| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
RESEARCH |
Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, N1G 2W1 Canada
Correspondence should be addressed to A C Hahnel; Email: ahahnel{at}ovc.uoguelph.ca
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
, ßI, ßII, 
, 
, 
, 
, 
, 
, 
/
) and a PKC-anchoring protein, receptor for activated C-kinase 1 (RACK1). Our results show that all of these isoforms of PKC are present between the two-cell and blastocyst stages of mouse preimplantation development, and that each has a distinct, dynamic pattern and level of expression. The data suggest that different complements of the isozymes are involved in various steps of preimplantation development, and will serve as a framework for further functional studies of the individual isozymes. In particular, there was a transient increase in the nuclear concentration of several isozymes at the early four-cell stage, suggesting that some of the PKC isozymes might be involved in regulation of nuclear organization and function in the early mouse embryo. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
, ßI, ßII, and and are activated by phosphatidylserine, diacylglycerol, Ca2+, and 4ß-phorbol 12-myristate 13-acetate (4ß-PMA). The novel PKCs (nPKCs) are , , , and , and are not Ca2+-dependent, but otherwise activated as the cPKCs. PKC µ is sometimes included in the nPKCs (Geiges et al. 1997), but it has homologies to both the PKC and calmodulin kinases, and is often placed into a PKC-like family (Hausser et al. 2002). The atypical PKCs (aPKCs) are and / and are lipid-dependent, but cannot be activated by either Ca2+ or 4ß-PMA (Newton 2001). Production of monoclonal antibodies and polyclonal antisera against peptides that distinguish the various isozymes has led to a number of experiments that indicate that individual PKC isozymes are localized differentially within cells before and after stimulation. Their restricted localization exposes them to different substrates and is believed to largely specify function (Schechtman & Mochly-Rosen 2001). It has been shown that phosphorylation of a group of conserved amino acids determines the activation state of PKC and affects the subcellular localization of each isozyme (Keranen et al. 1995). Also, binding of substrate and non-substrate proteins affects the ability of PKC isozymes to relocate and hence associate with particular substrates (Jaken & Parker 2000, Mochly-Rosen & Kauvar 2000, Schechtman & Mochly-Rosen 2001).
Several studies have established involvement of PKC in egg activation, fertilization, and progression through the meiotic cycles in mice and rats (Gallicano et al. 1995, 1997, Raz et al. 1998, Downs et al. 2001, Viveiros et al. 2001, 2003, Eliyahu & Shalgi 2002, Quan et al. 2003). However, there is less known of the role of the PKC during early cleavage. Activation with phorbol ester of the cPKC and nPKC blocks progression through the mitotic cycles (Quan et al. 2003). The synthesis of proteins that are required for embryonic gene activation is inhibited by H8 and H7, non-specific inhibitors of protein kinases A and C that act by competitive inhibition of their ATP-binding sites (Schultz 1993). During mouse preimplantation development, phorbol ester causes premature compaction of two-, four-, and early eight-cell-stage embryos (Ohsugi et al. 1993a, 1993b, Ohsugi & Yamamura 1993), and the non-specific PKC inhibitor D-sphingosine blocks induced and natural compaction (Winkel et al. 1990). PKC may be involved in compaction, since it redistributes to contact sites as compaction initiates and can phosphorylate ß-catenin (Pauken & Capco 1999). It has also been demonstrated that blastocoele formation is regulated through phospholipase C (an enzyme that is known to activate PKC-dependent pathways; Stachecki & Armant 1996), and that targeted disruption of mouse phospholipase Cß3 (the most widely expressed member of the phospholipase Cß family) results in early preimplantation death (Wang et al. 1998). Although all of these studies suggest a role for the PKC family in preimplantation development, the relative importance of each isozyme is unknown.
In rat oocytes, Raz et al.(1998) detected PKCs , ßI, ßII, , , , , and by Western blot and immunocytochemistry with isozyme-specific antibodies, but not PKCs or , and Eliyahu & Shalgi (2002) detected PKCs , ßI, and ßII by immunocytochemistry. PKCs , ßI, and have been detected in pig oocytes (Fan et al. 2002). The studies of mouse oocytes have given inconsistent results. Gangeswaran & Jones (1997) detected only PKCs and among nine isozymes using Western blot. Pauken & Capco (2000) detected PKCs , , , , µ, and by Western blot and immunocytochemistry, but not PKC ß. While Luria et al.(2000) found that PKCs , ßI and ßII were present by immunocytochemistry, and Downs et al.(2001) found that PKCs , ßI, , and , but not were present by Western blot. Luria et al.(2000), Eliyahu & Shalgi (2002) and Baluch et al.(2004) have supported some of the results with PKC activator/inhibitor studies. In preimplantation embryos, Pauken & Capco (2000) detected PKCs , , , , and but not isoform ß by Western blot and immunocytochemistry, but Eckert et al.(2004) detected PKCs ßI and ßII in freshly isolated inner cell masses of mouse blastocysts. Clearly neither Western blot nor immunocytochemistry has provided consistent results with PKC ß.
In this study, we used immunocytochemistry to examine the expression of 10 members of the PKC family (, ßI, ßII, , , , , , , /) between the two-cell and blastocyst stages of mouse development. We chose to use immunocytochemistry because it is very sensitive and provides the ability to localize individual PKC isozymes within cells. We included antibodies from two commercial sources that have been widely used in immunocytochemical detection of PKCs, and support the immunocytochemistry data with results from treatments with specific PKC activators. This study differs from previous ones in that we quantified the fluorescence resulting from binding of the antibody to each PKC isozyme. This was done in order to estimate concentrations of each isozyme in various subcellular compartments at various stages of cleavage. This is significant since it has been suggested that small changes in subcellular distribution can lead to functional changes in PKC action. It is difficult to obtain the numbers of preimplantation embryos that would be required for cell fractionation and biochemistry. Quantifying immunofluorescence is one way to circumvent this problem and has been used by others for live cell imaging of PKC translocations (Almholt et al. 1999, Schechtman et al. 2004).
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
To study the effects of activating PKC isozymes, two-cell embryos were flushed from oviducts at 45 h post-hCG with flushing-holding medium (Lawitts & Biggers 1993), and cultured in KSOM/AA medium at 37 °C under silicone oil in an atmosphere of 5% CO2/5% O2/90% N2 (Lawitts & Biggers 1993, Ho et al. 1995) with or without PKC activator. Activation was with 100 nM 4ß-PMA for 15 min (LC Laboratories, Woburn, MA, USA) or with 100 nM of the cPKC-activating peptide, pseudo-ßRACK (where RACK is receptor for activated C kinase) for 60 min (kindly donated by D Mochly-Rosen, Department of Molecular Pharmacology, Stanford University School of Medicine, Stanford, CA, USA). The 4ß-PMA was dissolved in ethanol at 30 µM (Watanabe et al. 2002), and used at 100 nM in medium as described for myocytes (Disatnik et al. 1994, Dorn et al. 1999) and similar to what has been used with oocytes (80, 100, and 160 nM; Gallicano et al. 1995, 1997, Raz et al. 1998, Luria et al. 2000). Control embryos were cultured in medium with 0.33% ethanol. We did not include inactive analogues of 4ß-PMA because others have shown that the analogues have non-specific activities (for example, see Doerner et al. 1990, Watanabe et al. 2002). The peptide activator pseudo-ßRACK is cross-linked to a dimer of Antennapedia carrier peptide (CRQIKIWFQNRRMKWKK). This conjugate has been used extensively by Mochly-Rosen’s group, and shown to enter cells and affect PKC distribution and cell function without compromising cell viability (Dorn et al. 1999). This group has also shown that pseudo-ßRACK does not affect distribution of other PKC isoforms at concentrations between 100 nM and 1 µM, and that scrambled peptide and peptides derived from other parts of the PKC molecule do not affect distribution of the PKC. Negative-control embryos were cultured in medium containing 100 nM carrier–carrier dimer (also kindly donated by D Mochly-Rosen). After culture in treatment or control media, the embryos were immediately fixed and processed for immunocytochemistry.
Immunocytochemistry
Monoclonal antibodies against seven isozymes of PKC (, , ,, , , and ) and RACK1 were purchased from Trans-duction Laboratories (Lexington, KY, USA). Polyclonal antibodies raised against specific peptides of nine isozymes of PKC (, ßI, ßII, , , , , , and ) were purchased from Research & Diagnostics Antibodies (Berkeley, CA, USA). These antibodies have been previously characterized and used in a number of studies on PKC in embryonic or adult cells (Disatnik et al. 1994, Johnson et al. 1996, Csukai et al. 1997, Miyamae et al. 1998, Dorn et al. 1999, Pauken & Capco 2000). Embryos were fixed in 2% paraformaldehyde in PBS for 30 min, and then permeabilized in 0.5% Triton-X 100 in PBS for 10 min. They were washed three times (20 min each) with 5 mM glycine in PBS, treated with 1% Tween-20 (10 min), and equilibrated in intracellular buffer (ICB; 100 mM KCl, 5 mM MgCl2, 3 mM EGTA, 1% BSA, and 20 mM Hepes, pH 6.8) for 5 min, as described by Pauken & Capco (1999, 2000). Groups of five embryos were transferred into 50 µl drops of ICB containing 10% serum matching the secondary antibody, 0.1% Tween-20, and antibody against the PKC isozyme (at a final concentration of 10 µg/ml for monoclonal antibodies and a final dilution of 1:100 for polyclonal antibodies), and incubated overnight at 4 °C. Embryos were washed four times (30 min each) with ICB and transferred into 50 µl drops of diluted secondary antibody at 20 µg/ml for 2 h at room temperature in the dark. Fluorescein-conjugated, affinity-purified F(ab’)2 fragment of sheep anti-mouse IgG (Boehringer Mannheim, Mannheim, Germany) was used to detect the monoclonal antibodies, and affinity-purified F(ab’)2 fragment of goat anti-rabbit IgG labeled with Rhodamine Red-X (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) was used to detect the polyclonal antibodies. The embryos were washed four times (30 min each at room temperature in the dark) with ICB and then mounted on 0.1% polylysine coated slides using Vectashield containing 4',6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA). Negative controls for immunocytochemistry were embryos exposed to medium rather than primary antibody. For each isozyme at each stage, there were at least nine embryos analyzed from three independent experiments.
Imaging was performed using a Bio-Rad MRC 600 confocal laser-scanning microscope attached to an Optiphot-II Nikon fluorescence microscope (with a 60 x , 1.4 numerical aperture plan-apochromat oil-immersion objective). Excitation was with 488 and 568 nm wavelengths from a krypton–argon laser.
Quantification of fluorescence intensity
We used the same confocal microscope settings for images from all embryos regardless of antibody, and for all trials. The settings were based on the fluorescence intensity of embryos immunostained for PKC ßII, because this antibody gave us the strongest signal (see Fig. 2C
, below). The settings were selected to ensure that the PKC ßII images were always less than 256 on the gray scale in the highest part of the linear range.
|
The mean fluorescence intensity for each subcellular area of each blastomere was calculated by dividing the sum of all gray values in the sub-regions by the total number of pixels enclosed. The background gray value in each image (a region outside of the embryo) was subtracted from the mean fluorescence intensity. Analysis of variance did not show a significant difference between the three trials, and thus we were able to treat the readings from each sub-cellular area of each blastomere at a given time point as a single population. The error bars in the graphs are the standard deviations of these readings, and represent the variations among blastomeres, embryos, and trials.
Difference in fluorescence intensity between the nucleus and cytoplasm was analyzed using a Student’s t-test (P < 0.05), and is indicated in Figs 2
–5 (see below) by an asterisk. Differences in fluorescence intensity among the apical, central, and basal regions of the cytoplasm were compared using ANOVA (P < 0.05), and the statistically different groups were identified by Tukey’s test (P < 0.05). Statistical difference is indicated by lower-case letters on Figs 2 and 5.
|
|
|
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
shows the staining of both an isozyme-specific monoclonal antibody and polyclonal antiserum from two manufacturers for PKCs , , , and . In each case the monoclonal antibodies and polyclonal antisera were raised against two distinct regions of the PKC isozyme, and both antibodies produced the same distribution patterns, although intensity of staining was usually less with the monoclonal antibodies (Fig. 1A). Hence, all further experiments used polyclonal antisera for PKCs , ßI, ßII, , , , , , and , and monoclonal antibodies for PKCs and , and RACK1. The monoclonal antibodies that recognize mouse PKCs and cross-react, and the proteins are 99% homologous (Transduction Laboratories). They are thought to be equivalent genes of human () and mouse (; Kazanietz 2000), so it was reassuring that they displayed almost identical patterns of binding on mouse embryos (see Fig. 5 below). Secondly, the evidence that the antisera against PKCs ßI and ßII detected these PKCs is that 4ß-PMA (a general activator of cPKCs and nPKCs) and pseudo-ßRACK (a specific peptide activator of cPKCs) affected expression of the molecules detected by these antisera at the two-cell stage. The changes were isozyme-specific, and the same with both types of activation (Fig. 1B); the distributions of PKCs ßI, ßII, and changed, but PKC staining remained more concentrated in nuclei, although the nuclear/cytoplasmic ratio decreased. The peptide pseudo-ßRACK was designed using PKC ß sequence, but from the constant (C2) region shared by all cPKCs. Thirdly, whereas all isozymes were detected during preimplantation development, not all isozymes were expressed at every stage. There were negative embryos for some isoforms with each of the two secondary antibodies, one that recognized the polyclonal antisera, and one that recognized the monoclonal antibodies. The stages that were negative for PKCs and (Figs 2A and 3A) were stained with the polyclonal secondary, and the stages that were negative for PKCs and and RACK1 (see Fig. 5 below) were stained with the monoclonal secondary. Finally, fluorescence intensity was above that of negative control embryos (no primary antibody) that were part of each trial.
|
–5). Whenever detected, PKCs ßII, , and and RACK 1 were more concentrated in the cytoplasm than in the nucleus (Figs 2 and 5). The opposite was true of PKC ; its nuclear concentration was consistently higher than the cytoplasmic concentration except at the late two-cell stage (Fig. 4). For the other PKCs, there were changes in the nuclear/cytoplasmic distribution between stages of preimplantation development. It is interesting that in early four-cell embryos there were more PKCs localized to the nuclei than at any other stage; the concentrations of seven out of 10 isozymes were higher in nuclei than in the cytoplasm.
Cytoplasmic versus nuclear distributions
At the two-cell stage (45 h post-hCG, G2), PKCs ßI, , , , , and were distributed equally between the cytoplasmic and nuclear compartments (Figs 2–4). Only PKC was more concentrated in the nucleus (P < 0.05), whereas PKCs ßII, , , and were more concentrated in the cytoplasm (P < 0.05).
At the four-cell stage (52 h post-hCG, G1/S), the distributions of PKCs , ßI, , , , , and changed, with six out of seven (all but PKC ) becoming more concentrated in nuclei (P < 0.05). PKC became equally distributed after being more concentrated in two-cell nuclei. PKC was below the detection limits of the assay.
At the eight-cell stage (69 h post-hCG), three out of six PKC isozymes with high nuclear concentrations at the four-cell stage became equally distributed between the cytoplasm and nucleus (ßI, , and ) or became more concentrated in the cytoplasm (; P < 0.05). Only PKCs and retained higher nuclear fluorescence intensity at this stage (P < 0.05). The concentration of PKC became higher in the cytoplasm from the equal distribution of the four-cell stage (P < 0.05).
At the morula stage (76 h post-hCG), the concentrations of PKC , ßI, and once again increased in nuclei (P < 0.05), whereas PKCs and became evenly distributed between the nucleus and cytoplasm. Immunostaining for PKCs , , and and RACK1 decreased to negative-control levels.
Of the eight PKC isozymes detectable at the blastocyst stage (93 h post-hCG), PKC was the only isozyme that was more concentrated in the nucleus (P < 0.05).
Cytoplasmic distributions
In the first photomicrographs, there seemed to be uneven distribution within the cytoplasm of some isozymes. Therefore, we separately measured fluorescence in the apical, basal, and central regions of the cytoplasm. At the two-cell stage, PKCs ßII, , , and were localized preferentially to the apical cytoplasm (P < 0.05; Figs 2, 4, and 5), and at the four-cell stage RACK1 was localized to the basal cytoplasm (P < 0.05; Fig. 5).
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
In most studies with adult cell types, PKC in different subcellular fractions has been detected by Western blots using 200 µg–1 mg protein, depending on isozyme abundance. Similar studies with preimplantation embryos certainly would be hampered by lack of material. A mouse oocyte has approximately 23 ng total protein (Hogan et al. 1994). However, live-cell imaging of translocation in individual cells, imaging for co-localization of intracellular signaling molecules, and photobleaching studies have quantified relative fluorescence intensity (Almholt et al. 1999, Ron et al. 1999, Echevarria et al. 2003, Schechtman et al. 2004). Thus whereas quantifying fluorescence intensity is not novel, it has not been used to localize signaling molecules in preimplantation mammalian embryos in quite this manner. The success of the approach is seen in the fact that differences were detected between the nucleus and cytoplasm and different areas of the cytoplasm. Thus, this approach should be effective for studying intracellular distribution and associations of molecules in preimplantation embryos where starting material is too limited for cell fractionation.
There are differences between this study and that of Pauken & Capco (2000) especially in the subcellular distribution of PKCs , , and . More recently, Quan et al.(2003) and Baluch et al.(2004) have shown PKC distribution similar to ours. However, discrepancies in distribution will likely remain since distribution of PKC is dynamic, and precisely when and how embryos are collected could affect results. In this study, we controlled timing of gonadotrophin administration and embryo collection as much as possible. The major difference between this study and that of Pauken & Capco (2000) is the presence of PKCs ßI and ßII. Pauken & Capco found no PKC ß in preimplantation mouse embryos. However, Eckert et al.(2004) detected both PKCs ßI and ßII in freshly isolated mouse inner cell masses. The expression of the PKC ß isoforms in mouse oocytes has been similarly controversial using both Western blots and immunofluorescence (Gangeswaran & Jones 1997, Luria et al. 2000, Pauken & Capco 2000, Downs et al. 2001). Because of this, we decided to use activation as a means to verify antibody specificity. The fact that both general and specific peptide activators of the cPKC caused translocation of the molecules recognized by our antibodies against PKCs ßI and ßII suggests that what is recognized are PKC ß, or at least PKC ß-like, molecules.
Localization of PKC isozymes to subcellular compartments can be mediated by binding to isozyme-specific anchoring proteins or through the cytoskeleton (Mochly-Rosen & Gordon 1998, Jaken & Parker 2000, Newton 2001, Schechtman & Mochly-Rosen 2001). Known anchoring proteins include substrates that interact with C kinases (STICKs) and isozyme-specific RACKs. Most known STICKs are located at interfaces between membranes and cytoskeletal structures, and phosphorylation by PKC modifies their function. They may or may not be multi-enzyme scaffold proteins. An example of the latter is AKAP 79, which binds to PKCs and ßII (Klauck et al. 1996), as well as at least two other signal transducers, protein kinase A and calcineurin (a Ca2+/calmodulin-dependent phosphatase). RACK1 is a non-substrate receptor for PKC ßII, although it also binds other PKC isoforms and other molecules (Jaken & Parker 2000, Rigas et al. 2003). Since RACK1 is involved in the translocation and association of PKC ßII with a normal substrate in cardiac myocytes (Ron & Mochly-Rosen 1995, Csukai et al. 1997), it is interesting that RACK1 and PKC ßII distributions differ in preimplantation embryos. In particular, RACK1 is concentrated in the basal region of cytoplasm in four-cell stage embryos, whereas PKC ßII is concentrated in the apical cytoplasm. Other RACKs, STICKs, or a receptor for inactivated C kinase (RICK) may govern PKC ßII localization at this stage. Further studies to find molecular partners of PKC isozymes are needed to elucidate their localization and function.
Although translocation of PKC is associated with activation in other cell types (Csukai & Mochly-Rosen 1999, Teruel & Meyer 2000), it must be noted that the state of activation of each isozyme during preimplantation development still needs to be determined. Also, there are several sources of change in the relative concentration of an isozyme observed in this study. First, there could be translocation of existing molecules between the cytoplasm and nucleus or of newly synthesized protein from the cytoplasm into the nucleus. Second, new synthesis of an isozyme without translocation would increase the relative cytoplasmic concentration. Or thirdly there could be differential degradation of an isozyme among subcellular compartments, which would decrease the relative concentration in that compartment. Further studies with individual isozymes to separate these possibilities are in order.
Functional implications of subcellular localization of PKC isozymes
This study defines the intracellular distribution of each PKC isozyme at specific times during cleavage, because it is sub-cellular location of PKC isozymes that correlates with different functions in a cell (Newton 2001). In adult cells, the PKC isozymes have a large and varied number of regulatory roles. Regulation of nuclear architecture (Hocevar et al. 1993, Maraldi et al. 1999), cell cycle (Livneh & Fishman 1997), and cellular polarization (Plant et al. 2003) are just a few examples. The PKC isozymes translocate to different subcellular areas during preimplantation mouse development, and likely are involved in multiple steps in preimplantation development. We found that nuclear localization of PKC (early two- and four-cell nuclei) overlaps with that of PKC (four- and eight-cell nuclei), whereas PKCs and are more concentrated in nuclei only at the early four-cell stage (Figs 2 and 3). Furthermore, we found that PKC localizes to the nuclei of eight-cell- to blastocyst-stage embryos (Fig. 4). Nuclear localization may indicate involvement in regulation of transcription. Hamatani et al.(2004) have shown increases in the number of genes transcribed at each stage of mouse preimplantation development. There is also evidence for chromatin remodeling in early mouse embryos (Thompson et al. 1998, Schultz et al. 1999). Recently, Santos et al.(2003) and Sarmento et al.(2004) have described constant modulation of histones during this period. It is very likely that some of the PKCs are involved in changing the relative lengths of the cell-cycle components during early cleavage (Pratt 1987, Livneh & Fishman 1997).
In conclusion, this study suggests fertile areas for further research. Activation state, effects of specific inhibitors and activators, co-localization with potential substrates and regulators, and temporal and spatial correlation of the isozymes with specific processes of mouse preimplantation embryogenesis are studies that can be undertaken in the future using this semi-quantitative approach. In the accompanying paper (Dehghani et al. 2005), we study the relationship between translocation of PKCs and into four-cell nuclei and transcription and nuclear morphology.
|
Acknowledgements |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
|
Footnotes |
|---|
(H Dehghani is now at Programme in Cell Biology, The Research Institute, The Hospital for Sick Children, Toronto, Ontario, M5G 1X8 Canada)
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionAcknowledgementsReferences |
|---|
Almholt K, Arkhammar PO, Thastrup O & Tullin S 1999 Simultaneous visualization of the translocation of protein kinase Calpha-green fluorescent protein hybrids and intracellular calcium concentrations. Biochemical Journal 337 211–218.
Baluch DP, Koeneman BA, Hatch KR, McGaughey RW & Capco DG 2004 PKC isotypes in post-activated and fertilized mouse eggs: association with the meiotic spindle. Developmental Biology 274 45–55.[CrossRef][Web of Science][Medline]
Csukai M & Mochly-Rosen D 1999 Pharmacologic modulation of protein kinase C isozymes: the role of RACKs and subcellular localisation. Pharmacological Research 39 253–259.[CrossRef][Web of Science][Medline]
Csukai M, Chen CH, De Matteis MA & Mochly-Rosen D 1997 The coatomer protein beta’-COP, a selective binding protein (RACK) for protein kinase Cepsilon. Journal of Biological Chemistry 272 29200–29206.
Dehghani H, Reith C & Hahnel AC 2005 Subcellular localization of protein kinase C and affects transcriptional and post-transcriptional processes in four-cell mouse embryos. Reproduction 130 453–465.
Disatnik MH, Buraggi G & Mochly-Rosen D 1994 Localization of protein kinase C isozymes in cardiac myocytes. Experimental Cell Research 210 287–297.[CrossRef][Web of Science][Medline]
Doerner D, Abdel-Latif M, Rogers TB & Alger BE 1990 Protein kinase C-dependent and independent effects of phorbol esters on hippocampal calcium channel current. Journal of Neuroscience 10 1699–1706.[Abstract]
Dorn GW 2nd, Souroujon MC, Liron T, Chen CH, Gray MO, Zhou HZ, Csukai M, Wu G, Lorenz JN & Mochly-Rosen D 1999 Sustained in vivo cardiac protection by a rationally designed peptide that causes epsilon protein kinase C translocation. PNAS 96 12798–12803.
Downs SM, Cottom J & Hunzicker-Dunn M 2001 Protein kinase C and meiotic regulation in isolated mouse oocytes. Molecular Reproduction and Development 58 101–115.[CrossRef][Web of Science][Medline]
Echevarria W, Leite MF, Guerra MT, Zipfel WR & Nathanson MH 2003 Regulation of calcium signals in the nucleus by a nucleoplasmic reticulum. Nature Cell Biology 5 440–446.[CrossRef][Web of Science][Medline]
Eckert JJ, McCallum A, Mears A, Rumsby MG, Cameron IT & Fleming TP 2004 PKC signalling regulates tight junction membrane assembly in the pre-implantation mouse embryo. Reproduction 127 653–667.
Eliyahu E & Shalgi R 2002 A role for protein kinase C during rat egg activation. Biology of Reproduction 67 189–195.
Fan HY, Tong C, Li MY, Lian L, Chen DY, Schatten H & Sun QY 2002 Translocation of the classic protein kinase C isoforms in porcine oocytes: implications of protein kinase C involvement in the regulation of nuclear activity and cortical granule exocytosis. Experimental Cell Research 277 183–191.[CrossRef][Web of Science][Medline]
Gallicano GI, McGaughey RW & Capco DG 1995 Protein kinase M, the cytosolic counterpart of protein kinase C, remodels the internal cytoskeleton of the mammalian egg during activation. Developmental Biology 167 482–501.[CrossRef][Web of Science][Medline]
Gallicano GI, McGaughey RW & Capco DG 1997 Activation of protein kinase C after fertilization is required for remodeling the mouse egg into the zygote. Molecular Reproduction and Development 46 587–601.[CrossRef][Web of Science][Medline]
Gangeswaran R & Jones KT 1997 Unique protein kinase C profile in mouse oocytes: lack of calcium-dependent conventional isoforms suggested by rtPCR and Western blotting. FEBS Letters 412 309–312.[CrossRef][Web of Science][Medline]
Geiges D, Meyer T, Marte B, Vanek M, Weissgerber G, Stabel S et al. 1997 Activation of protein kinase C subtypes alpha, gamma, delta, epsilon, zeta, and eta by tumor-promoting and nontumor-promoting agents. Biochemical Pharmacology 53 865–875.[CrossRef][Web of Science][Medline]
Hamatani T, Carter MG, Sharov AA & Ko MS 2004 Dynamics of global gene expression changes during mouse preimplantation development. Developmental Cell 6 117–131.[CrossRef][Web of Science][Medline]
Hausser A, Link G, Bamberg L, Burzlaff A, Lutz S, Pfizenmaier K & Johannes FJ 2002 Structural requirements for localization and activation of protein kinase C mu (PKC mu) at the Golgi compartment. Journal of Cell Biology 156 65–74.
Ho Y, Wigglesworth K, Eppig JJ & Schultz RM 1995 Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Molecular Reproduction and Development 41 232–238.[CrossRef][Web of Science][Medline]
Hocevar BA, Burns DJ & Fields AP 1993 Identification of protein kinase C (PKC) phosphorylation sites on human lamin B. Potential role of PKC in nuclear lamina structural dynamics. Journal of Biological Chemistry 268 7545–7552.
Hogan B, Beddington R, Costantini F & Lacy E 1994 In Manipulating the Mouse Embryo, a Laboratory Manual. New York: Cold Spring Harbor Press.
Jaken S & Parker PJ 2000 Protein kinase C binding partners. Bioessays 22 245–254.[CrossRef][Web of Science][Medline]
Johnson JA, Gray MO, Chen CH & Mochly-Rosen D 1996 A protein kinase C translocation inhibitor as an isozyme-selective antagonist of cardiac function. Journal of Biological Chemistry 271 24962–24966.
Kazanietz MG 2000 Eyes wide shut: protein kinase C isozymes are not the only receptors for the phorbol ester tumor promoters. Molecular Carcinogenesis 28 5–11.[CrossRef][Web of Science][Medline]
Keranen LM, Dutil EM & Newton AC 1995 Protein kinase C is regulated in vivo by three functionally distinct phosphorylations. Current Biology 5 1394–1403.[CrossRef][Web of Science][Medline]
Klauck TM, Faux MC, Labudda K, Langeberg LK, Jaken S & Scott JD 1996 Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science 271 1589–1592.[Abstract]
Lawitts JA & Biggers JD 1993 Culture of preimplantation embryos. Methods in Enzymology 225 153–164.[Web of Science][Medline]
Livneh E & Fishman DD 1997 Linking protein kinase C to cell-cycle control. European Journal of Biochemistry 248 1–9.[Web of Science][Medline]
Luria A, Tennenbaum T, Sun QY, Rubinstein S & Breitbart H 2000 Differential localization of conventional protein kinase C isoforms during mouse oocyte development. Biology of Reproduction 62 1564–1570.
Maraldi NM, Zini N, Santi S & Manzoli FA 1999 Topology of inositol lipid signal transduction in the nucleus. Journal of Cell Physiology 181 203–217.[CrossRef][Web of Science][Medline]
Miyamae M, Rodriguez MM, Camacho SA, Diamond I, Mochly-Rosen D & Figueredo VM 1998 Activation of epsilon protein kinase C correlates with a cardioprotective effect of regular ethanol consumption. PNAS 95 8262–8267.
Mochly-Rosen D & Gordon AS 1998 Anchoring proteins for protein kinase C: a means for isozyme selectivity. FASEB Journal 12 35–42.
Mochly-Rosen D & Kauvar LM 2000 Pharmacological regulation of network kinetics by protein kinase C localization. Seminars in Immunology 12 55–61.[CrossRef][Web of Science][Medline]
Newton AC 2001 Protein kinase C: structural and spatial regulation by phosphorylation, cofactors, and macromolecular interactions. Chemistry Reviews 101 2353–2364.
Ohsugi M & Yamamura H 1993 Differences in the effects of treatment of uncompacted and compacted mouse embryos with phorbol esters on pre- and postimplantation development. Differentiation 53 173–179.[CrossRef][Web of Science][Medline]
Ohsugi M, Ohsawa T & Semba R 1993a Similar responses to pharmacological agents of 1,2-OAG-induced compaction-like adhesion of two-cell mouse embryo to physiological compaction. Journal of Experimental Zoology 265 604–608.
Ohsugi M, Ohsawa T & Yamamura H 1993b Involvement of protein kinase C in nuclear migration during compaction and the mechanism of the migration: analyses in two-cell mouse embryos. Developmental Biology 156 146–154.[CrossRef][Web of Science][Medline]
Pauken CM & Capco DG 1999 Regulation of cell adhesion during embryonic compaction of mammalian embryos: roles for PKC and beta-catenin. Molecular Reproduction and Development 54 135–144.[CrossRef][Web of Science][Medline]
Pauken CM & Capco DG 2000 The expression and stage-specific localization of protein kinase C isotypes during mouse pre-implantation development. Developmental Biology 223 411–421.[CrossRef][Web of Science][Medline]
Plant PJ, Fawcett JP, Lin DC, Holdorf AD, Binns K, Kulkarni S & Pawson T 2003 A polarity complex of mPar-6 and atypical PKC binds, phosphorylates and regulates mammalian Lgl. Nature Cell Biology 5 301–308.[CrossRef][Web of Science][Medline]
Pratt HPM 1987 Isolation, Culture, and Manipulation of Preimplantation Mouse Embryos. In Mammalian Development: A Practical Approach, pp 13–42. Ed M Monk. Washington DC: IRL Press.
Quan HM, Fan HY, Meng XQ, Huo LJ, Chen DY, Schatten H, Yang PM & Sun QY 2003 Effects of PKC activation on the meiotic maturation, fertilization and early embryonic development of mouse oocytes. Zygote 11 329–337.[CrossRef][Web of Science][Medline]
Raz T, Eliyahu E, Yesodi V & Shalgi R 1998 Profile of protein kinase C isozymes and their possible role in mammalian egg activation. FEBS Letters 431 415–418.[CrossRef][Web of Science][Medline]
Rigas AC, Ozanne DM, Neal DE & Robson CN 2003 The scaffolding protein RACK1 interacts with androgen receptor and promotes cross-talk through a protein kinase C signaling pathway. Journal of Biological Chemistry 278 46087–46093.
Ron D & Mochly-Rosen D 1995 An autoregulatory region in protein kinase C: the pseudoanchoring site. PNAS 92 492–496.
Ron D, Jiang Z, Yao L, Vagts A, Diamond I & Gordon A 1999 Coordinated movement of RACK1 with activated betaIIPKC. Journal of Biological Chemistry 274 27039–27046.
Santos F, Zakhartchenko V, Stojkovic M, Peters A, Jenuwein T, Wolf E, Reik W & Dean W 2003 Epigenetic marking correlates with developmental potential in cloned bovine preimplantation embryos. Current Biology 13 1116–1121.[CrossRef][Medline]
Sarmento OF, Digilio LC, Wang Y, Perlin J, Herr JC, Allis CD & Coonrod SA 2004 Dynamic alterations of specific histone modifications during early murine development. Journal of Cell Science 117 4449–4459.
Schechtman D & Mochly-Rosen D 2001 Adaptor proteins in protein kinase C-mediated signal transduction. Oncogene 20 6339–6347.[CrossRef][Web of Science][Medline]
Schechtman D, Craske ML, Kheifets V, Meyer T, Schechtman J & Mochly-Rosen D 2004 A critical intramolecular interaction for protein kinase Cepsilon translocation. Journal of Biological Chemistry 279 15831–15840.
Schultz RM 1993 Regulation of zygotic gene activation in the mouse. Bioessays 15 531–538.[CrossRef][Web of Science][Medline]
Schultz RM, Davis W Jr, Stein P & Svoboda P 1999 Reprogramming of gene expression during preimplantation development. Journal of Experimental Zoology 285 276–282.
Stachecki JJ & Armant DR 1996 Regulation of blastocoele formation by intracellular calcium release is mediated through a phospholipase C-dependent pathway in mice. Biology of Reproduction 55 1292–1298.[Abstract]
Teruel MN & Meyer T 2000 Translocation and reversible localization of signaling proteins: a dynamic future for signal transduction. Cell 103 181–184.[CrossRef][Web of Science][Medline]
Thompson EM, Legouy E & Renard JP 1998 Mouse embryos do not wait for the MBT: chromatin and RNA polymerase remodeling in genome activation at the onset of development. Developments in Genetics 22 31–42.
Viveiros MM, Hirao Y & Eppig JJ 2001 Evidence that protein kinase C (PKC) participates in the meiosis I to meiosis II transition in mouse oocytes. Developments in Biology 235 330–342.
Viveiros MM, O’Brien M, Wigglesworth K & Eppig JJ 2003 Characterization of protein kinase C-delta in mouse oocytes throughout meiotic maturation and following egg activation. Biology of Reproduction 69 1494–1499.
Wang S, Gebre-Medhin S, Betsholtz C, Stalberg P, Zhou Y, Larsson C, Weber G, Feinstein R, Oberg K, Gobl A & Skogseid B 1998 Targeted disruption of the mouse phospholipase C beta3 gene results in early embryonic lethality. FEBS Letters 441 261–265.[CrossRef][Web of Science][Medline]
Watanabe H, Davis JB, Smart D, Jerman JC, Smith GD, Hayes P, Vreins J, Cairns W, Wissenbach U, Prenen J, Flockerzi V, Droogemans G, Benham CD & Nilus B 2002 Activation of TRPV4 channels (hVRL-2/mTRP12) by phorbol derivatives. Journal of Biological Chemistry 277 13569–13577.
Winkel GK, Ferguson JE, Takeichi M & Nuccitelli R 1990 Activation of protein kinase C triggers premature compaction in the four-cell stage mouse embryo. Developmental Biology 138 1–15.[CrossRef][Web of Science][Medline]
This article has been cited by other articles:
 |
|
 |
|
  H. Dehghani, C. Reith, and A. C Hahnel Subcellular localization of protein kinase C {delta} and {varepsilon} affects transcriptional and post-transcriptional processes in four-cell mouse embryos Reproduction, October 1, 2005; 130(4): 453 - 465. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
