ABSTRACT
Traditional histological staining techniques, as well as elastin-specific antibodies and electron microscopy, have been used to assess the distribution of elastin within the peripheral nerve. The location of the elastin identified by the VerHoeff-VanGiesen or Weigert stains has been shown to coincide with the unambiguous identification of elastin by immunospecific stains and electron microscopy. Elastin is located in all three connective layers of the peripheral nerve. Thick elastic fibres, consisting of amorphous elastin protein and microfibrils, are located consistently in the perineurium and, to a lesser extent, in the epineurium. The endoneurium contains small collections of elastic fibres widely distributed between the axons. Compared with collagen, the overall content of elastin, however, is small, suggesting that the visco-elastic properties of peripheral nerve may be due primarily to collagen.
Subject(s)
Elastic Tissue/cytology , Elastin/metabolism , Peripheral Nerves/chemistry , Peripheral Nerves/ultrastructure , Animals , Female , Histocytochemistry , Humans , Immunohistochemistry , Rats , Rats, Sprague-Dawley , Sciatic Nerve/chemistry , Sciatic Nerve/cytology , Sural Nerve/chemistry , Sural Nerve/cytologyABSTRACT
The temporal pattern of separation of the soft tissue between mouse digits was examined in an organ culture model system. Mouse limbs of different gestational age were cultured in vitro and the pattern of separation of the digits characterized. By gestational day 12.5 (E12.5) the limbs were committed to undergo separation of the soft tissue in the interdigital space when cultured in vitro. Prior to E12.5 digital separation did not occur and the limb tissues were not committed to this process. The addition of 10(-7) M retinoic acid (RA) to the media of E12 limbs was capable of inducing digit separation in the uncommitted limbs. Both the soft and hard tissue development of digits formed in vitro for either committed limbs or uncommitted limbs induced with RA was similar to the in vivo pattern.
Subject(s)
Extremities/embryology , Tretinoin/pharmacology , Animals , Bone and Bones/embryology , Gestational Age , In Situ Hybridization , Mice , Organ Culture Techniques , RNA, Messenger/analysis , Receptors, Retinoic Acid/analysis , Receptors, Retinoic Acid/geneticsABSTRACT
Mutations associated with genes of the EGF superfamily are implicated in limb malformations. To evaluate the potential role of EGF-mediated signal transduction in the control of early mammalian limb development, we developed a simple in vitro system which is permissive for morphogenesis and cytodifferentiation in serumless, chemically defined medium. Our experimental strategy was to ascertain if the EGF precursor gene was transcribed and translated into potentially bioactive growth factor. EGF mRNA transcripts are expressed in Swiss Webster mouse embryonic (42-44 somite pairs) forelimbs as determined by mRNA phenotyping. EGF transcripts are translated into precursor EGF polypeptides which were localized to limb covering epithelium and the chondrogenic mesenchymal cell lineages. EGF immunostaining patterns suggested a paracrine type of regulation for the cartilage blastema associated with forelimb development. To test whether EGF effects the timing and positional information required for limb-specific cartilage morphogenesis, we employed tyrphostin (RG 50864) which inhibits EGF receptor kinase activity in a concentration-dependent manner and severely retards limb development. These findings support our hypothesis that endogenous EGF or EGF-like proteins provide signaling for the size and shape of discrete forelimb cartilage formations during mouse embryonic morphogenesis.
