Daniel Zeve

White Fat Progenitor Cells Reside in the Adipose Vasculature

White adipose (fat) tissues regulate metabolism, reproduction, and life span. Adipocytes form throughout life, with the most marked expansion of the lineage occurring during the postnatal period. Adipocytes develop in coordination with the vasculature, but the identity and location of white adipocyte progenitor cells in vivo are unknown. We used genetically marked mice to isolate proliferating and renewing adipogenic progenitors. We found that most adipocytes descend from a pool of these proliferating progenitors that are already committed, either prenatally or early in postnatal life. These progenitors reside in the mural cell compartment of the adipose vasculature, but not in the vasculature of other tissues. Thus, the adipose vasculature appears to function as a progenitor niche and may provide signals for adipocyte development.

The developmental origins of adipose tissue

Adipose tissue is formed at stereotypic times and locations in a diverse array of organisms. Once formed, the tissue is dynamic, responding to homeostatic and external cues and capable of a 15-fold expansion. The formation and maintenance of adipose tissue is essential to many biological processes and when perturbed leads to significant diseases. Despite this basic and clinical significance, understanding of the developmental biology of adipose tissue has languished. In this Review, we highlight recent efforts to unveil adipose developmental cues, adipose stem cell biology and the regulators of adipose tissue homeostasis and dynamism.

The sexually dimorphic role of adipose and adipocyte estrogen receptors in modulating adipose tissue expansion, inflammation, and fibrosis

Our data demonstrate that estrogens, estrogen receptor-α (ERα), and estrogen receptor-β (ERβ) regulate adipose tissue distribution, inflammation, fibrosis, and glucose homeostasis, by determining that αERKO mice have increased adipose tissue inflammation and fibrosis prior to obesity onset. Selective deletion of adipose tissue ERα in adult mice using a novel viral vector technology recapitulated the findings in the total body ERα null mice. Generation of a novel mouse model, lacking ERα specifically from adipocytes (AdipoERα), demonstrated increased markers of fibrosis and inflammation, especially in the males. Additionally, we found that the beneficial effects of estrogens on adipose tissue require adipocyte ERα. Lastly, we determined the role of ERβ in regulating inflammation and fibrosis, by breeding the AdipoERα into the βERKO background and found that in the absence of adipocyte ERα, ERβ has a protective role. These data suggest that adipose tissue and adipocyte ERα protects against adiposity, inflammation, and fibrosis in both males and females.

Fighting Fat with Fat: The Expanding Field of Adipose Stem Cells

We are in the midst of a dire, unprecedented, and global epidemic of obesity and secondary sequelae, most prominently diabetes and hyperlipidemia. Underlying this epidemic is the most hated of cells, adipocytes, and their inherent dynamic ability to expand and renew. This capacity highlights a heretofore undefined stem compartment. Recent in vivo studies, relying upon lineage tracing and flow cytometry methods, have begun to unravel the identity of adipose stem cells, their niche, and the dynamism central to adipose expansion. Thus, the field is moving in a direction that may allow us to manipulate adipose stem cells to beneficial therapeutic ends.

Osteoclast Progenitors Reside in the Peroxisome Proliferator-Activated Receptor γ-Expressing Bone Marrow Cell Population

ABSTRACT Osteoclasts are bone-resorbing cells essential for skeletal development, homeostasis, and regeneration. They derive from hematopoietic progenitors in the monocyte/macrophage lineage and differentiate in response to RANKL. However, the precise nature of osteoclast progenitors is a longstanding and important question. Using inducible peroxisome proliferator-activated receptor γ (PPARγ)-tTA TRE-GFP (green fluorescent protein) reporter mice, we show that osteoclast progenitors reside specifically in the PPARγ-expressing hematopoietic bone marrow population and identify the quiescent PPARγ+ cells as osteoclast progenitors. Importantly, two PPARγ-tTA TRE-Cre-controlled genetic models provide compelling functional evidence. First, Notch activation in PPARγ+ cells causes high bone mass due to impaired osteoclast precursor proliferation. Second, selective ablation of PPARγ+ cells by diphtheria toxin also causes high bone mass due to decreased osteoclast numbers. Furthermore, PPARγ+ cells respond to both pathological and pharmacological resorption-enhancing stimuli. Mechanistically, PPARγ promotes osteoclast progenitors by activating GATA2 transcription. These findings not only identify the long-sought-after osteoclast progenitors but also establish unprecedented tools for their visualization, isolation, characterization, and genetic manipulation.

Oestrogen signalling in white adipose progenitor cells inhibits differentiation into brown adipose and smooth muscle cells

Oestrogen, often via oestrogen receptor alpha (ERα) signalling, regulates metabolic physiology, highlighted by post-menopausal temperature dysregulation (hot flashes), glucose intolerance, increased appetite and reduced metabolic rate. Here we show that ERα signalling has a role in adipose lineage specification in mice. ERα regulates adipose progenitor identity and potency, promoting white adipogenic lineage commitment. White adipose progenitors lacking ERα reprogramme and enter into smooth muscle and brown adipogenic fates. Mechanistic studies highlight a TGFβ programme involved in progenitor reprogramming downstream of ERα signalling. The observed reprogramming has profound metabolic outcomes; both female and male adipose-lineage ERα-mutant mice are lean, have improved glucose sensitivity and are resistant to weight gain on a high-fat diet. Further, they are hypermetabolic, hyperphagic and hyperthermic, all consistent with a brown phenotype. Together, these findings indicate that ERα cell autonomously regulates adipose lineage commitment, brown fat and smooth muscle cell formation, and systemic metabolism, in a manner relevant to prevalent metabolic diseases.

Host Resistance of CD18 Knockout Mice against Systemic Infection with Listeria monocytogenes

ABSTRACT Mice with targeted mutations of CD18, the common β2 subunit of CD11/CD18 integrins, have leukocytosis, impaired transendothelial neutrophil emigration, and reduced host defense to Streptococcus pneumoniae, a gram-positive extracellular bacterium. Previous studies using blocking monoclonal antibodies suggested roles for CD18 and CD11b in hepatic neutrophil recruitment and host innate response to Listeria monocytogenes, a gram-positive intracellular bacterium. We induced systemic listeriosis in CD18 knockout (CD18-ko) and wild-type (WT) mice by tail vein injection with Listeria. By 14 days postinjection (dpi), 8 of 10 WT mice died, compared with 2 of 10 CD18-ko mice (P < 0.01). Quantitative organ culture showed that numbers of Listeria organisms in livers and spleens were similar in both groups at 20 min postinfection. By 3, 5, and 7 dpi, however, numbers of Listeria organisms were significantly lower in livers and spleens of CD18-ko mice than in WT mice. Histopathology showed that following Listeria infection, CD18-ko mice had milder inflammatory and necrotizing lesions in both spleens and livers than did WT mice. Cytokine assays indicated that baseline interleukin-1β and granulocyte colony-stimulating factor (G-CSF) levels were higher in CD18-ko mice than in WT mice and that CD18-ko splenocytes produced higher levels of interleukin-1β and G-CSF than WT splenocytes under the same amount of Listeria stimulation. These findings show that CD18 is not an absolute requirement for antilisterial innate immunity or hepatic neutrophil recruitment. We propose that the absence of CD18 in the mice results in the priming of innate immunity, as evidenced by elevated cytokine expression, and neutrophilic leukocytosis, which augments antilisterial defense.

John B. Watson's Alleged Sex Research: An Appraisal of the Evidence.

In 1974, a story was published about clandestine research done by John B. Watson that was judged to be so reprehensible that it was offered as the real reason he was fired from his faculty position at Johns Hopkins University in 1920, at perhaps the peak of his academic career. Watsons dismissal from Johns Hopkins may have been the most important event in his career, and it almost certainly altered the history of American psychology. Thus, this story has great significance. The claims of the story, however, have never been validated or invalidated. This article examines the evidence for and against the existence of such research and discusses Watsons academic dismissal in light of that evidence.

Small at Birth, but How Small? The Definition of SGA Revisited

50 years of published discussion of the definition of SGA [3–5] . The 10th percentile was chosen as a cutoff for SGA in the 1960s as a result of multiple studies indicating that infants born at or below the 10th percentile have increased mortality compared to gestational age-matched controls [4] . It is worth noting that these studies were performed on neonates born at high altitudes, who tend to be smaller compared to those born at sea level [4, 5] . Defining SGA as 2 SDs below the mean was also suggested in the 1960s, as it would only define 4.6% of births as abnormal (approximately 2.3% SGA and 2.3% large for gestational age), as opposed to 20% (10% SGA and 10% large for gestational age). Additionally, the 2 SD cutoff roughly corresponded with an earlier study reporting that 2.3% of neonates were born at 25% below the mean weight when controlled for gestational age [5] . In 1995, the World Health Organization published recommendations defining SGA as less than the 10th percentile of weight for gestational age using localized anthropometric newborn curves [3] . In 2007, a consensus meeting that included representatives from seven international pediatric endocrinology societies, as well as a representation from obstetrics, perinatology and neonatology, pediatrics, epidemiology, and pharmacology, recommended that SGA be defined as more than 2 SDs below the mean for weight and/or length [2] . They also recBeing born small has been associated with many shortand long-term health sequelae. Perinatally, it is associated with increased mortality, lung disease, hypotension, necrotizing enterocolitis, poor thermoregulation, hypoglycemia, and polycythemia [1, 2] . Long-term, small infants are at risk for insulin resistance, type II diabetes mellitus, cardiovascular disease, chronic kidney disease, neurodevelopmental and cognitive impairments, developmental delays, behavioral problems, and adult short stature [1] . But how ‘small’ is ‘too small’? The term ‘small for gestational age’ (SGA) describes newborns who have lowerthan-expected weight, length, and/or head circumference when controlled for gestational age and sex. This is different from intrauterine growth retardation, which refers to poor growth in utero evidenced by at least two ultrasound measurements [2] , and prematurity, which is a broad term defining neonates born prior to 37 weeks gestation. Some, but not all, infants with intrauterine growth retardation and/or prematurity may be born SGA. In spite of potential significant health implications, the exact definition of SGA remains elusive. Multiple criteria have been used, including less than the 10th, 5th, and 3rd percentile in weight, length, or head circumference. Another definition of SGA is parameters more than 2 standard deviations (SDs) below the mean, or the 2.3rd percentile [1, 2] . These discrepancies are not novel, with over Received: August 11, 2016 Accepted: August 12, 2016 Published online: September 30, 2016 HORMONE RESEARCH IN PÆDIATRICS

Developmental expression of neuronal nitric oxide synthase in P/Q-type voltage-gated calcium ion channel mutant mice, leaner and tottering

Nitric oxide (NO) is a diffusible messenger molecule produced primarily by neuronal nitric oxide synthase (nNOS) in the central nervous system. Both nNOS expression and NO production are regulated by calcium ions. Leaner and tottering mice carry a mutation in the pore forming subunit (alpha1A) of P/Q-type voltage-gated calcium ion channels, which decreases calcium ion current through the affected channels and disrupts calcium homeostasis. We have previously shown that nNOS expression is altered in adult leaner and tottering cerebella. In addition, leaner and tottering mice have been shown to have abnormal cerebellar granule cell-Purkinje cell synapses and leaner cerebellar granule cells undergo abnormal apoptosis during early postnatal development. Since NO production has been linked to several developmental roles including neuronal cell death, synaptogenesis and neuronal cell survival, our objective was to evaluate the expression of nNOS in developing leaner and tottering cerebella. Our results show that nNOS is differentially expressed in leaner and tottering cerebella compared to wild type cerebella and compared to each other. In whole cerebella, Western blotting revealed a significant increase in nNOS expression at postnatal day 12 in tottering but not leaner or wild type cerebella. At the cellular level the NADPH-diaphorase marker for nNOS revealed a significant increase in nNOS expression in basket cell interneurons in both mutant mice. nNOS expression in granule cells in the internal granule cell layer in tottering mice was increased at P12, while granule cells of leaner mice exhibited decreased nNOS expression at P20. The changes in nNOS expression at P12 did not correlate with a change in overall NO production, but rather maintained wild type NO concentrations. These findings suggest that changes in nNOS expression in the leaner and tottering cerebella are compensatory in nature with NO most likely functioning as a calcium-regulated neuroprotective/neurotrophic factor in postnatal cerebellar development.