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New Players in the Mitophagy Game

Tuesday, March 3, 2020 - 11:03
Adenine nucleotide translocase 1 or ADP/ATP translocase (ANT) is expressed in the inner mitochondrial membrane where it exchanges free ATP and ADP from and into the mitochondrial matrix, respectively.

By Christina Towers, PhD

Mitochondrial turn over via the lysosome, otherwise known as mitophagy, involves engulfment of mitochondria into double membrane autophagosomes and subsequent fusion with lysosomes. Much is already known about this process including the canonical and non-canonical mechanisms of action, the critical machinery involved, as well as the...

Insulin signaling in adipocytes: Carbohydrate-signaling transcription factor ChREBP is the link between lipolytic enzyme Hormone-Sensitive Lipase and lipogenic enzyme ELOVL6

Tuesday, February 18, 2020 - 08:43
Fatty acid binding protein 4 (FAB4) is expressed in adipocytes where it activates hormone-sensitive lipase and helps regulate lipolysis.

By Jamshed Arslan, Pharm. D., PhD.

Insulin resistance in adipocytes is a major feature of metabolic syndrome   . Disrupted adipose tissue metabolism can lead to accumulation of lipid intermediates in insulin-sensitive tissues like liver and skeletal muscles, thereby diminishing insulin sensitivity. A key enzyme in adipocytes that mobilizes free fatty acids from adipose tissue into the bloodstream is...

The Gut-Brain Axis Effects in Autism Spectrum Disorders

Monday, February 3, 2020 - 11:43
Learn about different signaling pathways associated with autism spectrum disorder, a neurodevelopmental condition that affects how people learn, communicate and interact with others.

By Michalina Hanzel, PhD

The microbiota-gut-brain axis is a bidirectional relationship between the central and enteric nervous systems, which is heavily influenced by the microorganisms living in the gut. Accumulating evidence has pointed to the microbiome, composed mostly of bacteria, but also including archaea, fungi and viruses, as instrumental in maintaining human physiological functions.1 Importantly, the brain is influenced by various bacterial metabolites,...

Inhibiting incretin GIP hormone activity in mouse and monkey models to combat obesity

Monday, January 20, 2020 - 10:52
The release of insulin from pancreas beta-cells is controlled by glucose levels, Vagus nerve input, and GLP-1 from the intestines.

By Jamshed Arslan, Pharm. D., PhD.

We live in a world where 39% of adults are overweight. Our meals trigger the secretion of various gut-derived metabolic hormones called incretins. Fats and carbohydrates in our diet stimulate the release of an incretin from the duodenum called glucose-dependent insulinotropic polypeptide or gastric inhibitory polypeptide (GIP). GIP acts on its receptor (GIPR) in adipocytes and pancreas to promote fatty acid uptake and insulin...

Autophagy Research Update: What a difference a year makes!

Thursday, January 2, 2020 - 10:07
Autophagy Handbook provides an overview of molecular players and regulatory mechanisms involved in the process of autophagy.

By Christina Towers, PhD

Over the last two decades the field of autophagy has exploded! Innovative techniques, comprehensive analysis and disease-relevant models have yielded basic and clinical discoveries of consequence. The Nobel Prize for Physiology or Medicine was awarded to Dr. Yoshinori Ohsumi in 2016, for his ground-breaking work in yeast to identify many of the core autophagy proteins. Since then, the field has only expanded. In just the past year, the field has produced numerous...

Insulin signaling in brain’s subfornical organ is crucial for regulating cardiometabolic profile

Monday, December 16, 2019 - 09:54
Learn about cardiovascular biology resources from Bio-Techne

By Jamshed Arslan, Pharm. D., PhD.

The hypothalamus is an insulin receptor-rich brain region. Insulin receptor signaling in the CNS can regulate blood pressure, for example, by increasing sympathetic outflow to the cardiovascular organs. However, the cardiometabolic impact of deleting insulin receptors only in the hypothalamic region is different from deleting insulin receptors in the whole brain. In other words, insulin signaling outside the hypothalamus is also important for cardioregulatory activities. To explore this extra-hypothalamic insulin signaling, scientists from...

Gut-brain axis: microbiota influence behavior and mental well-being

Tuesday, December 3, 2019 - 09:44
Expression of Neurogranin in the rat brain hippocampus, a calmodulin-binding protein, and a neuron specific protein commonly found in postsynaptic terminals (dendritic spines).

By Jennifer Sokolowski, MD, PhD.

Gut-microbiome interactions

On and in our bodies, microbes outnumber our cells by about ten-to-one. Studies have revealed that the microbiome influences neurogenesis, cognition, and stress responses, leading to increased interest in identifying factors that affect the gut microbiota, and a drive to understand pathways involved in the gut-brain axis.1

The composition of microbiota in the gut affects the ...

Astrocytes: Diversity in type and function

Monday, November 18, 2019 - 09:30
In the CNS astrocytes have different functions including modulation of synaptic activity and synaptogenesis, communication with other astrocytes, oligodendrocytes and microglia, as well as modulation of the blood brain barrier.

By Michalina Hanzel, PhD

Astrocytes are a type of macroglia found in the central nervous system that regulate a vast array of essential brain functions, ranging from synaptogenesis, ion homeostasis and maintenance of the blood-brain barrier, to buffering of neurotransmitters and secretion of neuroactive agents. Considering the plethora of roles that astrocytes...

Muscle-specific UBE2O ablation requires activated AMPKα2 to protect against metabolic syndrome

Friday, November 8, 2019 - 10:45
Clearance of elevated glucose and fatty acids by skeletal muscle is affected by the development of insulin resistance.

By Jamshed Arslan, Pharm. D., PhD.

Generating energy (ATP) from nutrients is a recipe for life. One of the sensors of cellular energy levels is the serine/threonine kinase AMPK. This enzyme facilitates lipid oxidation and glucose uptake when energy levels fall, thereby promoting ATP production. AMPK comprises a catalytic alpha subunit with two regulatory subunits,...

Autophagy and Metastasis

Monday, November 4, 2019 - 10:08
EpCAM/TROP1 is localized by immunohistochemical analysis of human breast carcinoma tissue to the cell membrane and intercellular spaces.
 

By Christina Towers, PhD

The majority of cancer patients die from metastatic disease at secondary sites. The threshold to undergo metastasis is high. Only a minority of cancer cells acquire invasive phenotypes like motility and the ability to survive under detached conditions. Consistently, only few cancer cells acquire stem cell like properties, including a more mesenchymal morphology...

Metabolic Syndrome: Symptoms and associated disease states

Tuesday, October 29, 2019 - 09:23
The term, metabolism, refers to the totality of chemical reactions that occur in a biological system for the conversion of ingested nutrients into their basic constituents (i.e. proteins, lipids, nucleic acids and carbohydrates).

By Jamshed Arslan, Pharm. D., PhD.

What is metabolic syndrome?

It was 1988 when, after decades of research, Dr. GM Reaven    of the Stanford University, CA, explained the relationship between four diseases: insulin resistance, hypertension, diabetes and obesity....

Increased wild type FUS levels in ALS patients lead to a toxic microenvironment and motor neuron neurodegeneration

Tuesday, October 22, 2019 - 10:03
Dual expression of ALDH1L1-red and Vimentin-green identifies astrocytes which may be identified by their golden or red fluorescence. ALDH1L1 is expressed in the cell body and processes of astrocytes.

By Michalina Hanzel, PhD

FUS mutations in Amyotrophic Lateral Sclerosis

Fused in sarcoma (FUS) is a ribonucleoprotein that continuously shuttles between the nucleus and the cytoplasm to regulate pre-mRNA splicing, mRNA stability and mRNA transport. Mutations in FUS are found in 4% familial and 1% sporadic cases of amyotrophic lateral...

Detecting HIF alpha and beyond: Best controls for hypoxia Western blot analysis

Monday, October 14, 2019 - 09:17
Analysis of HIF-1 alpha expression in whole cell lysates from HeLa cells in culture treated or untreated with cobalt chloride and independent antibody validation where two different antibodies to HIF-1 alpha were used for immunoprecipitation and detection.

By Rosa Moreno, PhD.

Detecting HIF alpha and beyond: Best controls for hypoxia Western blot analysis

Physiological low levels of oxygen induce normal hypoxic events across biological systems. This hypoxic state activates hypoxia inducible factors (HIFs) to regulate transcription by binding to the hypoxia response element (HRE) region of...

Neurovascular signaling for repair enhances brain metastasis

Tuesday, October 8, 2019 - 09:02
High expression of GFAP (green) identifies astrocytes in rat cerebellum

By Jamshed Arslan, Pharm. D., PhD.

Stroke    is a leading cause of mortality and morbidity worldwide. Cellular players – neurons, astrocytes, endothelial and stromal cells – involved in post-stroke repair through angiogenesis...

Glypican 3 as a biomarker for gastro-esophageal adenocarcinoma

Tuesday, October 1, 2019 - 10:17
Expression of CD81 was detected by immunofluorescence with a mouse monoclonal antibody followed by secondary antibody labeled with DyLight488 and Phalloidin 568 staining for actin visualization

By Jamshed Arslan, Pharm. D., PhD.

Gastroesophageal adenocarcinoma originates from the glandular epithelium of the esophagus, gastroesophageal junction and stomach. The incidence of gastroesophageal adenocarcinoma is declining, but it is still one of the deadliest cancers worldwide. A key reason behind high mortality is that by the time gastroesophageal adenocarcinoma is diagnosed, the cancer has often already metastasized. The search for novel biomarkers underlying gastroesophageal...

Isolating Immune Cells From Peripheral Blood: How and When to Use Density Gradient Centrifugation

Tuesday, September 24, 2019 - 09:05
Adaptive immunity is mediated by lymphocytes (e.g., T cells and B cells) while innate immunity responses are mediated by several cellular mediators (e.g., Neutrophils, Natural Killer Cells and Eosinophils)

By Victoria Osinski

Lymphocytes, monocytes, and granulocytes, oh my!

There are many different immune cells (leukocytes) that are found in circulation along with other cellular components such as platelets and red blood cells. Red blood cells (RBCs) are the most abundant, followed by neutrophils and lymphocytes. The immune cells can be categorized as either mononuclear cells (monocytes and lymphocytes...

Suppressing breast cancer metastasis: The role of hypoxia-induced RhoB expression and activation

Tuesday, September 17, 2019 - 08:16
Expression of RhoA, B and C detected in the cytoplasm of A431 cells, a model of human epidermoid carcinoma, compared to negative control analyzed in the absence of RhoA Antibody (1A11-4G10) [NBP2-22528]

By Jamshed Arslan, Pharm. D., PhD.

The Ras homologous

Tips to Optimize Your Western Blot for Phosphorylated Protein Detection

Tuesday, September 10, 2019 - 10:48
Western blotting allows identifying and quantifying proteins from cell lysates for determination of protein expression, sub-cellular localization, post-translational modifications, protein processing, and protein-protein interactions.

By Jamshed Arslan, Pharm. D., PhD.

Protein phosphorylation refers to a reversible post-translational modification in which a protein kinase adds a phosphate group to an amino acid residue of a target protein. Protein phosphorylation, especially tyrosine phosphorylation, is one of the early events in signal transduction in all eukaryotic cells.

Once a cell is lysed, proteases and phosphatases are released that can degrade or...

The Ins and Outs of Survivin

Tuesday, September 3, 2019 - 08:40
Exosomes arise from the late endosomal compartment through inward budding of multivesicular bodies and may contain multiple proteins, lipids, DNA, RNAs and biomolecules of viruses/prions.

By Rachel M.A. Linger, Ph.D.

What is survivin?

Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of survivin occurs in several subcellular locations including the cytoplasm, nucleus,...

Explore IHC basics with this new guide: Immunohistochemistry Essential Elements and Beyond

Tuesday, August 27, 2019 - 09:07
3-color fluorescence IHC, nuclei-blue, cytoplasm-red and cytoskeleton-green, Immunohistochemistry Essential Elements and Beyond, Springer book

By Rosa Moreno, PhD.

This new introductory guide to methods and techniques in immunohistochemistry is written by Dr. Alexander E. Kalyuzhny, Bio-Techne, for the Springer series: Techniques in Life Science and Biomedicine for the Non-Expert   . This guide provides a comprehensive introduction along with conceptual and methodological basics for new users to ensure a clear understanding upon which researchers may build their expertise in ...

Chromatin reader domains of DNMT-targeting protein, UHRF1, are responsible for cancerous DNA hypermethylation

Tuesday, August 20, 2019 - 08:58
Expression of DNMT2 in human colon tissue shows cytoplasmic and nuclear staining in glandular cells, IHC

By Jamshed Arslan, Pharm. D., PhD.

DNA methylation represses transcription of many genes, including tumor suppressor genes. A protein called UHRF1 recruits DNA methyltransferases (DNMTs) to establish and maintain DNA methylation. UHRF1 has several domains, most notably:

  • N-terminal ubiquitin-like (UBL) domain
  • TTD (tandem Tudor domain) and PHD (ubiquitin-like, containing plant homeodomain) domains that...

Role of GFAP in astrocytes: Lessons from induced pluripotent stem cells in Alexander disease patients

Tuesday, August 13, 2019 - 10:52
GFAP-green and Neurofilament-red expression in astrocytes of mouse brain cerebellum, IHC

By Michalina Hanzel, PhD

Alexander disease is a progressive and fatal neurological disease with phenotypes ranging from myelination abnormalities, gait ataxia and megalencephaly to predisposition to seizures. It is an autosomal dominant disease with mutations in the glial fibrillary acidic protein (GFAP) gene that result in astrocytic cytoplasmic inclusions, termed Rosenthal fibers, which contribute to the global neurological...

Considerations for Quantitative Western blotting

Tuesday, August 6, 2019 - 09:11
GAPDH expression in human nucleus pulposus cell lysate, WB

By Jamshed Arslan, Pharm. D., PhD.

Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labelled with fluorescent/chemiluminescent probes has virtually removed the need for radioactivity. Western blotting has become a routine quantitative method in biological research and immunodiagnostics. The following primer discusses some considerations when quantitating Western blot data.


...

Antibody treatment can generate microglia-like cells from bone marrow

Tuesday, July 30, 2019 - 10:18
Sox2 antibody expression in induced pluripotent stem cells, ICC

By Jennifer Sokolowski, MD, PhD.

Microglia play important roles in the brain in both homeostatic and pathological conditions, acting to clear debris and dying cells. There is evidence to suggest that microglial dysfunction contributes to neurodegenerative diseases such as Alzheimer's and modulation of microglial activity may be a method to treat such diseases.1 Cell-based therapies represent a novel approach whereby the introduction of microglia that possess the desired phenotype could potentially be...

Optogenetic Control of Mitophagy: AMBRA1 based mitophagy switch

Tuesday, July 23, 2019 - 10:38
sAmbra1 expression and localization to mitochondria in human U-2 OS cell line, ICC

By Christina Towers, PhD

Mitophagy in the Brain

Selective autophagic degradation of damaged mitochondria, known as mitophagy, has been described as a cyto-protective process. Accordingly, defects in mitophagy have been associated with a number of diseases including muscle atrophy, cancer, and multiple neurodegenerative diseases. Defective mitophagy has been best described in neurons, where the accumulation of damaged mitochondria and the resulting increase in reactive oxygen species (...

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