Developmental Biology Commons^™

The 14-Day Rule For In Vitro Human Embryo Research Requires Adjustment: From A Comprehensive Perspective Of Science, Ethics And Policy Research, Xiao Lu, Ming Zhao, Huihui Liu, Tianqing Li, Tao Tan, Weizhi Ji

Bulletin of Chinese Academy of Sciences (Chinese Version)

Since it was proposed, in vitro human embryo research had been following the 14-day rule, which limited in vitro research on human embryos within 14 days after fertilization. With the advancement in embryo culturing technology, this ethical rule has been greatly challenged. In 2021, the International Society for Stem Cell Research (ISSCR) recommended a conditional extension of the existing time limit in ISSCR Guidelines for Stem Cell Research and Clinical Translation, hence the international scientific community has restarted the debate on the 14-day rule. This study systematically analyzed the historical background and controversies of the 14-day rule ethical regulation of …

Brown Anole (Anolis Sagrei) Hoxa5: Insights Into The Divergence Of Hoxa5 Gene Expression And Regulation Across Evolutionarily Divergent Gnathostome Vertebrates, Jennifer A. Lange, Amber L. Rittgers, Adam Davis

Georgia Journal of Science

Hox genes are evolutionarily conserved developmental regulatory genes that function, in part, to pattern the anterior-posterior (AP) axis of organs and organ systems during animal embryonic development. Hoxa5, specifically, is shown to be expressed in the spinal cord, somites, or transient compartments giving rise to the vertebrae and ribs, developing gut, lungs, and limbs of the mouse (Mus musculus). The cis-regulatory elements (CREs), or short DNA sequences, that direct Hoxa5 expression in these embryonic domains have been mapped and functionally tested in the mouse as well. Similar Hoxa5 expression patterns have been observed in chicken ( …

Trimeric Complex Interactions Of Antp-Tbp With Tfiieb And Exd Are Involved In The Genetic Control Of Drosophila Melanogaster, Gustavo Jiménez Mejía, Ruben De Jesus Montalvo Mendez, Claudia Dalila Altamirano Torres, Diana Reséndez Pérez

Research Symposium

Background: Homeoproteins are transcriptional factors (TFs) that shape animal body axes during development. These TFs are highly conserved and represent one of the most fascinating groups of regulatory molecules. Reports shown the multiplicity of interactions in hox proteins, as complexes trimeric involved to transcriptional activity. The study of trimeric complexes in Hox interactome will allow the better understanding of Hox genetic regulation during embryonic development.

Methodology: Using a new combination BiFCFRET approach performed in HEK293, the quantification was performed by FRETTY of ImageJ. Fly crosses were incubated at 25ºC on standard yeast-agar-cornmeal medium. Embryo cuticle preparations were carried out according …

Tata-Box Binding Protein Interacts With Antp, Scr, Ubx And Abdb Through Their N-Terminal Domains, Rubén Montalvo Méndez, Gustavo Jiménez Mejía, Diana Reséndez Pérez

Research Symposium

Background: Hox proteins are transcriptional factors (TFs) that define segment identity during embryonic development regulating specific target genes. These TFs interact with cofactors for DNA specificity and other TFs to regulate gene expression, which include basal transcriptional machinery members like BIP2, Med19, TFIIEβ, M1BP and TBP. Since TBP glutamine homopeptide (PolyQ) act as an interaction domain involved transcriptional regulation, we analyzed if TBP interact with Antp, Scr, Ubx and AbdB through its PolyQ region.

Methods: We used Bimolecular Fluorescent Complementation (BiFC) to determine TBP interaction with Antp, Scr, Ubx and AbdB as well as the implication of their homeodomain (HD) …

Trimeric Interaction Of Antp-Ubx With Tbp And Homeoprotein Exd In The Genetic Control Of Development In D. Melanogaster, Alely Cecilia Villarreal-Puente, Claudia Dalila Altamirano-Torres, Diana Resendez-Perez

Research Symposium

Homeoproteins specify body segments along the anteroposterior axis during embryo development; they bind DNA through the homeodomain (HD). Homeoproteins bind to similar and repetitive target sequences on DNA, raising the question of how they achieve functional specificity. Homeoproteins form complexes with proteins through the HDs or other regions. Antennapedia (Antp) and Ultrabithorax (Ubx) have an important role in conferring thorax identity; also they are involved in a repression transcriptional mechanism: Ubx binds to Antp promoter repressing its expression. We analyzed if Ubx and Antp perform protein-protein interaction as a regulation mechanism and if the complex Antp-Ubx are involved in trimeric …

Examining Pi3k-Signaling-Dependent Regulation Of Lens Organelle Free Zone Formation Via Immunolocalization And Immunoblotting In Chick Embryos, Rifah Gheyas, A. Sue Menko

Computational Medicine Center Faculty Papers

The elimination of lens organelles during development, required for mature lens function, is an autophagy-dependent mechanism induced through suppression of PI3K signaling. Here, we present a protocol for investigating the signaling pathways responsible for induction of the formation of this lens organelle free zone. We describe steps for preparation of lens organ culture and use of signaling pathway inhibitors. We then detail procedures for analyzing their impact using both confocal microscopy imaging of immunolabeled lens cryosections and immunoblot approaches. For complete details on the use and execution of this protocol, please refer to Gheyas et al. (2022).

Canine Testes Thin Sections Culture​, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Canine Testes Thin Sections Culture, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Appendage Abnormalities In Spiders Induced By An Alternating Temperature Protocol In The Context Of Recent Advances In Molecular Spider Embryology, Teresa Napiorkowska, Julita Templin, Pawel Napiorkowski, Mark A. Townley

Faculty Publications

In the literature there are numerous reports of developmental deformities in arthropods collected in their natural habitat. Since such teratogenically affected individuals are found purely by chance, the causes of their defects are unknown. Numerous potential physical, mechanical, chemical, and biological teratogens have been considered and tested in the laboratory. Thermal shocks, frequently used in teratological research on the spider Eratigena atrica, have led to deformities on both the prosoma and the opisthosoma. In the 2020/2021 breeding season, by applying alternating temperatures (14 °C and 32 °C, changed every 12 h) for the first 10 days of embryonic development, …

Muscle Defects Lead To Skeletal Deformities In A Zebrafish Model Of Distal Arthrogryposis, Emily A. Tomak

Electronic Theses and Dissertations

Distal Arthrogryposis Type 1 (DA1) involves mild muscle weakness and limb skeletal abnormalities thought to be caused by paralysis in utero. Why the limbs are particularly affected in DA1 and the degree of paralysis that leads to these skeletal deformities in utero remains unclear. Several muscle genes are known to cause DA1, including MYLPF (myosin light chain phosphorylatable), which encodes a myosin light chain protein that binds close to the force-generating head of myosin heavy chains. The zebrafish mylpfa^-/- mutant displays a phenotype consistent with DA1, including impaired myosin activity, reduced muscle force overall, and complete fin paralysis. I …

Exploring The Impact Of Pqn-75 And Glh-1/Vasa On Germline Development, Maintenance, And Gsc Reprogramming Using Caenorhabditis Elegans As A Model, Jesse D. Rochester

Electronic Theses and Dissertations

This thesis combines research on PQN-75 expression, functional motifs of GLH-1/Vasa, and germ granule components in Caenorhabditis elegans to provide a comprehensive understanding of germline development, maintenance, and reprogramming, while also examining the role of pharyngeal gland cells in stress resistance and thermotolerance. In C. elegans, pharyngeal gland cells secrete mucin-like proteins, such as PQN-75, with similarities to human PRB2. The expression of PQN-75 in gland cells confers stress resistance and thermotolerance but does not affect fertility, instead it plays a role in the organism's ability to adapt to varying environmental conditions. While, GLH-1/Vasa, an ATP-dependent DEAD-box helicase, plays …

Phenotyping Regression In A Female Mouse Model For Rett Syndrome Using Computational Neuroethology Tools, Michael J. Mykins

Doctoral Dissertations

Regression is defined as loss of acquired skills over time and is a key feature of many neurodevelopmental disorders such as Rett syndrome (RTT). RTT is caused by mutations in the X-linked gene Methyl CpG-Binding Protein 2 (MECP2) and is characterized by a period of typical development with subsequent regression of previously acquired motor and speech skills in girls. In human and animal models, it is clear syndromic phenotypes are dynamic over time but phenotyping regression over time in animal models has remained elusive. Lack of established timelines to study the molecular, cellular, and behavioral features of regression in female …

Functional Analysis Of The Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (Gabra1) Gene During Zebrafish Development., Nayeli Gabriela Reyes-Nava

Open Access Theses & Dissertations

The GABRA1 gene encodes for the alpha-1 (α1) subunit of the Gamma-Aminobutyric acid type A receptor (GABAAR), which are the primary modulators of synaptic inhibition in the central nervous system (CNS). Alpha-1 subunits are essential for maintaining the normal function of native receptors and contribute to over 60% of all GABAARs in the CNS. Remarkably, a broad spectrum of neurodevelopmental and epilepsy-associated disorders have been linked with mutations in the GABRA1 gene. However, the developmental, behavioral, and molecular mechanisms underlying GABRA1-associated epileptic disorders remain to be fully understood. Hence, the overarching goal of this dissertation is to investigate the behavioral …

A Novel Role For Enos In Regulating Lymphatic Valve Development During Embryogenesis, Drishya Iyer

USF Tampa Graduate Theses and Dissertations

Lymphedema is a disease that occurs when lymph flow is impaired, resulting in tissue swelling, fibrosis, chronic inflammation and recurrent secondary infections. Lymphatic valves play a critical role in maintaining unidirectional lymph flow and evidence for valve defects have been reported in lymphedema patients. The lack of drugs that can correct lymphatic valve defects warrants a better understanding of the molecular regulators of lymphatic valve development and maintenance. Lymphatic valves first develop during embryogenesis in response to mechanotransduction signaling pathways triggered by oscillatory lymph flow. Since eNOS (gene name: Nos3) is a well characterized mechanotransduction signaling molecule in blood vessels, …

Evaluation Of Nutrient Digestibility Of Weaned Calves From Early And Late Shedding Dams, Jennifer Keele

Theses and Dissertations

Researchers have investigated several factors that could alter fetal growth, including nutrient restriction (Valiente et al., 2021), hair shedding (Gray et al., 2011), and extreme hot and cold temperatures (Davidson et al., 2022). Hot temperatures and increased humidity percentages in the southeast United States caused researchers to investigate the hair coats of Angus cattle in the commercial production setting. An improvement in fiber digestibility and calf birth and weaning weights has been observed in Angus dams that shed 50% of the winter hair coat by May (Gray et al., 2011; Burnett et al., 2021). Our objective of this experiment was …

Mitochondrial Roles In Developmentally Programmed Heart Disease, Eli John Louwagie

Dissertations and Theses

Offspring of diabetic and obese mothers (ODOM) have greater risks of heart disease at birth and later in life. However, prevention is hindered because underlying mechanisms are poorly understood. Mounting studies in the Developmental Origins of Health and Disease field suggest that mitochondria play key roles in developmentally programmed heart disease similar to the roles they play in cardiomyopathy in adults with diabetes and obesity. However, whether mitochondria are responsible for the short[1]and long-term cardiac disease seen in ODOM remains unknown. Here, we sought to delineate the roles of mitochondria in the hearts of ODOM, determine whether mitochondria are playing …

Measuring Energy Demands Of Dynamic Remodeling Events During Embryogenesis In Live Drosophila Embryos, Grayson Young

Biological Sciences Undergraduate Honors Theses

Embryonic development is a complex process requiring dynamic cell movements to create complex tissue structures. We assume that these embryonic dynamic remodeling events are highly energy intensive and that this energy expenditure is mediated by mitochondria. However, this has been difficult to demonstrate in live embryos. Convergent extension of the Drosophila neuroectoderm is a promising system to study the bioenergetics of tissue remodeling because it changes from a static to a dynamically remodeling tissue with temporal precision. We hypothesize that the switch from static to dynamic development at the onset of convergent extension will be accompanied by an increase in …

Examining Transcriptional Regulators During Muscle Development In Drosophila Melanogaster, Chaamy Yapa

Student Theses and Dissertations

In Drosophila melanogaster embryos, a distinct approach to study the transcriptional regulation is to examine the larval somatic muscle development. Transcription factors are essential regulatory proteins that help to control gene expression and respond to signaling pathways and various cues. Today, there are at least twenty transcription factors that have been discovered to contribute to the development of the 30 distinct larval somatic muscles in each abdominal hemisegment of Drosophila melanogaster. Several studies have already been conducted on muscle regulatory transcription factors including midline and apterous. These transcription factors were shown to control the development of muscles through mutant …

Investigation The Plugging Behavior Of Virus Filters, Iman Abdulqader Saab

Graduate Theses and Dissertations

A virus filtration step is integral to the manufacturing of biopharmaceuticals to ensure viral safety. A virus filter is a single-use device that uses a size-based separation process and has a unique structure with minimal defects, so that contaminating virus particles cannot pass through the membrane pores, while therapeutic molecules can. The development of novel antibodies (Abs), including significant increases in product titers, is frequently challenged by virus filter fouling, making a better understanding of the underlying mechanisms essential. This thesis focused investigating of the effect of prefilter types, buffer type and salt content on virus filtration performance. The impact …

Genetic Regulation Of Müllerian Duct Regression, Malcolm Moses

Dissertations and Theses (Open Access)

Mammals, including humans, develop progenitor tissues for both male and female reproductive tract organs before they fully differentiate into a male or female tract. The progenitor tissue for the male reproductive tract is known as the Wolffian duct (WD), and the progenitor tissue for the female reproductive tract is the Müllerian duct (MD). The WD further differentiates into the vas deferens, epididymis, and seminal vesicle, while the MD differentiates into the oviduct, uterus and upper vagina. An essential step in sex differentiation for males is the regression of the MD. This regression initiates with anti-Müllerian hormone (Amh) transcription …