Expression of Six1 homeobox gene during development of the mouse submandibular salivary gland.
BACKGROUND
Members of the Six family of homeoproteins are expressed in numerous tissues during vertebrate embryogenesis, and are critical regulators of both cell proliferation and survival. Here we report the temporal and spatial expression of Six1 during maturation of the mouse submandibular salivary gland (SSG) from embryonic day 18.5 (E18.5) to postnatal day 28. Additionally, we examine the role of Six1 during SSG development using Six1-deficient mice.


METHODS
Six1 expression was assessed by reverse transcription-polymerase chain reaction, Western blot, and immunofluorescence. Proliferation was measured by bromodeoxyuridine (BrdU) incorporation index, and apoptosis was evaluated by TUNEL assay.


RESULTS
Six1 mRNA and protein levels are high in the epithelial SSG cells at E18.5 and decrease progressively in the postnatal maturing SSG. Although SSGs from Six1(-/-) embryos are significantly smaller than wild type SSGs, the histological structures of the SSG acini and ducts are similar. Six1(-/-) salivary epithelial cells exhibit an intrinsic defect in cell proliferation accompanied by a significant reduction in the Six1 target gene cyclin A1, previously shown to be a critical mediator of Six1-induced proliferation.


CONCLUSIONS
Our results suggest that the reduction in size of Six1(-/-) SSGs is result of a decrease in cell proliferation during development/maturation.
McCoy
E L
EL
Program in Molecular Biology, University of Colorado School of Medicine, Denver, CO, USA.
Kawakami
K
K
Ford
H L
HL
Coletta
R D
RD
D000818:Animals; D049109:Cell Proliferation; D019925:Cyclin A / Q000378:metabolism; D056750:Cyclin A1; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009929:Organ Size; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000378:metabolism*; D018709:Statistics, Nonparametric; D013363:Submandibular Gland / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D013997:Time Factors; D014018:Tissue Distribution
C503747:Ccna1 protein, mouse; D019925:Cyclin A; D056750:Cyclin A1; D018398:Homeodomain Proteins; D012471:Salivary Proteins and Peptides; C490947:Six1 protein, mouse