scholarly journals Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution

10.3791/57406 ◽  
2018 ◽  
Author(s):  
Tomoko Yamazaki ◽  
Wenling Li ◽  
Yoh-Suke Mukouyama
Keyword(s):  
Confocal Microscopy ◽  
Immune Cell ◽  
Branching Morphogenesis ◽  
Model System ◽  
Cell Distribution ◽  
Whole Mount ◽  
Vascular Branching

FGF10 is an inducer and Pax6 a competence factor for lacrimal gland development

Development ◽  
2000 ◽  
Vol 127 (12) ◽  
pp. 2563-2572 ◽  
Author(s):  
H.P. Makarenkova ◽  
M. Ito ◽  
V. Govindarajan ◽  
S.C. Faber ◽  
L. Sun ◽  
...  
Keyword(s):  
Lacrimal Gland ◽  
Normal Development ◽  
Branching Morphogenesis ◽  
Model System ◽  
Specific Marker ◽  
Fibroblast Growth ◽  
Secretory Function ◽  
Competence Factor ◽  
Branched Structure ◽  
Gland Development

We investigated the mechanism of tissue induction and specification using the lacrimal gland as a model system. This structure begins its morphogenesis as a bud-like outgrowth of the conjunctival epithelium and ultimately forms a branched structure with secretory function. Using a reporter transgene as a specific marker for gland epithelium, we show that the transcription factor Pax6 is required for normal development of the gland and is probably an important competence factor. In investigating the cell-cell signaling required, we show that fibroblast growth factor (FGF) 10 is sufficient to stimulate ectopic lacrimal bud formation in ocular explants. Expression of FGF10 in the mesenchyme adjacent to the presumptive lacrimal bud and absence of lacrimal gland development in FGF10-null mice strongly suggest that it is an endogenous inducer. This was supported by the observation that inhibition of signaling by a receptor for FGF10 (receptor 2 IIIb) suppressed development of the endogenous lacrimal bud. In explants of mesenchyme-free gland epithelium, FGF10 stimulated growth but not branching morphogenesis. This suggested that its role in induction is to stimulate proliferation and, in turn, that FGF10 combines with other factors to provide the instructive signals required for lacrimal gland development.

scholarly journals Analysis of chick somite myogenesis by in situ confocal microscopy of desmin expression.

1994 ◽  
Vol 42 (2) ◽  
pp. 265-272 ◽  
Author(s):  
W H Borman ◽  
D E Yorde
Keyword(s):  
Confocal Microscopy ◽  
Intermediate Filament ◽  
Intermediate Filament Protein ◽  
Chick Embryos ◽  
Whole Mount ◽  
Somite Formation ◽  
Filament Protein ◽  
The Relationship

We explored the relationship in chick embryos between somitogenesis and the onset of somite myogenesis by immunodetection of the muscle-specific intermediate filament protein desmin. Early somite desmin expression was detected by whole-mount in situ confocal microscopy. No detectable somite desmin was observed in embryos of 15 somites (Stage 12) or younger. In embryos having between 16 and 26 somites (Stages 12-15), desmin could be detected in somites positioned increasingly more caudal in the embryo. Finally, in embryos of 27 somites (Stage 16) and older, somite desmin expression was consistently present in all but the caudal-most six somites. Although the rate of somite formation is fairly constant, the rate of observed somite desmin expression progressing caudally in the embryo is greater initially than the rate of segmentation. After an embryo has formed about 27 somites, the rate of desmin appearance parallels the rate of segmentation at a distance of about six somites. This result suggests that very early somite myogenesis is not linked to somitogenesis.

scholarly journals Identification and Characterization of Novel Perivascular Adventitial Cells in the Whole Mount Mesenteric Branch Artery Using Immunofluorescent Staining and Scanning Confocal Microscopy Imaging

2012 ◽  
Vol 2012 ◽  
pp. 1-6
Author(s):  
Chandra Somasundaram ◽  
Rahul K. Nath ◽  
Richard D. Bukoski ◽  
Debra I. Diz
Keyword(s):  
Confocal Microscopy ◽  
Neurite Growth ◽  
Immunofluorescent Staining ◽  
Neural Cells ◽  
Microscopy Imaging ◽  
Stress Injury ◽  
Scanning Confocal Microscopy ◽  
Adult Rat ◽  
Calcium Sensing ◽  
Whole Mount

A novel perivascular adventitial cell termed, adventitial neuronal somata (ANNIES) expressing the neural cell adhesion molecule (NCAM) and the vasodilator neuropeptide, calcitonin gene-related peptide (CGRP), exists in the adult rat mesenteric branch artery (MBA) in situ. In addition, we have previously shown that ANNIES coexpress CGRP and NCAM. We now show that ANNIES express the neurite growth marker, growth associated protein-43(Gap-43), palladin, and the calcium sensing receptor (CaSR), that senses changes in extracellular Ca(2+) and participates in vasodilator mechanisms. Thus, a previously characterized vasodilator, calcium sensing autocrine/paracrine system, exists in the perivascular adventitia associated with neural-vascular interface. Images of the whole mount MBA segments were analyzed under scanning confocal microscopy. Confocal analysis showed that the Gap-43, CaSR, and palladin were present in ANNIES about 37 ± 4%, 94 ± 6%, and 80 ± 10% respectively, comparable to CGRP (100%). Immunoblots from MBA confirmed the presence of Gap-43 (48 kD), NCAM (120 and 140 kD), and palladin (90–92 and 140 kD). In summary, CGRP, and NCAM-containing neural cells in the perivascular adventitia also express palladin and CaSR, and coexpress Gap-43 which may participate in response to stress/injury and vasodilator mechanisms as part of a perivascular sensory neural network.

Sign in / Sign up

Export Citation Format

Share Document