caudal: Biological Overview | Evolutionary Homologs | Regulation | Developmental Biology | Effects of Mutation | References

Gene name - caudal

Synonyms -

Cytological map position - 38E5-6

Function - transcription factor

Keyword(s) - gap gene, plays a role in establishing the posterior domains of the embryo, which give rise to the posterior gut, regulates constitutive local expression of antimicrobial peptide genes in Drosophila epithelia

Symbol - cad

FlyBase ID:FBgn0000251

Genetic map position - 2-[54]

Classification - homeodomain

Cellular location - nuclear

NCBI link: Entrez Gene

caudal orthologs: Biolitmine
Recent literature
Shir-Shapira, H., Sharabany, J., Filderman, M., Ideses, D., Ovadia-Shochat, A., Mannervik, M. and Juven-Gershon, T. (2015). Structure-function analysis of the Drosophila melanogaster Caudal provides insights into core promoter-preferential activation. J Biol Chem [Epub ahead of print]. PubMed ID: 26018075
Regulation of RNA polymerase II transcription is critical for the proper development, differentiation and growth of an organism. The RNA polymerase II core promoter is the ultimate target of a multitude of transcription factors that control transcription initiation. Core promoters encompass the RNA start site and consist of functional elements such as the TATA box, initiator and downstream core promoter element (DPE), which confer specific properties to the core promoter. Previous studies have discovered that Drosophila Caudal, which is a master regulator of genes involved in development and differentiation, is a DPE-specific transcriptional activator. This study shows that the mouse Caudal-related Cdx proteins (mCdx1, mCdx2 and mCdx4) are also preferential core promoter transcriptional activators. To elucidate the mechanism that enables Caudal to preferentially activate DPE transcription, structure-function analysis was performed. Using a systematic series of deletion mutants (all containing the intact DNA-binding homeodomain) it was discovered that the C-terminal region of Caudal contributes to the preferential activation of the fushi tarazu (ftz). Caudal target gene. Furthermore, the region containing both the homeodomain and the C-terminus of Caudal is sufficient to confer core promoter-preferential activation to the heterologous GAL4 DNA-binding domain. Importantly, it was discovered that Drosophila CBP (dCBP) is a co-activator for Caudal-regulated activation of ftz. Strikingly, dCBP confers the ability to preferentially activate the DPE-dependent ftz reporter to mini-Caudal proteins that are unable to preferentially activate ftz transcription themselves. Taken together, it is the unique combination of dCBP and Caudal that enables the co-activation of ftz in a core promoter-preferential manner.
Verd, B., Clark, E., Wotton, K. R., Janssens, H., Jimenez-Guri, E., Crombach, A. and Jaeger, J. (2018). A damped oscillator imposes temporal order on posterior gap gene expression in Drosophila. PLoS Biol 16(2): e2003174. PubMed ID: 29451884
Insects determine their body segments in two different ways. Short-germband insects, such as the flour beetle Tribolium castaneum, use a molecular clock to establish segments sequentially. In contrast, long-germband insects, such as the vinegar fly Drosophila melanogaster, determine all segments simultaneously through a hierarchical cascade of gene regulation. Gap genes constitute the first layer of the Drosophila segmentation gene hierarchy, downstream of maternal gradients such as that of Caudal (Cad). This study used data-driven mathematical modelling and phase space analysis to show that shifting gap domains in the posterior half of the Drosophila embryo are an emergent property of a robust damped oscillator mechanism, suggesting that the regulatory dynamics underlying long- and short-germband segmentation are much more similar than previously thought. In Tribolium, Cad has been proposed to modulate the frequency of the segmentation oscillator. Surprisingly, the current simulations and experiments show that the shift rate of posterior gap domains is independent of maternal Cad levels in Drosophila. These results suggest a novel evolutionary scenario for the short- to long-germband transition and help explain why this transition occurred convergently multiple times during the radiation of the holometabolan insects.
Clark, E. and Peel, A. D. (2018). Evidence for the temporal regulation of insect segmentation by a conserved sequence of transcription factors. Development. PubMed ID: 29724758
Long-germ insects, such as the fruit fly Drosophila melanogaster, pattern their segments simultaneously, whereas short-germ insects, such as the beetle Tribolium castaneum, pattern their segments sequentially, from anterior to posterior. While the two modes of segmentation at first appear quite distinct, much of this difference might simply reflect developmental heterochrony. This study now shows that, in both Drosophila and Tribolium, segment patterning occurs within a common framework of sequential Caudal, Dichaete, and Odd-paired expression. In Drosophila these transcription factors are expressed like simple timers within the blastoderm, while in Tribolium they form wavefronts that sweep from anterior to posterior across the germband. In Drosophila, all three are known to regulate pair-rule gene expression and influence the temporal progression of segmentation. It is proposed that these regulatory roles are conserved in short-germ embryos, and that therefore the changing expression profiles of these genes across insects provide a mechanistic explanation for observed differences in the timing of segmentation. In support of this hypothesis it was demonstrated that Odd-paired is essential for segmentation in Tribolium, contrary to previous reports.
Wu, K., Tang, Y., Zhang, Q., Zhuo, Z., Sheng, X., Huang, J., Ye, J., Li, X., Liu, Z. and Chen, H. (2021). Aging-related upregulation of the homeobox gene caudal represses intestinal stem cell differentiation in Drosophila. PLoS Genet 17(7): e1009649. PubMed ID: 34228720
The differentiation efficiency of adult stem cells undergoes a significant decline in aged animals, which is closely related to the decline in organ function and age-associated diseases. However, the underlying mechanisms that ultimately lead to this observed decline of the differentiation efficiency of stem cells remain largely unclear. This study investigated Drosophila midguts and identified an obvious upregulation of caudal (cad), which encodes a homeobox transcription factor. This factor is traditionally known as a central regulator of embryonic anterior-posterior body axis patterning. This study reports that depletion of cad in intestinal stem/progenitor cells promotes quiescent intestinal stem cells (ISCs) to become activate and produce enterocytes in the midgut under normal gut homeostasis conditions. However, overexpression of cad results in the failure of ISC differentiation and intestinal epithelial regeneration after injury. Moreover, this study suggests that cad prevents intestinal stem/progenitor cell differentiation by modulating the Janus kinase/signal transducers and activators of the transcription pathway and Sox21a-GATAe signaling cascade. Importantly, the reduction of cad expression in intestinal stem/progenitor cells restrained age-associated gut hyperplasia in Drosophila. This study identified a function of the homeobox gene cad in the modulation of adult stem cell differentiation and suggested a potential gene target for the treatment of age-related diseases induced by age-related stem cell dysfunction.
Clark, E., Battistara, M. and Benton, M. A. (2022). A timer gene network is spatially regulated by the terminal system in the Drosophila embryo. Elife 11. PubMed ID: 36524728
In insect embryos, anteroposterior patterning is coordinated by the sequential expression of the 'timer' genes caudal, Dichaete and odd-paired, whose expression dynamics correlate with the mode of segmentation. In Drosophila, the timer genes are expressed broadly across much of the blastoderm, which segments simultaneously, but their expression is delayed in a small 'tail' region, just anterior to the hindgut, which segments during germband extension. Specification of the tail and the hindgut depends on the terminal gap gene tailless, but beyond this the regulation of the timer genes is poorly understood. This study used a combination of multiplexed imaging, mutant analysis, and gene network modelling to resolve the regulation of the timer genes, identifying 11 new regulatory interactions and clarifying the mechanism of posterior terminal patterning. It is proposed that a dynamic Tailless expression gradient modulates the intrinsic dynamics of a timer gene cross-regulatory module, delineating the tail region and delaying its developmental maturation.

During early embryogenesis in Drosophila, Caudal mRNA is distributed as a gradient with its highest level at the posterior of the embryo. This suggests that the Caudal homeodomain transcription factor might play a role in establishing the posterior domains of the embryo, which undergo gastrulation and give rise to the posterior gut. By generating embryos lacking both the maternal and zygotic mRNA contribution, caudal has been shown to be essential for invagination of the hindgut primordium and for further specification and development of the hindgut. Mature embryos lacking cad activity (maternal and/or zygotic contributions) were examined to assess the requirement for cad in establishing the structures that arise from the posterior ~15% of the blastoderm embryo, namely the posterior midgut, Malpighian tubules and hindgut. Embryos lacking both maternal and zygotic cad activity (referred to as cad m-z -, or cad-deficient), as well as embryos lacking maternal cad activity but carrying one zygotically active wild-type cad gene (cad m-z+) and embryos lacking only zygotic cad (cad m+z -) were generated. Staining with anti-Crumbs antibody, which labels the apical surface of the hindgut epithelium, reveals that cad activity, provided either maternally or zygotically, is essential for formation of the hindgut. When cad is provided only maternally (cad m+z-), the posterior gut appears essentially normal. When cad is provided only zygotically (cad m-z+), a hindgut tube is formed; this can range from almost normal to somewhat reduced in length and twisted. When cad activity is completely lacking (cad m-z-), the hindgut (as well as the anal pads and the eighth abdominal segment) is almost entirely absent. It is concluded that cad activity is essential for normal hindgut formation, and that while some of this activity must be provided maternally, most can be provided either maternally or zygotically. The Malpighian tubules and posterior midgut also arise from domains expressing cad, but form relatively normally in cad-deficient embryos. As revealed by their expression of forkhead (fkh), the tubule primordia form relatively normally in embryos lacking cad activity (Wu, 1998).

The stages of gastrulation can be observationally followed by using expression of brachyenteron byn as a marker for the hindgut primordium. In the wild-type embryo, byn is expressed in a ring at the circumference of the amnioproctodeal plate. The edges of this ring come together as the posterior midgut primordium invaginates during stages 6 and 7; the ring of the hindgut primordium then sinks inward during stage 8 and is completely internalized by the end of stage 9. The zygotically expressed cad stripe and the posterior wingless stripe are also expressed in the bordering ring (i.e., the hindgut primordium) of the invaginating amnioproctodeal plate. Strikingly, in cad-deficient embryos, the byn-expressing ring of hindgut primordium draws together, but fails to invaginate, remaining on the outside of the embryo. Thus, although internalization of the Malpighian tubule and posterior midgut primordia is normal in cad-deficient embryos, the gastrulation movements necessary for internalization of the hindgut primordium do not occur in embryos lacking cad activity (Wu, 1998).

The absence of the hindgut primordium from cad-deficient embryos suggests that Caudal regulates genes required for establishing and/or maintaining the hindgut primordium. tailless, forkhead, byn, bowl and wingless are likely targets for cad regulation, since all are required for some aspect of hindgut development: the hindgut is missing from both tll and fkh embryos, and severely reduced in wg, byn and bowl embryos. bowl, also called bowel, codes for a zinc finger transcription factor related to odd-skipped. Since maternally provided Caudal, which persists only through the blastoderm stage, is sufficient for essentially normal hindgut formation, the fact that all of these genes are expressed at the posterior of the embryo during the blastoderm stage means that they are potential targets for regulation by Caudal. The effect of absence of maternal and/or zygotic cad activity on the expression of these genes was assessed by in situ hybridization with appropriate probes. For tll, byn and bowl, absence of cad activity does not result in a detectable effect on expression. As described below, however, cad activity is essential for expression of fkh and wg. Both maternal and zygotic cad contributions are necessary for posterior wg expression. During early stage 5, just prior to its expression in 14 stripes that are required to establish the segmental pattern, wg is expressed in two domains at the anterior, and in a broad posterior stripe. This terminal wg stripe is located at approximately 8-12% EL, overlapping with the posterior of the zygotic cad stripe and with the position of the hindgut and Malpighian tubule primordia in the blastoderm fate map. Expression of the wg terminal stripe has been shown to be independent of other segmentation genes, but has not been otherwise characterized. All embryos from mutant cad germline mothers (even those expressing zygotic cad) fail to express the terminal stripe of wg. These results demonstrate that maternal cad activity is essential for the transcription of wg in the terminal stripe. Among embryos from wg heterozygous parents, approximately one-quarter (presumably those lacking only the zygotic component of cad expression) lack the terminal wg stripe. Thus both maternal and zygotic cad activities are required for expression of the terminal wg stripe (Wu, 1998).

The expression of the early cap of fkh also requires cad activity; approximately half of the embryos from mutant cad germline females mated to cad heterozygous males (i. e., cad m-z - embryos) show a dramatic reduction in both the size and intensity of the posterior cap of fkh expression. However, if cad is supplied either maternally or zygotically, fkh expression is normal. Thus expression of the posterior cap of fkh requires cad activity, which can be provided either maternally or zygotically. Later, by stage 10, fkh expression is as strong in cad-deficient as in wild-type embryos, indicating that this later expression is independent of cad activity. Since tll and hkb are also required to activate early fkh expression but are not themselves regulated by cad, cad must act combinatorially with these two genes to promote early fkh expression (Wu, 1998).

cad also regulates wg in combination with other genes. In addition to the demonstrated requirement for cad, expression of the posterior wg stripe requires positive input from fkh and tll, since the stripe is absent from the respective mutant embryos. Since embryos lacking either maternal or zygotic cad fail to express the posterior wg stripe, but still express fkh and tll, cad must act combinatorially with fkh and tll to promote formation of the posterior wg stripe. Expression of the terminal stripe thus requires the combinatorial action of cad, tll and fkh; the posterior limit of the stripe is known to be defined by repression by hkb (Wu, 1998).

The fact that tll, byn and bowl expression at the blastoderm stage are all apparently normal in cad-deficient embryos suggests that a hindgut primordium is established in the absence of cad activity. The lack of proper blastoderm stage expression of fkh and wg, however, indicates that this hindgut primordium is not properly specified. In byn mutant embryos one of the earliest phenotypic manifestations of an abnormally specified hindgut primordium is ectopic expression of the cell death gene reaper (rpr). To ask whether the extremely reduced hindgut in cad-deficient embryos might result from a similar course of programmed cell death, expression of rpr was examined in embryos lacking cad. A striking pattern of ectopic rpr expression is observed in cad minus embryos, beginning during stage 7 and continuing into stage 8 (gastrulation), in a ring at the circumference of the amnioproctodeal plate. However, the actual loss of cells that is presumably initiated by this ectopic rpr expression does not begin until after early stage 10, because the hindgut primordium is present at this stage in cad-deficient embryos, as indicated by its expression of byn and fkh. By stage 13, the cad-deficient embryo has a very short hindgut and no detectable anal pads; in sections of stage 13 embryos there are numerous apoptotic cells in the region of the hindgut remnant (Wu, 1998).

The amnioproctodeal plate of cad-deficient embryos, distinguished by the fact that it ectopically expresses rpr, becomes increasingly retarded in its anteriorward movements during stages 7 and 8. By early stage 10 (~4.5 hours AEL), when the germ band in the wild-type embryo is almost fully extended, the germ band in the cad minus embryo has extended only about 40% of the normal distance and the embryo appears twisted. This phenotype is particularly obvious when embryos are labeled for fkh expression (which is reestablished in cad-deficient embryos after the blastoderm stage): the hindgut primordium, which strongly expresses fkh, remains near the posterior of the embryo. Thus, in the absence of cad activity, there is a severe defect in germband extension (Wu, 1998).

The failure of the hindgut to become internalized in caudal-deficient embryos raises the question of whether cad might regulate a zygotically expressed gene required for the invagination of the amnioproctodeal plate. One gene known to be required for gastrulation is fog; fog mutant embryos lack not only the posterior midgut, but, as revealed by anti-Crb staining, the Malpighian tubules and hindgut as well. In the blastoderm stage embryo, fog expression is first activated in the region that will become the ventral furrow; shortly thereafter, expression is initiated in a posterior cap, in the region that will become the amnioproctodeal invagination. In cad-deficient embryos, fog expression in the prospective ventral furrow is normal, but is significantly reduced in the posterior cap. Thus, cad is required for the normal level of expression of fog in the prospective amnioproctodeal plate; decreased fog expression in cad-deficient embryos is likely responsible for the failure of the hindgut primordium to be internalized during gastrulation. Since fkh or wg mutant embryos do not display detectable defects in gastrulation, fog is the only gene presently known to mediate the effects of cad on gastrulation. In fog mutant embryos, none of the posterior gut primordia invaginate, while in cad-deficient embryos the posterior midgut and Malpighian tubule primordium do invaginate; thus, consistent with the in situ hybridization results, a low level of fog activity is present at the posterior of embryos lacking cad (Wu, 1998).

In addition to cad, three other genes (fkh, byn and wg,), which are required at the posterior of the Drosophila embryo for formation of the hindgut, are related to genes found throughout the metazoa, known as HNF-3 (alpha, beta, and gamma), Brachyury (also known as T) and Wnt, respectively. In many cases, these homologs are expressed in portions of the ‘blastopore equivalent’ at the posterior of the embryo, that overlap with domains of expression of cad (Cdx). In C. elegans, a Wnt homolog is expressed, and required for proper posterior development, in the same posterior blastomere where the cad homolog pal-1 functions. In sea urchin, HNF-3 and Brachyury homologs are expressed in the vegetal plate just prior to gastrulation. In fish and frog, Caudal, Brachyury and Wnt (Wnt8 and Wnt11) are initially expressed around most or all of the blastopore lip while HNF-3 expression is dorsally localized. As gastrulation proceeds, the expression of these genes becomes more restricted and non-overlapping, with HNF-3 and Brachyury expression becoming localized to the notochord and Wnt8 expression retreating from the dorsal position and becoming exclusively ventral. Patterns of expression of HNF-3 and Brachyury consistent with this general description have been found in ascidians (phylum Urochordata), amphioxus (phylum Chordata), chick and mouse. Required roles for some of these genes have been demonstrated by analysis of mutants: mouse HNF-3ß knockouts reveal requirements in the formation of the node, notochord and head process; fish no tail and mouse T mutants reveal a requirement for Brachyury in migration of mesoderm through the primitive streak and in formation of the notochord. There is thus a constellation of conserved genes -- cad (Cdx), fkh (HNF-3), wg (Wnt8 and Wnt11) and byn (Brachyury) -- whose overlapping expression patterns in the blastopore equivalent suggests function in a related process. The phenotypes of the available mutations in these genes suggest that the common function is to specify cell fate at the blastopore; in most cases, essential parts of this fate are internalization and forward migration, two of the cellular movements that occur during gastrulation (Wu, 1998 and references).

The striking conservation in expression (and likely in function) of cad suggests that the regulation of posterior terminal development in Drosophila by Caudal may represent a more ancient regulatory mechanism than the tor receptor and the two genes that it activates: tll and hkb. Of these three genes, a vertebrate homolog is known only for tll; the function of this vertebrate gene, Tlx, is related to that of Drosophila tll not in the posterior, but rather in the anterior, in the establishment of the brain. Thus the Torso receptor pathway and its activation of tll and hkb has probably been superimposed relatively recently (in evolutionary terms) upon a more ancient, Caudal-regulated network of gene activity controlling gastrulation and gut formation. The fact that the same four genes are expressed at both the blastopore equivalent of chordates and at the amnioproctodeal invagination of Drosophila suggests that these two highly dynamic domains are homologous. Given the regulatory hierarchy that is present in Drosophila, it is proposed that in embryos of the proximate ancestor to arthropods and chordates, the posterior was defined by a posterior-to-anterior gradient of Cad activity. Cad is thought to have then activated expression of downstream network of genes in control of invagination (gastrulation) and gut specification. Cad expression in the archenteron probably continued during evolution and played an essential developmental role, since this structure differentiated into the gut. Going beyond the bilaterian ancestor to chordates and arthropods, it is worth considering that this nexus of gene expression may have evolved even more basally in the metazoa. The foregoing, by homologizing the insect amnioproctodeal invagination with the echinoderm and vertebrate blastopore, does not fit with the classical definition of protostomes and deuterostomes. This view categorizes arthropods as protostomes, in which the mouth is derived from the primary invagination of gastrulation; chordates are categorized as deuterostomes, where the mouth arises from a secondary invagination. More recently, comparisons of gastrulation patterns in many different species, as well as construction of molecularly based cladograms, have called into question the utility of these classically defined groups. While there continues to be uncertainty in understanding of ‘protostome’ and ‘deuterostome’ phyla, the significant conclusion of the information presented here is that there may be a homology between the blastopore of vertebrates and the amnioproctodeal (posterior) invagination of insects (Wu, 1998 and references).

The actions of the homeotic gene caudal illustrate the significance of shifting maternal and zygotic gene expression, and the interaction of gene systems in the creation of evolutionary diversity among insects. caudal expression is driven from two promoters, each of which generates its own transcript. Maternally provided Caudal appears to activate fushi tarazu transcription in the embryo's posterior, and therefore is a strong factor in the essential segmentation [Images] of the posterior (Dearolf, 1989). Zygotic cad transcription under negative control of Hunchback appears to be important in refinement of the developmental process of the Drosophila tail. Zygotic cad may regulate Abd-B, a gene that acts to suppress some aspects of tail morphogenesis (Casanova, 1986).

The role of caudal in posterior segment formation is involved in the generation of diversity between insect species. Primitive insects develop segmentation by heterchronic gene activation. Each posterior segment forms independently in a time dependent manner. nanos activity eliminates Hunchback from the posterior precellular blastoderm region of primitive insects, and caudal, otherwise repressed by Hunchback, is able to function there at a later stage. Caudal proceeds to drive the addition of new segments over time. Thus, a gradient of Caudal would regulate the asynchronous segment development of primitive insects (Rivera-Pomar, 1995).

Drosophila pair-rule gene expression, in an array of seven evenly spaced stripes along the anterior-posterior axis of the blastoderm embryo, is controlled by distinct cis-acting stripe elements. In the anterior region, such elements mediate transcriptional activation in response to (1) the maternal concentration gradient of the anterior determinant Bicoid and (2) repression by spatially distinct activities of zygotic gap genes. In the posterior region, activation of hairy stripe 6 has been shown to depend on the activity of the gap gene knirps, suggesting that posterior stripe expression is exclusively controlled by zygotic regulators. The zygotic activation of hairy stripe 6 expression is preceded by activation in response to maternal caudal activity. Thus, transcriptional activation of posterior stripe expression is likely to be controlled by maternal and zygotic factors as has been observed for anterior stripes. To establish the potential of Cad and Kni to interact with the cis-acting DNA that mediates hairy stripe 6-like expression in the embryo, in vitro footprinting experiments were performed with the 532 bp hairy stripe 6-element DNA. Cad and Kni bind to thirty six in vitro binding sites, some of which overlap, throughout the element. The sequence of the Cad and Kni binding sites matches the consensus described for each of the two proteins. Most of the potential Cad and Kni binding sites are close to or overlapped by binding sites for Kruppel (eight sites), Hunchback (eight sites), and Tailless (five sites). Tests using fragments of the 532 bp enhancer and of another element, 284-HT, show that sequences mediating activation of reporter expression are not maintained within a minimal activation element but instead are dispersed throughout the enhancer (Hader, 1998).

The results suggest that activation and the expression level mediated by the hairy stripe 6-element depend on the number of activator binding sites; both activation and expression level are likely to involve additive rather than synergistic interactions. An identical transacting factor requirement is found for hairy stripe 6 and 7 expression. The arrangement of the corresponding binding sites for the common factors involved in the control of the two stripes share a high degree of similarity, but some of the factors exert opposite regulatory functions within the two enhancer elements (Hader, 1998).

The two opposing gradients of Bicoid and Cad provide a complementing transcriptional activator system along the entire axis of the preblastoderm embryo necessary for proper hairy stripe expression. The importance of Caudal as a posterior activator had been overlooked for some time. This is because Bicoid can partially compensate for the role of Cad as an activator of posterior gap genes. The activating role of Cad in the posterior region of the embryo, already suggested in the context of the fushi tarazu cis-acting control element, is substantiated by misexpression studies on hunchback, a regulator of zygotic caudal expression. The results of the misexpression studies imply that gene activation in response to Cad involves the combined action of maternal and zygotic caudal activities. These results indicate, however, that zygotic caudal activity cannot compensate for the lack of maternal caudal activity in the case of hairy. While Bcd is both necessary and sufficient for the activation of anterior genes and may require co-activations such as HB for establishing proper expression domains, maternal Cad seems to act by providing a basal level of activation on which other factors, such as Bcd, Kni or Kr, act to set the biologically relevant time and level of gene expression (Hader, 1998 and references).

Exposure to high-sugar diet induces transgenerational changes in sweet sensitivity and feeding behavior via H3K27me3 reprogramming

Human health is facing a host of new threats linked to unbalanced diets, including high-sugar diet (HSD), which contributes to the development of both metabolic and behavioral disorders. Studies have shown that diet-induced metabolic dysfunctions can be transmitted to multiple generations of offspring and exert long-lasting health burden. Meanwhile, whether and how diet-induced behavioral abnormalities can be transmitted to the offspring remains largely unclear. This study showed that ancestral HSD exposure suppressed sweet sensitivity and feeding behavior in the offspring in Drosophila. These behavioral deficits were transmitted through the maternal germline and companied by the enhancement of H3K27me3 modifications. PCL-PRC2 complex, a major driver of H3K27 trimethylation, was upregulated by ancestral HSD exposure, and disrupting its activity eliminated the transgenerational inheritance of sweet sensitivity and feeding behavior deficits. Elevated H3K27me3 inhibited the expression of a transcriptional factor Cad and suppressed sweet sensitivity of the sweet-sensing gustatory neurons, reshaping the sweet perception and feeding behavior of the offspring. Taken together, this study uncovered a novel molecular mechanism underlying behavioral abnormalities spanning multiple generations of offspring upon ancestral HSD exposure, which would contribute to the further understanding of long-term health risk of unbalanced diet (Yang, 2023).

Transgenerational behavioral change is present in many animal species. In C. elegans, exposure to pathogenic threats induces avoidance memories which can transmit for four generations via sRNA signaling. In fruit flies, exposure to predatory wasps leads to the inheritance of ethanol preference for five generations via maternal NPF repression (Bozler, 2019). Such a 'behavior memory' may be evolutionarily beneficial in a sense to pre-adapt offspring for changing environmental conditions. Nevertheless, it may also lead to devastating effects in human health. Individuals who were exposed to the Dutch Famine in early gestation exhibited deficits in metabolism, cardiovascular health, and mental health. Similarly, mice exposed to traumatic experiences and drugs induce depressive-like or autism-like behaviors in their progeny (Yang, 2023).

Globally, HSD has become a routine of modern lifestyle, which is linked to various human diseases, including obesity, type 2 diabetes, and neurobiological diseases. HSD also induces many behavioral disorders in animal models such as feeding abnormalities and addiction-like behaviors. This present study uncovers that HSD not only affects flies' overall physiology and metabolism within the same generation, but also affects their sweet sensitivity and feeding behavior in a manner spanning multiple generations of offspring. If similar observations hold true in human society, HSD exposure may lead to an additional layer of health risk that needs to be recognized and addressed (Yang, 2023).

Mechanistically, the data indicate that elevated H3K27me3 modifications upon ancestral HSD exposure are the key epigenetic factors underlying the transgenerational regulations of sweet sensitivity and feeding behavior. HSD exposure enhances genome-wide H3K27me3 but not H3K27ac or H3K9me2/3 modifications in early embryos. E(z) and Pcl, the key components of PCL-PRC2 complex, play a crucial role in catalyzing trimethylation of the repressive chromatin marker histone H3 lysine 27, thus maintaining this imprinting during fly development. Perturbation of PCL-PRC2 complex, via both genetic and pharmacological approaches, blocks the transmission of such repressive histone imprinting to offspring and eliminates the transgenerational modulation of sweet sensitivity and feeding behavior. Such epigenetic modulations are transmitted via the female germline, and exert a long-lasting effect on the expression of Cad (and possibly other transcriptional regulators) in the offspring. Cad, a transcription factor belonging to the Hox family, regulates the sensitivity of sweet-sensing gustatory neurons and plays a role in modulating PER responses to sucrose (Yang, 2023).

Feeding behavior is tightly regulated by various factors, such as internal nutritional needs, overall physiological status, and environmental cues. However, most of these regulations are quite dynamic in nature and do not last for long. For example, in fruit flies energy shortage can trigger foraging and feeding behavior in the timescale of hours, which can be rapidly suppressed upon acquisition of desirable food sources. These regulations are often mediated by rapid-acting molecular and cellular mechanisms such as ion channels, hormones, and neuropeptides. HSD can also modulate feeding behavior in a dynamic manner. Moreover, this mechanism is reversible within the same generation. For example, the number of PLCβ2+ taste bud cells in the fungiform papilla decreased within 4 weeks of HSD exposure in mice and was completely restored within 4 weeks upon the removal of HSD. However, the present study reveals a highly persistent and heritable mechanism of transgenerational modulation of sweet taste perception and feeding behavior that can transmit through multiple generations without re-exposure to the original HSD treatment. These findings suggest that feeding behavior can be modulated by different regulatory mechanisms with distinct timescales in response to various types of environmental and internal state changes. The relationship between these different mechanisms will be of great interest to further study. For example, it remains unclear whether and how hormonal and neurotransmitter changes that occur upon HSD exposure in flies within the same generation contribute to the reprogramming of H3K27me3 that can last for multiple generations (Yang, 2023).

Based on the present study, several important questions remain to be answered. The first question is how HSD evaluates H3K27me3 modifications. Previous work reported that some dietary bioactive compounds could regulate histone modifying enzymes. Thus, HSD may directly regulate the activity of histone methyltransferases or histone demethylases. Alternatively, HSD may affect H3K27me3 modifications via certain nutrient-sensing mechanisms such as O-GlyNAc transferase, which is known to facilitate H3K27me3 formation with PRC2. The second question lies in how elevated H3K27me3 modifications in specific gene loci are retained during gamete formation and embryo development, and how such epigenetic imprinting is removed after four to five generations on ND. Previous work reported that long-term memory of H3K27me3 depends on efficient copying of this mark after each DNA replication cycle in a PRC2-dependent manner (Coleman, 2017). It is therefore possible that PRC2-mediated maintenance of H3K27me3 imprinting can be regulated by dietary exposure. The third question is how H3K27me3 imprinting affects specific neurons in the adult via Cad signaling. This study showed that upregulated H3K27me3 modifications decreased Cad expression. Future studies are needed to examine how Cad signaling impacts the development and function of Gr5a+ gustatory neurons, and whether other transcription factors may also be involved. Previous work has reported that Cad regulates a network containing 119 candidate genes that were implicated in sensory perception of chemical stimulus, neuropeptide signaling pathways, signal transduction, and transcription factor activity, which could impact the sweet gustatory neurons (Vaziri, 2020) and play a role in modulating PER responses to sucrose (Yang, 2023).

This study focused on the function of histone modifications on transgenerational behavioral inheritance. It will also be of interest to explore other mechanisms that may also participate in the transgenerational inheritance of sweet perception. Especially, as the Piwi protein negatively regulates H3K27 trimethylation, future studies are needed to understand whether piRNAs, which have been reported to direct transient heterochromatin formation and stabilize maternal mRNAs during embryogenesis, are involved in the transgenerational inheritance identified in this study. Given HSD exposure imposes a systemic influence on flies' physiology and metabolism, other non-epigenetic factors, such as maternal nutritional conditions and their potential influences on the development and maturation of oocytes, may also play a role in the transgenerational inheritance of sweet perception upon ancestral HSD exposure. Furthermore, the identification of HSD exposure-induced transgenerational changes in sweet sensitivity and feeding raises the question of whether similar phenomena and mechanism can be extended to other behaviors. These findings highlight a novel and pivotal role of epigenetic modifications in preparing animals for the dynamic environment, which opens a new avenue of research to further uncover the interactions among prior experience, epigenetics, and behavioral modulations across generations (Yang, 2023).


Length of Transcript - 2.6 (zygotic); 2.4 maternal

Bases in 5' UTR - 460; maternal transcript is shorter at 5' end

Exons - 2; Intron is 10.5k bases

Bases in 3' UTR - 619; maternal transcript is shorter at 3' end


Amino Acids - 472

The CAD homeodomain shares 58% homology to FTZ homeodomain and 53% homology to ANTP homeodomain (Mlodzik, 1985 and Macdonald, 1986).

caudal: Evolutionary Homologs | Regulation | Developmental Biology | Effects of Mutation | References

date revised:  7 October 2021

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