Journal of Developmental Biology

14 pages, 1562 KiB

Open AccessArticle

Drosophila Males Differentially Express Small Proteins Regulating Stem Cell Division Frequency in Response to Mating

by Manashree S. Malpe, Leon F. McSwain, Heath M. Aston, Karl A. Kudyba, Chun Ng, Megan P. Wright and Cordula Schulz

J. Dev. Biol. 2025, 13(3), 21; https://doi.org/10.3390/jdb13030021 - 23 Jun 2025

Abstract

The germline stem cells (GSCs) in the male gonad of Drosophila can increase their division frequency in response to a demand for more sperm caused by repeated mating. However, the molecules and mechanisms regulating and mediating this response have yet to be fully [...] Read more.

The germline stem cells (GSCs) in the male gonad of Drosophila can increase their division frequency in response to a demand for more sperm caused by repeated mating. However, the molecules and mechanisms regulating and mediating this response have yet to be fully explored. Here, we present the results of a transcriptome analysis comparing expression from the testis tips from non-mated and mated males. An overlapping set of 18 differentially expressed genes (DEGs) from two independent wild-type (wt) strains revealed that the majority of the DEGs encode secreted proteins, which suggests roles for them in cell–cell interactions. Consistent with a role for secretion in regulating GSC divisions, knocking down Signal Recognition Particle (SRP) components within the germline cells using RNA Interference (RNAi), prevented the increase in GSC division frequency in response to mating. The major class of DEGs encodes polypeptides below the size of 250 amino acids, also known as small proteins. Upon reducing germline expression of small proteins, males no longer increased GSC division frequency after repeated mating. We hypothesize that mating induces cellular interactions via small proteins to ensure continued GSC divisions for the production of sperm. Full article

(This article belongs to the Special Issue Drosophila in Developmental Biology—Past, Present and Future)

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20 pages, 7139 KiB

Open AccessArticle

Cannabinoid Receptor 1 Regulates Zebrafish Renal Multiciliated Cell Development via cAMP Signaling

by Thanh Khoa Nguyen, Sophia Baker, Julienne Angtuaco, Liana Arceri, Samuel Kaczor, Bram Fitzsimonds, Matthew R. Hawkins and Rebecca A. Wingert

J. Dev. Biol. 2025, 13(2), 20; https://doi.org/10.3390/jdb13020020 - 17 Jun 2025

Abstract

Endocannabinoid signaling plays a significant role in neurogenesis and nervous system physiology, but its roles in the development of other tissues are just beginning to be appreciated. Previous reports have shown the presence of the key endocannabinoid receptor Cannabinoid receptor 1 (CB1 or [...] Read more.

Endocannabinoid signaling plays a significant role in neurogenesis and nervous system physiology, but its roles in the development of other tissues are just beginning to be appreciated. Previous reports have shown the presence of the key endocannabinoid receptor Cannabinoid receptor 1 (CB1 or Cnr1) in multiciliated (MCC) tissues and its upregulation in kidney diseases, yet the relationship between Cnr1 and renal MCC development is unknown. Here, we report that Cnr1 is essential for cilia development across tissues and regulates renal MCCs via cyclic AMP (cAMP) signaling during zebrafish embryogenesis. Using a combination of genetic and pharmacological studies, we found that the loss of function, agonism and antagonism of cnr1 all lead to reduced mature renal MCC populations. cnr1 deficiency also led to reduced cilia development across tissues, including the pronephros, ear, Kupffer’s vesicle (KV), and nasal placode. Interestingly, treatment with the cAMP activator Forskolin (FSK) restored renal MCC defects in agonist-treated embryos, suggesting that cnr1 mediates cAMP signaling in renal MCC development. Meanwhile, treatment with the cAMP inhibitor SQ-22536 alone or with cnr1 deficiency led to reduced MCC populations, suggesting that cnr1 also mediates renal MCC development independently of cAMP signaling. Our findings indicate that cnr1 has a critical role in controlling renal MCC development both via cAMP signaling and an independent pathway, further revealing implications for ciliopathies and renal diseases. Full article

(This article belongs to the Special Issue Feature Papers from Journal of Developmental Biology Reviewers)

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13 pages, 5576 KiB

Open AccessArticle

Ribosome Incorporation Transdifferentiates Chick Primary Cells and Induces Their Proliferation by Secreting Growth Factors

by Shota Inoue, Arif Istiaq, Anamika Datta, Mengxue Lu, Shintaro Nakayama, Kousei Takashi, Nobushige Nakajo, Shigehiko Tamura, Ikko Kawashima and Kunimasa Ohta

J. Dev. Biol. 2025, 13(2), 19; https://doi.org/10.3390/jdb13020019 - 1 Jun 2025

Abstract

Previously, we reported that mammalian cells, specifically human dermal fibroblasts (HDFs), could be transdifferentiated by lactic acid bacteria (LAB). Later, we observed that HDFs incorporated LAB-derived ribosomes, forming the ribosome-induced cell clusters (RICs) and transdifferentiating into cells derived from all three germ layers. [...] Read more.

Previously, we reported that mammalian cells, specifically human dermal fibroblasts (HDFs), could be transdifferentiated by lactic acid bacteria (LAB). Later, we observed that HDFs incorporated LAB-derived ribosomes, forming the ribosome-induced cell clusters (RICs) and transdifferentiating into cells derived from all three germ layers. Based on this insight, we hypothesized that incorporating ribosomes into non-mammalian cells could reveal the universality of this mechanism and open the door to commercial applications. Our current study demonstrates that ribosome incorporation can transdifferentiate chick primary muscle-derived cells (CMCs) into adipocytes, osteoblasts, and chondrocytes. Furthermore, the culture medium supernatant from ribosome-incorporated CMCs was found to significantly enhance CMC’s proliferation. RNA-seq analysis revealed that RICs-CMC exhibit increased expression of genes related to multi-lineage cell growth. In addition, we developed a novel technological shift in meat production—the “CulNet System”—which replicates organ interactions within mechanical systems for cell-cultured meat production. While significant efforts are still required to implement this technology in a cost-effective manner, we believe that combining the “CulNet System” with ribosome-incorporated multipotent cells that have prolonged culture capability could substantially improve the scalability and cost-effectiveness of cultured chicken meat production. This report highlights a promising approach for cell-culture-based meat production, offering a sustainable alternative to traditional methods. Full article

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12 pages, 1185 KiB

Open AccessArticle

Cornified Epithelial Teeth of Jawless Vertebrates Contain Proteins Similar to Keratin-Associated Proteins of Mammalian Skin Appendages

by Attila Placido Sachslehner, David A. D. Parry and Leopold Eckhart

J. Dev. Biol. 2025, 13(2), 18; https://doi.org/10.3390/jdb13020018 - 19 May 2025

Abstract

Keratins and keratin-associated proteins (KRTAPs) are the main components of mammalian nails and hair. Comparative genomics and gene expression studies have revealed that keratins are conserved in all vertebrates, whereas KRTAPs exist only in mammals. Recently, we found hair keratin-like cysteine-rich keratins in [...] Read more.

Keratins and keratin-associated proteins (KRTAPs) are the main components of mammalian nails and hair. Comparative genomics and gene expression studies have revealed that keratins are conserved in all vertebrates, whereas KRTAPs exist only in mammals. Recently, we found hair keratin-like cysteine-rich keratins in jawless vertebrates with confirmed expression in the cornified epithelial teeth of the sea lamprey (Petromyzon marinus). Here, we report that KRTAP-like proteins are also present in the horny teeth of the lamprey. Mass spectrometry-based proteomics identified proteins that share features with KRTAPs, such as high contents of cysteine and tyrosine residues, which support intermolecular interactions, and abundant glycine residues, which endow the proteins with flexibility. Genes encoding KRTAP-like proteins are arranged in a cluster in P. marinus, and the presence of at least one KRTAP-like protein is conserved in phylogenetically diverse species of lamprey, including Lampetra fluviatilis, Lethenteron reissneri, Geotria australis, and Mordacia mordax. The KRTAP-like genes of lampreys contain two exons, whereas mammalian KRTAPs have only a single exon. Although KRTAPs and KRTAP-like proteins are products of independent evolution, their common expression in cornified skin appendages suggests that they fulfill similar functions. Full article

(This article belongs to the Special Issue Feature Papers from Journal of Developmental Biology Reviewers)

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20 pages, 3146 KiB

Open AccessArticle

In Vitro Embryo Culture Impacts Heart Mitochondria in Male Adolescent Sheep

by Reza Amanollahi, Stacey L. Holman, Ashley S. Meakin, Monalisa Padhee, Kimberley J. Botting-Lawford, Song Zhang, Severence M. MacLaughlin, David O. Kleemann, Simon K. Walker, Jennifer M. Kelly, Skye R. Rudiger, I. Caroline McMillen, Michael D. Wiese, Mitchell C. Lock and Janna L. Morrison

J. Dev. Biol. 2025, 13(2), 17; https://doi.org/10.3390/jdb13020017 - 13 May 2025

Abstract

Assisted reproductive technology (ART)such as in vitro embryo culture (IVC), is widely used in human infertility treatments; however, its long-term effects on the cardiac health of offspring remain unclear. This study aimed to determine whether the effects of IVC on cardiac metabolism and [...] Read more.

Assisted reproductive technology (ART)such as in vitro embryo culture (IVC), is widely used in human infertility treatments; however, its long-term effects on the cardiac health of offspring remain unclear. This study aimed to determine whether the effects of IVC on cardiac metabolism and associated signaling pathways persist after birth into adolescence. Embryos were either transferred to an intermediate ewe (ET) or cultured in vitro in the absence (IVC) or presence of human serum (IVCHS) with methionine supplementation (IVCHS+M) for 6 days after mating. Naturally mated (NM) ewes were used as controls. Protein expression and hormone concentrations in the left ventricle (LV) were analyzed using Western blot and LC-MS/MS analyses, respectively. IVC was associated with sex-specific alterations in cardiac mitochondria, with males exhibiting reduced mitochondrial abundance. Cardiac protein expression of oxidative phosphorylation (OXPHOS) complexes 1 and 4 was reduced by IVC. Additionally, IVC reduced protein expression of PDK-4 and Mn-SOD in the IVCHS+M group, which may impact energy efficiency and defense against oxidative stress. These changes may predispose IVC offspring to cardiac oxidative stress and mitochondrial dysfunction, particularly in males. This study provides insights into the sex-dependent effects of IVC on cardiac health, emphasizing the importance of evaluating long-term cardiovascular risks associated with IVC protocols. Full article

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14 pages, 12403 KiB

Open AccessArticle

Dynamic Changes of Immunoreactive CD34, CD117, and CD41 Hematopoietic Stem Cells in Human Placentas of Different Gestational Ages

by Sanja Jovicic, Ivan R. Nikolic, Ljiljana Amidžić, Vesna Ljubojevic, Maja Barudzija and Ranko Skrbic

J. Dev. Biol. 2025, 13(2), 16; https://doi.org/10.3390/jdb13020016 - 9 May 2025

Abstract

Background: The process of prenatal hematopoiesis occurs in various anatomical locations, including the placenta. The placenta is not merely a temporary hematopoietic reservoir, but it is one of the key sites for the synthesis of hematopoietic stem cells (HSCs). This study aimed [...] Read more.

Background: The process of prenatal hematopoiesis occurs in various anatomical locations, including the placenta. The placenta is not merely a temporary hematopoietic reservoir, but it is one of the key sites for the synthesis of hematopoietic stem cells (HSCs). This study aimed to investigate the presence, distribution, and immunoprofiles of HSCs in the human placenta during different gestational periods. Materials and Methods: Placental samples of different gestational ages (first, second, and third trimesters) were analyzed using classical hematoxylin and eosin staining and immunohistochemical staining for CD34, CD117, and CD41 markers, with HSC quantification through numerical areal density (N_A). Results: Highly immunoreactive CD34 HSCs were present in placentas throughout gestation, while highly immunoreactive CD117 and CD41 HSCs were observed during the first two trimesters. In the first trimester, HSCs were found within the lumen of blood vessels and as individual cells in the mesenchyme of chorionic villi. With advancing gestation, the number of HSCs in the mesenchyme of chorionic villi increased. Conclusions: Immunoreactive CD34, CD117, and CD41 cells are present in significant proportions in various parts of the placenta throughout gestation, indicating that the placenta provides a substantial proportion of HSCs for hematopoiesis. Full article

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18 pages, 10507 KiB

Open AccessArticle

Probe Sequencing Analysis of Regenerating Lizard Tails Indicates Crosstalk Among Osteoclasts, Epidermal Cells, and Fibroblasts

by Darian J. Gamble, Samantha Lopez, Melody Yazdi, Toni Castro-Torres and Thomas P. Lozito

J. Dev. Biol. 2025, 13(2), 15; https://doi.org/10.3390/jdb13020015 - 3 May 2025

Abstract

Lizards are distinguished as the only amniotes, and closest relatives of mammals, capable of multilineage epimorphic regeneration. Tail blastemas of green anole lizards (Anolis carolinensis) consist of col3a1⁺ fibroblastic connective tissue cells enclosed in krt5⁺ wound epidermis (WE), both [...] Read more.

Lizards are distinguished as the only amniotes, and closest relatives of mammals, capable of multilineage epimorphic regeneration. Tail blastemas of green anole lizards (Anolis carolinensis) consist of col3a1⁺ fibroblastic connective tissue cells enclosed in krt5⁺ wound epidermis (WE), both of which are required for regeneration. Blastema and WE formation are known to be closely associated with phagocytic cell populations, including macrophages and osteoclasts. However, it remains unclear what specific phagocytic cell types are required to stimulate regeneration. Here, we explicitly assess the roles of osteoclast activity during blastema and WE formation in regenerating lizard tails. First, probe sequencing was performed at regenerative timepoints on fibroblasts isolated based on col3a1 expression toward establishing pathways involved in stimulating blastema formation and subsequent tail regrowth. Next, treatments with osteoclast inhibitor zoledronic acid (ZA) were used to assess the roles of osteoclast activity in lizard tail regeneration and fibroblast signaling. ZA treatment stunted lizard tail regrowth, suggesting osteoclast activity was required for blastema formation and regeneration. Transcriptomic profiling of fibroblasts isolated from ZA-treated and control lizards linked inhibition of osteoclast activity with limitations in fibroblasts to form pro-regenerative extracellular matrix and support WE formation. These results suggest that crosstalk between osteoclasts and fibroblasts regulates blastema and WE formation during lizard tail regeneration. Full article

(This article belongs to the Special Issue Skin Wound Healing and Regeneration in Vertebrates)

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12 pages, 907 KiB

Open AccessArticle

Follicular Fluid from Cows That Express Estrus During a Fixed-Time Artificial Insemination Protocol Promotes Blastocyst Development

by Audra W. Harl, Verónica M. Negrón-Pérez, Jacob W. Stewart, George A. Perry, Alan D. Ealy and Michelle L. Rhoads

J. Dev. Biol. 2025, 13(2), 14; https://doi.org/10.3390/jdb13020014 - 25 Apr 2025

Abstract

It is not yet understood why cows that exhibit estrus and ovulate are more likely to become pregnant than those that ovulate but do not exhibit estrus during a fixed-time artificial insemination (FTAI) protocol. The objective of this work was to determine whether [...] Read more.

It is not yet understood why cows that exhibit estrus and ovulate are more likely to become pregnant than those that ovulate but do not exhibit estrus during a fixed-time artificial insemination (FTAI) protocol. The objective of this work was to determine whether the follicular fluid from cows that exhibit estrus contributes to the increased likelihood of pregnancy. Lactating crossbred cows were subjected to an FTAI estrous synchronization protocol. Estrous behavior was observed and recorded prior to transvaginal follicle aspiration from cows that did (estrus, n = 7) or did not exhibit estrus (non-estrus, n = 6). Follicular fluid (25%) was then added to in vitro maturation media for the maturation of oocytes (n = 1489) from slaughterhouse ovaries. Cleavage rates were not affected by the estrous status of the cows from which the follicular fluid was collected. Blastocyst rates, however, were greater following maturation in the presence of follicular fluid from estrus cows compared to non-estrus cows (p ≤ 0.01). This difference in blastocyst rates was not related to blastocyst cell numbers (inner cell mass, trophoblast, and total), as they did not differ between estrus and non-estrus animals. This study demonstrates that the follicular fluid, and thus, the follicular environment just prior to ovulation does indeed contribute to improved pregnancy rates following FTAI. Full article

(This article belongs to the Special Issue Feature Papers in Journal of Developmental Biology 2025)

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16 pages, 1521 KiB

Open AccessPerspective

Origins of Aortic Coarctation: A Vascular Smooth Muscle Compartment Boundary Model

by Christina L. Greene, Geoffrey Traeger, Akshay Venkatesh, David Han and Mark W. Majesky

J. Dev. Biol. 2025, 13(2), 13; https://doi.org/10.3390/jdb13020013 - 18 Apr 2025

Abstract

Compartment boundaries divide the embryo into segments with distinct fates and functions. In the vascular system, compartment boundaries organize endothelial cells into arteries, capillaries, and veins that are the fundamental units of a circulatory network. For vascular smooth muscle cells (SMCs), such boundaries [...] Read more.

Compartment boundaries divide the embryo into segments with distinct fates and functions. In the vascular system, compartment boundaries organize endothelial cells into arteries, capillaries, and veins that are the fundamental units of a circulatory network. For vascular smooth muscle cells (SMCs), such boundaries produce mosaic patterns of investment based on embryonic origins with important implications for the non-uniform distribution of vascular disease later in life. The morphogenesis of blood vessels requires vascular cell movements within compartments as highly-sensitive responses to changes in fluid flow shear stress and wall strain. These movements underline the remodeling of primitive plexuses, expansion of lumen diameters, regression of unused vessels, and building of multilayered artery walls. Although the loss of endothelial compartment boundaries can produce arterial–venous malformations, little is known about the consequences of mislocalization or the failure to form SMC-origin-specific boundaries during vascular development. We propose that the failure to establish a normal compartment boundary between cardiac neural-crest-derived SMCs of the 6th pharyngeal arch artery (future ductus arteriosus) and paraxial-mesoderm-derived SMCs of the dorsal aorta in mid-gestation embryos leads to aortic coarctation observed at birth. This model raises new questions about the effects of fluid flow dynamics on SMC investment and the formation of SMC compartment borders during pharyngeal arch artery remodeling and vascular development. Full article

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15 pages, 14856 KiB

Open AccessArticle

Activation of Marck-like Genes and Proteins During Initial Phases of Regeneration in the Amputated Tail and Limb of the Lizard Podarcis muralis

by Lorenzo Alibardi

J. Dev. Biol. 2025, 13(2), 12; https://doi.org/10.3390/jdb13020012 - 14 Apr 2025

Abstract

Molecules involved in the activation of regeneration in reptiles are almost unknown. MARCK-like proteins are indicated to activate regeneration in some amphibians and fish, and it would be important to know whether this is a general process also present in other vertebrates. To [...] Read more.

Molecules involved in the activation of regeneration in reptiles are almost unknown. MARCK-like proteins are indicated to activate regeneration in some amphibians and fish, and it would be important to know whether this is a general process also present in other vertebrates. To address this problem, the present study reports the immunolocalization of a MARCK-like protein in injured tissues of a lizard. Bioinformatics and immunofluorescence after 5BrdU administration, and detection of MARCK-like proteins, have been performed on regenerating tail and limb of the lizard Podarcis muralis. Transcriptome data indicate up-regulation of MARCKS and MARCK-like1 expression in the initial regenerating tail and limb blastemas, supporting their involvement in the activation of regeneration in both appendages. Immunofluorescence for 5BrdU shows numerous proliferating cells in the blastemas of both appendages. Immunolocalization of a MARCK-like protein, using an antibody generated against a homologous protein from the axolotl, shows that the wound epidermis, nerves, and myotubes accumulate most of the protein in the limb and tail. MARCK-like immunolabeling is also detected in the regenerating spinal cord of the tail. The study indicates that, although the limb later turns into a scar, the MARCK-like protein is also up-regulated in this appendage, like in the regenerating tail. These results indicate that the initial reaction to an injury in lizards, an amniote representative, includes some triggering processes observed in amphibians and fish (anamniotes), with the activation of MARCK-like genes and proteins. This suggests that a MARCK-like-dependant mechanism for tissue repair is likely activated during the initial phases of vertebrate wound healing. Full article

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18 pages, 1690 KiB

Open AccessReview

Super-Enhancers in Placental Development and Diseases

by Gracy X. Rosario, Samuel Brown, Subhradip Karmakar, Mohammad A. Karim Rumi and Nihar R. Nayak

J. Dev. Biol. 2025, 13(2), 11; https://doi.org/10.3390/jdb13020011 - 9 Apr 2025

Abstract

The proliferation of trophoblast stem (TS) cells and their differentiation into multiple lineages are pivotal for placental development and functions. Various transcription factors (TFs), such as CDX2, EOMES, GATA3, TFAP2C, and TEAD4, along with their binding sites and cis-regulatory elements, have been studied [...] Read more.

The proliferation of trophoblast stem (TS) cells and their differentiation into multiple lineages are pivotal for placental development and functions. Various transcription factors (TFs), such as CDX2, EOMES, GATA3, TFAP2C, and TEAD4, along with their binding sites and cis-regulatory elements, have been studied for their roles in trophoblast cells. While previous studies have primarily focused on individual enhancer regions in trophoblast development and differentiation, recent attention has shifted towards investigating the role of super-enhancers (SEs) in different trophoblast cell lineages. SEs are clusters of regulatory elements enriched with transcriptional regulators, forming complex gene regulatory networks via differential binding patterns and the synchronized stimulation of multiple target genes. Although the exact role of SEs remains unclear, they are commonly found near master regulator genes for specific cell types and are implicated in the transcriptional regulation of tissue-specific stem cells and lineage determination. Additionally, super-enhancers play a crucial role in regulating cellular growth and differentiation in both normal development and disease pathologies. This review summarizes recent advances on SEs’ role in placental development and the pathophysiology of placental diseases, emphasizing the potential for identifying SE-driven networks in the placenta to provide valuable insights for developing therapeutic strategies to address placental dysfunctions. Full article

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15 pages, 3888 KiB

Open AccessArticle

Wound-Induced Regeneration in Feather Follicles: A Stepwise Strategy to Regenerate Stem Cells

by Ting-Xin Jiang, Ping Wu, Ang Li, Randall B. Widelitz and Cheng-Ming Chuong

J. Dev. Biol. 2025, 13(2), 10; https://doi.org/10.3390/jdb13020010 - 27 Mar 2025

Abstract

How to elicit and harness regeneration is a major issue in wound healing. Skin injury in most amniotes leads to repair rather than regeneration, except in hair and feathers. Feather follicles are unique organs that undergo physiological cyclic renewal, supported by a dynamic [...] Read more.

How to elicit and harness regeneration is a major issue in wound healing. Skin injury in most amniotes leads to repair rather than regeneration, except in hair and feathers. Feather follicles are unique organs that undergo physiological cyclic renewal, supported by a dynamic stem cell niche. During normal feather cycling, growth-phase proximal follicle collar bulge stem cells adopt a ring configuration. At the resting and initiation phases, these stem cells descend to the dermal papilla to form papillary ectoderm and ascend to the proximal follicle in a new growth phase. Plucking resting-phase feathers accelerates papillary ectoderm cell activation. Plucking growth-phase feathers depletes collar bulge stem cells; however, a blastema reforms the collar bulge stem cells, expressing KRT15, LGR6, Sox9, integrin-α6, and tenascin C. Removing the follicle base and dermal papilla prevents feather regeneration. Yet, transplanting an exogenous dermal papilla to the follicle base can induce re-epithelialization from the lower follicle sheath, followed by feather regeneration. Thus, there is a stepwise regenerative strategy using stem cells located in the collar bulge, papillary ectoderm, and de-differentiated lower follicle sheath to generate new feathers after different levels of injuries. This adaptable regenerative mechanism is based on the hierarchy of stem cell regenerative capacity and underscores the remarkable resilience of feather follicle regenerative abilities. Full article

(This article belongs to the Special Issue Skin Wound Healing and Regeneration in Vertebrates)

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14 pages, 3521 KiB

Open AccessArticle

Varanid Teeth Asymmetry and Correlation to Body Size

by Guy Sion and Domenic C. D’Amore

J. Dev. Biol. 2025, 13(1), 9; https://doi.org/10.3390/jdb13010009 - 10 Mar 2025

Abstract

Stressors such as injuries, embryonic instability during development, and higher levels of stress hormones such as testosterone can result in increases in fluctuating asymmetry in reptiles and other vertebrates. Digit asymmetry, digit ratio variability, and skull trait asymmetry such as eye and jaw [...] Read more.

Stressors such as injuries, embryonic instability during development, and higher levels of stress hormones such as testosterone can result in increases in fluctuating asymmetry in reptiles and other vertebrates. Digit asymmetry, digit ratio variability, and skull trait asymmetry such as eye and jaw size have been correlated with stress level in both snakes and lizards. Teeth asymmetry has also been used as a biomarker for stress and brain laterality. Body size is correlated with many potential stressors, yet there has been little research on how body size in reptiles relates to asymmetry. We investigate teeth asymmetry within the lizard family Varanidae, a clade with a diverse range of sizes consisting of the largest living lizard, Varanus komodoensis. Using a landmark/semi-landmark analysis, we derived Centroid Size for 671 pairs of teeth from 13 varanid species, and asymmetry was derived for each pair. Right-biased asymmetry was significantly greater in the upper tooth row, but breaking up tooth positions into further sections did not yield a significant difference. We found a significant positive linear correlation between body size and right-biased teeth directional asymmetry within Varanus, but only when excluding V. komodoensis. This significant correlation may result from fewer potential predators and more potential food items, thus resulting in less overall stress. When analyzed separately, V. komodoensis individuals with <180 mm head length demonstrated a positive, yet non-significant, trend along a similar trajectory to their congenerics with a high goodness of fit. On the other hand, individuals > 180 mm showed a high degree of scatter, with several specimens having pronounced left-biased asymmetry. We suspect that this dramatic change was due to a combination of ontogenetic niche shift, bigger home ranges, a greater susceptibility to negative anthropogenic influences, and/or a male bias in the bigger specimens sampled, but a larger sample size is required to determine if there is statistical significance in these intra-specific trends. Body asymmetry can reflect brain laterality, which may be a potential driver for the teeth asymmetry seen here. Full article

(This article belongs to the Special Issue Feature Papers from Journal of Developmental Biology Reviewers)

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22 pages, 3887 KiB

Open AccessArticle

Effects of Molybdenum Supplementation in the Form of Ammonium and Sodium Salts on Trophoblast Cell Physiology and Gene Expression In Vitro

by Vladimira Foteva, Joshua J. Fisher, Yixue Qiao and Roger Smith

J. Dev. Biol. 2025, 13(1), 8; https://doi.org/10.3390/jdb13010008 - 5 Mar 2025

Abstract

Molybdenum is an essential trace element sourced during pregnancy from the maternal diet. Studies regarding molybdenum have primarily focused on overexposure in animal and cell culture studies. The effects of molybdenum supplementation on placental function are unknown. An immortalised trophoblast cell line was [...] Read more.

Molybdenum is an essential trace element sourced during pregnancy from the maternal diet. Studies regarding molybdenum have primarily focused on overexposure in animal and cell culture studies. The effects of molybdenum supplementation on placental function are unknown. An immortalised trophoblast cell line was used to examine the placental cellular response to molybdenum in its bioavailable form as molybdate. Cells of the extravillous trophoblast first-trimester cell line HTR8-SVneo were cultured in complete cell media in the presence of 10 nM to 1 mM of ammonium molybdate or sodium molybdate. Following the addition of the molybdate salts, cell growth, viability, and several gene pathways were monitored. Sodium molybdate salt in doses from 10 nM to 1 mM did not affect cell growth or viability. Exposure to ammonium molybdate at a 1 mM concentration significantly decreased cell growth and viability (p < 0.05). Gene pathways involving molybdoenzyme expression, molybdenum cofactor synthesis, antioxidant response, and angiogenesis were affected following supplementation, although these effects differed depending on the dose and molybdate salt utilised. Molybdoenzyme activity was not affected by supplementation in a dose-dependent manner. The results indicate sodium molybdate is a more appropriate salt to use in vitro, as ammonium molybdate exposure reduced cell viability and growth and downregulated the expression of antioxidant genes NFE2L2 (p < 0.01), SOD1 (p < 0.001) and SOD2 (p < 0.001), suggestive of an inflammatory response. Sodium molybdate affected gene, protein, and activity levels of molybdoenzyme, antioxidant, and angiogenic molecules in vitro. This work demonstrates that sodium molybdate supplementation has pleiotropic effects in vitro and is well tolerated by placental cells at a range of nanomolar and micromolar concentrations. Full article

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5 pages, 159 KiB

Open AccessCorrection

Correction: Ko et al. Timing of Mouse Molar Formation Is Independent of Jaw Length Including Retromolar Space. J. Dev. Biol. 2021, 9, 8

by Daisy (Jihyung) Ko, Tess Kelly, Lacey Thompson, Jasmene K. Uppal, Nasim Rostampour, Mark Adam Webb, Ning Zhu, George Belev, Prosanta Mondal, David M. L. Cooper and Julia C. Boughner

J. Dev. Biol. 2025, 13(1), 7; https://doi.org/10.3390/jdb13010007 - 28 Feb 2025

Abstract

There was an error in the original publication [...] Full article

13 pages, 1356 KiB

Open AccessArticle

Changes in the Intracellular Composition of Macro and Microminerals After Cryopreservation of the Rabbit Stem/Progenitor Cells

by Jaromír Vašíček, Andrej Baláži, Mária Tirpáková, Marián Tomka and Peter Chrenek

J. Dev. Biol. 2025, 13(1), 6; https://doi.org/10.3390/jdb13010006 - 21 Feb 2025

Abstract

Cryopreservation is a widely used method for the long-term preservation of reproductive or somatic cells. It is known that this storage method may negatively affect cell viability, proliferation, differentiation, etc. However, there is a lack of information about whether cryostorage can alter the [...] Read more.

Cryopreservation is a widely used method for the long-term preservation of reproductive or somatic cells. It is known that this storage method may negatively affect cell viability, proliferation, differentiation, etc. However, there is a lack of information about whether cryostorage can alter the content of intracellular minerals. Therefore, we focused this study on the analysis of the mineral composition of living cells before and after long-term cold storage. Briefly, three different primary cell lines were established from rabbits as follows: endothelial progenitor cells from peripheral blood (EPCs), endothelial progenitor cells from bone marrow (BEPCs), and mesenchymal stem cells from adipose tissue (AT-MSCs), which were cultured until passage 3 prior to cryopreservation in liquid nitrogen. Samples from freshly cultured and frozen–thawed cells were mineralized and analyzed using inductively coupled plasma-optical emission spectroscopy (ICP-OES) for the content of minerals (macro: Ca, Na, K, and Mg, and micro: Zn, Fe, Cu, Al, Co, Mn, Sr, and Ni). After cryopreservation, we found significantly decreased content of K in frozen–thawed EPCs (p < 0.01) and BEPCs (p < 0.0001) and Ca in AT-MSCs (p < 0.05), while Na was increased in frozen–thawed BEPCs (p < 0.05). Concentrations of Fe and Al were reduced significantly in frozen–thawed EPCs (both p < 0.0001) and AT-MSCs (p < 0.001 and p < 0.0001, respectively). On the contrary, Fe and Al were elevated in frozen–thawed BEPCs (p < 0.0001 and p < 0.01, respectively) together with Ni (p < 0.0001). In addition, decreased Zn (p < 0.05) was observed in cryopreserved AT-MSCs. In conclusion, the ICP-OES technique might be used to analyze the basic elemental composition of animal cells in fresh or frozen–thawed conditions. Nevertheless, additional studies are needed to reveal the possible impact of cryopreservation on cell fate by changing the content of intracellular minerals. Full article

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24 pages, 19401 KiB

Open AccessArticle

CRISPR/Cas9-Targeted Myostatin Deletion Improves the Myogenic Differentiation Parameters for Muscle-Derived Stem Cells in Mice

by Mohamed I. Elashry, Victoria C. Schneider, Manuela Heimann, Sabine Wenisch and Stefan Arnhold

J. Dev. Biol. 2025, 13(1), 5; https://doi.org/10.3390/jdb13010005 - 11 Feb 2025

Cited by 1

Abstract

Skeletal muscle plays a pivotal role in physical activity, protein storage and energy utilization. Skeletal muscle wasting due to immobilization, aging, muscular dystrophy and cancer cachexia has negative impacts on the quality of life. The deletion of myostatin, a growth and differentiation factor-8 [...] Read more.

Skeletal muscle plays a pivotal role in physical activity, protein storage and energy utilization. Skeletal muscle wasting due to immobilization, aging, muscular dystrophy and cancer cachexia has negative impacts on the quality of life. The deletion of myostatin, a growth and differentiation factor-8 (GDF-8) augments muscle mass through hyperplasia and hypertrophy of muscle fibers. The present study examines the impact of myostatin deletion using CRISPR/Cas9 editing on the myogenic differentiation (MD) of C2C12 muscle stem cells. A total of five myostatin loci were targeted using guided RNAs that had been previously cloned into a vector. The clones were transfected in C2C12 cells via electroporation. The cell viability and MD of myostatin-edited clones (Mstn^−/−) were compared with C2C12 (Mstn^+/+) using a series of assays, including MTT, sulforhodamine B, immunocytochemistry, morphometric analysis and RT-qPCR. The clones sequenced showed evidence of nucleotides deletion in Mstn^−/− cells. Mstn^−/− cells demonstrated a normal physiological performance and lack of cytotoxicity. Myostatin depletion promoted the myogenic commitment as evidenced by upregulated MyoD and myogenin expression. The number of MyoD-positive cells was increased in the differentiated Mstn^−/− clones. The Mstn^−/− editing upregulates both mTOR and MyH expression, as well as increasing the size of myotubes. The differentiation of Mstn^−/− cells upregulates ActRIIb; in contrast, it downregulates decorin expression. The data provide evidence of successful CRISPR/Cas9-mediated myostatin deletion. In addition, targeting myostatin could be a beneficial therapeutic strategy to promote MD and to restore muscle loss. In conclusion, the data suggest that myostatin editing using CRISPR/Cas9 could be a potential therapeutic manipulation to improve the regenerative capacity of muscle stem cells before in vivo application. Full article

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38 pages, 3633 KiB

Open AccessReview

Utilizing C. elegans Spermatogenesis and Fertilization Mutants as a Model for Human Disease

by Sofia M. Perez, Helena S. Augustineli and Matthew R. Marcello

J. Dev. Biol. 2025, 13(1), 4; https://doi.org/10.3390/jdb13010004 - 25 Jan 2025

Abstract

The nematode C. elegans is a proven model for identifying genes involved in human disease, and the study of C. elegans reproduction, specifically spermatogenesis and fertilization, has led to significant contributions to our understanding of cellular function. Approximately 70 genes have been identified [...] Read more.

The nematode C. elegans is a proven model for identifying genes involved in human disease, and the study of C. elegans reproduction, specifically spermatogenesis and fertilization, has led to significant contributions to our understanding of cellular function. Approximately 70 genes have been identified in C. elegans that control spermatogenesis and fertilization (spe and fer mutants). This review focuses on eight genes that have human orthologs with known pathogenic phenotypes. Using C. elegans to study these genes has led to critical developments in our understanding of protein domain function and human disease, including understanding the role of OTOF (the ortholog of C. elegans fer-1) in hearing loss, the contribution of the spe-39 ortholog VIPAS39 in vacuolar protein sorting, and the overlapping functions of spe-26 and KLHL10 in spermatogenesis. We discuss the cellular function of both the C. elegans genes and their human orthologs and the impact that C. elegans mutants and human variants have on cellular function and physiology. Utilizing C. elegans to understand the function of the genes reviewed here, and additional understudied and undiscovered genes, represents a unique opportunity to understand the function of variants that could lead to better disease diagnosis and clinical decision making. Full article

(This article belongs to the Special Issue Caenorhabditis elegans – a Model for Understanding Development and Disease)

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17 pages, 5922 KiB

Open AccessArticle

Neuronal Populations Involved in Motor Function Show Prominent Expression of Sbno1 During Postnatal Brain Development

by Sunjidmaa Zolzaya, Dai Ihara, Munkhsoyol Erkhembaatar, Shinsuke Ochiai, Ayaka Isa, Mariko Nishibe, Jean-Pierre Bellier, Takahiro Shimizu, Satoshi Kikkawa, Ryo Nitta and Yu Katsuyama

J. Dev. Biol. 2025, 13(1), 3; https://doi.org/10.3390/jdb13010003 - 21 Jan 2025

Abstract

Human genome studies have suggested that strawberry notch homologue 1 (SBNO1) is crucial for normal brain development, with mutations potentially contributing to neurodevelopmental disorders. In a previous study, we observed significant developmental abnormalities in the neocortex of Sbno1 as early as [...] Read more.

Human genome studies have suggested that strawberry notch homologue 1 (SBNO1) is crucial for normal brain development, with mutations potentially contributing to neurodevelopmental disorders. In a previous study, we observed significant developmental abnormalities in the neocortex of Sbno1 as early as one week after birth. In the present study, we conducted an extensive analysis of Sbno1 postnatal expression in the brain of C57BL/6 mice using a newly developed in-house polyclonal antibody against Sbno1. We found that Sbno1 is expressed in all neurons, with certain neuronal populations exhibiting distinct dynamic changes (both temporal and spatial) in expression level. These findings suggest that the neuronal expression of Sbno1 is developmentally regulated after birth. They also indicate that while Sbno1 may play a general role across all neurons, it may also serve more specialized functions in certain neuronal types and/or for certain cellular activities related to particular neuronal pathways. Full article

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