Stage-specific Onset of Condensation and Matrix Deposition for Meckel's and Other First Arch Cartilages in Inbred C57BL/6 Mice

Condensation is a multistep process, involving cell recruitment, cell-to-cell contact by cell adhesions, and concomitant changes in cell shape. Condensation of prechondrogenic cells down-regulates and/or inactivates cell proliferation and enhances the activities of cartilage-specific genes. Timing of onset and duration of condensation are thus important regulatory processes mediating cellular and molecular events during chondrogenesis. The present study was undertaken to examine timing of onset and duration of condensation and onset of matrix formation for first arch cartilages in inbred C57BL/6 mice. Because timing can only be reliably assessed in very precisely staged embryos, mice were mated for only 2 hr, pregnancy was determined by weight increase, and embryos were assigned to substages and divisions of Theiler's [1972] stages on the basis of external development of the frontonasal area, eyes, vibrissae, mandibular and hyoid auricular hillocks (pinna) and some internal structures [Miyake et al., 1996]. Condensation and matrix formation were determined using PNA lectin histochemistry, type II collagen immunohistochemistry, Mallory's trichrome, Hall and Brunt's quadruple and toluidine blue stains on serially sectioned embryos, and 3-D reconstruction. A single, continuous, first arch chondrogenic condensation was identified. It consisted of three components: 1) a rostral component for the symphysis; 2) a core component for the major portion of Meckel's cartilage; and 3) a caudal component for the caudo-lateral area of Meckel's cartilage and the two ear cartilages, the malleus and incus. The caudal component was further divided into rostral and caudal subcomponents which gave rise to the malleus and incus, respectively. Cellular arrangement differed among the three components, probably reflecting major axes of matrix growth. The condensation was only weakly recognized by peanut agglutinin lectin (PNA), unless sections were pretreated with neuraminidase, showing the masking effects of sialic acid. The core and caudal components of the condensation appeared at stage 20.12 and ended at stage 21.14 and 21.32, respectively. The symphysial component began at stage 20.2 and ended at stage 21.32. Deposition of sulfated proteoglycans began at stage 21.14 in the core component of the condensation--preceding slightly deposition of type II collagen--and at stage 21.32 in the symphysial and caudal components.