Change in MASP1 Gene Expression Disrupts Cement Gland Formation in Xenopus laevis Embryos
Additional Funding Sources
The project described was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant No. P20GM103408.
Presentation Date
7-2021
Abstract
Mannan-binding lectin associated serine protease 1 (MASP1) is an alternative splice product of the MASP1/3 gene. MASP1 is essential for the activation of the lectin complement pathway within the innate immune system. Mutations in MASP1/3 have been linked to 3MC syndrome, a rare and complex developmental disorder that includes craniofacial phenotypes. Previous research in a zebrafish model has indicated that knockdown of total masp1/3 leads to disrupted craniofacial formation, suggesting a function for this gene in developmental processes. However, the individual role of MASP1 proteins in development remains unknown. We are currently conducting experiments in a Xenopus laevis (African clawed frog) model to determine the role of Masp1 in development. To better understand developmental roles of this protein, we are modulating masp1 expression via overexpression (mRNA microinjection) and knockdown (morpholino microinjection) in X. laevis embryos. Our initial experiments involving masp1 manipulation resulted in several complementary phenotypes. Notable changes occurred in the formation of the cement gland—a mucus-secreting organ that allows the embryo to attach to a solid surface prior to developing the ability to swim. Reducing Masp1 expression decreased cement gland size by 0.6-fold compared to the control. Conversely, overexpression of Masp1 increased cement gland size by 2-fold. This suggests that Masp1 is involved in regulating the region of cement gland formation. Several signaling molecules involved in cement gland formation are similarly involved in craniofacial development such that this system will allow us to identify the mechanisms by which masp1 is involved in human craniofacial development.
Change in MASP1 Gene Expression Disrupts Cement Gland Formation in Xenopus laevis Embryos
Mannan-binding lectin associated serine protease 1 (MASP1) is an alternative splice product of the MASP1/3 gene. MASP1 is essential for the activation of the lectin complement pathway within the innate immune system. Mutations in MASP1/3 have been linked to 3MC syndrome, a rare and complex developmental disorder that includes craniofacial phenotypes. Previous research in a zebrafish model has indicated that knockdown of total masp1/3 leads to disrupted craniofacial formation, suggesting a function for this gene in developmental processes. However, the individual role of MASP1 proteins in development remains unknown. We are currently conducting experiments in a Xenopus laevis (African clawed frog) model to determine the role of Masp1 in development. To better understand developmental roles of this protein, we are modulating masp1 expression via overexpression (mRNA microinjection) and knockdown (morpholino microinjection) in X. laevis embryos. Our initial experiments involving masp1 manipulation resulted in several complementary phenotypes. Notable changes occurred in the formation of the cement gland—a mucus-secreting organ that allows the embryo to attach to a solid surface prior to developing the ability to swim. Reducing Masp1 expression decreased cement gland size by 0.6-fold compared to the control. Conversely, overexpression of Masp1 increased cement gland size by 2-fold. This suggests that Masp1 is involved in regulating the region of cement gland formation. Several signaling molecules involved in cement gland formation are similarly involved in craniofacial development such that this system will allow us to identify the mechanisms by which masp1 is involved in human craniofacial development.