Abstract
Background: While data regarding expression of limb element and tissue markers during normal mouse limb development exist, few studies show expression patterns in upper and lower limbs throughout key limb development stages. A comparison to normal developmental events is essential when analyzing development of the limb in mutant mice models. Results: Expression patterns of the joint marker Gdf5, tendon and ligament marker Scleraxis, early muscle marker MyoD1, and blood vessel marker Cadherin5 (Cdh5) are presented during the most active phases of embryonic mouse limb patterning. Anti‐neurofilament staining of developing nerves in the fore‐ and hindlimbs and cartilage formation and progression also are described. Conclusions: This study demonstrates and describes a range of key morphological markers and methods that together can be used to assess normal and abnormal limb development. Developmental Dynamics 247:1217–1226, 2018. © 2018 The Authors. Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists
Highlights
Significant progress has been made in understanding the molecular and morphological mechanisms controlling limb development, just how neuronal innervation, cartilage formation and progression, vascular patterning, tendon development, and joint development are so intricately controlled and regulated is still unclear
We have analyzed the dynamic expression in mouse fore- and hindlimbs of Gdf5, MyoD1, and Cdh5 from E13.5 to E15.5; Scleraxis from E12.5 to E15.5; anti-neurofilament from E10.5 to E14.5; and cartilage/bone development and progression from E11.5 to E16.5
Mouse limb development begins around E9.0, the limb bud at this stage consists of a mass of undifferentiated cells, and the main limb structures—including tendons, skeleton, muscles, and vessels—are organized between
Summary
Significant progress has been made in understanding the molecular and morphological mechanisms controlling limb development, just how neuronal innervation, cartilage formation and progression, vascular patterning, tendon development, and joint development are so intricately controlled and regulated is still unclear. We have performed a detailed spatiotemporal analysis of markers of joint (Gdf5), tendon (Scleraxis), muscle (MyoD1), vascular (Cdh5), neuronal (anti-neurofilament), and cartilage (Alcian Blue/Alizarin Red staining) development. By E14.5, there was no longer continuous Gdf expression along cartilage condensations, and thick bands of Gdf expression were visible in carpometacarpal/tarsometatarsal joints, metacarpophalangeal/ metatarsophalangeal joints, and the developing distal interphalangeal joint in joint interzones (Fig. 1C white arrowhead, Fig. 1C–E). At E13.5, Scleraxis was expressed in the hand- and footplate (Fig. 2C white bracket) and in the digit rays, with thicker regions of expression at presumptive joint sites (compare to Fig. 1A–B). In E15.5 embryos, Scleraxis was expressed in distinct bands that extended along the digits, with strong areas of expression persisting around developing joint regions (Fig. 2H–I,K–L). Expression of Scleraxis corresponding to flexor tendons could be seen at the ventral side of section images (Fig. 2G,J, blue lines)
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