Labeling and sorting of avian primordial germ cells utilizing Lycopersicon Esculentum lectin.

Avian species are essential resources for human society, with their preservation and utilization heavily dependent on primordial germ cells (PGCs). However, efficient methods for isolating live PGCs from embryos remain elusive in avian species beyond chickens, and even in chickens, existing techniques have shown limited efficiency. In this study, we present a rapid, simple, and cost-effective method for labeling and sorting circulating-stage PGCs across various avian species, including Carinatae and Ratitae, using Lycopersicon Esculentum (Tomato) lectin (LEL).

Direct reprogramming of non-limb fibroblasts to cells with properties of limb progenitors.

The early limb bud consists of mesenchymal limb progenitors derived from the lateral plate mesoderm (LPM). The LPM also gives rise to the mesodermal components of the flank and neck. However, the cells at these other levels cannot produce the variety of cell types found in the limb.

LIN28 is essential for the maintenance of chicken primordial germ cells.

Understanding the mechanism of stem cell maintenance underlies the establishment of long-term and mass culture methods for stem cells that are fundamental for clinical and agricultural applications. In this study, we use chicken primordial germ cell (PGC) as a model to elucidate the molecular mechanisms underlying stem cell maintenance. The PGC is a useful experimental model because it is readily gene-manipulatable and easy to test gene function in vivo using transplantation.

Differential modularity of the mammalian Engrailed 1 enhancer network directs sweat gland development.

Enhancers are context-specific regulators of expression that drive biological complexity and variation through the redeployment of conserved genes. An example of this is the enhancer-mediated control of Engrailed 1 (EN1), a pleiotropic gene whose expression is required for the formation of mammalian eccrine sweat glands. We previously identified the En1 candidate enhancer (ECE) 18 cis-regulatory element that has been highly and repeatedly derived on the human lineage to potentiate ectodermal EN1 and induce our species' uniquely high eccrine gland density.

Prime editing in chicken fibroblasts and primordial germ cells.

CRISPR/Cas9-based genome editing technologies are revolutionizing developmental biology. One of the advanced CRISPR-based techniques is prime editing (PE), which enables precise gene modification in multiple model organisms. However, there has been no report of taking advantage of the PE system for gene editing in primordial germ cells (PGCs) thus far.

In ovo electroporation of chicken limb bud ectoderm: Electroporation to chick limb ectoderm: Electroporation to chick limb ectoderm.

Background: Deciphering how ectodermal tissues form, and how they maintain their integrity, is crucial for understanding epidermal development and pathogenesis. However, lack of simple and rapid gene manipulation techniques limits genetic studies to elucidate mechanisms underlying these events.

Results: Here we describe an easy method for electroporation of chick limb bud ectoderm enabling gene manipulation during ectoderm development and wound healing.

Repeated mutation of a developmental enhancer contributed to human thermoregulatory evolution.

Humans sweat to cool their bodies and have by far the highest eccrine sweat gland density among primates. Humans' high eccrine gland density has long been recognized as a hallmark human evolutionary adaptation, but its genetic basis has been unknown. In humans, expression of the () transcription factor correlates with the onset of eccrine gland formation.

Evaluation of a new apparatus combining a mobile device and hand-held roller for the detection of spinal deformities: association with the sum of the right and left angles of trunk inclination.

Purpose: To investigate the intra- and inter-observer reliabilities of the newly developed i-Scolioroller for scoliosis screening, and to determine the optimal i-Scolioroller measurement cutoff values for identifying adolescent scoliosis with a Cobb angle ≥ 20°.

Methods: The i-Scolioroller displays the right- and left-side maximum inclination angle (Rmax, Lmax) during the forward bending test (FBT), as well as the angle of trunk inclination (ATI, i.e.

L-type voltage-gated Ca channel Ca1.2 regulates chondrogenesis during limb development.

All cells, including nonexcitable cells, maintain a discrete transmembrane potential (), and have the capacity to modulate and respond to their own and neighbors' changes in Spatiotemporal variations have been described in developing embryonic tissues and in some cases have been implicated in influencing developmental processes. Yet, how such changes in are converted into intracellular inputs that in turn regulate developmental gene expression and coordinate patterned tissue formation, has remained elusive. Here we document that the of limb mesenchyme switches from a hyperpolarized to depolarized state during early chondrocyte differentiation.

Coordination between body growth and tissue growth: Wolffian duct elongation and somitogenesis proceed in harmony with axial growth.

During embryogenesis, different tissues develop coordinately, and this coordination is often in harmony with body growth. Recent studies allow us to understand how this harmonious regulation is achieved at the levels of inter-cellular, inter-tissue, and tissue-body relationships. Here, we present an overview of recently revealed mechanisms by which axial growth (tail growth) drives a variety of morphogenetic events, with a focus on the coordinated progression between Wolffian (nephric) duct elongation and somitogenesis.

Integration of Shh and Fgf signaling in controlling gene expression in cultured limb cells.

During embryonic development, fields of progenitor cells form complex structures through dynamic interactions with external signaling molecules. How complex signaling inputs are integrated to yield appropriate gene expression responses is poorly understood. In the early limb bud, for instance, Sonic hedgehog () is expressed in the distal posterior mesenchyme, where it acts as a mediator of anterior to posterior (AP) patterning, whereas fibroblast growth factor 8 () is produced by the apical ectodermal ridge (AER) at the distal tip of the limb bud to direct outgrowth along the proximal to distal (PD) axis.

Early formation of the Müllerian duct is regulated by sequential actions of BMP/Pax2 and FGF/Lim1 signaling.

The Müllerian duct (MD) and Wolffian duct (WD) are embryonic tubular tissues giving rise to female and male reproductive tracts, respectively. In amniote embryos, both MD and WD emerge in both sexes, but subsequently degenerate in the males and females, respectively. Here, by using MD-specific gene manipulations in chicken embryos, we identify the molecular and cellular mechanisms that link early MD specification to tubular invagination.

Clinical, physical, and radiographic analyses of lumbar degenerative kyphosis and spondylolisthesis among community-based cohort.

Purpose: To investigate longitudinal radiographic changes, and physical characteristics of lumbar degenerative kyphosis (LDK) and spondylolisthesis (DS).

Methods: Two-hundred eighty nine community-based female subjects were recruited from population register and studied longitudinally for a mean 12.3 years.

FGF8 coordinates tissue elongation and cell epithelialization during early kidney tubulogenesis.

When a tubular structure forms during early embryogenesis, tubular elongation and lumen formation (epithelialization) proceed simultaneously in a spatiotemporally coordinated manner. We here demonstrate, using the Wolffian duct (WD) of early chicken embryos, that this coordination is regulated by the expression of FGF8, which shifts posteriorly during body axis elongation. FGF8 acts as a chemoattractant on the leader cells of the elongating WD and prevents them from epithelialization, whereas static ('rear') cells that receive progressively less FGF8 undergo epithelialization to form a lumen.

Interepithelial signaling with nephric duct is required for the formation of overlying coelomic epithelial cell sheet.

In most organs of the body, epithelial tissues are supported by their own basement membrane and underlying stroma, the latter being regarded as a complex of amorphous cells, extracellular matrices, and soluble factors. We demonstrate here that an epithelial tube can serve as a component of stroma that supports the formation of epithelial cell sheet derived from a different origin. During development of the mesonephros in chicken embryos, the intermediate mesoderm (IMM), which contains the Wolffian duct (WD) and its associated tubules, is overlain by a sheet of epithelial cells derived from lateral plate (coelomic) mesoderm.

Transgenesis of the Wolffian duct visualizes dynamic behavior of cells undergoing tubulogenesis in vivo.

Deciphering how the tubulogenesis is regulated is an essential but unsolved issue in developmental biology. Here, using Wolffian duct (WD) formation in chicken embryos, we have developed a novel method that enables gene manipulation during tubulogenesis in vivo. Exploiting that WD arises from a defined site located anteriorly in the embryo (pronephric region), we targeted this region with the enhanced green fluorescent protein (EGFP) gene by the in ovo electroporation technique.

Epidemiology of degenerative lumbar scoliosis: a community-based cohort study.

Study Design: A 12-year prospective study of pre-existing and de novo degenerative lumbar scoliosis (DLS) among community-based female volunteers.

Objective: To investigate serial entire spine radiographs of healthy female volunteers and to clarify radiographical characteristics and predictors of pre-existing and de novo DLS.

Summary Of Background Data: DLS is among the most frequent spinal defomities in the aging spine; however, the onset or the natural course of this condition has not been elucidated.

Radiographic analysis of newly developed degenerative spondylolisthesis in a mean twelve-year prospective study.

Study Design: Prospective study of community-based female volunteers.

Objective: To investigate the incidence of newly developed degenerative spondylolisthesis (DS) among those without baseline deformity, and to clarify radiographic characteristics and predictors of DS.

Summary Of Background Data: There has been limited number of prospective studies of DS.

Cartilage degeneration is associated with augmented chemically-induced joint pain in rats: a pilot study.

Background: Osteoarthritis arising from cartilage degeneration is the most common cause of joint pain. However, the relationship between joint pain and cartilage degeneration is not well understood.

Questions/purposes: We asked whether the inflammatory mediators participate in the joint pain in the presence of cartilage degeneration.

Changes in the sagittal spinal alignment of the elderly without vertebral fractures: a minimum 10-year longitudinal study.

Background: Thoracic hyperkyphosis, or loss of lumbar lordosis, is often equated with osteoporosis because vertebral fractures are assumed to be a major causative factor. However, recent evidence suggests that up to one-half of the patients with hyperkyphosis have no evidence of underlying vertebral fracture. The shape characteristics of the intervertebral discs and their role in determining kyphotic curvature have been investigated.

Effects of dry needling at tender points for neck pain (Japanese: katakori): near-infrared spectroscopy for monitoring muscular oxygenation of the trapezius.

Background: Neck pain (katakori) is a common symptom in adult Japanese people. However, the pathophysiological aspect of this condition has not been well documented to date. The purpose of this study was to investigate the effects of tender point dry needling to the trapezius muscles and the resultant changes in muscular hemodynamics.

The effects of experimental neurolysis on ectopic firing in a rat chronic constriction nerve injury model.

Purpose: We studied the effects of experimental neurolysis on ectopic firing in a rat chronic constriction nerve injury (CCI) model.

Methods: Sixteen Wistar rats were used as CCI models; 8 in a neurolysis group and 8 in a sham operation group. Eight additional Wistar rats were used in a normal control group.

A prospective study of de novo scoliosis in a community based cohort.

Study Design: A 12-year prospective study of de novo scoliosis in a community based cohort. OBJECTIVE.: To investigate factors associated with development of de novo scoliosis.

Ectopic firing due to artificial venous stasis in rat lumbar spinal canal stenosis model: a possible pathogenesis of neurogenic intermittent claudication.

Study Design: An electrophysiologic analysis was performed on a chronic lumbar spinal stenosis model of rats. The effects of venous stasis on ectopic firing originating in the nerve root were investigated.

Objectives: To elucidate the mechanisms of neurogenic intermittent claudication in lumbar spinal canal stenosis.

Effect of sodium channel blocker (mexiletine) on pathological ectopic firing pattern in a rat chronic constriction nerve injury model.

We studied the efficacy of mexiletine as a sodium channel blocker for neuropathic pain by investigating the effect of mexiletine on the pathological ectopic firing pattern in a chronic constriction nerve injury (CCI) model. The experiment was conducted with 60 male Wistar rats. The CCI model was created by loosely ligating the sciatic nerve.