Receptor-type guanylyl cyclase Gyc76C is required for development of the Drosophila embryonic somatic muscle

Abstract

SummaryGuanylyl cyclases mediate a number of physiological processes, including smooth muscle function and axonal guidance. Here, we report a novel role for Drosophila receptor-type guanylyl cyclase at 76C, Gyc76C, in development of the embryonic somatic muscle. In embryos lacking function of Gyc76C or the downstream cGMP-dependent protein kinase (cGK), DG1, patterning of the somatic body wall muscles was abnormal with ventral and lateral muscle groups showing the most severe defects. In contrast, specification and elongation of the dorsal oblique and dorsal acute muscles of gyc76C mutant embryos was normal, and instead, these muscles showed defects in proper formation of the myotendinous junctions (MTJs). During MTJ formation in gyc76C and pkg21D mutant embryos, the βPS integrin subunit failed to localize to the MTJs and instead was found in discrete puncta within the myotubes. Tissue-specific rescue experiments showed that gyc76C function is required in the muscle for proper patterning and βPS integrin localization at the MTJ. These studies provide the first evidence for a requirement for Gyc76C and DG1 in Drosophila somatic muscle development, and suggest a role in transport and/or retention of integrin receptor subunits at the developing MTJs.

Receptor guanylyl cyclase Gyc76C is required for invagination, collective migration and lumen shape in the Drosophila embryonic salivary gland

“…We previously showed by whole mount in situ hybridization that gyc76C RNA is enriched in the CVM and FB surrounding the migrating salivary gland and in the mature gland (Patel et al, 2012). To determine whether gyc76C is required in the salivary gland or in the CVM/FB for gland invagination and migration, we expressed wild-type gyc76C , gyc76C WT in either the gland or mesoderm, with fkh -GAL4 and twi -GAL4, respectively, or in both.…”

“…To determine a role for gyc76C in salivary gland morphogenesis we analyzed gland invagination and migration in embryos mutant for a null allele of gyc76C , gyc76C 2388 , which is thought to lack the guanylyl cylcase domain (Patel et al, 2012). In gyc76C 2388 heterozygous embryos, all salivary gland cells invaginated to form a gland that turned posteriorly during stages 13 and 14 (Fig.…”

“…From a large-scale chemical mutagenesis screen designed to identify mutations affecting salivary gland and tracheal morphogenesis, we generated a novel allele of guanylyl cyclase at 76C, gyc76C 2388 (Myat et al, 2005; Patel et al, 2012). Guanylyl cyclases (GCs) catalyze the conversion of GTP to cGMP (guanosine 3′, 5′-cyclic monophosphate) in response to signals such as nitric oxide (NO), peptide ligands and changes in intracellular calcium.…”

“…We showed that in the Drosophila embryonic muscle, gyc76C is required for integrin receptor localization at sites of contact between the developing myotubes and tendon cells (Patel et al, 2012). Here, we report on a novel role for gyc76C in salivary gland invagination, migration and lumen shape, in part by regulating localization of the laminin matrix and talin.…”

Drosophila KASH-domain protein Klarsicht regulates microtubule stability and integrin receptor localization during collective cell migration

“…Stained embryos were mounted in Aqua Polymount (Polysciences, Inc., Warrington, PA) and thick (1 μm) fluorescence images were acquired on a Zeiss Axioplan microscope (Carl Zeiss) equipped with LSM 510 (Weill Cornell Medical College optical core facility, New York, NY) or LSM 710 (Medgar Evers College – CUNY, New York, NY). In situ hybridization was performed using myospheroid cDNA (Open Biosystems, GE Healthcare) as a template for generating antisense digoxygenin-labeled RNA probes as previously described (Patel et al, 2012). …”