Chapter 24 Imaging Neuronal Subsets and Other Cell Types in Whole-Mount Drosophila Embryos and Larvae Using Antibody Probes

Patel, Nipam H.

doi:10.1016/s0091-679x(08)60927-9

Cited by 499 publications

(360 citation statements)

References 53 publications

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“…Embryos were cultured for variable lengths of time, fixed in 4% formaldehyde in PIPES, EDTA, Magnesium (PEM) buffer 55 and dissected by hand. Immunohistochemistry was performed with anti-EN (4D9; 25 mg ml À 1 ; DSHB) and anti-fluorescein antibodies (Invitrogen; 1:100) to enhance the detection of the uncaged fluorescein.…”

Section: Methodsmentioning

confidence: 99%

Changing cell behaviours during beetle embryogenesis correlates with slowing of segmentation

Nakamoto

Hester²,

Constantinou

et al. 2015

Nat Commun

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Segmented animals are found in major clades as phylogenetically distant as vertebrates and arthropods. Typically, segments form sequentially in what has been thought to be a regular process, relying on a segmentation clock to pattern budding segments and posterior mitosis to generate axial elongation. Here we show that segmentation in Tribolium has phases of variable periodicity during which segments are added at different rates. Furthermore, elongation during a period of rapid posterior segment addition is driven by high rates of cell rearrangement, demonstrated by differential fates of marked anterior and posterior blastoderm cells. A computational model of this period successfully reproduces elongation through cell rearrangement in the absence of cell division. Unlike current models of steady-state sequential segmentation and elongation from a proliferative growth zone, our results indicate that cell behaviours are dynamic and variable, corresponding to differences in segmentation rate and giving rise to morphologically distinct regions of the embryo.

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Section: Methodsmentioning

confidence: 99%

Changing cell behaviours during beetle embryogenesis correlates with slowing of segmentation

Nakamoto

Hester²,

Constantinou

et al. 2015

Nat Commun

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“…All fluorescent double in situs were, therefore, carried out using reciprocal probe combinations (i.e., hunchback-DIG probe þ pangolin-BIO probe and hunchback-BIO probe þ pangolin-DIG probe). Gene fragments were amplified using the following primers: T. Protein immunostainings were carried out according to established protocols (Patel, 1994 Oegema et al, 1999). Factin staining was carried out using Phaloidin-FITC (Sigma, Cat.…”

Section: Experimental Procedures In Situ Hybridization and Antibody Smentioning

confidence: 99%

Early asymmetries in maternal transcript distribution associated with a cortical microtubule network and a polar body in the beetle Tribolium castaneum

Peel

Averof

2010

Developmental Dynamics

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The localization of maternal mRNAs during oogenesis plays a central role in axial specification in some insects. Here we describe a polar body-associated asymmetry in maternal transcript distribution in preblastoderm eggs of the beetle Tribolium castaneum. Since the position of the polar body marks the future dorsal side of the embryo, we have investigated whether this asymmetry in mRNA distribution plays a role in dorsal-ventral axis specification. Whilst our results suggest polar body-associated transcripts do not play a significant role in specifying the DV axis, at least during early embryogenesis, we do find that the polar body is closely associated with a cortical microtubule network (CMN), which may play a role in the localization of transcripts during oogenesis. Transcripts of the gene T.c.pangolin co-localize with the CMN at the time of their anterior localization during oogenesis and their anterior localization is disrupted by the microtubule-depolymerizing agent colcemid. Developmental Dynamics 239:2875-2887,

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“…Images were captured with the SPOT digital camera (Diagnostic Instruments) using differential interference contrast (DIC) optics on a Zeiss Axiophot microscope. Immunofluorescence was performed as described (Patel 1994). Mouse anti-fasciclin III (7G10) was used at 1:40 (Patel et al 1987).…”

Section: Staining and Imagingmentioning

confidence: 99%

Activation of the JNK pathway during dorsal closure in Drosophila requires the mixed lineage kinase, slipper

Stronach¹,

Perrimon²

2002

Genes Dev.

103

108

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The Jun kinase (JNK) pathway has been characterized for its role in stimulating AP-1 activity and for modulating the balance between cell growth and death during development, inflammation, and cancer. Six families of mammalian kinases acting at the level of JNKKK have emerged as upstream regulators of JNK activity (MLK, LZK, TAK, ASK, MEKK, and TPL); however, the specificity underlying which kinase is utilized for transducing a distinct signal is poorly understood. In Drosophila, JNK signaling plays a central role in dorsal closure, controlling cell fate and cell sheet morphogenesis during embryogenesis. Notably, in the fly genome, there are single homologs of each of the mammalian JNKKK families. Here, we identify mutations in one of those, a mixed lineage kinase, named slipper (slpr), and show that it is required for JNK activation during dorsal closure. Furthermore, our results show that other putative JNKKKs cannot compensate for the loss of slpr function and, thus, may regulate other JNK or MAPK-dependent processes.

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Chapter 24 Imaging Neuronal Subsets and Other Cell Types in Whole-Mount Drosophila Embryos and Larvae Using Antibody Probes

Cited by 499 publications

References 53 publications

Changing cell behaviours during beetle embryogenesis correlates with slowing of segmentation

Changing cell behaviours during beetle embryogenesis correlates with slowing of segmentation

Early asymmetries in maternal transcript distribution associated with a cortical microtubule network and a polar body in the beetle Tribolium castaneum

Activation of the JNK pathway during dorsal closure in Drosophila requires the mixed lineage kinase, slipper

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