Conditional Reverse Tet-Transactivator Mouse Strains for the Efficient Induction of TRE-Regulated Transgenes in Mice

Tetracycline or doxycycline (dox)-regulated control of genetic elements allows inducible, reversible and tissue specific regulation of gene expression in mice. This approach provides a means to investigate protein function in specific cell lineages and at defined periods of development and disease. Efficient and stable regulation of cDNAs or non-coding elements (e.g. shRNAs) downstream of the tetracycline-regulated element (TRE) requires the robust expression of a tet-transactivator protein, commonly the reverse tet-transactivator, rtTA. Most rtTA strains rely on tissue specific promoters that often do not provide sufficient rtTA levels for optimal inducible expression. Here we describe the generation of two mouse strains that enable Cre-dependent, robust expression of rtTA3, providing tissue-restricted and consistent induction of TRE-controlled transgenes. We show that these transgenic strains can be effectively combined with established mouse models of disease, including both Cre/LoxP-based approaches and non Cre-dependent disease models. The integration of these new tools with established mouse models promises the development of more flexible genetic systems to uncover the mechanisms of development and disease pathogenesis.     

Anthropometric profile, vertical jump, and throwing velocity in elite female handball players by playing positions

Women's handball is a sport, which has seen an accelerated development over the last decade. Although anthropometric and physical characteristics have been studied for male sports teams, in women's handball, studies are scarce. The aim of this study was twofold: first, to describe the anthropometric characteristics, throwing velocity, hand grip, and muscular power of the lower limbs in female handball players and second, to identify the possible differences in these parameters in terms of individual playing positions (center, back, wing, pivot, and goalkeeper). A total of 130 elite female Spanish handball players participated in the study (age 25.74 ± 4.84 years; playing experience 14.92 ± 4.88 years). Anthropometric assessment was performed for all the subjects following the International Society for the Advancement of Kinanthropometry protocols. Furthermore, all the subjects performed a vertical jump test (squat jump and countermovement jump). Hand grip and throwing velocity in several situations were also assessed. A 1-way analysis of variance and a Tukey post hoc test were used to study the differences among individual playing positions. Wings were less heavy, shorter, and showed a smaller arm span than did goalkeepers, backs and pivots (p ≤ 0.001). Additionally, pivots were heavier than centers. Backs and pivots exhibited higher muscular mass than did wings. Total players' somatotype was mesomorphy endomorphy (3.89-4.28-2.29). Centers showed higher throwing velocity levels than did wings in 9-m throws from just behind the line, with a goalkeeper. Backs exhibited higher hand-grip values than did wings. Statistical differences have been established between wings and other specific playing positions, especially with pivot and backs. Coaches can use this information to select players for the different specific positions.     

Spirostanic analogues of castasterone

The synthesis, spectroscopic characterization, and biologic activity of two bioactive spirostanic analogues of the naturally occurring brassinosteroid castasterone are described.     

c-Lysozyme from Senegalese sole (Solea senegalensis): cDNA cloning and expression pattern

Lysozymes are key molecules of innate immunity and proved high bactericidal activity in fish, thus becoming attractive as tools for enhancing fish defences. In this study, a full-length c-type lysozyme cDNA from Senegalese sole (Solea senegalensis) has been cloned and characterized. The cDNA sequence was inferred from two overlapping fragments obtained by RACE-PCR and consisting on 631bp coding for 143 aminoacids. Catalytic and other conserved residues required for lysozyme activity were identified. Pair wise alignments showed the higher identities with c-type lysozyme from other flatfish. Expression patterns under various conditions showed a basal level and a clear upregulation mostly in hematopoietic organs after stimulation with LPS or infection with Photobacterium damselae. This study represents a first step on the genetics and function of the c-lysozyme of Senegalese sole, though disclosing g-DNA structure, allelic variability and antibacterial activity must be requirements prior its immunological properties might have biotechnological applications.     

Interleukin-6 is an essential regulator of satellite cell-mediated skeletal muscle hypertrophy

Skeletal muscles adapt to increasing workload by augmenting their fiber size, through mechanisms that are poorly understood. This study identifies the cytokine interleukin-6 (IL-6) as an essential regulator of satellite cell (muscle stem cell)-mediated hypertrophic muscle growth. IL-6 is locally and transiently produced by growing myofibers and associated satellite cells, and genetic loss of IL-6 blunted muscle hypertrophy in vivo. IL-6 deficiency abrogated satellite cell proliferation and myonuclear accretion in the preexisting myofiber by impairing STAT3 activation and expression of its target gene cyclin D1. The growth defect was indeed muscle cell intrinsic, since IL-6 loss also affected satellite cell behavior in vitro, in a STAT3-dependent manner. Myotube-produced IL-6 further stimulated cell proliferation in a paracrine fashion. These findings unveil a role for IL-6 in hypertrophic muscle growth and provide mechanistic evidence for the contribution of satellite cells to this process.