumor samples lends credence to possible in vivo regulatory effects of COX-2 and ß- Catenin on the intracellular Survivin levels in mouse and human colon cancer.

Browsing by Subject "Mouse"

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